NUCIS Number 13. December 2006. 72 Pages (Full Version In

NUCIS NEWSLETTER Information Bulletin of the Research Network on Nuts (FAO-CIHEAM) Number 13 December 2006

CIHEAM IRTA Mas de Bover ● Coordination Centre of the Research Network on Nuts

FAO CIHEAM Nut Network

New almond plantings in Almería, Andalusia. The main Spanish almond growing region.

posium on Pistachio and Almond was tion programme experts to focus on only EDITORIAL held in Teheran, Iran. The corresponding one proposal. In the near future, COST Proceedings will be published in Acta pre-proposals should be presented on se- Horticulturae. veral tree nuts. This is regarded as a Activities 2004-2006 source of financial support to COST acti- During part of 2004, 2005 and 2006 a This year an European COST (Coopera- ons for our Research Network of COST number of activities were supported by tion in the field of Scientific and Technical Member States. An immediate challenge the FAO-CIHEAM Interregional Coopera- Resarch) preliminary proposal on walnut for the Network members is to propose tive Research Network on Nuts. In Nov- R&D work was prepared by network competitive R+D projects which address ember 2004 the V ISHS International members and submitted to the European relevant problems of the nut sector. The Walnut Symposium was held in Sorrento, Commission for approval. Several other coming 7th UE Framework Programme Italy. In November 2004 the III ISHS In- COST pre-proposals on almond, chest- for R+D can be an interesting opportunity ternational Symposium on Chestnut took nut, hazelnut and pistachio were also pre- for European member countries or asso- place in Chaves, Vila Real, Portugal. In pared but were not submitted finally follo- ciates to be explored. Once carried out May 2005 the IV ISHS International Sym- wing advice from the European COST ac- the projects, the generated information

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 1 has to be transferred to the corresponding agents of the nut sector.

Work carried out by our Nut Network has been reviewed and well assessed within FAO and CIHEAM. During several years a number of initiatives have been tried to rise funding but without much success. The scarce funding has limited activities. In recent years, the activities of the Nut Network have been affected as FAO is no longer providing specific budget for servi- cing the Networks and supporting activities. Thus activities to be carried out are hindered. However, the support given by some FAO services and external funding provided by the Spanish INIA, and the cosponsor and partner CIHEAM most planned activities could be finally developed like the publication of this Newsletter. In addition, some researchers from developing or transition countries have being supported to participate in different meetings and congresses. 1. Wood screw to apply pressure on side of nut. 2 & 3. Metal pincer to crack nut by pressure on side or end. 4. Metal screw to apply pressure sidewise. Nut Network Coordination Centre: 5, 8 & 12. Horizontal pluger with impact applied to end of nut by one stroke of mallet. IRTA Mas de Bover 6 & 10. Same as above with force furnished by tension of large rubber band. Since the start in 1990 of the Inter-regio- 7. Horizontal plunger activated by lever, to apply pressure on end of nut; adjusted to size of nut by screw-cylinder. nal Cooperative Research Network in tree 9. Same as seven except home-made. nuts IRTA Mas de Bover has been the 11. Vertical plunger activited by lever to apply pressure at end of nut; adjusted to nut size by notches Coordination Centre of the Network. Du- in upright staff. ring these years only two Coordinators have been in charge: J. Tasias (1990-92) From Woodroof (1967) and F.J. Vargas (1992-2005). After 15 years of coordination we feel that this CIHEAM Nut Network report. F.J. Vargas (2004, REU 66). These inventories pu- task should continue with a new Coordi- will still continue to work in close relation blished in the REU Technical Series nator. with our Network. (http://www.fao.org/regional/europe/PUB) are important compilations of the current- Change of Nut Network Coordinator The Nut Network’s future ly available species genetic resources FAO and CIHEAM have both agreed, af- Nut tree production, trade and industry and information on ongoing research pro- ter being asked by F.J. Vargas in Decem- are important economical activities rela- jects and bibliography. In addition, one ber 2005, that he will no longer carry on ted to sustainable agriculture, often in more inventory is being compiled and is coordinating our Nut Network and thus he marginal lands and under rainfed condi- close to completion. The inventory on resigned. Following FAO and CIHEAM tions, in both European and Near Eastern Pistachio is being collated by B.E. Ak and established protocols to change Network Regions. Nuts are of major importance N. Kaska. All these catalogues are being Coordination and after expert consultatio- and are typical components of the tradi- funded by FAO’s Regional Office for ns both organizations have agreed to tional and healthy Mediterranean diet. To Europe and the CIHEAM-IAMZ. name Dr. Mercè Rovira, from IRTA Mas accomplish sustainable development and de Bover, as new Nut Network Coordina- food security, a future combined effort in Also, a draft Descriptors List for Hazelnut tor. M. Rovira has been working on re- R&D, environmental management and has been developed by Network mem- search and development for more than 20 communication is needed. Looking ahead bers lead by A.I. Köksal and was submit- years in several aspects of nut trees, and thinking on our Nut Network financial ted to IPGRI (www.cgiar.org/ipgri/) for as- mainly in hazelnut and walnut but also support, FAO will not provide Çad hocÈ sessment. After CIHEAM agreed to co- with almond and pistachio. She has close funding but will be backing proposals to sponsor this Descriptors List it is time and strong relationships with many re- other funding Agencies like the EC now to be edited and published. searchers and R+D institutions world- through COST programmes or similar. In wide. We wish Dr. Rovira a successful addition, CIHEAM is fully committed with Proceedings of meetings and work- work ahead and we will provide all the the Nut Network and providing regular shops backing she will need to coordinate our funding since being a partner. Spain is Five publications related to Meetings and Network in the coming future. We thank supporting the Nut Network yearly through Workshops on Almond, Pistachio and F. J. Vargas for all the efforts made since INIA specific funding (Spanish Trust Fund). Economics of Nuts have been also edi- he took over in 1992 as Coordinator. The ted: Cahiers Options Méditerranéennes task of Network Coordinator is a time de- Genetic resources inventories and Volume 33 (X GREMPA) and Volume 56 manding activity which should be addres- descriptors (XI GREMPA), Options Méditerranéennes sed with enthusiasm and efficiency, as Regarding the Inventories on Germ- Série A/63 (XIII GREMPA). Options Médi- ÇPacoÈ has done over these 13 years. As plasm, Research and References, four terranéennes Série A/5 (Almond rootstoc- a result of his coordination a large num- have been already published: Almond ks) and Options Méditerranéennes Série ber of activities have being carried out (1997, RTS 51), Hazelnut (2000, RTS A/37 (Economics of nuts in the Mediterra- and are detailed in an inside overall FAO- 56), Chestnut (2001, RTS 65) and Walnut nean basin).

2 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 The FAO European Regional Office all contributors for their efforts and inter- CONTENTS Page and CIHEAM (IAMZ) est to produce and send me valuable in- Mrs. Karin Nichterlein, Research and Te- formation. The exchange of information chnology Officer from FAO Regional Offi- between Network members through the ce for Europe in Rome is our link. In rela- pages of this Newsletter is the basis for EDITORIAL ...... 1 tion to CIHEAM, Mr. Dunixi Gabiña, ba- developing collaboration. The editing sed in IAMZ of Zaragoza, Spain is our fo- task in the thirteen NUCIS issues already cal point. published has been huge (NUCIS 1, 9 ARTICLES AND REPORTS pages; 2, 20 pages; 3, 24 pages; 4, 28 The Nut Network on the web pages; 5, 36 pages; 6, 52 pages, 7, 44 Basic information is included in the REU pages, 8, 46 pages 9, 68 pages, 10, 48 • Farewell words by the former Network Coordinator. website for ESCORENA (see:www.fao. Welcome words by the Network Coordinator pages, 11, 48 pages 12, 52 pages and and Report of The FAO-CIHEAM Interregional org/world/regional/REU/content/escore- 13, 72 pages). In order to reduce the time Cooperative Research Network on Nuts ...... 4 na/index_en.htm). Additional information consuming formal editing we are asking about the Nut Network can be found at contributors who send articles, news, no- • Four new almond varieties released by IRTA: www.iamz.ciheam.org/en/pages/paginas/ tes, bibliographic references, etc., to the ‘Vayro’, ‘Marinada’, ‘Constantí’ and ‘Tarraco’ ...... 9 pag_investigacion2a.htm. different sections to provide them well or- • ‘Belona’ and ‘Soleta’, Two new autogamous ganized and elaborated. Information almonds ...... 12 NUCIS on the web should be sent in standard English. Con- • The almond cultivar ‘Francolí’ is self-compatible ...... 16 A full electronic version of each NUCIS tributions could be sent through Internet (from issue 1 to 13) is now available on using the Editor’s email. The alternative • New findings in the genetic control and expression the Internet web page of CIHEAM at is to provide them on diskette and also in of kernel bitterness in almond ...... 20 (www.iamz.ciheam.org/en/pages/pagi- printed format. This bulletin is reproduced • Tocopherol in almond kernels as a quality nas/pag_investigacion2a.htm). The con- in black and white only, including pictu- component ...... 22 tents of this Newsletter can be browsed res. Please send your contributions for through and also copied and printed. the next issue, number 14 by the end of • Long-term cryopreservation of almond germplasm: Readers will be able now to find the who- June 2007. We thank all who have contri- an update ...... 25 le set of NUCIS issues, some of which buted to this issue. I personally thank all • Flower drop by precocious abortion in walnut ...... 27 were already exhausted. the readers who are the focus of this Newsletter for their interest on this rewar- • California pistachio industry ...... 31 Contributions to NUCIS ding task. I should say thanks to all the • The low yield reasons and solutions for pistachio As in past NUCIS editorials, we again readers and also goodbye to you all as production in Turkey ...... 37 stress that this Newsletter should be an this is my last full NUCIS editing work. • Pistachio cultivation in Castilla-La Mancha: effective vehicle of communication for all From now on an Editorial Committee will twenty years later ...... 41 the Network members. The pages of this be established to undertake the editing bulletin are open to all readers who would and composition of NUCIS and I will be • Pistachio dieback in Australia: a case of multiple like to suggest ideas or to express their only the scientific editor. Finally, we wish inoculation events ...... 45 opinion about the work developed by the all Nut Network members and collabora- • Pistacia species genebank, conservation, Network (activities carried out and plan- tors a rewarding and Happy 2007. characterization and use at IRTA: current situation ned) or to publish short articles and re- and prospects in Spain ...... 47 ports on relevant horticultural subjects of • Domestication of the pistachio nut (Pistacia vera) .... 53 general interest. We receive generally a The Editor sufficient number of contributions from • Carob tree germplasm in Tunisia ...... 55 the Mediterranean Basin and overseas The designations employed and the for the articles and reports section. Howe- presentation of material in this informa- ver, the sections on news and notes and tion product do not imply the expression NOTES AND NEWS also on congresses and meetings are of any opinion whatsoever on the part usually difficult to cover due to the scarce of the Food and Agriculture Organiza- tion of the United Nations concerning • Walnut Research at East Malling Research ...... 59 information received and thus, contribu- the legal or development status of any tions are most welcomed. Otherwise, the country, territory, city or area or of its Editor has to report on the issues he is authorities, or concerning the delimita- aware of, but certainly there must be many tion of its frontiers or boundaries. CONGRESSES AND MEETINGS more issues on-going throughout the year This publication contains the collective which merit reporting. Also, the place for views of an international group of ex- • IV International Symposium on Pistachio ‘grey’ bibliography (references and docu- perts and does not necessarily repre- and Almond ...... 59 ments which are difficult to search like sent the decisions or the stated policy Masters or Ph Theses) is scarcely filled. of the Food and Agriculture Organiza- • Summary of the V International Walnut Congress .... 60 tion of the United Nations, the Interna- tional Centre for Advanced Mediterra- • Spanish-French workshop on walnut ...... 62 The NUCIS Newsletter is distributed nean Agronomic Studies nor of the Or- worldwide free of charge to 1.400 readers ganization for the Economic Coopera- • III International Chestnut Congress ...... 62 from over 60 countries. The dissemina- tion and Development. tion of information originated by the Net- Contributions should be written conci- TO BE HELD ...... 63 work is of paramount importance and sely in English. Please send contribu- through this bulletin has been largely suc- tions on paper and diskette (Microsoft¨ cessful. The first NUCIS was published in Word or Word Perfect¨). Authors are BIBLIOGRAPHY ...... 63 1993, this issue of the NUCIS Newsletter responsible for the content of their pa- is number 13 and during these thirteen pers. Reproduction of the articles is au- thorized, provided that the original BACKPAGE ...... 72 years a great editing effort has been source is clearly stated. made. I acknowledge and gratefully thank

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 3 ARTICLES AND REPORTS

FAREWELL WORDS BY THE FORMER NETWORK COORDINATOR

Dear Colleagues,

As l have recently left the job as coordinator of the FAO-CIHEAM Network on Nuts, which will be carried out by my colleague and friend Dr. Mercè Rovira, I wish to wri- te a few lines to all Network members.

I have had the task of being the Network on Nuts coordinator for a long time, perhaps too long, since 1992 (14 years). Se- veral circumstances, which are not relevant here, kept me much longer on this post than I had first planned. It has been Francisco José Vargas. Mercè Rovira. a very interesting experience for me. Of course I have had to face problems and some difficult periods, but I also achieved I wish to thank the institutions which sup- WELCOME WORDS goals which easily compensated some ported our activities since the Network BY THE NEW NETWORK was established in 1990 as an initiative of drawbacks. My personal overall assess- COORDINATOR ment is quite satisfactory. FAO (REU, RNE and AGPS). Wide- spread support from different national As coordinator, I have tried to accomplish and international institutions was recei- Dear Colleagues, the aims established when the network ved. In 1996 FAO and CIHEAM agreed to was founded in 1990, in Yalova, Turkey: cosponsor the Network, IAMZ providing a As many of you already know, F.J. Var- promotion of information exchange, joint steady and strong support. From the start gas has recently left his post as Nut Net- applied research, exchange of germ- of our Nut Network, consistent funding work Coordinator. After being asked to plasm and establishment of links among and support has been received from take over the coordination I was pleased researchers. Our activity has been mainly Spain, through INIA and IRTA institutes. to accept his offer in this new and cha- focussed on fostering cooperation in the llenging undertaking and responsibility. frame of research on nut tree crops Many people have collaborated closely As many of you already know, we are among Network members. with the Network and have contributed to both working in Mediterranean Fruit Trees ease my job as coordinator: Subnetwork at IRTA Mas de Bover. My research acti- In this issue of NUCIS a report about the Liaison Officers, FAO and CIHEAM Offi- vity is mainly focussed on hazelnut and activities carried out by FAO-CIHEAM cers, researchers, symposium conve- walnut crops, but I also develop work on Network on Nuts since its start is inclu- ners, authorities, editors, administratives, almond and pistachio, mainly in floral bio- ded: a) organization of technical mee- etc. They cannot be all mentioned, as the logy aspects. tings, b) promotion of R&D activities and list would be too long. Nevertheless, I support of training grants for young re- would like to name some people, unfortu- I hope to coordinate the Network with the searchers c) edition of proceedings and nately some of whom are not longer same enthusiasm as F.J. Vargas has reports, d) edition of inventories of germ- among us, in key stages of its establish- shown over all these years. I am sure that plasm and research lines, e) edition of the ment or development: H. Olez, U. Menini, with your help, we will carry on develo- NUCIS Newsletter and f) organization of J. Tasias, J. Boyazoglu, D. Gabiña, E. ping collaboration on different topics. In international courses. These activities Germain, F. Monastra, N. Kaska and I. the near future we will focuss our efforts were developed with the financial support Batlle. I wish to express my gratitude to on finishing two publications: the Hazel- of several institutions and the efforts of all of them. nut IPGRI descriptors and the inventory many colleagues. We have had very acti- of Pistachio research, germplasm and re- ve periods and times with less activity, My best wishes to the new coordinator ferences. Most of the work has been mostly related to budget availabilities. I Mercè Rovira. She has a demanding and made on these two useful publications would like to highlight the importance of exciting task ahead. In the near future, I during the last years, and it is time to fi- the edition of 4 inventories of germplasm will carry on helping her, fostering almond nish both of them. resources and research activities (al- activities. mond, hazelnut, chestnut and walnut), 13 I would like to encourage you to contact issues of NUCIS Newsletter and many me if you have any suggestion and ideas Proceedings of Congresses and Meetings F. J. Vargas about new possible activities. They all will and Reports and the organization of two FAO-CIHEAM Former Nut Network Coordinator be carefully assessed. international courses. In my opinion, we IRTA Mas de Bover should be rather satisfied about the work [email protected] We have also planned, with the help of we have been able to make. CIHEAM, to organise the ÇIII International

4 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 industry. These activities depend on several factors which need to be integrated in order to succeed. Some of these factors evolve quickly and it is necessary to adapt them to the changing conditions. The adaptation ability can make the difference between profit and loss.

Regarding the importance of the nut sector, FAO (REU, RNE and AGPS) called in 1990 an Expert Consultation with the basic aim of fostering cooperation between countries in Europe, North Africa and Near East on R&D issues in tree nuts. From the consultation the ÇInterregional Cooperative Research Network on NutsÈ was established. Promotion of information exchange, joint applied research, exchange of germplasm and establishment of links between researchers were identified as the main objectives. Member institutions and researchers come from the 41 European and 30 Near East Region FAO member nations. Some Network members also come from outside of the two Regions. In 1996, FAO and CIHEAM agreed to cosponsor the Network. The Young almond orchard growing on the slopes on Sierra de Maria, Andalucia, Spain. Network is part of the ESCORENA (Eu- ropean System of Cooperative Resear- Course on Economics and Nut produc- motion of information exchange, joint ch Networks in Agriculture) scheme of tionÈ. You will be timely informed about it applied research, exchange of germ- FAO (REU). Regarding NUCIS, our information bulle- plasm and establishment of links between tin within the FAO-CHEAM Network, I researchers were identified as the main It is interesting to note that before the es- would like to encourage you to send rele- objectives. In 1996, FAO and CIHEAM tablishment of this Network CIHEAM was vant contributions (articles, reports, notes agreed to cosponsor the Network. CI- already involved in related activities re- and news, congresses and meetings and HEAM was already involved in fostering garding nut trees. Thus GREMPA bibliography) to the editor to be publis- research nut tree activities. Since the («Groupe de Recherches Méditerranéen- hed. As you know, the NUCIS Bulletin is start of its activities, the Network received nes pour l’Amandier et le Pistachier» / the main vehicle of information among us widespread support from different natio- Mediterranean Research Group for Al- within the Network. Thus, the contributi- nal and international institutions. Particu- mond and Pistachio) is a Working Group ons that you may provide are essential to lar support has been received from the coordinated by CIHEAM since the seven- publish new issues. Spanish authorities, through INIA and ties. During the eighties and the begin- IRTA institutes. The main activities ca- ning of the nineties, part of these activi- M. Rovira rried out so far were: a) organization of ties were carried out in collaboration with FAO-CIHEAM Nut Network Coordinator specific meetings and workshops; b) pro- the AGRIMED programme of the DG VI of IRTA Mas de Bover motion of R+D activities and support of the European Commission. During the [email protected] training grants for young researchers; c) years 1990-96, a research project coordi- edition of inventories of germplasm and nated by CIHEAM was financed within research lines, proceedings and reports; the framework of the CAMAR programme d) edition of the NUCIS Newsletter and e) of the European Commission. organization of two international training REPORT OF THE FAO-CIHEAM courses on ÇNut Production and Eco- From the start, the Nut Network has re- INTERREGIONAL nomyÈ. Unfortunately, since 2002, due to ceived wide support from research orga- COOPERATIVE RESEARCH a strong cut in the financial support recei- nizations of several European and non NETWORK ON NUTS ved by FAO (REU), the activity of the Net- European countries. Particularly, two work on Nuts has had to be unavoidably Spanish institutes have supported consis- and considerably reduced. tently the Network: the Instituto Nacional FAO CIHEAM de Investigación y Tecnología Agraria y Nut Network AIM Alimentaria (INIA) and the Institut de Re- In this overall report from the start of our cerca i Tecnologia Agroalimentàries (IRTA). Network in 1990 to 2006, an account of In addition, several international institu- objectives, background, organization and tions like the International Plant Genetic activities of the Network on Nuts is pre- Resources Institute (IPGRI) has provi- sented. ded wide support in its subject over the years. Also, close collaboration with the SUMMARY BACKGROUND AND AIMS ISHS (International Society for Horticul- The Network on Nuts was established in Most countries in the Mediterranean re- tural Sciences) has been carried out du- 1990, after an expert consultation organi- gion have important economic activities ring the organization of several Sympo- zed by FAO (REU, RNE and AGPS). Pro- related to nut crop production, trade and sia.

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 5 Table 1. Technical Meetings.

Date Venue Meeting

1990, June Yalova (Turkey) Expert Consultation on the Promotion of Nut Production in Europe and Near East Regions. Nut Network establishement 1991, October Tarragona (Spain) International Walnut Meeting Meetings of the Subnetworks on Walnut and Pecan Meeting of the Coordinaton Board 1992, September Alba (Italy) Third International Congress on Hazelnut Meeting of the Subnetwork on Hazelnut 1993, May Agrigento (Italy) First International Almond Congress Meeting of the Subnetwork on Almond 1993, May Sciacca (Italy) IX GREMPA Pistachio Meeting Meeting of the Subnetwork on Pistachio 1993, October Spoleto (Italy) International Congress on Chestnut Meeting of the Subnetwork on Chestnut 1993, November Antalya (Turkey) Second Pecan Subnetwork Meeting 1994, September Adana (Turkey) First International Symposium on Pistachio Nut Meeting of the Subnetwork on Pistachio Meeting of the Coordinaton Board 1995, June Zaragoza (Spain) Meeting of the Subnetwork on Economics 1995, June Alcobaça (Portugal) Third International Walnut Congress Meetings of the Subnetworks on Walnut and Genetic Resources Meeting of the Coordinaton Board 1995, June Palermo (Italy) IPGRI-FAO Workshop on Genetic Resources on Pistacia 1995, September Rafsanján (Iran) First National Workshop on Pistachio Nut 1995, November Madrid (Spain) Meeting of the Subnetwork on Stone Pine 1996, July-August Ordu (Turkey) Fourth International Hazelnut Congress Meetings of the Subnetworks on Hazelnut and Genetic Resources 1996, October Meknes (Morocco) X GREMPA Meeting (Almond and Pistachio) Meetings of the Subnetworks on Almond and Pistachio Meeting of the Coordinaton Board First Technical Consultation of the Network 1996, December Zaragoza (Spain) Meeting of the Subnetwork on Economics 1997, October Roma (Italy) Meeting of the Coordinaton Board 1998, June Braunschweig (Germany) IPGRI-ECP/GR-FAO European Symp. on Plant Gen. Res. for Food and Agric. 1998, October Bordeaux (France) Second International Symposium on Chestnut Meeting of the Subnetwork on Chestnut 1998, December Irbid (Jordan) IPGRI Workshop Genetic Resources on Pistacia 1999, September Sanliurfa (Turkey) XI GREMPA Meeting (Almond and Pistachio) Meetings of the Subnetworks on Almond and Pistachio 1999, September Bordeaux (France) Fourth International Walnut Symposium Meeting of the Subnetwork on Walnut 2000, February Valladolid (Spain) First International Stone Pine Symposium Meeting of the Subnetwork on Stone Pine 2001, April Zaragoza (Spain) Meeting of the Coordinaton Board 2001, May Zaragoza (Spain) III Intern. Symposium on Pistachios and Almonds and XII GREMPA Meeting Meetings Subnetworks on Almond, Pistachio, Gen. Resources and Economics 2003, May Mirandela (Portugal) XIII GREMPA Meeting (Almond and Pistachio) 2004, June Tarragona (Spain) Fifth International Hazelnut Congress 2004, October Chaves-Vila Real (Portugal) Third International Symposium on Chestnut 2004, November Sorrento (Italy) Fifth International Walnut Congress 2005, May Tehran Fourth International Symposium on Pistachio and Almonds

6 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Around 2001, due to budgetary restrictio- ns, FAO (REU) started a revision process of its policy of support to the networks included in ESCORENA, analysing the activities done and their interest. In a report presented in 2003, during the 33rd Ses- sion of the European Commission on Agriculture, the interest of the activities developped by the Network on Nuts was acknowledged and the Network was included among those considered to have a good potential for continuity also in the future. However, the budgetary restrictio- ns from FAO (REU) in relation to ESCO- RENA, have not been solved yet. This fact has had a very negative influence on the normal functioning of the Network on Nuts the last years.

STRUCTURE AND ORGANIZATION The Network structure is based in a Coor- dination Centre (Coordinator and Secre- tary), supported by different Subnetworks (Working Groups) having the mission of fostering and coordinate specific activities. IRTA Mas Bové has been the Coor- dination Centre from the start of the Net- work activities. Until April 2001 there were 9 Subnetworks (7 tree crop species and 2 disciplinary): Almond, Hazelnut, Walnut, Pistachio, Pecan, Chestnut, Sto- Almond nursery near Baza, Granada, Spain. ne Pine, Genetic Resources and Econo- mics. From this date, and due mainly to budget limitations, the number of Subnet- A short account of the cooperative activi- Publications works were reduced to 6, including al- ties and achievements is presented. The Network publishes directly or has ac- mond and stone pine activities within the tively participated on the edition of the fo- general coordination. Two representati- Technical Meetings llowing publications (Annex 1): ves, one from each supporting institution There has been collaboration with diffe- (FAO and CIHEAM) are also integrated in rent institutions for the organization of in- NUCIS Newsletter managing the Network. ternational meetings (Congresses, Sym- The Coordination Centre publishes year- posia, Meetings and Workshops). Since ly, since 1993, the NUCIS Newsletter The general activities of the FAO-CI- 1990, over 30 meetings on nut tree which is distributed world-wide for free to HEAM Research Nut Network are propo- crops, genetic resources and economics 1.400 readers from over 60 countries. sed, discussed, agreed and planned in have been organized, alone or jointly This information bulletin presents the the Technical Consultations (participation with other institutions like ISHS, IPGRI, Network activities and planning, plus arti- of representatives of the member coun- etc (Table 1). Financial support was gi- cles and reports on nut research in the tries) and at the Coordination Board mee- ven to some experts to participate in world. In addition, it includes sections on tings (FAO and CIHEAM Officers, Net- them. Close collaboration in editing most news and notes, congresses and mee- work Coordinator and Subnetwork Liai- of the corresponding proceedings has tings, events to be held and recent biblio- son Officers). Two Technical Consultatio- been made. graphy. ns (Turkey, 1990 and Morocco, 1996) and six Coordination Board Meetings Promotion of R&D activities Apart from the printed version, the whole (Spain 1991, Turkey 1994, Portugal 1995, A major issue of the Nut Network is the collection of the 13 issues of the NUCIS Morocco 1996, Italy 1997 and Spain work carried out on genetic resources: Newsletter are placed, in digital version, 2001) have already been held. conservation, characterization and use. in the website: Apart from the exchange of plant material http://www.iamz.ciheam.org/en/pages/ ACTIVITIES between several institutions in various paginas/pag_investigacion2a.htm The main activities carried out during the species, a number of activities listed be- fifteenth running years have been: low have been developed. Another major Proceedings and Reports ¥ Organization of specific meetings issue of the Network is its Subnetwork on A list of the main references is presented and workshops Economics. This working group analyses in Annex 1. ¥ Promotion of R&D activities issues related to science/society on pro- ¥ Edition of proceedings and reports duction, marketing, trade and consump- ¥ Edition of inventories of germplasm tion. It can be pointed out also, in the Inventories of Germplasm Resources and and research lines field of joint applied research, that one Research Activities ¥ Edition of the NUCIS Newsletter project CAMAR CE/DG.VI («Amélioration A reference list is given in Annex 1. ¥ Organization of two international d’espèces à fruits à coque: noyer, aman- courses on ÇNut Production and EconomyÈ dier, pistachierÈ) was succesfully carried Species descriptors list ¥ Training grants for young researchers out. Main contributions are listed in Annex 1.

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 7 Training Advanced Courses Aletà, N.; Girona, J.; Tasias, J. (eds). ISHS. Acta Horticulturae 494, 528 pp. On the initiative of the Research Network 1993. Proceedings of the ÇInternational (Number of papers: 78). Walnut MeetingÈ. Tarragona, Spain, on Nuts, two Advanced International 1991. ISHS. Acta Horticulturae 311, 315 Catalán, G. (ed). 2000. Proceedings of Courses on ÇNut Production and Econo- pp. (Number of papers: 39). the «Primer Simposio del Pino Piñonero micsÈ have been organized by CIHEAM- (Pinus pinea L.)È. Valladolid, Spain, IAMZ, FAO, and some national institutio- Tuzcu, O.; M. Kaplankiran, M. (eds.). 2000. Published by Junta de Castilla y ns. Both courses were designed for gra- 1994. Proceedings of the ÇSecond Pe- León, Valladolid. Tomo I, 340 pp. and can Subnetwork MeetingÈ. Antalya, Tomo II, 438 pp. (Number of papers: duates working in R&D and professionals Turkey, 1993. Published by FAO, Uni- 80). of the sector. Each course was attended versity of Çukurova and MAFRA, 115 by some 30 students from over 15 coun- pp. (Number of papers: 12). Germain, E. (ed). 2001. Proceedings of tries of the Mediterranean Basin, Near the ÇFourth International Walnut Sym- Kaska, N.; Kuden, A. B.; Ferguson, L.; posiumÈ. Bordeaux, France, 1999. East and Central and South America. Michailides, T. (ed). 1995. Proceedings ISHS. Acta Horticulturae 544, 615 pp. Students gave a good grade to the overall of the ÇFirst International Symposium (Number of papers: 84). outcome of both courses. The first course on Pistachio NutÈ. Adana, Turkey, was held at Reus, Spain in November 1994. Acta Horticulturae 419, 425 pp. Ak, B.E. (ed). 2001. Proceedings of 1994 and the Second was held at Adana, (Number of papers: 71). the ÇXI GREMPA Seminar on Pista- chios and AlmondsÈ. Sanliurfa, Tur- Turkey in May 1998. A third course is be- Catalan, G. (ed.). 1996. Proceedings of key, 1999. Cahiers Options Méditerra- ing planned. the «I Reunión de la Red de Frutos Se- néennes, vol 56, 414 pp. (Number of cos de La FAO sobre el Pino Piñonero: papers: 69). Fellowships El Pino Piñonero como árbol productor de frutos secos en los países mediterrá- Two CIHEAM fellowships for short stays Oliveira, M.M. and Cordeiro, V. (eds). neosÈ. Madrid, Spain, 1996. Published 2005. Proceedings of the ÇXIII GREM- (3 months) at IRTA Mas de Bover (Cons- by INIA, Madrid, 148 pp. (Number of pa- PA Meeting on Almonds and Pista- tantí, Spain) were granted to students pers: 12). chiosÈ, Mirandela (Portugal), 2003. Op- from Adana University (Turkey, 1998) and tions Méditerranéennes, Série A, 63, from Ecole National d’Agriculture (Mek- Germain, E. (ed). 1997. Amélioration 408 pp. (Number of papers: 57). d’espèces à fruits à coque: noyer, nes, Morocco, 1999) for training on al- amandier, pistachier. (Résultats de re- mond breeding. Inventories of Germplasm Resources cherches menées au titre du Program- and Research Activities me CAMAR CE/DG.VI, contrat 8001- CT90-0023). Options Méditerranéen- Monastra, F; Raparelli, E. 1997. Inven- F. J. Vargas nes, Serie B 16, 147 pp. (Number of pa- tory of almond research, germplasm FAO-CIHEAM Former Nut Network pers: 8). and references. Published by FAO and Coordinator CIHEAM, Roma. REUR Technical Se- IRTA Mas de Bover Gomes Pereira, J.A., Martins, J.M.S., ries 51, 232 pp. Crta. Reus-El Morell, Km 3,8 Pinto de Abreu, C. (eds). 1997. Procee- E-43120 Constantí (Tarragona), Spain dings of the ÇThird International Walnut Koksal, A.I. 2000. Inventory of hazel- Tel: (34) 977 32 84 24 / 977 32 66 91 Congress». Alcobaça, Portugal, 1995. nut research, germplasm and referen- Fax: (34) 977 34 40 55 ISHS. Acta Horticulturae 442, 445 pp. ces. Published by FAO and CIHEAM, E-mail: [email protected] (Number of papers: 70). Roma. REU Technical Series 56, 129 pp. Koksal, A. Ï., Okay, Y. and Gunes, N.T. (eds). 1997. Proceedings of the ÇFourth Bounous, G; Barrel, A.; Beccaro, G.; International Symposium on HazelnutÈ. Lovisolo, C.; Gomes Pereira, J.A. Ordu, Turkey, 1996. ISHS. Acta Horti- 2001. Inventory of chestnut research, culturae 445, 556 pp. (Number of pa- germplasm and references. Published pers: 72). ANNEX 1. PUBLICATIONS by FAO and CIHEAM, Roma. REU Te- chnical Series 65, 174 pp. CIHEAM (ed). 1998. Proceedings of the NUCIS Newsletter ÇX GREMPA SeminarÈ. Meknes, Moro- Germain, E. 2004. Inventory of walnut cco, 1996. Cahiers Options Méditerra- research, germplasm and references. Batlle, I (ed.). NUCIS Newsletter num- néennes 33, 237 pp. (Number of pa- Published by FAO and CIHEAM, ber 1, April 1993, 12 pages. Number 2, pers: 36). Roma. REU Technical Series 66, 264 April 1994, 20 pages. Number 3, Janua- pp. ry 1995, 24 pages. Number 4, Decem- Albisu, L.M. (ed). 1999. Proceedings of ber 1995, 28 pages. Number 5, Decem- the ÇSeminar of the Subnetwork on Currently in preparation: Ak, B.E and ber 1996, 36 pages. Number 6, Decem- Economics of the FAO-CIHEAM Net- Kaska, N. Inventory on pistachio re- ber 1997, 52 pages. Number 7, Decem- work on NutsÈ. Economics of nuts in search, germplasm and references. ber 1998, 44 pages. Number 8, Decem- the Mediterranean basin. Zaragoza, ber 1999, 48 pages. Number 9, Decem- Spain, 1996. Options Méditerranéen- ber 2000, 68 pages. Number 10, Dec- nes, Serie A 37, 135 pp. (Number of Descriptors ember 2001, 48 pages. Number 11, De- papers: 10). cember 2002, 48 pages. Number 12, Barone, E., Padulosi, S., Van Mele, I. September 2004, 52 pages. Number 13, Batlle, I.; Monastra, F.; Koksal, A.I.; (eds.). 1997. Descriptors for Pistachio December 2006, 72 pages. Germain, E.; Kaska, N.; Tuzco, Ö.; (Pistacia vera L.). Published by IPGRI, Gomes Pereira, J. A.; Catalan, G.; Roma, Italy, 51 pp. (Contribution: Pista- Proceedings and Reports Vargas F.J. 1999. Nut tree genetic re- chio and Genetic Resources Subnet- sources conservation and documenta- works). Menini, U.G.; Olez, H.; Buyukilmaz, M.; tion in Europe and the Mediterranean Ozelkok, S. (eds.). 1991. Proceedings region. European symposium on plant Kaska, N., Van Mele, I. Padulosi, S. of the ÇExpert Consultation on the Pro- genetic resources for food and agri- (eds). 1998. Descriptors for Pistacia motion of Nut Production in Europe and culture, Braunschweig (Germany), spp. (excluding Pistacia vera L.). Pu- the Near East RegionsÈ. Nut Production 1998. Published by IPGRI, Rome, blished by IPGRI, Roma, Italy, 48 pp. and Industry in Europe, Near East and Italy: 343. (Contribution: Pistachio and Genetic North Africa. Yalova, Turkey, 1990. Pu- Resources Subnetworks). blished by FAO and MAFRA. FAO, Salesses, G. (ed). 1999. Proceedings of REUR Technical Series B, 411 pp. the ÇSecond International Symposium Currently in preparation: Descriptors for (Number of papers: 31). on ChestnutÈ. Bordeaux, France, 1998. Hazelnut, IPGRI-CIHEAM.

8 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 FOUR NEW ALMOND Table 1. Almond growing surface (ha) per regions in Spain in 2003. VARIETIES RELEASED Regions Non-irrigated Irrigated Total BY IRTA: ‘VAYRO’, ‘MARINADA’, ‘CONSTANTÍ’ Andalusia 178.779 10.391 189.170 AND ‘TARRACO’ Valencia’s region 104.344 9.278 113.622 Aragón 70.926 3.299 74.225 Murcia 66.420 7.586 74.006 Balearic Islands 62.252 380 62.632 INTRODUCTION Catalonia 58.936 2.790 61.726 An almond scion breeding programme by Castilla-La Mancha 44.944 2.347 47.291 controlled crosses was started at IRTA La Rioja 9.615 172 9.787 Mas de Bover in 1975 and it has been ac- Navarra 2.996 856 3.852 tive since. The main aims are to obtain Extremadura 2.500 450 2.950 new varieties having late blooming (to Castilla and León 1.685 6 1.691 avoid spring frost damage), self-fertility Others 735 1 736 (to reduce pollination drawbacks), high TOTAL 604.132 37.556 641.688 producing capacity, kernel quality, easy training and pruning, vigour, and resistan- Source: MAPA, Anuario de Estadística Agroalimentaria (2004). ce to diseases and droughtness. Some 35.000 seedlings have been raised and a few cultivars selected mostly using early Table 2. Origin. selection techniques (Vargas et al., 2.005). In 1992, the three first released Variety Origin Crossing year cultivars were registered, ‘Masbovera’, Glorieta’ and ‘Francol’’ (Vargas and Ro- ‘Vayro’ ‘4-665’ x ‘Lauranne’ 1991 mero, 1994), which have been widely ‘Marinada’ ‘Lauranne’ x ‘Glorieta’ 1994 planted in Spain and in other Mediterra- ‘Constantí’ ‘FGFD2’ (op) 1993 nean countries. Recently, four new promi- ‘Tarraco’ ‘FLTU18’ x ‘Anxaneta’ 1991 sing cultivars ‘Vayro’, ‘Marinada’, ‘Cons- tantí’ and ‘Tarraco’ have been selected, and are in the process of being registered, having a range of outstanding featu- Almond production in Spain was establis- ric Islands, Catalonia and Castilla-La res (Figures 1 to 8). Apart from the three hed in Roman times and it is based in Me- Mancha (Table 1). Almond orchards are previous cultivars, the four new varieties diterranean areas mainly: Andalusia, mostly placed in marginal land (small fa- are already commercially available. Valencia’s region, Aragón, Murcia, Balea- rms). Spanish’s current growing surface

Tabla 3. Blooming date, self-fertility, S incompatibility genotypes and production (yield potential and precocity).

Variety Blooming date1 Self-fertility S genotype2 Yield potential Precocity

New:

‘Vayro’ 28 Yes S9Sf Very high Early

‘Marinada’ 37 Yes S5Sf Very high Very early

‘Constantí’ 29 Yes S3Sf High-very high Early

‘Tarraco’ 38 No S1S9 Very high Very early Reference:

‘D. Largueta’ 0 No S1S25 Mid-high Mid-late

‘Ferragnes’ 30 No S1S3 High-very high Mid

‘Guara’ 29 Yes S1Sf High-very high Early 1 Average blooming date at Mas de Bover, as number of days from ‘Desmayo Largueta’ full bloom (mean: 30 January). Mean 8 years dates (1998-2005). 2 López et al. (2006).

Table 4. Tree vigour, growth habit, branching density and training and pruning ease.

Variety Vigour Growth habit Branching density Training and pruning

New: ‘Vayro’ Very strong Medium Mid Very easy ‘Marinada’ Mid Medium-upright Mid Very easy ‘Constantí’ Strong Medium-upright Mid Very easy ‘Tarraco’ Mid Medium-upright Mid Very easy Reference: ‘D. Largueta’ Mid Spreading Mid-high Medium ‘Ferragnes’ Strong Medium-upright Mid Very easy ‘Guara’ Mid Drooping Scarce Difficult

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 9 Figure 1. ‘Constantí’ tree. Figure 2. ‘Constantí’ fruiting habit.

Figure 3. ‘Marinada’ tree. Figure 4. ‘Marinada’ fruiting habit. is 641.000 ha, most being under rainfed BAS-CSIC and IRTA), mainly ‘Guara’, blooming. IRTA’s parents are vigorous, conditions (300-650 mm per year) and ‘Masbovera’, ‘Glorieta’, ‘Francol’’, ‘Anto– have good growth habit, are productive only 6% of this surface is irrigated (Var- eta’ and ‘Marta’, have been widely plan- and show good kernel appearance. ‘Lau- gas et al., 2006). However, recently a ted. Currently, there is a trend towards ranne’ is self-compatible, high yielding large improvement of: varieties and using recently bred late blooming and and shows good growth habit. ‘FLTU18’ rootstocks, orchard management, har- self-compatible cultivars. Almond growers is self-fertile and the tree has good sha- vest and postharvest conditions is taking have a rising interest in newly released pe. ‘FGFD2’ flowers very late. place. cultivars by IRTA, like self-fertile ‘Vayro’, ‘Constantí’, ‘Marinada’ or self-incompati- On Tables 3, 4 and 5 some important Almonds produced in Spain are all hard ble ‘Tarraco’. agronomic and commercial traits are gi- shelled having kernel percentages from ven. Three widely grown reference culti- 25 to 40%. The two major traditionally ORIGIN AND DESCRIPTION vars are included ‘Desmayo Largueta’, grown almond cultivars are early flowe- In the Table 2 their origin of the four new ‘Ferragnes’ and ‘Guara’. ring: ‘Marcona’ and ‘Desmayo Largueta’. cultivars is presented. As genitors there More recently, in the last 25 years, some are three parents from IRTA (‘Anxaneta’, ‘Marinada’ y ‘Tarraco’ are very late late flowering French cultivars, like ‘Fer- ‘Glorieta’ and ‘4-665’) and three from blooming (some 8 days after ‘Guara’ at ragnes’, and some Spanish releases from INRA (‘Lauranne’, ‘FLTU18’ and ‘FGFD2’). Mas de Bover) and, ‘Vayro’ and ‘Constan- public breeding programmes (CITA, CE- All six genitors are late or very late tí’ flower late (similar to ‘Guara’). In addi-

10 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Figure 5. ‘Tarraco’ tree. Figure 6. ‘Tarraco’ fruiting habit.

Figure 7. ‘Vayro’ tree. Figure 8. ‘Vayro’ fruiting habit. tion, ‘Vayro’, ‘Marinada’ and ‘Constantí’ nut removal). ‘Vayro’ and ‘Tarraco’ seem ‘Vayro’, ‘Marinada’ and ‘Constantí’, com- are self-compatible with a high level of to be tolerant to Phomosis amygdali (Çfu- pared with ‘Guara’ and ‘Lauranne’ (consi- self-fertility (high fruit set in bagged bran- sicoccumÈ). dered highly productive). It is outstanding ches). Instead ‘Tarraco’ is self-incompati- the producing consistency of ‘Vayro’ and ble and thus needs cross pollination All All four cultivars show good nut featu- ‘Marinada’ and the prococity of ‘Marina- four cultivars shown high production ca- res: hard shell, with near absence of da’. pacity. ‘Marinada’ and ‘Tarraco’ are pre- double kernels and having attractive cocious (Table 3). ‘Vayro’ is early matu- kernel (Table 5). ‘Tarraco’ has a big ker- USE ring and the others are mid-season har- nel size. The almond shells are hard ‘Vayro’, ‘Marinada’, ‘Constantí’ and ‘Ta- vesting. and are well suited to the European in- rraco’ have shown good and consistent dustry, based on hard shelled cultivars. results in experimental trials. About to ‘Vayro’ outstands for being vigourous. All It is also interesting to point out that the make new almond plantings and design four show mid to upright tree vegetative almond hard shell trait is reducing worm accordingly to flowering time and vigour growth, except ‘Vayro’ which is medium, damage and thus preventing aflatoxin (related to tree spacing), two cultivar and mid branching density, bearing nuts contamination. pairs can be considered: ‘Marinada’ and mainly on spurs (Table 4). The four culti- ‘Tarraco’, very late blooming and mid vi- vars are easy to train and prune and, well Table 6 shows the productive performan- gour and, Vayro’ and ‘Constantí’, late adapted to mechanical harvesting (easy ce of a trial about self-fruitful cultivars: blooming and strong vigour.

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 11 Table 5. Nut characteristics. Mean values of samples analyzed during (7-30 years), ‘BELONA’ AND ‘SOLETA’, TWO kernel yield (shelling percentage, %), double kernels (%) and note on kernel NEW AUTOGAMOUS ALMONDS appearance (scale 1-9, with 9 highest mark).

Variety Kernel Kernel Double Kernel weight yield kernels appearance INTRODUCTION ‘Belona’ and ‘Soleta’ are two new almond New: (Prunus amygdalus Batsch) cultivars ‘Vayro’ 1,24 28,8 0,1 7,1 from the breeding program of the Unidad ‘Marinada’ 1,34 31,2 0,4 6,7 de Fruticultura, CITA de Aragón, charac- ‘Constantí’ 1,22 26,7 1,6 6,0 terized by their kernel quality and by the ‘Tarraco’ 1,71 31,6 0,1 6,8 possibility of becoming a commercial al- Reference: ternative to the two traditional almond cultivars in the Spanish market, ‘Marco- ‘D. Largueta’ 1,34 27,2 1,4 6,7 na’ and ‘Desmayo Largueta’, to which ‘Ferragnes’ 1,49 33,8 0,1 6,4 they are comparable in their aspect and ‘Guara’ 1,31 34,7 11,9 6,2 industrial quality, but differing from them in their late blooming date, tolerance to Table 6. Mean and accumulated production of kernel (kg/tree). frosts and, mainly, self-compatibility. Trial at Corbins, Lleida, under deficit irrigation conditions. Trees planted in 1995 and regrafted in 2000. The selection of a new fruit cultivar is a Randomized blocs design, 3 repetitions and 5 trees per plot. long process where many traits must be studied in order to consider any seedling Variety 2002 2003 2004 2005 Accumulated for further evaluation and final release as production (2002-2005) a cultivar. Since the beginning of almond research in Zaragoza in 1966, the most New: important problem detected in almond ‘Vayro’ 0,61 4,03 5,39 5,53 15,56 growing in Spain was its low productivity, ‘Marinada’ 1,67 5,15 2,50 5,23 14,54 very often not reaching 125 kg/ha, which, ‘Constantí’ 0,58 3,73 2,24 4,60 11,14 evidently, does not cover the minimum growing costs (Felipe, 1984). This low Reference: productivity is mainly due to three cau- ‘Guara’ 0,32 3,61 1,98 5,22 11,14 ses: the occurrence of frosts at blooming ‘Lauranne’ 0,47 3,98 3,90 5,30 13,66 time or later, deficient pollination and drought (Socias i Company, 2001), as almond is mostly grown in non irrigated conditions (Felipe, 2000). REFERENCES

López, M., Vargas, F. J. and I. Batlle, Frost incidence is important in all inland 2006. Self-(in) compatibility almond geno- growing regions, but also has relevance types: a review. Euphytica, 150: 1-16. in some coastal zones. As a consequence, late blooming was considered a very MAPA 2006, Anuario de Estadística Agroalimentaria (2004). www.mapa.es interesting trait in almond, a species characterized by early blooming time. As Vargas, F.J., and M.A. Romero. 1994. most Spanish cultivars are early ‘Masbovera’, ‘Glorieta’ and ‘Francolí’, blooming, the first task was the introduc- three new almond varieties from IRTA. tion of late blooming cultivars, mainly Acta Horticulturae 373: 75-82. from France, Italy and the then Soviet Figure 1: Almond producing areas in Spain, Vargas, F.J., M.A. Romero, J. Clavé, I. Union. Late blooming has two main ad- importance according to colour intensity Batlle 2005. Early selection in the al- vantages: to avoid early frosts and for mond breeding programme at IRTA blooming to happen when temperatures AVAILABILITY Mas Bové. Options Méditerranéennes, Série A, 63: 17-22. are higher and, thus, more favourable for The four varieties bred by IRTA are in the pollination and fruit set. process to be registered by the Oficina Vargas, F.J., M.A. Romero, I. Batlle. EspaÐola de Variedades Vegetales 2006. Almond and its uses in Spain. The deficient pollination was due to the (OEVV) belonging to the Spanish Minis- (Editors: D. Avanzato and I. Vasallo). absence or low presence of pollinating Following Almond Footprints (Amygda- tery of Agriculture, Fisheries and Food lus communis L.) across Sicily. Cultiva- insects, to bad weather conditions distur- (MAPA). The varieties can only be propa- tion and Culture, Folk and History, Tra- bing bee flight, to an incorrect distribution gated under royalty agreements with ditions and Uses (in press), Italy. of cultivars in the orchard, or to a low IRTA. IRTA has granted a multiplication overlapping of cultivars blooming, as ob- license for the four new varieties to served in many orchards where the two ALMERIPLANT (www.almeriplant.com). F. Vargas, M. Romero, J. Clavé, main Spanish cultivars, ‘Marcona’ and J. Vergés, J. Santos, I. Batlle ‘Desmayo Largueta’, were planted without ACKNOWLEDGEMENTS IRTA, Mas de Bover considering their low bloom overlap (Fig. Part of this scion breeding programme Mediterranean Fruit Trees 1). As a consequence, the main aims of was funded by three INIA (Spanish Mi- Ctra. Reus-El Morell, km 3.8, E-43120 Constantí,Tarragona, Spain the breeding program (Felipe and Socias nistry of Science and Technology) pro- E-mail: [email protected] i Company, 1985) was the development jects (SC97-049, RTA01-081 and of late-blooming and self-compatible cul- RTA04-030). tivars, at the same time that a rootstock

12 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 breeding program was attempting to obtain new rootstocks that were well adapted to irrigated and non-irrigated conditions (Gómez Aparisi et al., 2001).

The first three cultivars released were ‘Aylés’, ‘Guara’ and ‘Moncayo’ (Felipe and Socias i Company, 1987), ‘Guara’ having represented more than 50% of the new almond orchards in recent years (MAPA, 2002). Later three more cultivars were registered in 1998, ‘Blanquerna’, ‘Cambra’ and ‘Felisia’ (Socias i Company and Felipe, 1999), ‘Blanquerna’ being of very good productivity and kernel quality, and ‘Felisia’ of very late blooming time (Fig. 1). Breeding, however, has continued to increase the number of favourable traits in the new selections, although no single cultivar can be considered as the one covering all the requirements in all conditions (Socias i Company et al., Young almond orchards growing in Almería, Spain. 1998). A main requirement, as in most almond breeding programmes (Socias i Company, 2002) is autogamy. Kernel peatedly utilized in the breeding crosses, fruit and kernel quality, ripening time and, quality, aiming at defining the best end- but their best offspring, as with Selection finally, kernel composition and industrial use of each cultivar (Socias i Company et E-5-7, rated as outstanding during eva- utilization. al., 2006), has been included in the last luation and was thus chosen as a parent evaluation steps taking into account the in this cross, but not released as the culti- AUTOGAMY preference of the Spanish market for var ‘Blanquerna’ (Socias i Company y Fe- Self-compatibility was tested as soon as ‘Marcona’ and ‘Desmayo Largueta’. lipe, 1999) until nine years later than the the original seedlings produced the first cross so far described. flowers, but autogamy was studied on the ORIGIN grafted trees over several years due to Both cultivars were obtained from artifi- Nuts were stratified, seedlings were plan- the large variability found between years cial pollinations following the steps of a ted in a bedding nursery and finally trans- in field trials for fruit set (Socias i Com- traditional breeding programme (Socias i planted to the field where the first evalua- pany et al., 2005). Table 1 shows the re- Company y Felipe, 1987). Both come tion was undertaken. Due to seedling sults of a single year for three pollination from the cross done in 1988 of Selection mortality in this field, the plants showing treatments: self-pollination, cross-pollina- E-5-7 (seedling of open pollinated ‘Gen- the best kernels were grafted on a peach tion with cross-compatible pollen of ‘Mar- co’, a self-compatible Italian cultivar) by x almond hybrid and plant at the edge of cona, and bagging, although the same the French cultivar ‘Belle d’Aurons’, cha- the almond germplasm collection repre- pattern of results was observed over all racterized by its kernels of excellent qua- sented by three trees, where evaluation years. lity. This cross was made with the aim of was deepened. The first traits considered utilizing a self-compatibility source other were self-compatibility, blooming time, Selections F-4-25 and F-4-43 showed a than ‘Tuono’, so far the mostly utilized pa- tree morphology, and fruit traits. Later very low level of self-compatibility, as rent in almond crosses (Socias i Com- other traits were included, such as auto- shown by the low fruit sets after self-pol- pany, 2002). Normally parents are not re- gamy, branching habit, flower density, lination and bagging, in agreement with

Figure 1: Average blooming dates of the new cultivars in comparison with the mostly grown. Percentages indicate the amount of open flowers.

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 13 pect, without double kernels. Shell is hard, adapted to the Spanish industry. Shape has been deeply considered, as ‘Soleta’ kernel is very similar to that of ‘Desmayo Largueta’, although not the nut (Fig. 2). After roasting, the tegument is very easily removed, a trait very peculiar of ‘Desmayo Largueta’. ‘Belona’ nut is very similar to that of ‘Marcona’, but the kernel is less round and thinner (Fig. 3), which gives very good results at blanching. Industrial cracking has shown good results, with very few broken kernels, with an outstanding behaviour of Figure 2. ‘Belona’ almonds. Figure 3. ‘Soleta’ almonds. ‘Belona’. In spite of the hard shell, kernel percentage is not so low because the ker- the previous observations on pollen tube 30%. This fact must be related to their nels fill the inner shell space. In 2005, growth, whereas the other six selections blooming time, which in Zaragoza takes when a small nut size was observed in showed a good self-pollen tube growth. place on average on March 2, a few days most growing regions, kernel size reduc- Thus, genetic self-compatibility, as shown before ‘Guara’, on March 5 (dates refer- tion was lower than nut size decrease. by a good pollen tube growth, also gave a red as 50% of open flowers). good result in the field, with fruit sets for The chemical analysis of the kernels of these six selections ranging between 22 Ripening time is later than in ‘Guara’, whi- the two new cultivars has shown very and 55%, resulting in acceptable yields ch allows the succession of harvest of the high contents in fatty acids, higher than in due to their high bloom density. different cultivars. The average ripening ‘Marcona’ and ‘Desmayo Largueta’, a date in Zaragoza is August 23 for ‘Guara’, very interesting trait for nougat produc- Selections F-4-9 and F-4-43 showed a September 8 for ‘Belona’ and September tion. The percentage of oleic acid, that of good fruit set after self-pollination, but not 17 for ‘Soleta’, following the same order higher quality in the lipid fraction, is espe- after bagging, due to their flower morpho- as ‘Marcona’ and ‘Desmayo Largueta’. cially high in ‘Belona’, over 75%. This cul- logy not allowing natural autogamy. The Nut fall before harvest was very low, but tivar also shows a very high content of to- four other selections continued their eva- nuts fell easily when shaken. copherols, although not so high as in luation due to their sufficient level of auto- ‘Marcona (Kodad et al., 2006), indicating gamy (Grasselly et al., 1981) which, toge- Tree formation has been easy. Adult a good storage quality as tocopherols ther with their bloom density, may ensure trees show a good equilibrium between have an important role in avoiding ranci- a commercial crop. As a consequence, vegetative growth and production, thus dity and in nutrition for their vitamin E ac- these two selections, as well as the pre- pruning may be reduced, more in ‘Soleta’ tivity. vious two, were eliminated from the eva- than in ‘Belona’. ‘Soleta’ is slightly sus- luation process, which only maintained ceptible to Polystigma, but ‘Belona’ is qui- DESCRIPTION the other four. te tolerant. Belona (Fig. 2) Origin: ‘Blanquerna’ x ‘Belle d’Aurons’. FIELD BEHAVIOUR Some external experimentations have Selection number: F-4-12. Clone 502. One important point considered was re- shown their good adaptation to different Number of register in OEVV 20054615 sistance to frosts. Especially important growing conditions, both in very cold cli- (27 October 2005). was the observation in 2003, with a frost mates (800 m at Aniñón, Zaragoza) and Tree: Drooping growth habit. Intermediate of -2,5¼C for five hours on March 18. milder ones (550 m at El Pinós, Alacant). vigour. Whereas cultivars considered as resistant Blooming and ripening dates measured in Flower: Late blooming time, 2-3 days be- to frosts such as ‘Guara’ (Felipe, 1988) these locations are earlier in El Pinós fore ‘Guara’. White flowers, of intermedia- suffered a yield reduction rated at 50%, than in Zaragoza, but later in Aniñón. te to large size, mainly on spurs and less the four selections under study suffered on one-year shoots. High bloom density. different levels of losses: in F-3-34 and F- KERNEL QUALITY Pollination: Autogamous, does not requi- 3-35 the decrease was similar to the le- Kernel quality has been a decisive crite- re cross-pollination, but may cross-polli- vels observed in ‘Guara’, but in ‘Belona’ rion in the selection process. Nuts and nate simultaneous blooming cultivars. and ‘Soleta’ the reduction was only of 20- kernels show a very good size and as- Fruit: Hard shell, without layers, heart- shaped. Shelling percentage 27-35 %. Kernel: Heart-shaped. Mean weight of 1.3 Table 1. Fruit sets obtained in eight almond selections depending g. Very sweet taste. Easy blanching. on the pollination treatment Observations: Flower morphology allows self-pollination. Easy tree formation. Pru- Selection Fruit set (%) ning requires some rejuvenation. Interes- Self-pollination Cross-pollination Bagging ting for the quality and composition of the kernel. The intermediate ripening time F-3-34 55.3 44.0 22.2 may allow a progressive harvesting of F-3-35 31.0 55.8 27.0 ‘Guara’, ‘Belona’ and ‘Soleta’. F-4-9 33.8 34.6 5.7 ‘Soleta’ 22.4 34.8 11.4 Soleta (Fig. 3) ‘Belona’ 38.4 48.1 10.9 Origin: ‘Blanquerna’ x ‘Belle d’Aurons’. F-4-25 1.8 11.5 0.0 Selection number: F-4-10. Clone 503. F-4-35 54.3 53.7 5.0 Number of register in OEVV 20054616 F-4-43 0.3 29,7 0.5 (27 October 2005).

14 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Almond orchards growing under rainfed conditions in Almería, south of Spain.

Tree: Drooping growth habit. Intermediate J.L. Espada and P. Castañer (Centro de vigour. Técnicas Agrarias de la DGA) and G. Val- terion in almond breeding. J. Hort. Sci. Flower: Late blooming time, 2-3 days be- dés (Estació Experimental Agrària, Elx) in Biotechnol. (in press). fore ‘Guara’. White flowers, of intermedia- the experimental orchards. MAPA, 2002. Web page of the Spanish te to large size, mainly on spurs and less Minsitry of Agriculture, Fisheries and on one-year shoots. High bloom density. Food. http//www.mapya.es/agric/pags/ Pollination: Autogamous, does not requi- semillas/vivero/almendro.pdf. REFERENCES re cross-pollination, but may cross-polli- Socias i Company, R., 2001. Almendro. nate simultaneous blooming cultivars. Felipe, A.J., 1984. Profitability of al- In: F. Nuez and G. Llácer (eds.): La hor- Fruit: Hard shell, without layers, elliptical. monds orchards in Spain. Acta Hort. ticultura española. SECH - Ed. Horticul- Shelling percentage 27-35 %. 155, 287-290. tura, Reus, pp. 271-274. Kernel: Elliptical. Mean weight of 127 g. Felipe, A.J., 1988. Observaciones so- Socias i Company, R., 2002. Latest ad- Very good taste. Easy peeling after roas- bre comportamiento frente a heladas vances in almond self-compatibility. ting. tardías en almendro. Rap. EUR 11557, Acta Hort. 591, 205-212. Observations: Flower morphology allows 123-130. self-pollination. Easy tree formation and Socias i Company, R., Felipe, A.J., Felipe, A.J., 2000. El almendro. I. El ma- pruning. Interesting for the quality of the 1987. La mejora genética del almendro. terial vegetal. Integrum, Lleida, 461 pp. Frutic. Prof. 11, 64-66. kernel and easy peeling when roasted. The medium-late ripening time may allow Felipe, A.J., Socias i Company, R., Socias i Company, R,, Felipe, A.J., a progressive harvesting of ‘Guara’, ‘Be- 1985. L’amélioration génétique de 1999. ‘Blanquerna’, ‘Cambra’ y ‘Felisia’: lona’ and ‘Soleta’. l’amandier à Saragosse. Options Médi- tres nuevos cultivares autógamos de al- terr. CIHEAM/IAMZ 85/I, 9-14. mendro. Inf. Técn. Econ. Agrar. 95V (2), 111-117. REGISTER AND AVAILABILITY Felipe, A.J., Socias i Company, R., These two cultivars have been presented 1987. ‘Aylés’, ‘Guara’, and ‘Moncayo’ Socias i Company, R., Felipe, A.J., Gó- to patent on 27 October 2005 at the Spa- almonds. HortScience 22(5), 961-962. mez Aparisi, J., García, J.E., Dicenta, nish Registry of Protected Cultivars and F., 1998. The ideotype concept in al- Gómez Aparisi, J., Carrera, M., Felipe, mond. Acta Hort. 470, 51-56. are available to nurseries though provi- A.J., Socias i Company, R., 2001. ‘Gar- sional licenses by GESLIVE. nem’, ‘Monegro’ y ‘Felinem’: nuevos pa- Socias i Company, R., Gómez Aparisi, trones híbridos almendro x melocotone- J., Alonso, J.M., 2005. Year and enclosu- ACKNOWLEDGMENTS ro resistentes a nematodos y de hoja re effects on fruit set in an autogamous para frutales de hueso. Inf. Técn.. almond. Scientia Hort. 104, 369-377. The long-term work to develop these cul- Econ. Agrar. 97V (3), 282-288. tivars has been funded by successive re- Socias i Company, R., Kodad, O., Alon- search projects of the Spanish INIA and Grasselly, C., Crossa-Raynaud, P., Oli- so, J.M., 2006. Mejora de la calidad en CICYT, most recently AGL2004-06674- vier, G., Gall, H., 1981. Transmission el almendro. III Congreso de Mejora du caractère d’aitocompatibilité chez Genética de Plantas, Valencia, 597-611. C02-01. We appreciate the technical work l’amandier. Options Méditerr. CIHEAM/ of the auxiliary personnel of Fruticultura, IAMZ 81/I, 71-75. mainly of J.M. Ansón, J. Búbal and A. Es- R. Socias i Company and A.J. Felipe cota, as well as the collaboration of the Kodad, O., Socias i Company, R., Unidad de Fruticultura, CITA de Aragón, industries «Frutos Secos Alcañiz» and Prats, M.S., López Ortiz, M.C., 2006. Apartado 727, Zaragoza, Spain Variability in tocopherol concentration in [email protected] ÇCastillo de LoarreÈ, the growers of the almond oil and its use as a selection cri- external fields and the collaboration of

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 15 THE ALMOND CULTIVAR Figure 1: Stylar ribonuclease interpretative diagram alfer NEpHGE migration ‘FRANCOLÍ’ IS conditions of eight almond cultivars. Lanes 1 to 8: 1. ‘Gabaix’ (S S ), 2. ‘Glorieta’ (S S ), 3. ‘Garbí’ (S S ), 4. ‘Cristomorto’ (S S ), SELF-COMPATIBLE 10 25 1 5 1 5 1 2 5. ‘Falsa Barese’ (S1S1), 6. ‘Tuono’ (S1S1), 7. ‘Francolí’ (S1S1) and 8. ‘Genco’ (S1S1). Dotted lines represent secondary bands.

INTRODUCTION Almond [Prunus amygdalus Batsch or Prunus dulcis Miller (D.A. Webb)] is widely grown in the Mediterranean region, Cali- fornia and Australia. As most cultivars are self-incompatible, cross-compatible cultivars having overlapping blooming times or self-fertile cultivars are planted in commercial orchards to ensure good fruit set and thus economic yields. The introduction of the self-compatibility trait, conferred by the dominant Sf allele, into commercial cultivars is pursued in the scion almond breeding programme of IRTA Mas de Bover and several others. A wide review of cultivar S genotypes can be found (101 seedlings) from the cross ‘Cristo- cultivar collection of IRTA Mas de Bover in López et al. (2006). morto’ x ‘Gabaix’ made at Mas de Bover, at Constantí and from trials of the IRTA from which ‘Francolí’ was believed to de- Estació Experimental of Lleida at Gime- The cultivar ‘Francolí’, released from rive, bloomed in average approximately nells were used. Tissue samples of six IRTA’s breeding programme in 1994 (Var- 17 days after ‘Cavaliera’, except for only cultivars with known S alleles were inclu- gas and Romero, 1994; Brooks and Olmo, 4 seedlings that flowered around 28 days ded for comparison in RNase and PCR 1997), is widely planted as pollinator of after ‘Cavaliera’, ‘Francolí’ being one of analysis (‘Cristomorto’, ‘Falsa Barese’, ‘Masbovera’ and ‘Glorieta’ in Spain. ‘Fran- them, early named as selection 3-361 ‘Garbí’, ‘Genco’, ‘Glorieta’ and ‘Tuono’). colí’ is a late blooming cultivar and heavy (Vargas et al., 1984). Second, consistent cropper producing hard shelled nuts. Cu- high fruit set of ‘Francolí’ in the field un- Pollination tests. Six field test crosses rrently, ‘Francolí’ is under trial in several der adverse climatic conditions during and pollination tests in the laboratory countries (USA, Chile and Australia) and blooming supported the possibility of be- were made to check some of the possible is being commercially propagated in Fran- ing self-compatible. In an experimental S genotypes according to ‘Francolí’ re- ce and Portugal. The cultivar ‘Francolí’ plot at Corbins, Lleida, ‘Francolí’ growing ported crossing origin (Table 1). Pollen was putatively raised from the cross along with other self-incompatible almond was collected from closed buds at balloon cultivars in 1998, was the only cultivar ‘Cristomorto’ (S1S2) x ‘Gabaix’ (S10S25) stage (Felipe, 1977) from male parents, made at IRTA Mas de Bover in 1976 (Var- that cropped in the absence of beehives. dried and stored at 4¼C. Pollen viability gas and Romero, 1994). The S incompati- Also, in a cultivar trial at Borges Blan- was checked before pollination using the bility genotype of ‘Cristomorto’ was first ques, Lleida, ‘Francolí’ trees gave good fluorochromatic test procedure (FCR) assigned by Crossa-Raynaud and Gras- nut production, after frosty conditions du- (Heslop-Harrisson et al., 1984). ‘Francolí’ selly (1985) using test crosses and later ring blooming time in 2003, as it happe- was selfed in the field and in the labora- confirmed by Boskovic et al. (1997) using ned with other self-compatible cultivars tory to test self-compatibility. Test cros- stylar ribonuclease analysis. ‘Gabaix’ was under trialling like ‘Guara’ or ‘Lauranne’. ses between cultivars were made one first genotyped as S5S10 (Boskovic et al., year (2003), except ‘Francolí’ which was 2003), but it has recently been relabelled This work, based on López et al., (2005), selfed in 2002 and 2003 (at two sites: as S10S25 (López et al., 2004b). As both reports the new self-compatible genotype Constantí and Gimenells). Both senses of parents were self-incompatible the possi- assigned to ‘Francolí’, an early conside- cross were tested for two of the crosses bility of ‘Francolí’ being self-fertile was ne- red self-incompatible cultivar obtained at (Table 1) in order to check the possibility ver tested. Recently, work by López et al., IRTA, using three different techniques of unilateral incompatibility, as reported 2005 re-classified it as self-compatible. In and the possible crossing origin is also for the cultivar ‘Jeffries’ (Kester et al., agreement with its reported origin and pa- discussed after microsatellites analysis. 1994b). rental genotypes, ‘Francolí’ was first sug- The stylar ribonuclease analysis raised at gested to be S1S5 by PCR analysis with first the question about ‘Francolí’ propo- Cross-compatibility assessment in the the conserved AS1II/AmyC5R primer sed S genotype and its reported origin. field and in the laboratory followed López (Martínez-Gómez et al., 2003). However, This indication was confirmed later by et al. (2004b). To assess ‘Francolí’ self- only the S1 allele was clearly and consis- controlled pollination tests and PCR and compatibility in the field, at least 100 flo- tently detected using the stylar ribonuclea- microsatellites analysis. In this article we wer buds were bagged before anthesis se analysis at IRTA, indicating the possibi- present the discovery of ‘Francolí’s self- with white cotton bags. Fruit set in the lity of ‘Francolí’ being self-compatible. In compatibility as a useful trait. In addition, field (number of fruits/number of pollina- addition, breeding records of two early the sketch of IRTA’s breeding plot, ted flowers) was recorded 60 days after crosses, ‘Francolí’ x ‘Garbí’ (S1S5) and showing the position of the ‘Cristomorto’ full bloom; when fruit set percentages

‘Francolí’ x ‘Glorieta’ (S1S5), set nuts indi- mother tree, is included. were above 4%, cultivars were conside- cating a different S genotype for ‘Francolí’ red cross-compatible. In the laboratory, and causing us to speculate that the re- MATERIALS AND METHODS the number of pistils compatible with pol- corded parentage would be incorrect. Plant material. Leaves, pollen or styles len from the male parent was divided by from cultivars (‘Achaak’, ‘Ardechoise’, the total number of pistils tested, and if The cultivar ‘Francolí’ shows some striking ‘Desmayo Largueta’ and ‘Francolí’) were two or more of the twelve pistils observed agronomic features. First, the offspring collected. Trees from both the almond (>17%) showed pollen tubes reaching the

16 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 ovary, cultivars were considered cross- Figure 2: PCR agarose gels of S alleles from eight almond cultivars. compatible. Genomic DNA was amplified using primer pair AS1II/AmyC5R (Tamura et al., 2000) (Fig. A) and primer pair SfF/R (Channuntapipat et al., 2003) (Fig. B). Stylar ribonuclease analysis. During A DNA ladder was used for size determination. three years (from 2001 to 2003), pistils were collected from flowers at balloon stage (Felipe, 1977) of trees growing in the field and immediately stored at -20¼C until use. Extraction of stylar proteins followed the method described by Boskovic et al. (1997). Stylar ribonucleases were separated applying Non Equilibrium pH Gradient Electrofocusing (NEpHGE) electrophoresis conditions, and gels were stained for ribonuclease activity following

Boskovic et al. (1999). The Sf allele is not detectable on the zymograms due to the absence of RNase activity (Boskovic et A. Primer pair AS1II/AmyC5R. Lanes 1 to 5: B. Primer pair SfF/R. Lanes 1 to 8: 1. 123 bp Ladder, 2. ‘Gabaix’ (S10S25), 1. PUC-Mix Ladder, 2. ‘Francolí’ (S1S1), al., 1997) and thus, provided fair resolu- 3. ‘Glorieta’ (S S ), 4. ‘Garbí’ (S S ), 3. ‘Gabaix’ (S S ), 4. ‘Cristomorto’ (S S ), tion, suggesting self-compatibility. 1 5 1 5 10 25 1 2 5. ‘Francolí’ (S1S1). 5. ‘Tuono’ (S1S1), 6. ‘Glorieta’ (S1S5),

7. ‘Falsa Barese’ (S1S1), 8. ‘Genco’ (S1S1). DNA isolation and S allele specific PCR. Leaves were collected from trees and stored at -80¼C until genomic DNA Microsatellites analysis. Seven Prunus RESULTS AND DISCUSSION was extracted by the CTAB extraction persica microsatellites isolated from a CT After stylar ribonuclease analysis, ‘Fran- method of Doyle and Doyle (1987). The enriched library (BPPCT007, 008, 011, colí’ showed only the S1 band with a faint PCR reaction, programme conditions, 017, 023, 025 and 037) (Dirlewanger et secondary one anodal to the S5 band po- electrophoretic separation of PCR pro- al., 2002) were used for parentage tests sition in the zymograms (Figure 1). The ducts and DNA band visualization follo- involving ‘Francolí’, ‘Gabaix’, ‘Tuono’ and secondary band associated to the S1 wed López et al. (2004a). The primer ‘Cristomorto’. DNA extraction, amplifica- band was observed at similar position as

AS1II (forward, 5'-TATTTTCAATTTGTG- tion, separation in acrylamide gels and the S25 band, causing difficult phenotype CAACAATGG-3') and AmyC5R (reverse, radioactive labelling and detection were interpretation and thus genotype assign- 5'-CAAAATACCACTTCAT GTAACAAC- as in Mnejja et al. (2004). Approximate ment. The same secondary band was amplicon sizes were determined with early observed in the self-compatible cul- 3') was used to amplify the S1, S5, S10 and standard DNA ladders. The shared band tivars ‘Falsa Barese’ and ‘Tuono’ (Figure S25 alleles (Tamura et al., 2000; López et al., 2004b). The Sf allele was amplified by similarity (Lynch, 1990) coefficients were 1). The possible S5, S10 or S25 bands were the primer SfF (forward, 5'-GTGCCCTAT- calculated on SSR data and the UPGMA not detected in the zymograms. CTAATTTGTTGAC-3') and SfR (reverse, clustering method (Sneath and Sokal, 5'-GACATTTTTTTAG AAAGAGTG-3') 1973) was applied on the similarity ma- After the PCR analysis using the conser- (Channuntapipat et al., 2003). Main trix. All these calculations were perfor- ved AS1II/AmyC5R primer, useful to re- bands visible on the gels were associated med by the NTSys-pc v. 2.11j (Rohlf, veal the S1, S5, S10 and S25 bands, a DNA with the S alleles. 2003) statistical package. fragment of approximately 900 bp corres-

Table 1. Result of six pollination tests (field and laboratory) made at IRTA Mas de Bover involving ‘Francolí’ for the determination of cross-compatibility (CC) and incompatibility relationships (CI).

Cross-incompatibility relationships (CI) Crossing Laboratory Deduced Field test Cross year test CI1,3 Nr. Fruit CC 1 1 flowers set CI (%)2 CI (%)

‘Francolí’ x ‘Desmayo Largueta’ (S1S25) 1990 182 25.8 C - C 2003 - - - 100 C

‘Desmayo Largueta’ (S1S25) x ‘Francolí’ 2003 120 5.0 C 20.0 C C

‘Francolí’ x ‘Ardechoise’ (S1S25) 2003 130 49.2 C 71.4 C

‘Ardechoise’ (S1S25) x ‘Francolí’ 2003 158 22.2 C 100 C C

‘Francolí’ x ‘Achaak’ (S2S25) 2003 - - 81.8 C C 2002 - - - 33.3 C ‘Francolí’ x ‘Francolí’ 20034 197 11.2 C 18.2 C C 20035 220 26.8 C - -

1 C = cross-compatible. 2 (Nr. flowers compatible with male parent / Total Nr. flowers tested) x 100. 3 Consensus between field and laboratory results. 4 Gimenells, Lleida. 5 No data available. Source: López et al., (2005)

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 17 Figure 3: Cluster analysis based on shared band similarity coefficients ved AS1II/AmyC5R primer agreed with for ‘Francolí’, its female parent (‘Cristomorto’) and two possible male parents the results from the ribonuclease analy-

(‘Gabaix’ and ‘Tuono’) constructed from SSR data. sis, discarding the S5, S10 and S25 alleles from its genotype (according to its early reported origin).

Since the S1 allele of ‘Francolí’ was clearly detected after RNase and PCR

analysis, only the S1S10 and the S1S25 genotypes would be theoretically possible, according to the new genotype assigned to ‘Gabaix’ (López et al., 2004b). The five test crosses made and records of crosses from our breeding programme confirmed that the S genotype of

‘Francolí’ is different from S1S5, S1S25

and S2S25. The other possible genotypes

S1S10 and S2S10 were untested as suitable cultivars or selections are not longer

available, since the S1S10 genotype of

‘Ardechoise’ and S2S10 of ‘Achaak’ and ‘Ferrastar’ first reported (Boskovic et al., 1999; Ortega, 2002) were later reassig- ponding to the S allele was successfully self-fertile in the field showing a good le- 1 ned to S1S25 and S2S25 respectively by amplified in ‘Francolí’ on the agarose gels vel of autogamy (19% mean fruit set) af- López et al. (2004b). Unilateral incom- (Figure 2a). In addition, the results obtai- ter selfing 417 flowers at two different lo- patibility was discarded, after observing ned after using the specific SfF/R primer cations (Constantí, Tarragona and Gime- that results obtained from crossing clearly showed that ‘Francolí’ carried the nells, LLeida) (Table 1). In addition, po- ‘Francolí’ by ‘Ardechoise’ (S1S25), ‘Des- Sf allele (Figure 2b). llen tubes reached the ovary after self-po- mayo Largueta’ (S1S25) and ‘Glorieta’ llinating 37 ‘Francolí’’s flowers in the la- (S1S5) were the same disregarding the The outcome of the crosses between boratory. crossing side. ‘Francolí’ and the cultivars ‘Ardechoise’

(S1S25) and ‘Desmayo Largueta’ (S1S25) All seven microsatellites amplified al- The possibility of ‘Francolí’ having the (Table 2) were consistent with ‘Francolí’s mond DNA and were polymorphic as it of- S1Sf genotype was envisaged after the genotype being different from S1S25. Mo- ten occurs with peach SSRs in almond RNase analysis and controlled pollinatio- reover, ‘Achaak’ (S2S25) was successfully (Aranzana et al., 2003). Table 2 shows ns. The result from ‘Francolí’ self-pollina- crossed by ‘Francolí’ in the laboratory to that five of the eight SSR loci studied tion strongly supported that ‘Francolí’ is test this originally possible genotype (Ta- (BPPCT008 detected two loci) discarded self-compatible, offering additional infor- ble 1). Pollen viability was optimal, as ran- ‘Gabaix’ as one of the parents of ‘Franco- mation on its autogamy level. Later, the ged from 39% to 87% for all the cultivars lí’. Seven discarded ‘Gabaix’ considering detection of the Sf allele by PCR using used as male parents. The results of other that ‘Cristomorto’ was the female parent, the specific SfF/R primer confirmed the three crosses made in the field were avai- whereas all of them were in agreement genetic self-compatibility of ‘Francolí’. lable from records of the breeding progra- about ‘Tuono’ (with self-compatible geno- The specific SfF/R primer was designed mme: ‘Glorieta’ (S1S5) x ‘Francolí’ (50.0% type S1Sf) and ‘Cristomorto’ being the from intron sequences of DNA regions of fruit set in 1992), ‘Francolí’ x ‘Glorieta’ parents of ‘Francolí’. (Channuntapipat et al., 2003), thus it is (S S ) (21.6% of mean fruit set for 4 1 5 specific for the Sf allele. The usefulness years), and ‘Garbí’ (S S ) x ‘Francolí’ In the polyacrylamide gels, the absence 1 5 of the Sf specific PCR technique applying (34.0% of fruit set in 1987). of the expected S5 band reported by Mar- this primer was demonstrated in two se- tínez-Gómez et al. (2003) using PCR was gregating almond progenies (López et al., ‘Francolí’ was selfed in the field and in the first explained as low RNase activity, as 2004a). laboratory to check its cross-compatibility observed before in other almond culti- and was observed to be unexpectedly vars. The PCR results using the conser- As ‘Francolí’ carries the S1 and the Sf allele and is self-fertile in the field and in the laboratory, we reassign to ‘Francolí’ the S S genotype instead of the earlier Table 2. Amplified fragments (in base pairs) of seven peach SSRs used 1 f reported S S (Martínez-Gómez et al., to test the ‘Francolí’ pedigree. 1 5 2003). In addition, the segregating population of the cross ‘Stelliete’ (S S ) x SSRs ‘Cristomorto’ ‘Gabaix’ ‘Tuono’ ‘Francolí’ 3 f ‘Francolí’ (S1Sf) made in our almond bree- BPPCT007 150/152 146 148/126 150/126 ding programme in 1991 matched the ex- BPPCT008A 133 139/125 127/123 133/127 pected ratio 3 self-compatible: 1 self-in- BPPCT008B 118 116/100 116 118/116 compatible (unpublished results), confir-

BPPCT011 172/158 158/150 170/156 172/170 ming the S1Sf genotype for ‘Francolí’. BPPCT017 154 160 156 156/154 BPPCT023 202 210 202 202 At IRTA’s almond breeding programme, BPPCT025 179 169 179/169 179 hand pollinated flowers are not bagged BPPCT037 136 136 136/132 136 due to the low cross-contamination risk detected by isozyme analysis (Arús et al., Source: López et al., (2005) 1994), since insects are not attracted by

18 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Figure 4. Drawing of the almond breeding plot at Mas de Bover where the cross ‘Cristomorto’ x ‘Tuono’ originating ‘Francolí’ was produced in 1976. diterranées: l’amandier et le pistachier. Rapport EUR 14081 FR : 201-207.

Boskovic, R., Tobutt, K. R., Batlle, I., and H. Duval, 1997. Correlation of ribonuclease zymograms and incompatibility genotypes in almond. Euphytica 97: 167-176.

Boskovic, R., Tobutt, K. R., Duval, H., Batlle, I., Dicenta, F., and F. J. Vargas, 1999. A stylar ribonuclease assay to de- tect self-compatible seedlings in almond progenies. Theor. Appl. Genet., 99: 800-810.

Boskovic, R., Tobutt, K. R., Batlle, I., Duval, H., Martínez-Gómez, P., and T. M. Gradziel, 2003. Stylar ribonucleases in almond: correlation with and predic- tion of incompatibility genotypes. Plant Breed., 122: 70-76.

Brooks, R.M. and H.P. Olmo, 1997. Re- gister of fruit and nut varieties. 3rd Ed. ASHS Press, Alexandria: 744 pp.

Broothaerts, W., 2003. New findings in apple S-genotype analysis resolve pre- 'Cristomorto' mother tree originating 'Francolí'. vious confusion and request the re- numbering of some S-alleles. Theor. Appl. Genet. 106: 703-714. emasculated flowers and the wind has We conclude that the S genotype of Channuntapipat, C., Sedgley, M., and low influence in almond pollen transport ‘Francolí’ is S S , and thus it is self-com- G. Collins, 2001. Sequences of cDNAs 1 f and genomic DNAs encoding the S1, (Kester and Asay, 1975). As the reported patible. This finding clarifies the genotype S7, S8 and Sf alleles from almond, Pru- crossing origin of ‘Francolí’, ‘Cristomorto’ of this almond cultivar and explains its nus dulcis. Theor. Appl. Genet., 103: agronomic field behaviour. The sequen- 1115-1122. (S1S2) x ‘Gabaix’ (S10S25), could not give self-compatible seedlings, self-fertility cing of the Sf allele from ‘Francolí’ would ensure the origin of this self-compatibility Channuntapipat, C., Wirthensohn, M., was never tested before, as it is usually Ramesh, S. A., Batlle, I., Arús, P., checked in seedlings selected from self- allele from ‘Tuono’. In addition, ‘Francolí’ Sedgley, M., and G. Collins, 2003. Iden- compatible segregating progenies. In the will be useful for breeding programmes to tification of incompatibility genotypes in case of ‘Francolí’, probably artificial polli- transfer self-compatibility together with its almond (Prunus dulcis Mill.) using spe- nation failed and one ‘Cristomorto’ flower outstanding agronomic features. Since cific primers based on the introns of the S-alleles. Plant Breed., 122: 164-168. was pollinated with pollen of nearby rows this cultivar is largely recommended to be of the self-compatible ‘Tuono’, which has planted in new orchards it will have the Crossa-Raynaud, P., and C. Grasselly, overlapping flowering time and it was the added value of its known self-compatibility. 1985. Existence de groupes d’inters- only self-compatible cultivar at IRTA’s re- térilité chez l’amandier. Options Médi- terranéennes 1985-I : 43-45. ferred breeding plot (Figure 4). ACKNOWLEDGEMENTS The authors are grateful to M. Rovira Doyle, J. J., and J. L. Doyle, 1987. A ra- The cluster analysis based on the shared (IRTA-Mas de Bover) and P. Arús (IRTA- pid DNA isolation procedure for small band similarity coefficients for ‘Francolí’ Cabrils) for their valuable comments and quantities of fresh leaf tissue. Phyto- and its possible parents (Figure 3) revea- to M. Mnejja (IRTA-Cabrils) and S. Alegre chem. Bull. 19: 11-15. led that ‘Francolí’ is closer to ‘Tuono’ than (IRTA-EE Lleida) for laboratory and field Felipe, A. J., 1977: Estados fenológicos to ‘Gabaix’. In addition, leaf isoenzymatic help respectively. This research was con- del almendro. Información técnica y records of 13 loci supports that ‘Francolí’ ducted under three INIA (Spanish Minis- económica agraria. ITEA 8 (27): 8-9. could have been raised from the cross try of Science and Technology) Projects ‘Cristomorto’ x ‘Tuono’ (Arús et al., 1990, (SC97-049, RTA01-081 and RTA04-030). Heslop-Harrison, J., Heslop-Harrison, Y., and K. R. Shivanna, 1984. The eva- 1994 and unpublished results). Also work Mercè López gratefully acknowledges a luation of pollen quality, and a further by Viruel (1995) comparing almond culti- grant from the INIA postgraduate appraisal of the fluorochromatic (FCR) vars by RFLP analysis found that ‘Fran- programme. test procedure. Theor. Appl. Genet., 67: colí’ was closer to ‘Cristomorto’ (81% si- 367-375. milar) and ‘Tuono’ (83% similar) than to REFERENCES Kester, D. E. and R. A. Asay, 1975. Al- ‘Gabaix’ (69% similar). Thus, in agree- monds. In: J. Janick and J.N. Moore ment with our results and the position of (eds.), Advances in Fruit Breeding: 387- the ‘Cristomorto’ mother tree placed next Arús, P., Olarte, C., Romero, M. A. and 419. Purdue University Press, Indiana. F. J. Vargas, 1994. Linkage analysis of to ‘Tuono’ in the original breeding plot (Fi- ten isozyme genes in F1 segregating al- Kester, D. E., Gradziel, T. M., and W. C. gure 4), we could suggest that ‘Francolí’ mond progenies. J. Amer. Soc. Hort. Micke, 1994a. Identifying pollen incom- derives from the cross ‘Cristomorto’ Sci., 119 (2): 339-344. patibility groups in California almond cultivars. J. Am. Soc. Hort. Sci 119: (S1S2) x ‘Tuono’ (S1Sf) and not from ‘Cristomorto’ (S S ) x ‘Gabaix’ (S S ) as Arús, P., Vargas, F.J. and M. A. Rome- 106-109. 1 2 10 25 ro, 1990. Linkage analysis of isozyme considered until now and, thus ‘Tuono’ genes in almond. Amélioration généti- Kester, D. E., Micke, W. C., and M. Vi- conferred the self-compatibility to ‘Fran- que de deux espèces de fruits secs mé- veros, 1994b. A mutation in ‘Nonpareil’ colí’.

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 19 NEW FINDINGS Grasselly and Crossa Raynaud (1980) almond conferring unilateral incompati- IN THE GENETIC CONTROL found highly variable sweet:bitter ratios bility. J. Amer. Soc. Hort. Sci 119 (6): which could indicate a more complex ge- 1289-1292. AND EXPRESSION OF KERNEL netic control than the Heppner hypothe- BITTERNESS IN ALMOND López, M., Mnejja, M., Romero, M. A, sis. Nevertheless, recently different stu- Vargas, F. J., and I. Batlle, 2004a. Use dies confirmed Heppner’s hypothesis and of Sf specific PCR for early selection of provided new information regarding the self-compatible seedlings in almond genotype (homozygous or heterozygous) breeding. Options Méditerranéennes A INTRODUCTION 63 : 269-274. The almond [P. dulcis (Mill.) D.A. Webb; for this characteristic in many sweet ker- syn. P. amygdalus Batsch] is a species of nel cultivars (Grasselly and Crossa-Rayn- López, M., Mnejja, M., Rovira, M., Co- genus Prunus subgenus Amygdalus (Ro- aud, 1980; Dicenta and García, 1993; llins, G., Vargas, F.J., Arús, P. and I. saceae, subfamily Prunoideae) which is Vargas et al., 2001). Batlle, 2004b. Self-incompatibility genotypes in almond re-evaluated by PCR, grown commercially worldwide, being its stylar ribonucleases, sequencing analy- edible seed (the kernel) the trading pro- GENETIC MAPPING OF KERNEL sis and controlled pollinations. Theor. duct. However, almond species presents BITTERNESS IN ALMOND USING SSRS Appl. Genet, 109: 954-964. in many genotypes a bitter kernel due to PCR-based, simple sequence repeat the cyanoglucoside amygdalin. In these López, M., Romero, M. A, Vargas, F. J., (SSR) markers (microsatellites) are beco- Mnejja, M., Arús, P. and I. Batlle, 2005. bitter kernelled trees, amygdalin is syn- ming the main chosen marker for finger- ‘Francolí’, a late flowering almond culti- thesized from prunasin in a well known printing and genetic diversity studies for a var reclassified as self-compatible. metabolic pathway (McCarty et al., 1952; wide range of plants. Because of their Plant Breeding, 124: 502-506. Conn, 1980; Frehner et al., 1990) (Figure 1). high polymorphism, abundance, and co- López, M., Vargas, F. J. and I. Batlle, dominant inheritance, they are well suited 2006. Self-(in) compatibility almond ge- The cultivated almond is thought to have for the assessment of genetic variability notypes: a review. Euphytica, 150: 1- been originated in the arid mountainous within crop species, and for genetic rela- 16. regions of Central Asia (Grasselly, tionships among species (Powell et al., 1976a). In addition, several wild species, Martínez-Gómez, P.; López, M., Alonso, 1996; Wünsch and Hormaza, 2002). The- J. M., Ortega, E., Batlle, I., Socías and described in some cases as ancestors of se SSR markers have been used in al- Company, R., Dicenta, F., Dandekar, A. the cultivated species, are found growing mond for molecular characterization and M., and T. M. Gradziel, 2003: Identifica- in these mountainous areas from Tian identification of cultivars (Martínez-Gó- tion of self-incompatibility alleles in al- Shan in western China through the de- mond and related Prunus species using mez et al., 2003a) and related Prunus PCR. Acta Horticulturae, 622: 397-401. serts of Iran and Iraq (Grasselly, 1976b; species (Martínez-Gómez et al., 2003b); Kester and Gradziel, 1996).These ances- and the elaboration of genetic maps (Jo- Sonneveld, T., Tobutt, K. R., and T. P. tors of cultivated almonds are, in most ca- obeur et al., 2000; Bliss et al., 2002; Robbins, 2003. Allele-specific PCR de- ses, bitter-kernelled (Grasselly, 1976a; Aranzana et al., 2003). tection of sweet cherry self-incompatibi- 1976b; Gradziel et al., 2001), and the lity (S) alleles S1 to S16 using consensus and allele-specific primers. Theor. sweet taste seems to be the consequen- The cosegregation analysis facilitated the Appl. Genet. 107: 1059-1070. ce of a mutation of an originally bitter al- linkage analysis between markers and mond (Heppner, 1923) which has been qualitative or quantitative loci controlling Tamura, M., Ushijima, K., Sassa, H., Hi- maintained in the cultivated almond rano, H., Tao, R., Gradziel, T. M., and horticultural important traits, as kernel bit- A. M. Dandekar, 2000. Identification of through selection by men. terness. In this sense, the major gene for self-incompatibility genotypes of al- kernel taste (Sk) has been placed in mond by allele-specific PCR analysis. Seed sweet taste continues to be the first linkage group 5 (G5) by Joobeur (1998) Theor. Appl. Genet. 101: 344-349. selection feature in almond breeding pro- and Bliss et al. (2002), being bitterness grammes. The efficiency of breeding pro- Vargas, F. J., Romero, M. A., Rovira, recessive. More recently, this major gene M., and J. Girona, 1984. Amélioration grammes depends on the information was also identified in the G5 in a map de- de l’amandier par croisements des va- available about the transmission of traits veloped with SSR markers by Sánchez- riétés. Résultats préliminaires à Tarra- to be improved. In addition, the develop- Pérez (2006) (Figure 2). Its peak was lo- gone (Espagne). Options Méditerra- ment of molecular markers to use in the cated close to the locus of the SSR mar- néennes, IAMZ 84/II: 101-122. early selection of traits, as sweet kernel, ker PceGA025, and the percentage of Vargas, F. J., and M. A. Romero, 1994: in almond breeding programmes, could trait variability explained by this character ‘Masbovera’, ‘Glorieta’ and ‘Francolí’, be of great interest in the selection of new was up to 81.3%. three new almond varieties from IRTA. cultivars (Sánchez-Pérez et al., 2004). Acta Horticulturae, 373:_ 75-82. Selection by molecular markers is also Viruel, M. A., 1995. Construcción de un GENETIC CONTROL OF KERNEL particularly useful in fruit breeding as mapa genético con marcadores molecu- BITTERNESS IN ALMOND most species showed a long juvenile pe- lares en almendro (Prunus amygdalus Heppner (1923; 1926) indicated long time riod. Marker-assisted selection (MAS) is Batsch.). Estudio de la variabilidad in- ago that kernel bitterness in almond is a emerging as a very promising strategy for traespecífica y aplicación a la identifica- ción varietal. Barcelona University. PhD monogenic trait with a qualitative or Men- increasing selection gains. Marker loci thesis. delian inheritance and being the bitter linked to major genes can be used for se- allele recessive. In addition, most of the lection, which is more efficient than direct commercial cultivars have been characte- selection for the target gene (Arús and rized as heterozygous for this trait (Gras- Moreno-González, 1993). I. Batlle, M. López, M. A. Romero selly and Crossa-Raynaud, 1980; Dicenta and F. J. Vargas and García, 1993; Vargas et al., 2001). Currently, one of the most effective IRTA Mas de Bover However, some authors suggested a Mediterranean Fruit Trees approaches for the analysis of marker- Ctra. Reus - El Morell, km 3.8 more complex mechanism of inheritance. trait association in Prunus is the use of E-43120 Constantí, Tarragona, Spain Spiegel-Roy and Kochba (1974) indicated these mapping populations segregating E-mail:[email protected] the possibility of three genes involved in for the characters of interest. The diffe- the expression of this trait. In addition, rent linkage maps developed could inclu-

20 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Figure 1. Schematic representation of the biosynthesis and the hydrolysis Suelves and Puigdomenech (1998) iden- of amygdalin in Prunus. Description of the scientific works (identification tified and sequenced a gene highly of the enzyme activity, location of the enzymes, identification of EST, etc.) expressed in the floral organs of almond developed for the establishment of the genetic control of this biosynthesis pathway. and coding for the cyanogenic enzyme (R)-(+)- mandelonitrile lyase. However, the gene responsible for this mandelonitrile lyase was located in the linkage group 1 (G1) instead of in the linkage group 5 as was located the gene responsible of the bitterness in almond (Joo- beur, 1998; Bliss et al., 2002; Sánchez- Pérez, 2006). These results indicate the independence of the activity of this enzyme with the expression of kernel bitterness in almond.

In addition, Frehner et al. (1990) indicated that the responsible for the accumulation in the seed could be an undescribed prunasin glucosyltransferase. In this sense, Vezvaei et al. (2004) developed a strategy for discovering the glucosyltransferase gene responsible for producing bitterness taste in almond using degenerate primers based on consensus regions of glucosyltransferase genes for other plants. Nevertheless, these studies were not conclusive regarding the identification Figure 2. Map of the ‘R1000’ x ‘Desmayo Largueta’ F1 almond population obtained of the concrete glucosyltransferase res- with 56 SSRs, with the approximate location of the major gene for kernel taste (Sk). ponsible for producing bitter kernel in almond.

More recently, Sánchez-Pérez (2006) analyzed several candidate genes involved in the biosynthesis and hydrolysis of amignalin including ESTs from mandelonitrile lyase, amygdalin hydrolase and prunasin hydrolase isolated in Prunus, and three glucosyltransferase identified in Arabidopsis thaliana. The results of the study of cosegregation of these candidate genes in several sweet and bitter almond cultivars also indicated a lack of correlation between these candidate genes and the kernel bitterness expression in almond cultivars. In addition, these candidate genes were located in the linkage group 1 (G1) instead of in the linkage group 5 also confirm the unrelated activity de markers associated with horticultural mond based on cDNA libraries has been of these enzymes with the expression of traits as kernel bitterness. In our work, released in the Clemsom University (South kernel bitterness in almond. the PceGA025 SSR marker was linked to Caroline, USA), and more than 3,800 pu- the gene responsible for kernel bitterness tative unigenes have been detected In addition, Dicenta et al. (2002) indicated in almond at an estimated distance of 2 (http://www.mainlab.clemson.edu/gdr/). that prunasin and amygdalin production CM. The association of this marker with This work is complementary to the other occur by different processes. Prunasin is the kernel bitterness trait is being confir- works regarding EST development in Pru- present in the vegetative part of most al- med in different sweet (homozygous or nus performed by different research groups mond genotypes, although only some ge- heterozygous) and bitter (homozygous) in other European countries presenting a notypes are able to store amygdalin in the almond cultivars in order to develop effi- collection of 6,817 ESTs prepared from seed. Therefore bitter taste would be re- cient strategies of marker assisted selec- four cDNA libraries obtained from peach lated to the ability to transform the pruna- tion for kernel bitterness in almond. as part of the work of the Italian National sin of the vegetative parts into amigdalin Consortium for Peach Genomics (http:// in the kernel. CANDIDATE GENE ANALISIS LINKAGE www.itb.cnr.it/ESTree). From these data TO KERNEL BITTERNESS IN ALMOND bases, different ESTs have been identi- ACKNOWLEDGMENTS Besides the data base of the National fied from genes of several enzymes invol- This work has been supported by the re- Center for Biotechnology Information ved in the biosynthesis or the hydrolysis search projects AGL2001-1054-C03-01 (NBCI; http://www.nbci.nlm.nih.gov/entrez/), of amygdalin including glucosyltransfera- and AGL2004-06674-C02-02/AGR of the a collection of ESTs (Expressed sequen- se, mandelonitrile lyase, amygdalin hydro- Spanish Ministry of Science and Educa- ce targets) specifically from peach and al- lase, and prunasin hydrolase (Figure 1). tion.

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 21 TOCOPHEROL IN ALMOND BIBLIOGRAPHY nick, J. (eds.). Fruit Breeding. New York: Wiley. p. 1-97. KERNELS AS A QUALITY Aranzana M.J., Pineda A., Cosson P., COMPONENT Dirlewanger E., Ascasibar J., Cipriani G., Martínez-Gómez P., Arulsekar S., Pot- Ryder C.D., Testolin R., Abbott A., King ter D., T.M. and Gradziel T.M. 2003a. G.J., Iezzoni A.F. and P. Arús. 2003. A An extended interspecific gene pool set of simple-sequence (SSR) markers available to peach and almond breeding covering the Prunus genome. Theor. as characterized using simple sequence INTRODUCTION Appl. Gent. 106: 819-825. repeat (SSR) markers. Euphytica 131: The definition of almond kernel quality in 313-322. international markets is currently based Arús P. and J. Moreno-González. 1993. on physical parameters such as size and Martínez-Gómez P., Arulsekar S., Pot- Marker-assisted selection. In: Hayward, shape, and on sample homogeneity, dis- M.D., Bosemark, N.O., and Romagosa, I. ter D., and T.M. Gradziel. 2003b. Rela- (eds.). Plant Breeding. Principles and tionships among peach and almond and regarding the kernel’s chemical composi- Prospects. London, United Kingdom: related species as detected by SSR tion; however, the best end-use of each Chapman & Hall. p. 314-331. markers. J. Amer. Soc. Hort. Sci. 128: cultivar is a function of its chemical com- 667-671. position (Berger, 1969). The high nutritive Bliss F.A., Arulsekar S., Foolad M.R., value of almond kernels arises mainly Becerra V., Gillen A., Warburton M.L., McCarty C.D., Leslie J.W. and Frost H.B. Dandekar A.M., Kocsine G.M. and K.K. 1952. Bitterness of kernels of almond x from their high lipids content, which cons- Mydin. 2002. An expanded genetic linka- peach hybrids and their parents. Proc. titutes an important caloric source, but ge map of Prunus based on an interspe- Am. Society Hort. Sci. 59: 254-258. does not contribute to cholesterol forma- cific cross between almond and peach. tion in humans, due to their high level of Genome 45: 520-529. Powell W., Morgante M., Andre C., Ha- nafey M., Vogel J., Tingey S., and A. unsaturated fatty acids, mainly mono-un- Conn E.E. 1980. Cyanogenic compounds. Rafalski. 1996. Comparison of RFLP, saturated fatty acids (Sabaté and Hook, Ann. Rev. Plant Physiol. 31: 433-451. RAPD, AFLP and SSR (microsatellite) 1996). markers for germplasm analysis. Mol. Breed. 2: 225-238. Dicenta F. and García J.E. 1993. Inheri- Most vegetable oils contain tocopherols tance of the kernel flavour in almond. Heredity 70: 308-312. Sánchez-Pérez R. 2006. Desarrollo y in differing amounts, especially oils that aplicación de marcadores moleculares contain high levels of unsaturated fatty Dicenta F., Martínez-Gómez P., Grané a la mejora genética del almendro. PhD acids. Tocopherols are natural mono- N., Martín M.L., León A., Cánovas J.A. Thesis dissertation. Universidad de phenols with anti-oxidant activities (Reis- and Berenguer V. 2002. Relationship Murcia, Murcia, Spain. between cyanogenic compounds in ker- che et al., 1998), with several isomers de- nels, leaves and roots of sweet and bitter Sánchez-Pérez R., Dicenta F., Gradziel pending on the position and number of kernelled almonds. J. Agr. Food Chem. T.M., Arús P., and Martínez-Gómez P. methyl groups. Their main biochemical 50: 2149-2152. 2004. Application of molecular markers function is believed to be the protection of in almond breeding programmes. Nucis- poly-unsaturated fatty acids against pe- Frehner M., Scalet M. and Conn E.E. Newsletter 12: 9-12. 1990. Pattern of the cyanide-potential in roxidation (Kamal-Eldin and Appelqvist, developing fruits. Plant Physiol. 94: 28-34. Spiegel-Roy P. and Kochba J. 1974. 1996). The inheritance of bitter and double ker- Gradziel T.M., Martínez-Gómez P., Di- nel characters in the almond. Z. Pflan- Tocopherol concentrations have been de- centa F., and D.E. Kester. 2001. The uti- zenzuchtg 71: 319-329. lization of related almond species for al- termined in oils from several tree species: mond variety improvement. J. Am. Po- Suelves M., Puigdomenech P. 1998. hazelnut, Corylus avellana L. (Özdemir et molog. Soc. 55: 100-109. Molecular cloning of the cDNA coding al., 2001), walnut, Juglans regia L., and for the (R)-(+)-mandelonitrile lyase of olive, Olea europea L. (Kamal-Eldin and Prunus amygdalus: temporal and spa- Grasselly C. 1976a. Origine et évolution Andersson, 1997), and have been corre- de l’amandier cultivé. Options Méditerr. tial expression patterns in flowers and 32: 45-50. mature seeds. Planta 206: 388-393. lated with their anti-oxidant activity. In almond, tocopherol concentration plays an Grasselly C. 1976b. Les espèces sauva- Vargas F.J., Romero M.A. and Batlle I. important role in protecting lipids against ges d’amandier. Options Méditerr. 32: 28-44. 2001. Kernel taste inheritance in al- oxidation and thus lengthening their sto- mond. Options Méditerr. 56: 129-134. rage time (García-Pascual et al., 2003; Grasselly C. and Crossa-Raynaud P. 1980. L’amandier. Maisonneuve et Laro- Vezvaei A, Parfitt DE, Chan HM, Gra- Senesi et al., 1996; Zacheo et al., 2000). se. Paris. 446 pages. dziel TM (2004) Isolation of a gene con- Furthermore, Fourie and Basson (1989) trolling bitterness in almond. Internatio- studied variation in tocopherol concentra- nal Rosaceae Genome Mapping Confe- Heppner J. 1923. The factor for bitterness tions in several nuts, and found that al- in the sweet almond. Genetics 8: 390-392. rence. Clemsom, USA. http://www.geno me.clemson.edu/gdr/conference/ mond kernels, with a higher tocopherol Heppner J. 1926. Further evidence on program.html. concentration than other nuts, had longer the factor for bitterness in the sweet al- storage ability. mond. Genetics 11: 605-606. Wünsch A. and J.I. Hormaza. 2002. Cul- tivar identification and genetic finger- Information on tocopherol concentrations Joobeur T. 1998. Construcción de un printing of temperate fruit tree species mapa de marcadores moleculares y aná- using DNA markers. Euphytica 125: 56-67. is important to determine the end-use of lisis genético de caracteres agronómicos the kernels and for predicting their stora- en Prunus. PhD Thesis dissertation. Uni- ge-life. Several studies have shown that versitat de Lleida, Lleida, Spain. 1R. Sánchez-Pérez, 2W. Howad, 2P. Arús, whole almond nuts can be stored for 9 1P. Martínez-Gómez and 1F. Dicenta. months and maintain quality (García-Pas- Joobeur T., Periam N., de Vicente M.C., 1 King G.J. and P. Arús. 2000. Develop- CEBAS-CSIC. Departamento de Mejora cual et al., 2003; Zacheo et al., 2000). Si- ment of a second generation linkage y Patología Vegetal, Apdo 4195, 30.080, milarly, Rizzolo et al. (1994) concluded Murcia, Spain. map for almond using RAPD and SSR 2 that almond storage for ≥ 1 year, without markers. Genome 43: 649-655. CSIC-IRTA. Genética Vegetal. Crta. Cabrils, km 2, E-08348 Cabrils, quality loss, can only be attained with cul- Barcelona, Spain. Kester D.E. and T.M. Gradziel. 1996. Al- tivars that possess high concentrations of E-mail: [email protected] monds (Prunus). In: Moore, J.N. and Ja- natural anti-oxidants such as ␣-tocopherol, suggesting that this isomer is the

22 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Kernel samples of 'Marcona' (left) and 'Masbovera' (right).

Table 1. Oil and composition of different tocopherol isomers in almond cultivars Given the importance of all aspects related to almond kernel quality, and its main- Cultivar Oil content ␣-tocopherol ␥-tocopherol ␦-tocopherol Total tenance during storage, our objective was (% kernel DW) (mg kg-1 oil) (mg kg-1 oil) (mg kg-1 oil) tocopherol to determine the concentrations of diffe- (mg kg-1 oil) rent isomers of tocopherol in a series of almond cultivars. ‘Belona’ 65.4 418.4 18.5 18.4 455.3 ‘Soleta’ 61.8 214.0 13.3 15.0 242.3 MATERIALS AND METHODS ‘Marcona’ 58.0 463. 18.5 18.7 500.6 Kernels of nine cultivars of almond were ‘Moncayo’ 57.6 412. 18.8 21.2 452.5 analysed. Included were two traditional ‘Bertina’ 56.3 241.6 15.0 16.6 273.2 Spanish cultivars (‘Marcona’ and ‘Desma- ‘D. Largueta’ 55.6 304.3 15.3 16.6 336.2 yo Largueta’), a French cv. (‘Ferragnès), ‘Felisia’ 55.5 250.6 18.2 17.3 286.2 a Spanish local selection (‘Bertina’) and ‘Ferragnès’ 63.0 377.5 18.7 18.4 414.6 five releases from the CITA breeding pro- ‘Guara’ 55.8 385.4 15.7 17.6 418.8 gramme (‘Belona’, ‘Felisia’, ‘Guara’, ‘Moncayo’ and ‘Soleta’). main component protecting kernel quality Consequently, kernel quality may be in- Nuts were harvested at maturity, when (Kamal-Eldin and Appelqvist, 1996) and creased by higher levels of ␣-tocopherol, the fruit mesocarp had fully dried and split prolonging storage (Senesi et al., 1996), due to their lipid stability function and to along the fruit suture, and peduncle as has been shown in olive oil (Deiana et its nutritive value as vitamin E, taking into abscission was complete (Felipe, 1977). al., 2002). Fatty acid oxidation becomes account present consumer trends for Two replicates of 20 fruits of each cultivar only significant after a latent period, du- foods without synthetic additives (Krings were collected at random around the ca- ring which kernel anti-oxidants are deple- and Berger, 2001). However, quantifica- nopy and dried for 2 d. The shells were ted, as the total concentration of tocophe- tion of other tocopherols is also impor- cracked and the kernels blanched in rols decreases during storage (Sun et al., tant. In many cases, ␥-tocopherol is belie- water at 100¼C. After drying in air at room 2001; Zacheo et al., 2000), due to their ved to play a role in protecting the body temperature for 24 h, kernels were anti-oxidant activity (Zacheo et al., 2000). from damage by free radicals, because ␥- ground in a domestic electric grinder. tocopherol can be even more important A high concentration of tocopherol has than ␣-tocopherol in preventing the perni- Oil was extracted from 5 g ground al- also been shown to be very important in cious effects of some specific radicals monds kernels in a commercial fat-extrac- the human diet, due to its vitamin E activi- such as peroxynitrite (Hoglen et al., tor for 2 h using petroleum ether as sol- ty (Kamal-Eldin and Appelqvist, 1996). 1997). vent with the heating source at 135 ¼C.

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 23 Oil content was expressed as a percenta- copherol concentration in almond. It could ge of kernel DW. Saponification was per- be suggested that early blooming culti- Kamal-Eldin, A., Appelqvist, L.A., 1996. formed according to an adaptation of the vars, with an earlier kernel filling period, The chemistry and antioxidant properties of tocopherols and tocotrienols. Li- EU official method (DOCE L174/39, 13 when temperatures are lower, could have pids, 31: 671-701. July 2000), to determine, simultaneously, lower tocopherol concentrations in their tocopherols and sterols in oils and fatty kernel oil. However, it is unquestionable Kodad, O., Gracia Gómez, M.S., Socias products (Kodad et al., 2006). that the cultivar with the highest tocophe- i Company, R., 2005. Fatty acid composition as evaluation criterion for kernel rol concentration, ‘Marcona’, is of early quality in almond breeding. Acta Hortic., RESULTS AND DISCUSSION blooming time. 663: 301-304. Oil content varied between cultivars, from 55.5% of the kernel DW in ‘Largueta’ and The present research related to the nutri- Kodad, O., Socias i Company, R., Prats, ‘Felisia’ to 65.4% in ‘Belona’ (Table 1). tional and antioxidant benefits of ␣- and M.S., López Ortiz, M.C., 2006. Variabili- ␥ ty in tocopherol concentration in almond The oil contents were in general higher -tocopherol must thus be taken into ac- oil and its use as a selection criterion in than previous results from other almond count when developing new breeding almond breeding. J. Hort. Sci. Biotech- cultivars from California (Abdallah et al., strategies in almond in order to modify nol. (in press). 1998) and Australia (Vezvaei and Jack- the level of these important components son, 1996). The high amount of fatty of vitamin E. Krings, U., Berger, R.G., 2001. Antioxi- dant activity of some roasted foods. acids in ‘Belona’ may be considered in Food Chem., 72: 223-229. the production of some almond confectio- ACKNOWLEDGEMENTS nery, mainly nougat. This research was conducted under pro- Özdemir, M., Açkurt, F., Kaplan, M., Yil- ject and AGL2004-06674-C02-01 of the diz, M., Löker, M., Gürcan, T., Biringen, ␣ G., Okay, A., Seyhan, F.G., 2001. Eva- The concentration of -tocopherol was Spanish CICYT. OK acknowledges a luation of new Turkish hybrid hazelnut the highest, followed by ␥ and ␦ (Table scholarship grant from INIA. Technical (Corylus avellana L.) varieties: fatty acid 1). The content of a-tocopherol was the assistance from J.M. Ansón is highly composition, ␣-tocopherol, mineral com- most variable between cultivars, from 214 appreciated. position and stability. Food Chem., 73: mg kg-1 in ‘Soleta’ to 453.3 in ‘Marcona’, 411-415. ␥ whereas -tocopherol ranged from 13.3 Reische, D.W., Lillard, D.A., Eitenmiller, in ‘Soleta’ to 18.8 in ‘Moncayo, and d-to- R.R., 1998. Antioxidants. In: Food Li- copherol from 15.0 in ‘Soleta’ to 21.2 in pids. Chemistry, Nutrition, and Biotech- REFERENCES ‘Moncayo’. Thus, ‘Soleta’ showed the lo- nology (Akoh, C.C. and Min, D.B., Eds.). Marcel Dekker, New York, USA. Pp. west amount of all tocopherol isomers Abdallah, A., Ahumada, M.H., Gradziel, 423-448. whereas ‘Marcona’ was distinguished by T.M., 1998. Oil content and fatty acid the high amount of the ␣ isomer and of composition of almond kernels from dif- Rizzolo, A., Senesi, E., Colombo, C., the total tocopherol. ferent genotypes and California produc- 1994. Studies on the storage of shelled tion regions. J. Amer. Soc. Hortic. Sci., and in-shelled almonds. Acta Hortic., 123: 1029-1033. 373: 259-261. These results have confirmed the variability in the concentration of tocopherol iso- Berger, P., 1969. Aptitude à la transfor- Sabate, J., Hook D.G., 1996. Almonds, mers between the cultivars studied, as mation industrielle de quelques variétés walnuts, and serum lipids. In: Lipids in was already reported for other almond d’amandier. Bull. Techn. Inf., 241: 577- Human Nutrition. (Spiller, G.A., Ed.). 580. CRC Press Inc., Boca Raton, FL, USA, cultivars. However, the number of almond pp. 137-144 cultivars studied has been always very Deiana, M., Rosa, A., Falqui Cao, C.,. small: five by Fourie and Basson (1989) Pirisi, F.M, Bandino, G., Dessi, M.A., Senesi, E., Rizzolo, A., Colombo, C., and four by Zacheo et al. (2000) and Gar- 2002. Novel approach to study oxidative Testoni, A., 1996. Influence of pre-pro- cía-Pascual et al. (2003), with the only stability of extra virgin oilve oils: impor- cessing storage conditions on peeled almond quality. Ital. J. Food Sci., 8: 115- exception of the 54 genotypes studied by tance of a-tocopherol concentration. J. Agric. Food Chem., 50: 4342-4346. 125. Kodad et al. (2006), who observed a continuous distribution in the concentrations Felipe, A.J., 1977., Almendro: estados Sun, W., Kawano, Y., Shiomori, K., Yo- of the three isomers of tocopherol with a fenólogicos. Inf. Tecn..Econ. Agrar., 27: nekura, M., Mitani, H., Hatate, Y., 2001. 8-9. Auto-oxidation rate of linoleic acid and wide range of variability. effect of antioxidants on the oxidation. Fourie, P.C., Basson, D.S., 1989. Pre- Kagaku Kogaku Ronbun-shu, 27: 76-84. The differences found for the concentra- dicting occurrence of rancidity in stored tions of the three isomers between the nuts by means of chemical analyses. Vezvaei, A., Jackson, J.F., 1996. Al- cultivars studied showed that they de- Lebens.-Wissens. Technol., 22: 251- mond nut analysis. In: Modern Methods 253. of Plant Analysis, vol. 18, Fruit Analysis pend on each one and may be conside- (Linskens, H.F. and Jackson, J.F., Eds.. red a cultivar trait, as it also happened for García-Pascual, P., Mateos, M., Carbo- the fatty acid composition of these same nell, V., Salazar, D.M., 2003. Influence Springer-Verlag, Berlin, Germany. Pp. cultivars (Kodad et al., 2005). The high of storage conditions on the quality of 135-148. concentrations of a-tocopherol and the shelled and roasted almonds. Biosyst. Engineer., 84: 201-209. Zacheo, G., Cappello, M.S., Gallo, A., medium-high concentrations for the other Santino, A., Cappello, A.R., 2000. Chan- isomers found in ‘Marcona’ may have Hoglen, N.C, Waller, S.C., Sipes, I.G., ges associated with postharvest ageing been a basic reason, although ignored Liebler, D.C., 1997. Reactions of pe- in almond seeds. Lebens.-Wissens. until now, of its very high estimation in roxynitrite with ␥-tocopherol. Chem. Technol., 33: 415-423. the Spanish market for use in almond Res. Toxicol., 10: 401-407. confections, especially nougat. Kamal-Eldin, A., Andersson R., 1997. A multivariate study of the correlation O. Kodad and R. Socias i Company An increase of tocopherol content with between tocopherol and fatty acid in ve- Unidad de Fruticultura, CITA de Aragón higher temperatures has been found (Ko- getable oils. J. Amer. Oil Chem. Soc., Apartado 727, 50080 Zaragoza, Spain 74: 375-376. E-mail: [email protected] dad et al., 2006), but blooming time does not seem to have any relationship with to-

24 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 LONG-TERM CRYOPRESERVA- TION OF ALMOND GERM- PLASM: AN UPDATE

INTRODUCTION Germplasm conservation is vital for almond improvement as plant breeding options depend on genetic variability, without which there is no heterosis from recombination and genetic assortment. As almond is vegetatively propagated, in vitro methods have been employed for germplasm conservation. Cryopreserva- tion allows the opportunity to preserve valuable germplasm at a much-reduced cost compared to ex situ germplasm collections in the field. This is due to the labour- Figure 1. Shoots on MS multiplication media. saving and associated costs of land use, 1a: shoots from cryopreserved shoot tips (2-2.5mm); 12 weeks after thawing. as well as providing a contamination-free, 1b: shoots from non-cryopreserved shoot tips (.7-1.0cm); 4 weeks after culturing. low maintenance environment. Plant cryopreservation has moved forward from the initial work on freeze-induced dehydration to multiplication media for a further 1 or 2 (Bright’s hybrid). Mother trees of ‘Nonpa- to new techniques of vitrification using weeks, after which they were either put reil’ and ‘Ne Plus Ultra’ were between 10 concentrated solutions to protect plant or- onto rooting media or micrografted onto and 15 years old. Bright’s hybrid control ganelles prior to immersion in liquid nitro- cultured rootstocks to generate whole trees were in pots. All in-shell and kernel gen. Tissue culture, however, can gene- plants (Channuntapipat et al., 2003). measurements were made with electronic rate genetic variation, termed somaclonal Mean survival of shoot tips was 80% for scales and calipers. Data were analysed variation (Scowcroft, 1984). ‘Ne Plus Ultra’, 54% for ‘Nonpareil’, and using GenStat 6 and ANOVA. 78% for the hybrid rootstock, and there Previously shoot tips of ‘Nonpareil’, ‘Ne were no significant differences in survival Table 1 shows that there were significant Plus Ultra’ and an almond-peach hybrid between 3 days and 2 years. The whole differences between the controls and the rootstock, Prunus dulcis ‘Titan’ x P. persi- plants (tissue cultured only and cryopre- treatments for nearly all characters mea- ca ‘Nemaguard’ were successfully cryo- served) were grown in a shade house and sured. With ‘Nonpareil’, the control trees preserved for 6 months using a one-step finally planted out in 2001 in the almond (C) had significantly higher kernel weights, vitrification technique in our laboratory. breeding cage close to the mother trees shelling percentage, kernel width, and That work has since been extended to at the Waite Campus. kernel thickness, but significantly lower test for plant viability and genetic integrity fruit weight. Trees from cryopreserved after storage under liquid nitrogen for up GENETIC AND MORPHOLOGICAL material (LN) had significantly higher fruit to 2 years. At that stage it was unknown COMPARISONS weights. The same trend was seen with whether the treatments would damage The possibility of genetic change during ‘Ne Plus Ultra’ except that trees from tis- DNA and lead to unwanted mutations, long-term preservation was analysed by sue cultured material (TC) had significan- however the regenerated shoot tips did comparing DNA of the cryopreserved ma- tly higher fruit weight than C and LN not show any apparent morphological ab- terial with authentic non-preserved mate- trees. In both cases the control trees had normalities. This article is an update on rial. Changes in DNA could be of a struc- the lowest in-shell weights but the hea- the trueness to type of the regenerated tural kind, or changes in the placement or viest kernels. This may be due to their trees from those shoots and their genetic numbers of methyl groups on the DNA. age and size, however, as the control status. Changes in methylation can occur at high trees had a heavier crop load, and there frequencies during tissue culture, thus is an inverse relationship between crop CRYOPRESERVATION OF ALMOND causing genetic variation. The results of density and average kernel size (Hill et GERMPLASM the analysis showed that for ‘Nonpareil’ al., 1987), we would expect the control Shoot tips of two almond scion cultivars, and ‘Ne Plus Ultra’ there were some trees to have smaller kernels than the ‘Nonpareil 15-1’ and ‘Ne Plus Ultra’, and changes in DNA structure and methyla- treatments but we see the opposite effect an almond/peach hybrid rootstock, tion but this possibly occurred during the so the differences seen may be due to Bright’s Hybrid, were successfully cryo- process of tissue culture, which then per- the treatments. When we re-analysed the preserved using a one-step vitrification sisted through cryopreservation. For data without the controls (data not technique. This involved cold hardening, Bright’s hybrid, which originally arrived at shown) the significant differences betwe- preculturing and dehydration in a vitrifica- the Waite as tissue culture, the tests en the two treatments remained the same tion solution, followed by storage in liquid showed no detectable differences due to as in Table 1. In the case of ‘Nonpareil’ nitrogen (-196oC) for up to 2 years. the cryopreservation process. the LN treatment was significantly higher than TC for fruit weight, kernel weight, REGENERATION OF TISSUE To assess morphological characters, fruit shelling percentage and kernel width. After storage in liquid nitrogen the shoot was harvested from tissue-cultured only The opposite trend was observed for ‘Ne tips were removed after 3 days and then and from cryopreserved ‘Nonpareil’, ‘Ne Plus Ultra’ except for shelling percenta- at intervals of 3 months up to 24 months, Plus Ultra’ and Bright’s hybrid trees. This ge, which was not significant, and the then thawed rapidly at 30oC, washed and was compared to the original mother addition of kernel thickness, which was transferred to a growth medium and cultu- trees (in the case of ‘Nonpareil’ and ‘Ne significantly greater. In the case of red at 25oC. Shoots were then transferred Plus Ultra’) or untreated control trees Bright’s hybrid, the control trees were

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 25 Shoot tips of ‘Nonpareil’, thawed from LN, and micrografted Six-weeks old after hardening-off. on to almond x peach rootstock ('Bright').

Table 1. Comparison of fruit characteristic means

Trait ‘Nonpareil’ 1 ‘Ne Plus Ultra’ ‘Bright’s hybrid LN TC C LN TC C LN TC C2

Fruit weight (g) 2.06a 1.96b 1.83c 2.57b 2.77a 2.17c 5.7b 6.19a 3.99c Kernel weight (g) 1.17b 1.08c 1.25a 1.26c 1.37b 1.49a 0.85b 0.94a 0.41c Shelling percentage 56.5b 54.9c 68.4a 49.1b 49.6b 68.4a 14.8a 15.2a 10.0b % double kernels 6 2 0 22 20 20 0 0 0 % twin kernels 2 12 2 0 0 0 0 0 0 Kernel width (mm 12.6b 12.2c 13.2a 12.3c 12.9b 13.2a 13.8b 14.3a 12.2c Kernel length (mm) 24.4a 24.5a 24.3a 27.2a 26.7a 26.7a 23.5a 23.8a 18.9b Kernel thickness (mm) 7.49b 7.48b 7.80a 7.57c 7.94b 8.37a 6.40a 6.30a 5.05b Number 50 50 50 50 50 50 50 48 41

1Within cultivars, numbers in the same row with different letters are significantly different, l.s.d. P=0.05 LN = trees produced from cryopreserved material TC = trees produced from tissue cultured material only C = original mother trees except for Bright’s hybrid 2 ‘Bright’s hybrid control trees were in pots smaller and in pots, which probably resul- nels were present in Bright’s hybrid. Per- al., (1993). Environmental conditions ted in the observed significantly smaller centage double kernels appears to be were the same for each tree. fruit size, so the analysis was redone on genetically determined and varies greatly the treatments only. The results showed with cultivar (Egea & Burgos, 2000), but Percentage twin kernels were higher in that the TC trees were significantly higher may fluctuate due to physiological, envi- TC ‘Nonpareil’ than LN or control trees. for fruit weight, kernel weight and kernel ronmental or sampling conditions. The There were no twin kernels in ‘Ne Plus width, a similar trend to ‘Ne Plus Ultra’. small difference between ‘Nonpareil’ LN Ultra’ or Bright’s hybrid. The occurrence kernels and the others may be attributed of twin embryos is known to occur in Percentage double kernels were higher to some of these factors, e.g. sampling ‘Nonpareil’ (Kester & Gradziel, 1996) and for LN trees than TC or control trees in size was small due to the age and size of is strongly influenced by environment. ‘Nonpareil’, the same also for ‘Ne Plus Ul- the trees, however this was the same Martínez-Gómez et al., (2001) have sug- tra’ but to a lesser extent. No double ker- sampling size as was used by Dicenta et gested a sexual origin of twin embryos

26 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 that is under genetic control. The increa- In the future, other morphological charac- se seen here could be due to genetic al- teristics will be measured as the trees pact on germplasm conservation and terations brought about during tissue cul- mature such as, date of budburst, date of utilization. IPBGR Secretariat, Rome. ture. flowering, fruiting percentage, shell seal, Towill, L.E., Forsline, P.L., Walters, C., and biennial bearing. We will also be eva- Waddell, J.W. and Laufmann, J. 2004. When tree size and architecture were luating change at functional loci using mi- Cryopreservation of Malus germplasm compared between the two treatments, crosatellites and quantitative traits. using a winter vegetative bud method: the TC trees for both cultivars and the hy- results from 1915 accessions. Cryolet- ters 25:323-334. brid were larger than LN trees, however ACKNOWLEDGEMENTS the level of branching was similar bet- The authors are grateful to the Almond ween the treatments. Board of Australia, Horticulture Australia M. Wirthensohn1, C. Channuntapipat2, G. Limited and the Australian Research Collins1 and M. Sedgley3 Changes in methylation can occur at high Council for financial assistance. One of 1School of Agriculture & Wine, University of Adelaide, Waite Campus, PMB 1 frequencies during tissue culture, thus the authors (C.C.) was supported with a Glen Osmond, SA 5064 Australia. causing genetic variation. The results scholarship from the Thai government. 2Department of Biology, Silkaporn University, from our previous work showed that for Sanamchandrara Palace Campus, Nakhon ‘Nonpareil’ and ‘Ne Plus Ultra’ there were Pathom 73000, Thailand. 3 some changes in DNA structure and me- Faculty of The Sciences, University of New LITERATURE CITED England, Armidale, NSW 2350, Australia thylation but this possibly occurred during E-mail: [email protected] the process of tissue culture, which then Channuntapipat, C., Sedgley, M. and persisted through cryopreservation. Ge- Collins, G. 2003. Micropropagation of netic integrity often depends on tissue almond cultivars ‘Nonpareil’ and ‘Ne type used in tissue culture, which is the Plus Ultra’ and the hybrid rootstock Ti- reason meristems were chosen as these tan x Nemaguard. Sci. Hortic. 98:473- FLOWER DROP 484. usually retain developmental integrity. BY PRECOCIOUS ABORTION For 'Bright’s hybrid, which originally arri- Dicenta, F., Garcia, J.E., and Carbonell, IN WALNUT ved at the Waite as tissue culture, the E.A. 1993. Heritability of fruit characters tests showed no DNA structural changes in almond. J. Hortic. Sci. 68: 121-126. due to the cryopreservation process - Egea, J. and Burgos, L. 2000. Ovule dif- only methylation changes. Our current ferences between single-kernelled and INTRODUCTION work has shown that there are cultivar di- double-kernelled fruits in almond (Pru- As occurs in many other fruit and nut fferences in the response to this genetic nus dulcis). Ann. Appl. Biol. 136:291- trees, walnut presents different fruit variation caused by tissue culture. Other 295. drops, which usually are associated to workers have also found differences in Engelmann, F. 1997. In vitro conserva- competition phenomena or lack of pollina- the level of genetic instability generated tion methods. p.119-162. In: B.V. Ford- tion. Such drops usually stabilize through during culture between cultivars of a gi- Lloyd, H.J. Newburry and J.A. Callow appropriate orchard design and manage- ven species. Schwartz et al., (1981) (eds.), Biotechnology and Plant Genetic ment. However, the Persian walnut suf- using strawberry (Fragaria ananassa) Resources. Conservation and Use. fers a completely different fruit drop to the CABI, Wallingford, UK. stolon tips found cultivar differences in previous and which origin is still unk- the frequency of off-types. Towill et al., Forsline, P.L., Towill, L.E., Waddell, nown. This is a flower drop called Pistilla- (2004) showed an effect of cultivar on the J.W., Stushnoff, C., Lamboy, W.F. and te Flower Abscission (PFA) by Catlin el success rate of recovery after cryopreser- McFerson, J.R. 1998. Recovery and al. (1987). vation of Malus germplasm. Engelmann longevity of cryopreserved dormant apple buds. J. Amer. Soc. Hort. Sci. (1997) however, had found no modifica- 123:365-370. At the end of the 1970s, in California tions at the morphological, biochemical or some spectacular production decreasing molecular level in plants regenerated Helliot, B., Madur, D., Dirlewanger, E. of the cultivar ‘Serr’ was associated with from cryopreserved samples. Similar fin- and De Boucaud, M.T. 2002. Evaluation a flower drop which had not been obser- of genetic stability in cryopreserved dings have been made by other groups Prunus. In vitro Cell. Dev. Biol., Plant ved until then in walnut orchards. The in- working with Malus and Prunus (Forsline 38:493-500. cidence of this phenomenon varied ac- et al., 1998; Helliot et al., 2002). cording cultivars and orchard location. Hill, S.J., Stephenson, D.W. and Taylor, The economic importance of this pheno- B.K. 1987. Almond yield in relation to Though some changes manifested in fruit tree size. Sci. Hortic. 33:97-111. logic dysfunction led to search its origin. characteristics, it is difficult to determine whether they are the result of somaclonal Kester, D.E. and Gradziel, T.M. 1996. In 1990, in Hungary a low walnut orchard variation or other external factors. Never- Almonds. p.1-97. In: J. Janick and J.N. production was associated to the flower theless it is better to encourage conser- Moore (eds.) Fruit Breeding, Vol. 3, drop described in California as PFA, but, Nuts. John Wiley & Sons, New York. vation of valuable germplasm by as many they added an important information: PFA methods as possible. Cryopreservation Martínez-Gómez, P., Arulsekar, S. and were always higher on the trees placed can also be employed for uses other than Gradziel, T.M. 2001. Characterization of close to pollinators. These results allo- germplasm conservation, such as cryose- twin embryos in almond. Acta Hort. wed to suppose that an excess of pollen 591:257-262. lection; the selection through freezing of could cause a premature flower drop as samples with special properties, or cryo- Schwartz, H.J., Galleta, G.J. and Zim- already was referred to in some forest therapy; the elimination of viruses from merman, R.H. 1881. Field performance species (Pór and Pór, 1990). Afterwords, infected plants through apex cryopreser- and phenotypic stability of tissue cultu- several studies carried out by the Univer- vation. Because of its high potential, it is re-propagated strawberries. J. Amer. sity of California, in Davis and by IRTA - Soc. Hort. Sci. 106:667-673. expected that cryopreservation will beco- Mas de Bover in Catalonia, Spain, corro- me more frequently employed for long- Scowcroft, W.R. 1984. IBPGR report. borated that weighted load of pollen on term conservation of plant genetic resour- Genetic variability in tissue culture: Im- the stigma stimulated PFA. Many other ces. observations made in different countries

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 27 sition of the scar insertion on the stem shows the occurence of PFA on the tree, it is rare to observe mummified flowers still on the shoot. - Competition within flowers of the same female inflorescence. Occasionally, some flowers develop slower in relation to their nears and, finally, the most disadvanta- ged flowers drop. This drop is different from PFA, as in this case the flower’s de- tachment occurs at the basis of the ovary, flowers drop without peduncle (Figure 3). - Unfertilized flowers. Flowers can reach an advanced development stage without fecundation; they reach an appearance of small fruits, up to 8-10 mm of diameter. Those floral organs mostly detach about 3 weeks to one month after full blooming, - Young fruit drop. Recently fertilized nut drops are usually attributed to nutrients competition, fruit size is about 12-18 mm in diameter.

All of these flower and young fruit drops Figure 1. Flower drop due to PFA. Flowers of the same fruit set drop together linked to the peduncle. decrease the final harvest, but, the production loss corresponding to PFA is the highest. In commercial orchards an Table 1. PFA Incidence in 19 cultivars of different origen, during two years of study. amount of dropping up to 10-20% is considered as normal. Origen Cultivar/selection PFA (%)* 1998 1999 1998-99 ASPECTS INFLUENCING THE EXPRESSION OF PFA California ‘Chandler’ 4.7 2.2 3.0 IN AN ORCHARD ‘Chico’ 15.7 2.2 3.0 Plant material ‘Hartley’ 12.6 3.0 7.8 The phenomenon of PFA has been spe- ‘Serr’ 33.7 10.9 22.3 cially studied in Californian orchards of ‘Vina’ 17.0 9 13.0 cultivar ‘Serr’, where the problem was observed for the first time. Some orchards Chile ‘AS-0’ 10.1 1.0 5.9 of ‘Serr’ had up to 90% of flowers drop- ‘AS-1’ 12.0 1.6 6.8 ped due to PFA. Other cultivars like ‘AS-5’ 26.9 - - ‘Chandler’, ‘Chico’, ‘Franquette’ and ‘AS-7’ 12.1 3.6 7.8 ‘Hartley’ show PFA too, but the affection never reaches such important losses as Spain ‘MBT-31’ 64.0 - - described in ‘Serr’. ‘MBT-40’ 84.9 91.9 88.6 ‘MBT-119’ 86.6 60.2 73.4 At IRTA Mas de Bover this phenomenon ‘MBT-247’ - 45.2 - has been studied since the 1990s. The first works were focussed on the assess- France ‘Franquette’ 5.1 0.0 2.7 ment of the PFA«s incidence level on cul- ‘Lara’ 1.1 - - tivars of different origins: California, Chi- ‘Marbot’ 13.2 14.0 13.6 le, France and Spain. Four or five culti- ‘Mayette’ 13.7 - - vars per origin were studied. On each ‘Parisienne’ 11.7 16.9 14.1 tree, 200 flowers were labelled all around the canopy and two trees per cultivar - no data available were controlled. Records were taken, * percentage over the 200 flowers controlled every year from the beginning of female blooming (stage Ff: appearance of the primary pistillate flower), until fruit set (stage Gf). like Chile, France, Hungary, Spain and nomic incidence, the following can be Twice per week flowers were observed USA, highlighted that this early drop of considered: and drop due to PFA was recorded. Con- pistillate flowers exists in nearly all com- - Flower drop due to PFA. Flowers evolve trols were taken during two consecutive mercial cultivars. normally until stigmas start diverging; at years. Incidence of PFA was quantified this time, ovary stops growing and very as the percentage of female flowers dried HOW CAN PFA BE DISTINGUISHED quickly flowers get brown and end mum- in relation to the total number of initially FROM OTHER FLOWER DROPS mificated, finally they drop from the marked flowers. The results are shown in IN WALNUT TREES? shoot. Flowers have a diameter of 3-5 Table 1. In common walnuts, Juglans regia L., di- mm and drop together with the peduncle; fferent flower drops take place after the this fact distinguishes clearly the PFA The group of Spanish selections was the pollination period. According to their eco- from other drops (Figures. 1, 2). The po- most affected by PFA, showing 73,4% of

28 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 flower loss. The comparison with the behaviour of Chilean selections, which only had 6,8% flower drop due to PFA, resulted very surprising. The Californian and French cultivars showed medium values, but closer to the Chilean selections (Ta- ble 2). This table also contains fruit set values, which are opposite to those of PFA. The strong negative correlation between PFA and fruit set, confirms the economic importance of losses due to PFA compared to other types.

The location of the plots In the environment of walnut collections at Mas de Bover (Constantí-Tarragona), the PFA level in ‘Serr’ as described in different orchards in California has never been found. Observations made in Fran- ce show results more similar to the Spa- nish results too. The same occurs considering other Californian assessed cultivars (‘Chandler’, ‘Chico’, ‘Hartley’) and French (‘Franquette’): the damage by PFA in Spanish conditions has always Figure 2. Flower drop due to PFA. Both pistillate flowers drop together, linked to their peduncle. been less than in Californian conditions. The influence of the orchard’s location on the PFA incidence level is clear and has been referred to in several countries and cultivars.

The amount of pollen The first suggestion that an excess of pollen load increased flower drops by PFA, come from observations made in Hunga- ry, as already mentioned in the introduction, where tree rows next to pollinators had less yield than any other, more dis- tant rows. These observations were the Californian starting point and works were focussed mainly on this aspect of floral walnut biology. McGranaham et al. (1994) saw that an important amount of pollen on the pistillate flower stigma increased PFA in the cultivar ‘Serr’.

At Mas de Bover, some trials started to show the effect of an artificial supply of pollen in flowers, considering material from different origins: cultivars ‘Serr’ and ‘Hartley’ from California, and the selec- Figure 3. Drop due to competition in one floral set. The flower detached by the ovary basis, tion ‘AS-1’ from Chile and ‘MB-T-119’ not by its peduncle. from Spain. Data recording were made following the same process followed to assess PFA on different cultivars. Half of The orchard age made as a necessary step and annual the marked flowers were artificially polli- When an orchard reaches its adult phase, growths get shorter over time. Such situa- nated, placing pollen on the stigma using trees produce more reproductive organs: tion leads to a high increasement of sta- a brush, at the maximum pistillate flower pistillate and staminate flowers. A high minate flowers in relation to only one lo- receptivity (stage Ff1), and the others amount of pistillate flowers is always wis- cation of pistillate flowers: the terminal were open pollinated. All of them were hed: production increases. Trees usually shoot. This important catkin quantity pro- surveyed until fruit set (Gf stage), recor- have enough harvest mechanisms, such duces an unnecessary amount of pollen ding flower drops due to PFA twice a as flower drop or young fruit drop due to in the orchard which causes a considera- week. Results were clear: all cultivars in- competition, to maintain a productive ba- ble rise of PFA. creased the percentage of PFA (Fig. 4) if lance in adult phase. However, such con- flowers received a supplementary pollen trol does not exist for regulating the cat- The year load on the stigma (artificial pollination). kin production and the aging process Climatic conditions during the year have In the case of ‘Hartley’ artificially pollina- supposes a continuous increase of male an influencing aspect in the amount of flo- ted pistillate flowers dropped more than 6 flowers in the orchard. In an adult walnut wer drop due to PFA. The leafing out and times of the open pollinated flowers. orchard, production pruning is not usually blooming period of different walnut culti-

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 29 Table 2. PFA Incidence and fruit set Figure 4.- Percentage of PFA affection in 4 materials of different origin, according in 5 Californian cultivars, 4 selections pollination type (free or applying supplementary pollen with brush). from Chile, 5 French cultivars and 5 Spanish selections (two years average 1998-99).

Origen PFA (%) Fruit set (%)

California 11.0 82.8 Chile 6.8 88.0 France 11.9 79.0 Spain 73.4 21.6

vars and their dichogamy (time gap between male and female flowering period) possesses a strong genetic component. However, as the leafing out may change some days from one year to another, the homogamy of one cultivar (period in which both male and female flowering periods overlap) is affected directly by the previous autumn and winter temperatures. As female flowering overlaps mostly with the male flowering of the same cultivar in an adult orchard, an excess of pollen occurs at the time when the pistillate flower stigmas are receptive. This fact will originate a higher flower drop due to PFA. blem should be solved. In some orchards two years lasting assays, results are so in Spain (Extremadura), California and promising, that in Chile the process to re- Conclusions concerning PFA Chile, trials were made to eliminate cat- quest the registration of Retain(r) for wal- As a result of studies made in different kins, at the start of stigma receptivity, by nut was started, as it seems that the solu- countries and from the knowledge gathe- shaking trees. Results have shown a po- tion to the problem of flower drop due to red from flower drop by PFA, the follo- sitive effect in all cases. However, al- PFA could have been found. wing can be concluded: though pollen amount decreases in the - PFA affects all walnut cultivars in more orchard, the system is not fully understo- or less degree, no matter their origin. od and there are still a few important BIBLIOGRAPHY - Different degrees of PFA losses have questions to be answered as: When been observed, according to different cul- would be the best time, the most effecti- Catlin, P.B.; Ramos, D.E.; Sibett, G.S.; tivars. Selections from Chile are outstan- ve, for tree shaking?; How often should Olson, W.H.; Olson, E.A. 1987. Pistilla- ding for their reduced flower loss due to such catking elimination be made?, etc. te flower abscission of the Persian wal- PFA in comparison with the Spanish local Although these trials were made in ‘Serr’ nut. HortScience, 22 (2): 2012-205. introductions which can not be cultivated orchards and have shown good results, Gamalier Lemus, S. 2005. Control de la due to the high PFA levels detected. other more practical solutions should be caída de flores en Nogal ‘Serr’. Tierra - PFA has a reducing effect on produc- found. Adentro:18-21. tion. - An excess of pollen in the pistillate flo- Currently, different studies have confir- McGranaham, G.H.; Voyiatzis, D.G.; Catlin, P.B.; Polito, V.S. 1994. High pol- wer stigmas increases flower drop due to med that an excess of pollen generates len loads can cause pistillate flower PFA more than the average ethylen amount in abscission in walnut. J. Amer. Soc. - Climatic conditions during the year, the the floral organs, which origins the abs- Hort. Sci., 119 (3): 505-509. place and environmental conditions, may cission of female flowers. Recently, in influence the affection level due to PFA in Chile and in California, research is car- Pór A.; Pór, J., 1999. The effect of the excess of pollen on the fruitset of wal- some cultivars. ried out with blocking substances of the nuts in Balatonboglar. Acta Horticultu- ethylen synthesis. Thus the use of Çami- rae, 284: 253-256. MEASURES TO BE ADOPTED no ethoxy vinyl glicineÈ (AVG) is studied, IN A WALNUT ORCHARD TO AVOID which commercial product is called Rovira, M.; Aletà, N. 1997. Pistillate flower abscission in walnut cultivars. Acta OR PALLIATE FLOWER DROP DUE Retain(r). This product is already registe- Horticulturae, 442: 231-234. TO PFA red and used in apple to avoid preharvest To avoid PFA in an orchard it will be im- drop. The application of the product in or- Rovira, M.; Ninot, A.; Aletà, N. 1999. portant to chose cultivars with reduced chards of ‘Serr’ in Chile, gave harvest va- Pistillate flower abortion in walnut (J. re- flower drop by precocious abortion, but lues about four times more than a ‘Serr’ gia L.). Acta Horticulturae, 544: 287-293. also, orchard management is essential in orchard without treatment, which means order to control its aging with appropriate that the orchard production was nearly M. Rovira, N. Aletà pruning interventions and fertilization. four times bigger. Results from assays IRTA Mas de Bover. Mediterranean Fruit Trees made in California are less surprising, Ctra. Reus - El Morell, km 3.8 If the PFA is detected in an adult orchard however, trees treated with Retain(r) out- E-43120-Constantí (Tarragona) Spain or observations show that losses through numbered in about 50% the non-treated E-mail: [email protected] flower drop increases every year, the pro- trees. Although it only concerns one or

30 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 CALIFORNIA PISTACHIO 3,027 kg of dry in shell split nuts, or mar- 1974, and it harmed only trees less than INDUSTRY ketable nuts, per hectare. The total crop three years of age. Further, the southern value was US$ 573.27 million. The avera- San Joaquin Valley’s lack of spring rain ge return to the grower was US$ 4.48 per and low relative humidity, generally less kilo of marketable, dried inshell split nuts. than 75% RH, deter the foliar fungal di- INTRODUCTION The average value per bearing hectare seases that plague pistachio production Much of the history and production deve- was US$ 13,548.00. As it can be seen in north of 38¼ N latitude. The deep even loped for the California pistachio industry Table 2, planting and prices have been ri- slightly alkaline soils with good drainage is available in the online Pistachio Pro- sing since 2001. The California Pistachio and high boron concentrations supply the duction Manual: Fourth Edition 2005 Commission, (CPC), produces the Cali- conditions favored by the deeply and (Ferguson et al., 2005) available at https: fornia Pistachio Industry Annual Report, widely rooted Pistacia species used as //FruitsandNuts.UCDavis.edu. This article available for purchase through their web- rootstocks in California. The international will focus on recent cultivar and rootstock site, www.pistachios.org, which gives an production data now available demons- developments, and recent physiological excellent annual update of industry size, trates California has the most productive research on the mechanism of alternate production and value per acre, and irrigated pistachio industry, per unit of bearing, using irrigation to enhance nuts- grower return. cultivated ground, in the world. hell splitting and pistachio cultivar chill requirements. CALIFORNIA PISTACHIO CALIFORNIA PISTACHIO PRODUCTION AREAS PRODUCTION ECONOMICS CALIFORNIA PISTACHIO INDUSTRY Over 93% of the California’s pistachio There are currently about 600 pistachio STATISTICS production, and the world’s most produc- growers in California. Much of the crop is Table 1 demonstrates California is cur- tive orchards, are in the southern half of grown by large farming companies. The rently the second largest pistachio indus- the San Joaquin Valley, generally on the high establishment costs for pistachio, try in the world, ranking second only to west side between 33 - 35¼ N latitude and due primarily to an extended juvenility, Iran and surpassing Turkey, Syria, Gree- 118 - 120¼ W longitude. Within this longi- are high. Currently average establish- ce and Italy. Since 2001 California has tude and latitude the accumulated heat ment costs through the 6th year are US$ produced 28% of the world’s total pista- units during the growing season are well 27,397.00 per ha, exclusive of land costs. chio production. within the 2200-2800 range that produce Due to the increasing popularity of me- good shoot growth and fruit ripening. Oc- chanical hedging, and the total reliance California produces over 99% of the pis- casionally, lack of dormant chill is a pro- on mechanical harvesting, both generally tachios produced in the United States. blem, particularly if it has been a winter done by contract labour, an economically Pistachios have been a commercial crop without the fogs that prevent solar hea- feasible orchard should be at least 40 ha in Californian since 1977 and are now the ting of the dormant fruit buds. Data is limi- with 38.5 ha in production, spaced 5.2 m third largest nut crop in California, after ted but it appears 750 - 850 accumulated in the row and 5.8 m between rows. Or- almonds at 223,000 ha and walnuts at hours below 7.2¼ C is sufficient if the chill chards begin producing in the 6th year 91,000 ha. Currently there are 42,312 ha is not interrupted from December through and reach full bearing in the 8th to 10th bearing and 13,070 ha non-bearing ha, a February by clear sunny days with sustai- year. A more complete analysis of costs total of 55,382 ha, of pistachios in Califor- ned temperatures over 7.2¼ C. Freezes can be obtained at http://coststudies.uc nia. In 2005 the California industry produ- are less frequent, thus far there has been davis.edu. Generally an orchard at matu- ced 128.1 million kg with an average of only one since the industry started in rity will need to produce an average an-

Table 1. World pistachio production Mt, 2001-2005.

Year Iran* Total Iran** (RPPC) California Turkey Syria Greece Italy (w/o RPPC)

2001 114.99 69.99 72.71 35.02 39.92 6.49 0.09 269.21 2002 219.99 179.99 137.17 35.02 52.84 8.48 1.86 455.36 2003 184.88 165.11 53.52 89.9 49.99 8.98 2.00 389.36 2004 190.01 130.00 157.31 29.98 40.01 9.48 2.40 429.19 2005 190.01 179.98 128.10 60.01 60.01 9.48 2.40 450.00

Sources: United States Department of Agriculture and California Pistachio Commission. *Iran’s Ministry of Agriculture; figures cannot be verified. **Rafsanjan Pistachio Producers Cooperative (RPPC); figures cannot be verified.

Table 2. California pistachio crop area, yield and value, 2001-2005.

Surface (ha) Yield Crop Value Non- New Bearing Production Avg. Total Value/ Year Bearing bearing Total Plantings yield/ha (million kg)1 return value Bearing kg2 US $ ha US$

2001 31565 9853 41418 3248 2803 72.7 2.23 161.90 10649 2002 33589 9528 43107 1001 4409 137.1 2.43 332.64 9904 2003 35612 9273 44886 1220 1503 53.5 2.69 143.96 4023 2004 37636 10009 47645 2960 4180 157.3 2.952 464.71 11611 2005 42312 13070 55382 4640 3027 128.2 4.48 573.27 13548

Sources: California Agricultural Statistics Service, CPC Processors’ Producer Delivery Reports and Acreage Survey. 1 Calculated on CPC assessed-weight basis. 2 Weighted average which includes shelling stock. Revised by CASS periodically.

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 31 nual crop of 2,806 kg/ha at US$ 3.30 per kg return on dry, inshell split marketable nuts to the grower to generate a reasona- ble return per ha above total cash costs.

CALIFORNIA PISTACHIO PRODUCTION PRACTICES The California Pistachio Production Ma- nual: Fourth Edition 2005 is available online at http://FruitsandNuts.UCDavis.edu. There the current California pistachio practices can be reviewed in detail. Californian pistachio orchard. 'Kerman' pistachio nuts.

While many of the world’s major fruit and nut industries often have the same production practices, pests and diseases, and harvest and processing methods as their major international competitors, the California pistachio industry has developed in strikingly different ways from the world’s largest pistachio producer, and their major competitor, Iran. Iran, a much older, more traditional, industry, has a wealth of diverse germplasm, a more ex- treme climate, severely limited supplies of poor quality water, limited labor, an in- Pistachio nuts in a transport container. Seeds from P. vera, P. chinensis, P. palaestina and P. atlantica. sect pest complex developed over centuries, no effective extension service, poor cooperation within the industry, and an unstable government. While all of these things could produce major differences between any two industries, it is the available germplasm, water and labour resources, and insect pest complex that have produced the greatest differences. Generally the California pistachio industry has had the advantage of sufficient, good quality water, available labor and mecha- nization for the two most expensive production factors, pruning and harvesting, Tree and nuts of 'Golden Hills'. Tree and nuts of 'Lost Hills'. and relative freedom from, or quick solutions to, truly devastating pests. And only now is California exploring limited cultivar diversification.

The California pistachio industry has adapted and modified practices from all the previously established California fruit and nut industries. The average pistachio orchard is established on cheaper, less desirable ground, often with soil and drainage limitations that must be physically or chemically amended prior to planting and Tree and nuts of 'Kerman' from years 2003 Flowers and tree of pollinator 'Randy' . during production. Increasingly, marginal and 2004. quality water is used for irrigation. The orchards are planted with unbudded, 8-14 applied foliarly. Orchard floors are kept the nuts after shell hardening inflict nut month old seedling rootstocks field clear with mowed cover crops, herbicides damage that must be separated from the budded within the first season in the or discing. Dormant tree pruning is in- final product, and therefore are more da- ground. Tree training for the first two creasingly done mechanically with hed- maging. Foliar and fruit fungal diseases years is done during both the growing gers and toppers. Insect pests are con- have been less frequent, and less of a and dormant seasons. Thereafter, in trolled with insecticides, there is little problem than insect pests. However, they years three through six, training is done biocontrol. The major insect pests are have been devastating when inoculum during the dormant season only. Or- generally divided into those which harm buildup was sufficient and exacerbating chards are budget irrigated with drip or the nuts prior to shell hardening and tho- weather conditions, particularly cold, microsprinkler systems that allow fertiga- se that harm the nuts after shell harde- wet springs, coincided. The major foliar tion. The trees are not luxury consumers ning. Insects that harm the crop before fungal diseases, Botryosphaeria dothi- of macronutrients and require only nitro- shell hardening, in effect, thin the crop dea, Alternatia alternata, and Botryotinia gen and potassium fertilization. Only the within the cluster, and therefore can be fuckeliana (Botrytis cinerea) have been micronutrients boron, zinc and copper are somewhat tolerated. Insects that harm controlled thus far with pruning and pro-

32 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Figure 1. California pistachio rootstocks replicated field trials. CALIFORNIA CULTIVARS AND ROOTSTOCKS Within the genus Pistacia there are eleven species. Of the eleven only one, Pis- tacia vera, produces nuts large enough to be considered edible. Among the other ten species, several are used as rootstocks.

CALIFORNIA FEMALE CULTIVARS If the California pistachio industry has an Achilles heel it is the industry’s reliance on a single female cultivar; Pistacia vera cv. ‘Kerman’. Collected in Iran in 1929 by W.E. Whitehouse and released to the industry in 1957 after selection trials by Lloyd Joley at the United States Depart- ment of Agriculture (USDA) Chico Experi- ment Station in northern California. ‘Ker- man’ was selected primarily for its size, split percentage and yield. As California Cold Tolerance: Dec. 1990 Production Recommendation: Salinity Tolerance has had virtually no other accessions ri- ¥ Soil water extract: gorously tested since there is little infor- ¥ 11 nights at 4-12 ¼F ¥ Pistachios on all three rootstocks produce mation about ‘Kerman’s performance re- equally well up to: lative to other cultivars under commercial -> UC Berkeley I: UCBI ¥ 8 dS/m ECe production conditions. No death ¥ 6720 ppm TDS ¥ 8 mmho/cm In 2005, after eight years of replicated -> Pioneer Gold I: PGI trials, two females, ‘Lost Hills’ and ‘Gol- 41% died ¥ Limiting factors: den Hills’ were released by Parfitt et al. ¥ soil saturation (2005), Department of Plant Sciences, ¥ osmotic pressure UC Davis. Both cultivars appear to have ¥ Cl and Na uptake with Integerrima the potential for higher yields, a better percentage of split marketable nuts and an earlier harvest than ‘Kerman’. ‘Golden Tolerance Ranking of Pistachio Rootstocks in SJV 1989 - 2002 Hills’ flowers one to two weeks before ‘Kerman’ and ripens two to four weeks Limiting Factor (1) best worst (4) earlier. This fact alone confers an advan- Rootstock Frost Yield Salinity Vert. tage in more efficient use of harvest ma- chinery and processing plants, as well as P. integerrima 4231the ability to avoid the third flight of navel P. atlantica 1413orange worm, Amyelosis transitella. Con- PGII 3 3 2 4 trolling this insect requires insecticides UCB1 2 1 2 3 with 14 to 21 day pre harvest intervals, which often results in shell staining hull Alternate bearing average = for all four rootstocks: scale of 0 - 1 = 0,67 degradation and lower grades. Though nut size and weight are similar to ‘Ker- man’ the percentage of split nuts was 5% phylactic sprays. Verticillium dahliae, the CALIFORNIA PISTACHIO INDUSTRY higher, the percentage of blanks lower, only soil borne disease of any importance ORGANIZATION and total yields 46% higher. The tree is is controlled by tolerant and resistant Industry affairs are governed through the smaller and denser than ‘Kerman’. rootstocks. Nematodes present no pro- grower and processor organization, the blem. Specific information on all aspects California Pistachio Commission (CPC); ‘Lost Hills’ has the same bloom and har- of California pistachio pests and diseases http://www.pistachios.org. Established by vest time advantages as ‘Golden Hills’. is available at http://www.ipm.ucd.edu. As a grower majority vote, supported by gro- ‘Lost Hills’ has larger and heavier nuts with any newly introduced crop, insect wer production levies, and run by elected than ‘Kerman’ and produces 7% higher pests and diseases build to a threshold grower members and a hired staff, the split nuts and 26% higher yields than before becoming evident. The pistachio CPC sets the grower production levies and ‘Kerman’. Both new cultivars flower better insect pest complex in Iran provides nut quality standards, generates industry under marginal chill conditions; chill accu- California with a valuable warning of statistics, and supports generic market mulations of less than 750 hours below potential insect pests. Pistachio har- promotion, production research, and gro- 7.2¼C with frequent clear, sunny days. vesting in California is totally mechani- wer education. The CPC serves as a fo- This suggests determining when to har- cal. The harvested product is sold at the rum for grower and processor concerns vest these new cultivars for maximum processor gate and the average grower though it is prohibited by law from political inshell split yield and minimum shell stain does not market his product. Virtually all activity. Multiple departments within the will be easier than with ‘Kerman’. the product is sold as dried, inshell, and University of California, California State salted as a snack item. In striking con- University and United States Department CALIFORNIA MALE CULTIVARS trast to walnuts and almonds there are of Agriculture have strong cooperative re- ‘Kerman’ is pollinated almost exclusively few ingredient or cooking uses for pista- lationships with the CPC primarily for ge- by the ‘Peters’ male, a California selec- chios. nerating research and other information. tion found in the early 1900s by A.B. Pe-

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 33 ters in Fresno, CA. Occasionally the two Russian selections, the early 02-16 and the late 02-18, are used to compliment ‘Peters’ bloom period. Parfitt et al. (2005) have released a new male cultivar, ‘Ran- dy’. ‘Randy’ blooms one to three weeks before ‘Peters’, and, like ‘Peters’ sheds pollen for two weeks. Pollen from ‘Randy’ is more durable, declining from 75% viability to 35% after 29 days of storage versus 45% to 15% for ‘Peters’ after 27 days. While ‘Randy’ will probably be the primary pollinater for «Lost Hills’ and ‘Gol- den hills’ it may also be useful for pollinating ‘Kerman’ under marginal chilling conditions when ‘Peters’ often blooms too Figure 2. Canopy of scion grown on UCBI Figure 3. Canopy of scion grown on P. atlantica late for effective pollination of ‘Kerman’. rootstock. (Photo courtesy of T.M. Spann). rootstock. (Photo courtesy T.M. Spann).

These three new patented cultivars, the females ‘Lost Hills’, and ‘Golden Hills’ hworth, Department of Plant Pathology, control treatments. The trials used diffe- and the male ‘Randy’, were limited indus- University of California Berkeley. The re- rent factors to measure salinity tolerance; try releases in 2005 and 2006. Hopefully, sulting rootstock, the only commercial in- in the greenhouse trial it was growth. In with these patented releases the Califor- terspecific hybrid, was named University the field trial it was yield. The trials produ- nia pistachio industry will, in the future, of California Berkeley I, UCBI. ced different rankings. Because the accept the responsibility for ascertaining greenhouse trial was more replicated, the budwood provenance. Until this time Figure 1 summarizes the results of multi- and controlled, the results from this trial the origin of unpatented buds of ‘Kerman’ ple, long term, replicated field trials (Eps- were used to develop the rankings in this and ‘Peters’, ’02-16', and ’02-18' have tein et al., 2005; Ferguson et al. 2002; table. Generally, trees on Atlantica root- been largely unaccounted for and unre- Ferguson et al. 2005; Sanden et al. 2004) stocks are the most salinity tolerant follo- corded by both the grower and contract into relative rootstock rankings for the wed by trees on the hybrid rootstocks. budder. Thus far this lack of budwood ac- major limiting factors in California pista- Trees on PGI rootstocks were the least countability has caused no problem, but, chio rootstock selection. Generally, the li- tolerant of salinity. However, in the green- any horticulturist familiar with the citrus miting factors determining rootstock se- house and field trial trees on all the root- industry knows the devastation bud borne lection in pistachio production are, in or- stocks showed no decrease in growth or viruses can wreak in an industry with di- der, freeze, Verticillium and salinity tole- yield when the ECe of the soil water ex- versified cultivars, much less in an indus- rance. The effect of rootstock on yield, a tract was 8 dS/m or less. This ranks pista- try with a single female and male cultivar. horticultural enhancement, is considered chios as the most salinity tolerant tree In 1990 this author observed a mildly vi- after these limiting factors have been fruit crop after dates. rulent pathogen in a young orchard in met. Figure 1 demonstrates that while Australia. The pathogen caused stem-pit- trees on Atlantica rootstocks tolerate cold Interestingly, despite significant differences ting and flagging of the distal branch. The and salinity significantly better than trees in yield, trees on the four different root- pathogen was successfully transmitted in on the other three rootstocks they are stocks had no consistent, significant diffe- a limited demonstration of Koch’s postu- susceptible to Verticillium and have signi- rences in the percentage of non-split, or lates in 1990. Buds from a symptomatic ficantly lower yields. Trees on PGI root- blank nuts within a crop year, and no sig- branch produced the same symptoms in a stocks have the best tolerance of Vertici- nificant differences in the calculated alter- previously asymptomatic greenhouse bud- llium, but are more susceptible to cold nate bearing index. Severity of alternate ded seedling. and salinity, and produce lower yields bearing, alternating high and low crops, is than trees on UCBI rootstocks. Trees on indicated by an alternate bearing index CALIFORNIA PISTACHIO ROOTSTOCKS PGII rootstocks possess no great advan- (ABI). On a scale of 0 to 1, 0 means even As with most tree fruit industries the intro- tages and are the most susceptible to crops annually and 1 indicates crop on al- duction of new rootstocks is primarily a Verticillium. For this reason alone PGII is ternate years. Among the four rootstocks function of diseases that affect the cur- no longer commercially available. Howe- in the three trial locations the calculated rent rootstocks. The California industry ver, in the trials individual trees on PGII alternate bearing indexes ranged, insigni- was established on the horticulturally har- rootstocks were stellar performers. This ficantly, from 0.55 to 0.79. Therefore, while dy rootstock, P. atlantica. However, Atlan- suggests that when better discriminatory rootstock can produce strong effects in ab- tica’s susceptibility to the soil borne fun- tools are available this hybrid should be solute crop load of the ‘Kerman’ scion, gus, Verticillium dahaliae (Kleb) precipita- more thoroughly investigated. Trees on the degree of alternate bearing appears to ted the mid 1980s introduction of the UCBI rootstocks are more cold tolerant be primarily a function of the scion cultivar. more productive, but less cold hardy, P. than trees on all but Atlantica, tolerate integerrima, now marketed as Pioneer Verticillium as well as trees on PGI, and As a result of the above trials, the Califor- Gold I, PGI. This species selection was produce significantly higher yields than nia pistachio industry is now being deve- also used to produce the hybrid rootstock trees on all the other rootstocks. loped on two rootstocks; PGI and UCBI. P. atlantica X P. integerrima, Pioneer Due to established reputation, and grea- Gold II, PGII. In the late 1980s a hybrid of Salinity tolerance among these rootsto- ter availability, PGI rootstocks dominate. an import from Chios, Greece, either P. cks was developed in two separate trials; Under California conditions both perform atlantica, (D. Parfitt, personal communi- a budded but non bearing, well replicated equally well. The major difference to be cation) or P. lentiscus (J. Doyle, personal greenhouse trial with excellent control expected between the two is the higher communication) was crossed with the pol- and a long term field trial with only three yields, approximately 15%, the rootstock len parent, P. integerrima, by Lee As- replicates per treatment and less ability to trial results indicate will be produced by

34 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Figure 4. California pistachio production from 1985 through 2005. The alternate creasing soil salinity levels, combined bearing pattern is obvious. The aberrations in 1993 through 1995, and 2005 may with the losses in growth and yield obser- have been produced by new orchards entering bearing. ved as soil water salinity increased, suggests the primary mechanism of salinity damage to pistachios is osmotic pressure, not specific ion toxicity. Future research should focus on this question and the ability of pistachios to adjust osmoti- cally.

EFFECT OF ROOTSTOCK ON ‘KERMAN’ SCION GROWTH HABIT, YIELD AND ALTERNATE BEARING In the rootstock trial mentioned above Kerman scions grown on rootstocks with Integerrima parentage, either PGI or UCBI, Figure 2, versus trees grown on P. atlantica rootstocks, Figure 3, had a growth pattern that produced the higher yields. The significant differences in yield among the trees on the different rootstocks were not a function of nut weight, or number of nuts per cluster. Rather the increase in yield was a function of more clusters per tree. Interestingly, recent trees on UCBI rootstocks. However, the- Within this trial trees on PGII rootstocks work by T.M. Spann, (unpublished data) se trials were only conducted through the were highly infected, had high mortality, demonstrated these increased clusters 13th leaf. As the trees age and are pru- and such marked declines in vigor that were not borne differentially on the longer ned to an equivalent canopy this UCBI they produced significantly lower yields shoots within the Kerman scion canopy of rootstocks yield advantage may diminish. than trees on P. altantica rootstocks. In trees grown on the Integerrima parentage all the other trials trees on PGII rootsto- rootstocks. Thus, even though it was the CALIFORNIA INDUSTRY ROOTSTOCK cks had significantly higher yields than effect of rootstock on the growth and frui- TRENDS trees on P. atlantica rootstocks. This dif- ting habit of the ‘Kerman’ scion that pro- Within the industry, one nursery is produ- ference in reaction to Verticillium sug- duced the differences in yields, the most cing a tissue culture clone of UCBI. There gests UCBI and PGII are not the same obvious difference, longer shoots with are no replicated trials comparing perfor- hybrid cross of P. atlantica X P. integerri- both preformed and neoformed growth mance of this clone to a UCBI seedling ma, as has been suggested. The mater- were not factors in the increased number population. However, thus far there is no nal parent in UCBI may not be P. atlantica. of clusters per tree. reason to suspect trees on this rootstock will perform better, or worse, than trees EFFECT OF ROOTSTOCK RECENT RESEARCH: ALTERNATE on seedling rootstocks. Thus far orchards ON ION UPTAKE, TRANSLOCATION BEARING, IRRIGATION EFFECTS developed with the clones appear uniform. AND SPECIFIC ION DAMAGE ON NUT SPLITTING AND CHILL The rootstock salinity trials demonstrated REQUIREMENTS ROOTSTOCK EFFECTS strikingly different uptake of chloride, so- ON VERTICILLIUM INFECTION, ION dium and boron among the rootstocks. Alternate Bearing UPTAKE, SCION GROWTH AND YIELD Generally, trees on PGI rootstocks took Among the interesting physiological phe- In the rootstock trials some interesting dif- up, and translocated to the scion, signifi- nomena germane to pistachios is the ferences among the rootstocks emerged. cantly more chloride, sodium and boron phenomenon of alternate bearing, alter- These differences were primarily in how to the scion than trees on UCBI rootsto- nating high and low crops. The alternate the trees on different rootstocks tolerated cks. Trees on UCBI rootstocks took up sig- bearing history of the California pistachio Verticillium, and salinity, and how growth nificantly less sodium, boron and chloride industry can be seen in Figure 4. and yields were affected by rootstock. and sequestered it primarily in the rootstock. This differentially higher ion uptake, Pistachios are strongly apically dominant EFFECTS OF ROOTSTOCK and translocation to scion, coincided with and bear their crop on one year old wood. ON VERTICILLIUM INFECTION significantly less growth of trees on PGI Further embryo, or nut, development is PGI and UCBI, the rootstocks that produ- rootstocks, but if the two phenomena are sequential; the nutshell forms, increases ce best in Verticillium infested conditions linked is unknown (Ferguson et al. 2002). in size, and hardens, before the embryo, appear to have different modes of infec- or nut, grows. Simultaneously, the sub- tion. Trees on PGI are generally heavily Further, all the specific ion toxicities in tending distal shoot growth is setting and infected but appear to tolerate the disea- the ‘Kerman’ scion leaves in both the developing the buds for the next year’s se, and not manifest symptoms. Trees on greenhouse and field trials were produ- crop. Thus, as the crop on one year old UCBI rootstocks are infected much less ced by boron. And the appearance of wood is producing the strongest pho- frequently, but, when infected manifest specific ion toxicity produced by boron tosynthetic demand of the season, filling with lower vigor. However, even with in- was consistent among the rootstocks or the nutshells that have formed, current fection and some loss of vigor, trees on with increasing salinity level. Sodium and shoot growth is simultaneously develo- UCBI rootstocks still produced significant- chloride produced no specific ion toxici- ping the buds for the next year’s crop. It ly higher yields than trees on PGI root- ties and were equally distributed between is during this late June early July period stocks. Thus it appears PGI is tolerant and necrotic and normal tissue. This lack of that the apparent mechanism of alternate UCBI is resistant (Epstein et al., 2004). consistent specific ion toxicities with in- bearing manifests; the newly set and de-

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 35 veloping fruit buds on current year’s III, the result would be a higher percenta- In a series of experiments Beede et al. growth abscise. The heavier the subten- ge of split nuts. (2006) have demonstrated dormant oil ding crop the heavier the bud abscission. applications for dormant scale control, no It has long suggested that the reason for The experimental results in 2003 and later than two weeks before the end of the bud abscission was a simple carbo- 2004 supported this hypothesis (Goldha- the chill accumulation period at the end hydrate competition, mediated by a mer et al., 2006.) To produce stage I of February, can sometimes compensate growth regulator signal. Endogenous abs- water stress requires either withholding for insufficient chill. However, in particu- cisic acid levels, exogenous benzyl ade- irrigation, or extracting winter and spring larly poor chill years such as the winter of nine and 2, 4-D applications, and girdling rainfall, with a cover crop. Using these 2005 - 2006 even dormant oil applica- work support this theory. However, until methods stage I water stress was succes- tions were not sufficient to produce good recent work by Tim Spann (personal com- sfully produced in spring 2003 and 2004. bloom and fruit set. munication) nobody had supported this In those two seasons stage I water stress theory with experimental data. His data reduced non splitting by 45% and increa- suggests that the photosynthates produ- sed split nut yields by 21%. However, the LITERATURE CITED ced by the immediately subtending leaves resulting individual nut weight was also are not sufficient to support both shoot reduced by 6.5%. Also, the stage I stress Beede, R.H., L. Ferguson, K. Beinhorn, and bud growth. His data demonstrates increased the incidence of early split nuts, T. Thomphson, G. Driever and T.J. Mi- carbohydrates are mobilized from the sto- their infestation by Amyelois transitella chailides. 2006. Comparison of three rage reserves of the tree shoots, and that (Walker) and potential for colonization by Chill hour accumulation methods for in the competition for these, the strongest Aspergillus flavus and parasiticus. In monitoring rest in Pistachio; Final Re- port. California Pistachio Industry: An- sink, the developing fruit, is favored. In- 2005, due to heavy winter and spring ra- nual Report Crop Report 2005 - 2006. terestingly, after bud abscission, and by ins and low evaporative demand during California Pistachio Commission. 1318 season’s end, the carbohydrate reserve stage I water stress could not be induced East Shaw Ave, Suite 420, Fresno, Ca. status of the fruit and inflorescence bea- and there were no differences among the 93710-7912. USA. Pages 98-99. ring shoot is restored. Thus, to observe treatments. Thus, while rainfall and wea- Epstein, L., R,. Beede, S. Kaur, and L. this carbohydrate competition and remo- ther may sometimes prevent this method Ferguson. 2004. Rootstocks effects on bilization sampling requires in season of increasing nutshell split, if irrigation can pistachio trees grown in Verticillium shoot sampling. By the end of the be manipulated to achieve the correct ba- dahliae-infested Soil. Phytopathology growing season, after leaf abscission, the lance between enhancing nutshell split, 94 (4): 388-395. carbohydrate status of the shoots has and minimizing early split incidence and Ferguson, L., J.A. Poss, S.R. Grattan, been restored to the same level as a non- effects on nut size, this could prove to be G.M. Grieve, D. Wang, C. Wilson, T.J. bearing shoot. This is the first data that a valuable technique which also conser- Donovan, and C.T. Chao. 2002. Pista- elucidates the cause of alternate bearing. ves water. This research is continuing. chio rootstocks influence scion growth While it does not suggest any method of and ion relations under salinity and boron stress. Jour. Amer. Soc. Hort. Sci. control it is the first experimental data to Chill Hour Accumulation Requirements 127 (2): 194-199. support the carbohydrate competition for ‘Kerman’ and ‘Peters’ theory of alternate bearing. As pistachio, a deciduous tree of tempe- Ferguson, L., H. Reyes, B.L. Sanden, rate origins continues to be planted in S. R. Grattan, L. Epstein, and W.H. Irrigation Effects on Shell Splitting Krueger. 2005 Pistachio Rootstocks. In subtropical climates areas the question of Pistachio Production Manual: Fourth D. Goldhamer, Land, Air and Water Re- chill accumulation requirements, and how Edition 2005. Ed. L. Ferguson. pages sources Department (LAWR), UC Davis, to best assess them is important. Three 62-65. earliest work on irrigation effects on the years of chill assessment work with ‘Ker- yield components of pistachio demonstra- man’ and ‘Peters’ in California suggests Goldhamer, D.A., R.H. Beede, T. J. Mi- chailides, M. Salinas, M.A. Doster. ted water stress at different stages of the that the climate of the San Joaquin Valley 2006. Effects of regulated deficit irriga- nut’s sigmoid growth curve could produce does not always meet the chill accumulation on shell splitting and nut quality differences in nut size and shell splitting. tion requirements of either cultivar and (third year report). California Pistachio Stage I, early April to mid May is the pe- that dormancy breaking treatments are Industry: Annual Report Crop Report riod of nutshell enlargement and shoot sometimes necessary. Research thus far 2005 - 2006. California Pistachio Com- mission. 1318 East Shaw Ave, Suite elongation. Stage II, mid May through suggest the male ‘Peters’ is more sensiti- 420, Fresno, Ca. 93710-7912. USA. pa- early June is the period of nutshell thicke- ve to marginal chilling then the female ges 104 - 105. ning, or hardening, and further develop- ‘Kerman’. Using total accumulated hours ment of next season’s inflorescence bud below 7.2¼C with no correction for higher Parfitt, D.E., C.E. Kallsen, J. Maranto. 2005. Pistachio Cultivars. In Pistachio development on the current years shoot daytime temperatures, the male ‘Peters’ Production Manual: Fourth Edition growth. Stage III is the period of nut fill requires a minimum 850 hours below 2005. Ed. L. Ferguson. pages 62-65. and nutshell splitting. Goldhamer’s earlier 7.2¼C to provide minimal flowering. Below work demonstrated Stage I water stress this level of accumulated chill the ‘Peters’ Sanden, B. L. Ferguson, H. Reyes, and decreased nut size but also increased often blooms after the full bloom period of S.R. Grattan. 2004 Effect of salinity on evapotranspiration and yield of San nutshell splitting. He found stage II water the ‘Kerman’ female, which appears to Joaquin Valley Pistachios. 2004 Acta stress had no effect on nut shell splitting. bloom well with 850 accumulated chill Horticulturae 4th International Sympo- He further found stage three water stress hours below 7.2¼C. The results thus far sium on Irrigation of Horticultural decreased nutshell splitting. Work by V. suggest the chill accumulation monitoring Crops. XX.XXX. In press. Polito, Plant Sciences, UC Davis has sug- methods need to account for higher dayti- gested nutshell splitting is a purely me- me temperatures, and perhaps include a chanical phenomenon, that the force of method of monitoring sunlight hours and L. Ferguson the developing nut forces the shell open its effects on bud temperatures. It also Department of Plant Sciences, University along the nutshell suture. This suggests suggests, that as pistachios continue to of California Davis that if the nutshells could be made sma- be planted in subtropical areas, the chill 1 Shields Ave. Davis CA 95616 USA ller by stage I irrigation stress, and the requirements of the cultivars should be E-mail: [email protected] nuts larger by optimal irrigation in stage well known.

36 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 THE LOW YIELD REASONS AND Pistachio is also grown in Iran, Syria, Tur- SOLUTIONS FOR PISTACHIO key and USA. The other pistachio produ- Pollination occurs by the transport of po- PRODUCTION IN TURKEY cing countries are in the Near East, North llen from male to female trees by the Africa (Tunisia, etc.) and Southern Euro- wind. Pistachio flowers have no petals pe (Greece, Italy and Spain). According which attract insects. Therefore pistachio to six years average (1999-2004), the orchards should have male trees and the ABSTRACT production values in Iran are over ratio of male to female trees planted Pistachio can only be grown in some 261.000 t (46,2 %) which is nearly half of should be 1/8 or 1/11 (Kaska, 1990, Ak, countries due to its continental climatic the world production. USA with 86.000 t 1992) (Table 2). It is necessary to have requirements. Pistacia spp. are not widely represents 23.2 %, and Turkey with a enough male trees to insure adequate po- naturally spread in the world. The leading yield of 58.000 t keeps 11,5 % of total llination. pistachio producing country is Iran with world production (Table 1). A surprising 46,2 % of the world production, second is pistachio producing country is China. Chi- The orchard should possess a sufficient the USA with 23,2 % and Turkey is third nese scientists reached amazing figures. number of male trees and pollen shed- producing approximately 58.000 t and They claim that Chinese production is ding time should overlap with female flo- this is 11.5 % average of 6 years total around 1.000 t (personal communication wering time. In addition, a good male tree world production. Only Iranian and Cali- of researchers working in Kashgar Pista- should have some of the following featu- fornian pistachio orchards are irrigated. In chio Research Station). res: Turkey pistachio grows under rainfed conditions as well as on poor soil. If these Table 1 shows that Pistachio production 1. Growth should be strong and upright three important pistachio producing coun- is very low when the area and producing 2. Flowering period of the male should be tries are compared, the largest pistachio surfice are compared. The reasons are long enough to overlap the flowering pe- planted areas belong to Iran with 420.000 explained below. riod of females ha. The USA has 80.0000 ha and Turkey 3. The number of clusters must be high has 220.000 ha of pistachio surfice. Tur- INSUFFICIENT POLLINATION 4. Flower clusters must be big sized key shows a very low pistachio nut yield Male and female flowers of pistachio are 5. The amount of pollen production in among the producing countries. The main produced on separate trees. This trait each cluster must be high crop limitations are: pollination, irrigation has been recognized since 1697, (White- 6. Yield potential should be high and fertilization. house and Stone, 1941) which means 7. Germination rate of pollen must be that pistachio trees (Pistacia vera L.) are high INTRODUCTION dioecious. 8. Pollen’s viability "in vivo" should be The pistachio nut belongs to the genus long Pistacia of the family Anacardiacea. Pis- The male trees in Turkey were cutted or 9. It should not show alternate bearing tacia genus have 11 species. Pistacia top worked to females due to unfruitful- (Ak et al., 1998; Ak, 2001). vera is mainly commercially grown at 30¼- ness in the 1950s. It was thought that 40¼ latitude North and suited to microcli- male trees were not necessary for pro- LACK OF IRRIGATION matic areas in the world. The biggest production. This situation was carried out for In Turkey pistachio orchards were esta- duction areas belong to Iran (420.000 a very long time. Later on farmers were blished in dry and unirrigated land where ha). The second country is Turkey but its well informed by researchers. Pollination soils are poor, unfertile and calcareous. yield is very low when compared to pro- and fertilization are necessary to get see- The farmers still believe that pistachio duction surfice (Arpaci et al., 2005). ded fruits. should be grown without irrigation. Their

Table1. World Pistachio production (Mt).

COUNTRIES YEARS AVER RATE AGE (%) 1999 2000 2001 2002 2003 2004 2005

Iran 304.000 112.000 249.000 220.000 184.000 190.000 190.000 207.000 46,20% U.S.A. 55.790 110.220 73.030 137.440 53.980 157.397 140.000 103.479 23,20% Turkey 40.000 75.000 30.000 35.000 90 000 30.000 60.000 51.429 11,48% Syria 30.133 39.923 37.436 52.840 50.000 40.000 60.000 44.333 9,90% China 29.000 22.000 26.000 28.000 30.000 32.000 34.000 28.714 6,40% Greece 9.027 9.536 9.540 8.500 9.000 8.500 9.500 7.724 1,70% Italy 2.649 2.768 1.762 1.877 1.993 2.400 2.400 2.264 0,50% Tunisia 1.200 1.600 1.100 800 800 800 800 1.014 0,23% Uzbekistan 1.100 1.000 1.000 1.000 1.000 1.000 1.000 1.014 0,23% Pakistan 194 209 196 195 200 200 200 199 0,04% Madagascar 270 160 160 160 160 160 160 176 0,04% Kyrgyzstan 100 100 100 100 100 100 100 100 0,022% Morocco 50 50 30 30 50 50 50 44 0,0098% Mexico 36 31 21 7 7 7 7 17 0,004% Cyprus 18 15 15 15 15 15 15 15 0,003% Mauritius 5 5 5 5 5 5 5 5 0,001% WORLD 473.572 374.61 429.395 287.969 421.310 462.634 498.237 448.027 100 %

Source: http://www.fao.org

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 37 grandfathers raised them with this idea. In order to catch and store the water from fertile soil conditions. According to Ak These old men suffered a bad experience the winter and spring rains and snows, and Agackesen (2003) pistachio trees with irrigation. After they had irrigated the soils are ploughed in autumn. On the should be established under irrigated and their pistachio trees, the trees had died. other hand, in order to prevent evapora- fertile or good soil conditions to get high And they passed this information to their tion, soils are cultivated by chisel ploughs yield and quality. Comparing both condi- sons. But this situation occurred due to over the whole spring and summer mon- tions, the yield of ‘Kirmizi’ pistachio culti- wrong irrigation methods. However in the ths. This type of soil workings destroy the var either fresh weight or dried weight un- last decades Research Institutes depen- weeds as well (Kaska, 1995). As it is well der good conditions (irrigation and fertile ding of the Ministry of Agriculture, and known, some expected changes due to soils) was higher than under rainfed con- Universities started research making irri- irrigation like leaf size, number of current ditions. In addition, fertile soil without irrigation experiments on pistachio trees. year’s shoot and length of shoot will be gation is better than poor soil with irriga- Currently, farmers are informed about the increased. Irrigation results in more yield, tion. The results obtained are summari- possitive effects of irrigation. The region bigger nut size, high splitting percentage, zed below: where pistachio is growing, the orchards low blank nut rate and less alternate bea- have no irrigation facilities until the ring. Arpaci et al., (1995) determined that 1. Pistachio fresh nut weight of 21.8 kg/ 1980’s. Now the Southeast Anatolia Pro- under irrigated conditions ‘Siirt’ cultivar tree was recorded in arid soil and irriga- ject (GAP) provides irrigation facilities. yield was 262 kg/da while it was 60-80 ted conditions, while 15.50 kg/tree under Water is distributed through a network of kg/da under rainfed conditions. This ex- rainfed conditions. A yield of 31.00 kg channels. In the future new and irrigated periment shows that yield increased at was obtained in fertile soil irrigated condi- pistachio nut orchards in the region will least three or four times when pistachio tions, 25.87 kg in unirrigated conditions, expand Turkey’s pistachio nut areas. It is orchards were irrigated. respectively. One kg fresh fruit resulted in expected that when the project is comple- 559.63 g after sun drying. According to ted the Turkish pistachio nut production As it is mentioned above, pistachio or- this calculation, the highest dry weight will be at least doubled. As it is well known, chards were established under dry and was determined with 17.043 kg per tree, irrigation is an important factor in obtaining very poor soil conditions. But, high yiel- grown on a fertile soil and under irrigated high yields and good kernel quality. ding can be obtained under irrigated and conditions.

2. Pistachio nut shell splitting rate was Table 2. Distribution of Male And Female Trees In The Orchard. determined analyzing 300 fruits per tree. The records under arid soil irrigated con- oooo oo oo oo o oooo o oo ooo o ooo ditions were 58.89 % and under rainfed oxoo xo ox oo x ooxo o xo oxo o xoo conditions 37.45 %, on fertile soil they oooo oo oo oo o oooo o oo ooo o ooo were 61.43 % obtained under irrigated oooo oo oo oo o oooo o oo ooo o ooo conditions and 61.10 % under rainfed oxoo xo ox oo x ooxo o xo oxo o xoo conditions. The blank fruit rate was lo- oooo oo oo oo o oooo o oo ooo o ooo west, shell splitting and total filled fruit oooo oo oo oo o oooo o oo ooo o ooo number rate increased with irrigation. oxoo xo ox oo x ooxo o xo oxo o xoo oooo oo oo oo o oooo o oo ooo o ooo 3. One hundred pistachio nut weight de- oooo oo oo oo o oooo o oo ooo o ooo termined the highest value under fertile oxoo xo ox oo x ooxo o xo oxo o xoo soil irrigated conditions in splitted nuts. oooo oo oo oo o oooo o oo ooo o ooo 100 nut weight was determined statistica- oooo oo oo oo o oooo o oo ooo o ooo lly different from each other. Fruit weight oxoo xo ox oo x ooxo o xo oxo o xoo and kernel weight increased with irriga- oooo oo oo oo o oooo o oo ooo o ooo tion. On average 86.64 g were obtained oooo oo oo oo o oooo o oo ooo o ooo in nut and 44.37 g in kernel. Blank or empty nuts may be due to lack of pollina- 8 female 1 Male orchard design (x: male tree, o: female tree). tion and fertilization. But mainly irrigation can affect nut filling. Under irrigated con- oooo oo oo oo o oooo o oo ooo o ooo ditions blank nut rate was 17 % while on oooo oo oo oo o oooo o oo ooo o ooo rainfed trees reached an average rate of oooo oo oo oo o oooo o oo ooo o ooo 30 %. Likewise on poor soils the blank nut rate was 29.71 % while on fertile soils oxoo xo ox oo x ooxo o xo oxo o xoo it was 17.70 %. oooo oo oo oo o oooo o oo ooo o ooo oooo oo oo oo o oooo o oo ooo o ooo oooo oo oo oo o oooo o oo ooo o ooo LACK OF FERTILIZATION oxoo xo ox oo x ooxo o xo oxo o xoo Fertilization is a very important factor to oooo oo oo oo o oooo o oo ooo o ooo obtain high quality and high yield from the oooo oo oo oo o oooo o oo ooo o ooo trees. But pistachio trees suffer from sali- oooo oo oo oo o oooo o oo ooo o ooo nity and alkalinity. Fertilization depends oxoo xo ox oo x ooxo o xo oxo o xoo on irrigation and soil pH. Some minerals oooo oo oo oo o oooo o oo ooo o ooo can not be translocated because of high oooo oo oo oo o oooo o oo ooo o ooo pH. The availabity of nutrients and pH cri- oooo oo oo oo o oooo o oo ooo o ooo tical levels are given below. oxoo xo ox oo x ooxo o xo oxo o xoo oooo oo oo oo o oooo o oo ooo o ooo Most soils under pistachio production in oooo oo oo oo o oooo o oo ooo o ooo Turkey are lacking N, P, K and organic matter. A survey conducted in 30 pista- 11 female 1 Male orchard design (x: male tree, o: female tree). chio orchards in Southeast Anatolia, Te-

38 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Figure 1. Pistachio orchard at low planting density. Figure 2. Female inflorecence of pistachio. kin et al., (1985) revealed that in many but these are not absorbed by the trees ‘Kirmizi’, ‘Uzun’, ‘Halebi’ and ‘Siirt’ are the orchards the trees were markedly defi- due to antagonism. major cultivars. cient in phosphorus and zinc and slightly deficient in nitrogen, iron and mangane- CULTIVARS AND ALTERNATE PRUNING se. The level of potassium was found BEARING Pruning pistachio trees, grown under dry adequate in many orchards though there Alternate bearing is one of the most im- and poor soil conditions, should be ca- were some districts where the trees portant features of pistachio production rried out carefully. Otherwise trees can be showed slight deficiencies. In this region, behaviour. Alternate bearing cultivars damaged seriously. In Turkey and Syria soil pH varies between 7.5 and 9.3 and produce heavy crops in ÇonÈ years and the pruning is done in two stages, after the organic matter content is very low. little or no crop in the ÇoffÈ years. In con- harvest and before flowering time. During Especially zinc deficiency affects nut set. trast to other fruit species such as olive, the first stage, after harvest, dried shoots Crane and Maranto (1988) claimed that apple, etc. (Ak and Kaska 1992), this si- must be cutted out. The second stage pistachio is not a luxury N consumer tuation may differ from one cultivar to consists of a standard pruning of the tree. when it is widely available in the soil. The another. For example the cultivar ‘Siirt’ is In all pistachio producing countries pru- chemical form of nitrogenous fertilizers less alternating than others (Ak, 1998). ning is done by hand using labour. But in should be chosen depending on the soil This means the tendency of alternate pis- California pruning is made mechanically pH. For instance, in the GAP (Southeast tachio cultivar bearing is different. It may due to the very good vegetative growth Anatolian Region) area only ammonium be reduced by irrigation and fertilization. under irrigation. sulfate is recommended as a source of N because of the alkaline soils. Nitroge- The most important characteristics of the Pruning is another effective factor to ob- nous fertilizers should be applied at the pistachio nuts requiered by markets are tain high yields. Turkish farmers do not end of February or the beginning of Mar- the following: usually prune pistachio trees and if so ch at the rate of 1.5 to 4 kg per tree (Kas- - large size they apply wrong pruning systems. The ka, 1995). In California boron plays a uni- - high percentage of shell splitting research was made on three pruning le- que role in pistachio. Responses to foliar - low percentage of blank nuts vels. The shoot lenghts were 14 cm on Boron application include increased po- - high oil and protein content the heavy pruned trees, 12 cm on the lig- llen viability and germination rate, increa- - regular bearing ht pruned trees and 10 cm on the location sing nut set and yield, decreased blan- - high percentage of green kernels (farmer) pruned trees. On the heavy pru- king percentage and increased leaf bo- ned trees less canopy volume was deve- ron concentration (Brown, 1995a). Gene- In pistachio, a green kernel is one of the loped (25 %) than on light pruned trees. rally nutrient statutes of pistachios should most desired characteristics. Green ker- Although, the light pruned trees had given be assessed regarding leaf analysis. nel nuts are always a premium. Although more yield (12 %) than the cutting place Leaf samples should be taken 16-31 July the green kernel is a varietal characteris- in the pruned trees (Arpaci et al., 2001). according to Tekin et al. (1990). In addi- tic, it is also related to altitude and har- tion, Tekin (2002) determined the levels vest time. Generally, the nuts of early PESTS AND DISEASES of nutrients for pistachio trees grown un- harvested trees and trees growing on high Pest and diseases management is very der dry conditions (Table 3). plateaux produce greener kernels than important to get high yield every year. those of late harvested and low land gro- Some pests damage next years’s flower Also Brown (1995b) claimed critical le- wing trees. On the other hand greenness buds. vels of nutrient in Kerman cultivars grown is affected by pollen source. According to under irrigated conditions (Table 4). observation Pistacia terebinthus pollen The main pests in Turkish orchards are causes metaxenia and xenia (Ak, 1992). Agonoscena spp. (Yaprak Psyllasõ), Idio- When the nurient levels fall to minimum cerus stali Fieb., (Pistachio Leaf hopper), values deficiency, above these nutrient There are major cultivars in each main Anapulvinirai pistaciae (Torbalõ Ko_nil), levels toxicity will start according to producing country. According to Sheibani Suturaspis pistaciae (Beyaz Kabuklu Bit), scientists. According to some soil analy- (1995) there are 60 named cultivars in Kermania pistacaiella, Ams. (pistachio sis in pistachio growing lands, there are Iran. But main production cultivars are twig borer), Thaumatopoea solitaria enough elements or nutrients in the soil ‘Ohady’ and ‘Kaleh Ghochi’. In Turkey, (Fıstık Gözkurdu), Recurvaria pistacicola,

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 39 Table 3. Critical Nutrition Levels of Pistachio Leaves Grown Under Dry Conditions. Ak, B.E. and N. Agackesen, 2003. An investigation on determination of yield Macro Nutrients % Micro Nutrients ppm and quality on pistachio grown at Bire- cik province (In Turkish with English Nitrogen (N) 1.80 - 2.40 Iron (Fe) 43 - 170 summary). University of Harran, Project Phosphorus (P) 0.06 - 0.14 Zinc (Zn) 10 - 25 No: 58 Final Report, 59 pages. Potassium (K) 0.80 - 1.20 Manganese (Mn) 25 - 50 Calcium (Ca) 2.30 - 3.00 Copper (Cu) 6 - 90 Arpaci, S., F. Akkök, and H.Tekin, 1995. Determination of yield and quality chan- Magnesium (Mg) 0.50 - 0.90 Boron (B) 100 - 180 ges under dry and irrigated conditions (In Turkish with English summary). Tür- Also Brown (1995b) claimed critical levels of nutrient in Kerman cultivars grown under kiye II. Ulusal Bahçe Bitkileri Kongresi. irrigated conditions (Table 4). Adana. Cilt I. S.: 429-433.

Table 4. Nutrition Levels of Kerman Pistachio Leaves Under Irrigated Conditions. Arpaci, S., H.S. Atli and H.Tekin, 2001. Improvement of pruning techniques for bearing pistachio nut trees (in Turkish Macro Nutrients % Micro Nutrients ppm with English summary). GAP II. Tarim Kongresi, Harran Universitesi, 24-26 Nitrogen (N) 2.50 - 2.90 Zinc (Zn) 7 - 15 Ekim 2001, Sanliurfa: 31-38. Phosphorus (P) 0.14 - 0.17 Manganese (Mn) 30 - 80 Potassium (K) 1.00 - 2.00 Copper (Cu) 4 - 10 Arpaci, S., I. Acar, H.S.Atli and S. Kara- Calcium (Ca) 1.30 - 4.00 Boron (B) 90 - 250 dag, 2005. Comparison of Pistachio Pro- duction in Turkey and World (õn Turkish, Magnesium (Mg) 0.60 - 1.20 summary in English). GAP IV. Tarim Kon- gresi, Harran Üniversitesi Ziraat Fakül- tesi, 21-23 Eylül 2005, Sanliurfa: 238-243. Danil. (Pistachio fruit moth), Hylesinus modernizing. That means Turkish pista- Brown, P., 1995a. Nutrition of pistachio. vestitus (Karagöz Kurdu), Capnodis ca- chios are aflatoxin free and possess high Acta Horticulturae, 419: page 77 riosa (Pistachio root beetle), Megastig- quality in relation to flavour and taste. mus pistaciae (Meyve çkurdu) etc. Turkish cultivars are preferred in many Brown, P., 1995b. Diagnosing and Cor- European and USA markets due to their recting nutrient deficiencies. Pistachio Production, s.: 95-100. The main diseases are: Verticillium wilt, good taste and uniformly green kernels. Root and crown rot, Septoria leaf spot, st- Research carried out showed that the Crane, J.C. and Maranto, J., 1988. Pis- gmatomycosis, Powder mildew, Phyto- effects of irrigation amounted 70 %, fertili- tachio Production. Univ. of California, phtora parasitica etc. zation 50 %, variety 45 % and light pru- Publication 2279: 15 pages. ning 17 % on yield of the pistachio trees. Kaska, N., 1990. Pistachio research CONCLUSION In the future, when these limitations will and development in the Near East, Pistachio nut culture is centuries old in be solved, Turkish pistachio will gain im- North Africa and Southern Europe. Nut Iran, Turkey and Syria. In these coun- portance in the world, similar to hazelnut. Production Industry in Europe, Near tries, the pistachio producing area is the East and North Africa. Reur Technical Series 13: 133-160. largest and the number of trees is the highest in the world. In spite of these facts REFERENCES Kaska, N., 1995. Pistachio nut produc- the production is very low in Turkey and tion growing in Turkey. Acta Horticultu- Syria. Kernel taste is good but the nuts rae, 419: 161-164. Ak, B.E. 1992. Effects of different Pista- are small and their splitting percentage is cia spp pollens on the fruit set and qua- Sheibani, A., 1995. Pistachio production low. However, one should bear in mind lity of Pistachios. (In Turkish with an En- in Iran. Acta Horticulturae, 419: 165-168. that Turkey is the only country in the glish summary) Cukurova University - world where pistachio nuts are grown in Institute of Natural and Applied Scien- Tekin, H., Genc, C., Kuru, C. and Akkok, ce. Department of Horticulture, PhD. F., 1985. Investigations in the determi- such marginal lands with dry climate and Thesis, Adana, 210 p. poor, rocky and calcareous soils. nations of nutrient contents of pistachio nuts (in Turkish). Bahce, 14 (1-2): 47-57. Ak, B.E., 1998. The Yield and Fruit Turkey is full of other types of Pistacia Quality of Pistacia vera cv. Siirt Grown Tekin, H., 2002. Deficiencies of nu- spp regarded as wild pistachios. Topwor- at The Ceylanpõnar State Farm. Acta trients and fertilization in pistachio (in Horticulturae, 470: 510-515. king wild trees was done in the 1980s. Turkish). Pistachio Research Institute Publication N¼14: 33 pages. These trees are on the hill or mountains Ak, B.E., 2001. Selection Criteria Of as irregular orchards. Some are very The Best Pistachio Male Trees. Nucis, Tekin, H., Ca_lar, G., Kuru C., and small and some do not receive any cultu- 10: 24-25. Akkök F., 1990. Determination of Nu- trients for Pistachio trees and the best ral practices. This situation makes the to- Ak, B.E. and Kaska, N., 1992. Alternate tal yield very low. But these areas are not leaf samples collection periods (in Tur- bearing problem in pistachios and its kish with an English summary). First suitable for other agricultural crops. The- reasons, and the sitiuation in different National Symposium on Pistachio Nut, refore in this type of areas pistachio profit varieties (in Turkish). First Turkish Na- 11-12 September, 1990: 120-138. is valued. Also due to this situation regu- tional Horticultural Congress. 13-16 Oc- tober, 1992. Vol I (Fruit): 67-72. lar orchards will be supported by the Tur- Whitehouse, W.E. and Stone, C.L., 1941. Some aspects of dichogamy and kish government and growing techniques Ak, B.E., Acar, I. and Kaska, N., 1998. pollination in Pistacho. Proc. Amer. will change to modern system. On the An Investigation on The Male Determi- Soc. Hort. Sci., 39: 95-100. other hand, Turkey has a very important nation For Some Female Varieties Throughout Five Years (1992-1996) advantage about aflatoxin. The Turkish Grown at Ceylanpinar State Farm in environment, with low humidity and high B. E. Ak and H. Parlakci Sanliurfa Conditions. Proceedings of University of Harran, Faculty of Agriculture temperature during growing and harves- The X. GREMPA Seminar, 14-17 Octo- Department of Horticulture ting season, provides nuts free from afla- ber 1996, Meknes (Morocco). Cahiers 63200 Sanliurfa/Turkey toxin. They can be contaminated during Options Mediterraneennes, Vol 33: 99-104. E-mail: [email protected] processing. Processing systems are also

40 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Figure 1. Villahermosa pistachio orchard. Figure 2. Chaparrillo's R&D team (from left to right F. Couceiro, J. Guerrero, A. Moriana and Mª. C. Gijón).

PISTACHIO CULTIVATION CLIMATE AND SOILS CULTIVATION SYSTEM IN CASTILLA-LA MANCHA: According to the Koeppen classification, The crop is mostly placed under rainfed the most common climate in the region is TWENTY YEARS LATER farming although there are few irrigated Csa. By the Papadakis method, the most orchards. Irrigation in this region is redu- prevalent climate in Castilla-La Mancha is ced to a specific amount of water. Gene- the Continental Mediterranean climate. rally, it may not exceed 1,500 - 2,000 m3/ INTRODUCTION ha/year for large surface areas and 3,500 Castilla-La Mancha is a Spanish region The most frequent soil textures are clay - 3,800 m3/ha/year for small orchards. that covers about 4,000,000 ha, of which loam and sandy loam. Depth is generally This is because a large proportion of the 1,700,000 ha are under cereal, 570,000 between 40 and 80 cm. In most cases water used in the region for irrigation co- ha vines, 360,000 ha olives, 200,000 ha there is a high proportion of total carbo- mes from over-exploited aquifers. industrial crops, 180,000 ha pulses, nates and active limestone, especially in 50,000 ha other fruit trees and about the provinces located to the south. Soils The most widely-used planting space is 7 1,000,000 ha are left fallow. in the region are rich in potassium and m x 6 m under both dry and irrigated far- poor in phosphorus and organic matter. ming. The main limiting factors for crop diversification in the region are spring frosts and water shortage. These are the factors Table 1. Surface, year of planting, grafting and number of buds supplied. which led to the proposal in 1986 of research into pistachio production as an al- Year of grafting Year of planting Number of buds supplied Surface area (ha) ternative to the traditional cereal and vine crops (which bring very low returns) and 2003 2001 180,000 350 as a complement to others such as olive 2004 2002 120,000 343 and almond. 2005 2003 115,000 322

In 1988, a network of trials was established for the whole region, with an experi- Graph 1. Percentage of accumulated grafting take over three years. mental plot in each of the five provinces. Cv. ‘Kerman’. Different letters in the same colour indicate significant differences Between 1992 and 1997 a study on the (p<0.05. Tukey test). soil and climate adaptability of this species was carried out for the province of Ciudad Real (W 3¼ 56«; N 38¼ 59«), concluding that the pistachio tree suited to most of this territory. The first private plantations were set up in 1996 by a large number of farmers who decided in favour of this crop.

CURRENT SITUATION In Castilla-La Mancha there are more than 3,000 ha planted with pistachio although only part of them have reached full production. The average surface area per farmer is around 6 ha, varying between 1 and 50 ha. Current production is between 100,000 and 150,000 kg per year. The average annual increase in orchards is about 300 ha and interest in the crop increases daily.

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 41 Graph 2. Annual production and differences between the P. terebinthus L. - Good affinity with the different cultivars and P. atlantica Desf. rootstocks. No significant differences were found between studied rootstocks (Tukey Test). - Robustness - Very low cost of rootstock plants

CULTIVARS Although the most widely-used cultivar is ‘Kerman’ because of its fruit quality (size) and late flowering, there are others with excellent behaviour such as ‘Larnaka’, ‘Kastel’, ‘Mateur’ and ‘Avdat’.

The ‘Kerman’ cultivar is recommended for areas where irrigation is possible and where spring frosts are early. The other cultivars are recommended for dry farming and for areas where there is a reduced risk of spring frosts.

POLLINATION Orchards are laid out with a male su- rrounded by eight females (12%). This male proportion is more than enough to Orchards establishment starts with the same rootstock per year if the bud is not achieve satisfactory pollen density and purchase of the Pistacia terebinthus L. attached (dried) after 15 or 20 days. Nor- thus nut set. rootstock at the nursery. mally, grafting of the whole plantation is completed in about two years. Low springtime temperatures (spring The farmers purchase the rootstocks with frost) are the deciding factor for the one or two growths, and graft them after ROOTSTOCKS farmer’s choice of cultivar. In most cases, one year in the orchard. In Castilla La Mancha the most widely- late-flowering cultivars are chosen toge- used rootstock is Pistacia terebinthus for ther with pollenizers having the same flo- PROPAGATION the following reasons: wering time. After one year growing in the orchard, - It is a native species grafting is carried out from July to Sept- - Well adapted to soil and climate conditions PESTS AND DISEASE ember. Grafting is by shield budding, and - Good production and high yield (g/cm2) To date, the number of pests and/or di- 2 or 3 grafts are usually made on the in dry-farming seases observed in both experimental

Graph 3. Annual production (g/tree) for different cultivars.

Tukey Test. Significance = .05.

42 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Figure 3. Chaparrillo pistachio collection. Figure 4. Snow fall in a pistachio orchard in Ciudad Real, Spain. plots and commercial orchards is limi- different water provision. The male culti- RESULTS ted. New pests can be expected to ap- var is ‘Peter’ and the female ‘Kerman’. pear over time because of natural adap- The rootstocks used are P. terebinthus PROPAGATION tation of the native fauna to this new and P. atlantica. Since it is difficult to obtain quality plant crop. But the appearance and propaga- material (there are no grafted plants in tion of new pests or diseases wil have to - Rootstock experimental plot used for the market), the CMA 'El Chaparrillo' su- overcome several factors including the the production of seeds of P. terebinthus, pplied farmers, free of charge, with all the following: P. atlantic, P. intergerrima, P. vera and necessary material for grafting their or- various hybrids obtained by controlled chards. Table 1 shows the list of buds su- - Orchards are placed separated from crossing. Plant density is 7 m x 6 m with a pplied and the average approximate sur- each other. Castilla-La Mancha has a lar- total surface area of 1 ha. face area grafted every year. ge surface area, with many geographical barriers and different soil and climate - A total of five experimental plots run by ROOTSTOCKS conditions which would prevent crop ex- farmers to carry out various tests on fruit The main drawback regarding the roots- pansion. quality control in relation to water supply. tock (P. terebinthus L.) which is the most

- The low relative humidity and high temperatures during the vegetative period Table 2. Data on fruit quality in different cultivars. would also be unfavorable for proliferation. Cultivar % split % blank Size (g/kernel) EXPERIMENTAL PLOTS ‘Aegina’ 66.8808 b 8.1807 b 1.5275 bc The results of the various studies carried ‘Ashoury’ 70.5457 b 12.7811 c 1.5829 c out on this crop were obtained from the ‘Avdat’ 70.7730 b 7.2888 b 1.6861 d following trial plots: ‘Avidon’ 82.2585 c 6.8912 b 1.2947 a ‘Kerman’ 45.1957 a 12.2639 c 1.9623 e - Experimental plot without irrigation me- ‘Larnaka’ 71.4382 b 3.5836 a 1.5789 bc asuring 2.8 ha, with a plant density of 6 m ‘Mateur’ 68.2194 b 8.8198 b 1.5744 bc x 7 m, goblet-shaped trees with the crotch ‘Napoletana’ 37.6368 a 15.6205 c 1.4989 b at between 80 and 100 cm in the case of female plants and 180-200 cm in the case Tukey Test. Significance = .05. of pollenizers. The design is completely randomized for eight female cultivars (‘Kerman’, ‘Mateur’, ‘Larnaca’, ‘Aegina’, Table 3. Period of full flowering of different female cultivars studied at the CMA. ‘Napoletana’, ‘Ashoury’, ‘Avidon’ and ‘Av- dat’) and eight pollenizers (‘Peters’, ’02- Early flowering Mid-term flowering Late flowering 18', ‘Mateur’, ‘C special’, ‘Egino’, ‘Nazar’, Status B Status B Status B Status B ‘Askar’, ‘M-38’). Four rootstocks were 3rd week March 3rd week March 4th week March 4th week March used (P. terebinthus, P. atlantica, P. inte- Status D Status D Status D Status D gerrima and P. vera.). 1st week April 2nd week April 2nd week April 3rd week April

- Cultivar experimental plot with drip irri- ‘Mateur’ gation and a plant density of 6 m x 7 m. ‘Aegina’ ‘Lathwardy’ Today this plot has over 50 cultivars from ‘Batoury’ ‘Bronte’ ‘Napoletana’ ‘Kastel’ different locations. The rootstock used is ‘Iraq-2’ ‘Boundoky’ ‘Joley’ ‘Kerman’ P. atlantica. ‘Ashoury’ ‘Sfax’ ‘Avidon’ ‘Larnaka’ ‘Ajamy’ ‘Ouleimy’ - Irrigation trial plot, with a total surface ‘Avdat’ area of 1 ha and a plant density of 5 m x 5 m and divided into 5 sectors, each with STATUS B: Start of flowering; STATUS D: Full flowering

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 43 widely-used in Castilla La Mancha is its Table 4. Period of full flowering in male cultivars studied at the CMA. variable vigour. This makes it essential to choose carefully the location for collecting EARLY FLOWERING MID-TERM FLOWERING LATE FLOWERING seeds and then selecting them for vigour. ‘M-B’ Since 1989, studies have been carried ‘M-C’ out to compare the following rootstocks: ‘M-502’ ‘Egino’ Pistacia terebinthus, P. integerrima S., P. ‘M-1’ ’02-18' atlantica and P. vera. Over ten years, the ‘C Especial’ ‘M-38’ ‘Peters’ following observations were made: ‘M-36’ ‘M-11’ ‘C-16 (CMA)’ ‘Mateur’ ‘K-13 (CMA)’ - No significant differences were noted re- ‘Askar’ garding graft take and affinity in relation ‘Peter-1’ to the rootstocks studied (Graph 1). ‘Nazar’

Grafting take can be considered acceptable, reaching more than 70% in the third CULTIVARS The cultivars with the highest level of al- year after planting. In this study, P. tere- The behaviour of ‘Kerman’ was observed ternate bearing were ‘Kerman’ and ‘Avi- binthus and P. atlantica rootstocks stood to be similar to that obtained in traditional don’ and those with the least were ‘Lar- out, with 90% average take. Regarding producing areas regarding both production naka’, ‘Ashoury’ and ‘Avdat’. The other cul- graft affinity, P. vera showed the least and vegetative development. The same tivars were in an intermediate position. affinity with the ‘Kerman’ cultivar (almost can be considered about the rest of the 30% of stocks in the third year did not re- cultivars. (Graph 3). Regarding fruit quality in the cultivars sult in graft taking). studied, the following results were obtai- Good production was achieved in dry far- ned in the non irrigated plot (Graph 4 and - In the studies on production, the conclu- ming during the seven years studied (1999- Table 2). sion is that the main significant differen- 2005) with ages between 6 and 12 years ces arise between varieties, without respectively. Cultivars such as ‘Mateur’ and POLLINATION clear differences between rootstocks ‘Larnaka’ exceeded averages of 1,100 kg/ha, The most usual pollinators were ‘Peters’, (Graph 2). with a yield of over 1,900 kg/ha some years. ‘Egino’ and ’02-18', although recently two

Table 5. Possible incidence of some pests and diseases in plantations in Castilla-La Mancha several years after their establishment.

Associated Hosts flora Common Experimental Pest or disease with Castilla-La Mancha Castilla-La Mancha Plots P. terebinthus

Plodia interpunctella Hb. + + + + Nezara viridura L. + + + + Saisetia oleae Bern. + + + + Clytra spp.. - + - + Phytocoris spp. - + - + Acanthosma spp. - + - + Teleia humeralis Zel. + + - - Megastimus pistaciae Milk. + + - - Brevipalpus lewisi McG. - + + - Coeliodes ruber Marsh. - + - - Botriosphaeria spp. - + - - Pileolaria terebinthi Cast. + + - + Verticillum dahliae -+++

Table 6. Data of epicarp injuries on nuts of five commercial cultivars.

Year Cultivar Average* Standard error Lower limit Upper limit

‘Kerman’ 9.869 c 0.630 8.633 11.104 September 2003 ‘Mateur’ 6.148 ab 0.664 4.846 7.451 ‘Larnaka’ 4.742 a 0.575 3.614 5.870 ‘Aegina’ 6.748 b 0.575 5.620 7.876 ‘Kerman’ 9.823 c 0.414 9.010 10.636 ‘Mateur’ 1.545 ab 0.407 0.746 2.344 September 2004 ‘Larnaka’ 2.648 b 0.378 1.906 3.391 Aegina’ 1.180 a 0.385 0.425 1.935 ‘Avdat’ 2.598 b 0.378 1.856 3.340

Tukey Test. Significance = .05.

44 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Figure 5. Pistachio orchard in Motilla del Palancar, Cuenca, Spain. male cultivars have been obtained at the Graph 4. Fruit quality (% split, % blank and weight) in different cultivars. CMA which flower later - ‘C-16’ and ‘K-13’ (Tables 3 and 4). The combination of these five males in the orchard guarantee optimum pollination of cultivars such as ‘Kerman’ or ‘Kastel’ (suitable for cold areas).

In new orchards, growers are advised to increase the number of cultivars, not only to diversify the possible risk of low spring temperatures but also to increase the supply on the market.

PEST AND DISEASE Table 5 shows the pests and diseases which might arise in the medium term in a pistachio plantation in Castilla-La Mancha based on samples taken in experimental orchards and on data provided by growers.

Table 6 has been drawn up from data observed during 2003 and 2004 on injuries to the nut epicarp of five cultivars. It shows that ‘Kerman’ is significantly the - All the cultivars studied performed well, large-scale demand in a large number of variety that is most damaged by sucking with ‘Kerman’ standing out because of its European Union countries. insects. The causes of this higher inci- commercial interest and phenological de- dence are unknown although they may be velopment. Of special interest among - Pistachio cultivation can therefore be related to the plant phenology which di- earlier cultivars are ‘Mateur’, ‘Larnaka’ considered a potencial alternative for lar- ffers from that of the other cultivars. and ‘Avdat’. ge areas of the Castilla-La Mancha region. CONCLUSIONS - Castilla-La Mancha is an ideal location - All of the combinations (cultivar-roots- for pistachio production with good health tock) studied showed good adaptability quality because of the climate which can Guerrero J.; Gijón, Mª. C.; Moriana, for the Castilla-La Mancha environmental be expected to prevent excessive develo- A.; Couceiro J. F. conditions. pment of pests and diseases during the Centro de Mejora Agraria El Chaparrillo vegetative period (low relative humidity (Ciudad Real)-Junta de Comunidades - The crop has become consolidated not and high summer temperatures). Markets de Castilla-La Mancha. E-mail: [email protected] only with the P. terebinthus combination are obviously likely to start demanding but also with P. atlantica and P. integerrima. this type of product as there is already

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 45 PISTACHIO DIEBACK We studied the rate of spread of the bac- We have also set up a field trial where IN AUSTRALIA: A CASE teria within the vascular tissue and the young trees (not bearing yet) have been OF MULTIPLE INOCULATION effect of the infection on water movement inoculated with a suspension of Xtp (or through the xylem. We inoculated young sterile distilled water) every 3 months sin- EVENTS potted trees with Xtp via pruning wounds, ce January 2004. Shoots are assessed assessed the presence of the bacteria at for staining and presence of bacteria re- distances from the inoculation point over gularly. Internal staining has been obser- BACKGROUND time, recorded the onset of discolouration ved in treated trees but no external Symptoms of pistachio dieback include (staining) in the vascular tissue, and mea- symptoms have been observed yet. Ino- decline, dieback, internal staining, trunk sured the water conductivity of the main culations and monitoring continue. and limb lesions and, in some instances, stems. Xtp spread more than 50 cm be- tree death. The causal agent is Xantho- low the inoculation point in 1 week. The IMPLICATIONS FOR POSSIBLE monas translucens (Xtp) (1). Bacteria are onset of discolouration of vascular tissue CONTROL normally found in the sapwood of trunks, coincided with leaf production and was We tested if it was possible to control the limbs and twigs, and less frequently in confined in the tissue present at the time spread of the pathogen into new tissue by current season growth and leaves. of inoculation. The water conductivity of drastically pruning severely disease trees infected stems was reduced by 60% com- with simultaneous application of bactericide. The main mode of transmission of the pared to controls. However, we did not bacteria from tree to tree appears to be observe external symptoms on any of the Severely diseased trees (Figure 2a) were through pruning tools (2). Many aspects infected trees. drastically pruned in September 2004 (Fi- of the disease cycle and epidemiology re- gure 2b). The control comprised a set of main unknown. These observations generated two hypo- trees that was not pruned. Bactericide theses: 1. The bacteria are localised to was sprayed on the cut surface at pruning. A the vascular tissue active in the year of We labelled each cut branch and assessed inoculation. 2. Diseased trees have sus- the excised wood for the presence of bac- tained several inoculation events. We set teria. New shoots were assessed for the up experiments in the glasshouse and in presence of bacteria in autumn 2005. st the field to test these hypotheses. Yields of non-pruned trees and drastically pruned trees will be compared when pru- pith TESTING HYPOTHESES - GLASS- ned trees resume production of nuts. HOUSE AND FIELD EXPERIMENTS We inoculated young potted trees with The incidence of the symptoms was redu- Xtp via pruning wounds in July 2003. Ste- ced in pruned trees compared to the July 2003. rile distilled water was used for the con- same trees before pruning (Figure 2c). trols. A sub-set of the inoculated trees Bacteria were recovered from new tissue B was inoculated again in July 2004. Perio- in only 6% of the twigs (~ 300 twigs) that dic assessment for presence of bacteria were sampled from the previously infected (destructive harvest) was conducted from stumps. November 2003 until January 2005. SUMMARY pith st Main stems from trees inoculated once The localisation of the stain to the vascu- had one distinct ring of discoloured vas- lar tissue active in the year of infection st cular tissue (Figure 1a), whereas main and the scarcity of bacteria found in new stems from trees that were inoculated tissue produced in young trees with only July 2003 and July 2004. twice had two rings of stain or the stain one inoculation event support our hypo- had covered the whole stem cross-sec- theses. Without another inoculation event Figure 1: Sections of main stems of young potted tion (Figure 1b). In January 2005, bacte- the new tissue may remain free from bac- trees inoculated once (a) or twice (b), in July ria were isolated more often from new tis- teria. Accordingly, we are testing the idea 2003, and July 2003 and July 2004 respectively. sue (shoots, leaves) of trees inoculated that drastic pruning with simultaneous In a, there is one distinct ring of stain (st). In b, there are two rings. twice than from trees inoculated once. application of bactericide would allow the

A 64-61 Mar 04 B C

¥ 45% dieback ¥ 0% dieback ¥ 100% stunted growth Drastic pruning Sept. 04. ¥ 20% stunted growth

Figure 2: a) Severely diseased tree (N¼ 64-61) prior to pruning in March 2004. b) Severely diseased tree drastically pruned with simultaneous application of bactericide in September 2004. c) Tree 64-61 6 months after drastic pruning.

46 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 recovery of severely disease trees by rootstocks, their potencial for reforesta- controlling the spread of the pathogen tion of areas under process of desertifica- among branches of the same tree and tion is being considered. Different Pista- among trees. Preliminary results are pro- cia tree parts have been used for several mising. aims, more in ancient times than currently. Pistacia seed oil, leaves and resin ACKNOWLEDGEMENTS have been used traditionally for medicine. We thank Horticulture Australia Ltd and the Pistachio Growers Association for su- It is believed that the cultivated pistachio pport. (P. vera L.) was first introduced in Spain from Asia by the Romans. The crop was developed during the Middle Ages and la- REFERENCES ter disappeared for unknown reasons, as perhaps unbearing male trees were cut, Facelli, E. et al. (2002). Bacterial dieback of pistachio in Australia. Australa- preventing nut production due to lack of pollination. This crop was reintroduced in sian Plant Pathology 31: 95-96. Figure 1. 'Aegina' tree. Spain in the early 1980s and some 3.700 Taylor, C. et al. (2005). Spread and ha of young orchards are currently plan- control of pistachio dieback in Australia. In: Oliveira, M. M. and Cordeiro, V. ted, in both dry and drip irrigated conditio- (eds). XIII GREMPA Meeting on Al- ns, mainly in Castilla-La Mancha, Catalo- monds and Pistachios. Options Médite- nia, Andalusia and Extremadura regions. rranénnes Série A, No. 63. Zaragoza. Around 2.200 ha are currently bearing and another 1.500 ha planted are yet to bear nuts. The oldest orchards were planted at Lleida province (250 ha) which are E. Facelli1, C. Taylor2, E. Scott1, M. Escoto- Rodriguez3, N. Nancarrow2, R. Emmett2 around 24 years old and thus full bearing. and M. Sedgley4 The main producing region is Castilla-La 1 School of Agriculture and Wine, University Mancha (Guerrero et al., 2006). Interest of Adelaide, PMB 1, Glen Osmond, SA, 5064 on this crop is rising although two main 2 Department Primary Industries Victoria, drawbacks are restraining its develop- PO Box 905, Mildura, Victoria, 3502 3 School of Earth and Environmental Sciences, ment: lack of available trees in the nurse- University of Adelaide, Adelaide, SA, 5005 ries (scarce and short cultivar range) and 4 Faculty of The Sciences, The University crop management unexperience. Inland of New England, Armidale, NSW, 2351 dry or semidry continental like areas Figure 2. 'Ashoury' tree. growing low return cereals or vineyards producing poor quality wine or areas un- suited to almond growing due to high risk of late spring frost are suitable for new PISTACIA SPECIES pistachio planting (Vargas et al., 1999; GENEBANK, CONSERVATION, Batlle et al., 2002). CHARACTERIZATION AND USE AT IRTA: CURRENT SITUATION In most Spanish orchards, the Californian AND PROSPECTS IN SPAIN model regarding plant material has been largely adopted. Thus P. atlantica as rootstock and ‘Kerman’ and ‘Peters’ as female and male cultivars respectively INTRODUCTION are mainly planted. Different aspects of Three wild Pistacia species are native to the Californian pistachio technology and Spain: P. atlantica Desf., P. lentiscus L. management are not appropriate for most and P. terebinthus L. and their geogra- of the Spanish orchards growing on sha- phic distribution is given in Table 1. The llow and poor soils and with scarce water interspecific hybrid P. x saportae (P. len- resources (Vargas et al., 1999; Batlle et Figure 3. 'Bianca' tree. tiscus x P. terebinthus) appears occasio- al., 2002. Apart from the risk of genetic nally in places where both species thrive vulnerability due to the use of only a fe- together. Species detailed distribution in male cultivar, in areas with Mediterranean reduced number of female cultivars in the mainland and the Balearic and Cana- continental climate, ‘Kerman’ is perfor- new plantings: ‘Aegina’ in Greece, ‘As- ry islands was reported already by Batlle ming very well while it is not adapted to houry’ in Syria, ‘Bianca’ in Italy, ‘Sirora’ in et al., (1996). The wild populations of P. milder areas where its high chilling requi- Australia, ‘Larnaka’ in Cyprus, ‘Mateur’ in lentiscus (warm areas) and P. terebinthus rements are not fully covered. In coastal Tunisia and ‘Kerman’ in California and (cool areas) are not considered to be en- zones, there are other cultivars better Spain (Figures 1-7). In Iran and Turkey dangered however, P. atlantica due to its adapted like ‘Aegina’, ‘Larnaka’ and ‘Ma- however, major pistachio producers world- restricted distribution range in the Canary teur’ (Vargas et al., 1995b; Vargas et al., wide, a larger range of cultivars are still Islands, can be regarded as an endange- 1997; Vargas et al., 2001). being used like ‘Ohady’, ‘Kaleh Ghochi’ red species. Some survey and collecting and ‘Fandoghi’ in Iran and ‘Unzun’, ‘Kir- work of P. terebinthus in continental Pistachio cultivar characteristics from di- mizi’ and ‘Siirt’ in Turkey. IRTA is actively Spain has been carried out and it is cu- fferent countries differ markedly and their fostering collaboration for the exchange rrently under evaluation (Guerrero et al., cultivation range overlaps with the coun- of information, methodologies and their 2002). Apart from the interest of some try land. There is widespread concern implementation, on an international basis wild Pistacia species or their hybrids as about this tendency towards the use of a through the FAO-CIHEAM Nut Network

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 47 Table 1. Geographical distribution of the three Pistacia species native to Spain

Species Iberian Peninsula Balearic Islands Canary Islands

P. atlantica x P. lentiscus xxx P. terebinthus xx

Table 2. IRTA’s Pistacia genebank, number of accessions and origin

Species Number of accessions Origin

P. atlantica 65 Syria, USA P. integerrima 35 USA P. khinjuk 33 Turkey P. palaestina 32 Greece, Syria P. terebinthus 54 Greece, Spain P. vera 77 (clons) 13 different countries (1) Figure 4. 'Sirora' tree. (1): Australia, Cyprus, France, Greece, Iraq, Iran, Israel, Italy, Spain, Syria, Turkey, and IPGRI, to ensure the sustainable Tunisia, and USA conservation and utilization of the genus Pistacia (IPGRI, 1997 and 1998). Table 3. Pistachio cultivar collection held at IRTA Mas de Bover, indicating cultivar’s origin We present this review paper on the Pis- tacia genetic resources held at IRTA Mas Females Origin Males Origin de Bover and on activities being carried out regarding their conservation, charac- ‘Aegina’ Greece ‘A’ Greece terization and evaluation. However we ‘Ajamy’ Syria ‘Ask’ Israel would like also to stress that we consider ‘Ashoury’ Syria ‘B’ Greece genetic resources available and the ways ‘Avdat’ Israel ‘C’ Greece in which it can be best used to enhance ‘Avidon’ Israel ‘C-Especial’ Greece agriculture production and sustainable ‘Batoury’ Syria ‘Chico’ USA development. In Spain, there is also a ‘Beyaz Ben’ Turkey ‘Enk’ Israel ‘Bianca’ Italy ‘M-1’ Italy Pistacia germplasm field collection at AIC ‘Bianca Regina’ Italy ‘M-11’ Syria 'El Chaparrillo' in Ciudad Real (Couceiro ‘Boundoky’ Syria ‘M-2’ Italy et al., 1998). ‘Bronte’ USA (from Italy) ‘M-3’ Italy ‘Cakmak’ Turkey ‘M-25’ Tunisia CONSERVATION ‘Cappuccia’ Italy ‘M-3’ Italy Genetic resources conservation requires ‘Cavaillon’ France ‘M-36’ Syria characterization of the existing diversity ‘Cerasola’ Italy ‘M-37’ Syria in both the genepools and the gene- ‘Degirmi’ Turkey ‘M-38’ Syria banks. We have studied only ex situ va- ‘El Guetar’ Tunisia ‘M-47’ Syria riation by description of morphological ‘Ghiandolara’ Italy ‘M-5’ Italy traits on field collections, mainly on P. ‘Gialla’ Italy ‘M-502’ Italy ‘Halebi’ Syria ‘M-57’ Syria vera, following IPGRI descriptors. ‘Insolia’ Italy ‘M-6’ Italy However this approach has limitations ‘Iraq’ Iraq ‘M-7’ Italy as highly heritable traits often show little ‘Joley’ USA (from Iran) ‘M-8’ Italy variation over much of the material stu- ‘Kastel’ Israel ‘M-9’ Italy died. In addition, quantitative trait expres- ‘Kerman’ USA (from Iran) ‘M-P3’ Spain (from Italy) sion is subject to environmental variation ‘Keten Gomlegi’ Turkey ‘M-P9’ Spain (from Italy) and is difficult to measure. Complementa- ‘Kirmizi’ Turkey ‘Nazar’ Israel ry characterization by biochemical tech- ‘Larnaka’ Cyprus ‘Peters’ USA niques was also considered. No work on ‘Lassen’ USA ‘Túnez’ Tunisia Pistacia cryopreservation is in progess ‘Latwhardy’ Syria ’02-18' USA ‘Marawhy’ Syria ’25-A’ Tunisia in Spain. ‘Mateur’ Tunisia ’40-A’ Tunisia ‘Muntaz’ Iran Genebanks ‘Ouleimy’ Syria Pistacia species ‘Pignatone’ Italy Some 250 seedling trees belonging to six ‘Red Aleppo’ Syria Pistacia species are planted in groups on ‘Red Jalap’ Syria isolated plots to prevent hybridization at ‘Safeed’ Iran IRTA. Currently, the Spanish Pistacia ‘Sfax’ USA (from Tunisia) species genebank holds accessions of P. ‘Silvana’ Italy atlantica, P. integerrima Stewart, P. khin- ‘Sirora’ Australia juk Stocks, P. palaestina Boiss and P. te- ‘Sultani’ Turkey ‘Tard. Serra di Falco Italy rebinthus (Table 2). This represents a ‘Uzun’ Turkey good sample of the Pistacia genus diver- ‘White Ouleimy’ Syria sity present in the Mediterranean and

48 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Table 4. Vigour, growth habit and branching of female and male pistachio cultivars at IRTA Mas de Bover (1)

Cultivar Tree Growth Branching Cultivar Tree Growth Branching vigour1 habit2 habit3 vigour1 habit2 habit3 ‘Aegina’ 5 2 5 ‘A’ 5 1 7 ‘Ajamy’ 7 2 5 ‘Ask’ 5 2 3 ‘Ashoury’ 5 1 3 ‘B’ 7 3 7 ‘Avdat’ 5 2 5 ‘C’ 5 4 5 ‘Avidon’ 3 3 3 ‘C-Especial’ 7 2 3 ‘Batoury’ 5 4 5 ‘Chico’ 5 3 3 ‘Bianca’ 5 3 5 ‘Enk’ 3 4 3 ‘Bianca Regina’ 5 2 5 ‘M-1’ 5 1 3 ‘Boundoky’ 7 2 7 ‘M-11’ 5 4 5 ‘Bronte’ 3 3 5 ‘M-2’ 5 3 5 ‘Capuccia’ 3 2 5 ‘M-3’ 7 4 3 ‘Cavaillon’ 5 2 5 ‘M-36’ 3 3 5 ‘Cerasola’ 3 3 5 ‘M-37’ 5 3 7 ‘El Guetar’ 3 3 3 ‘M-38’ 3 3 5 ‘Ghiandolara’ 5 3 7 ‘M-47’ 3 2 3 Figure 5. 'Larnaka' tree. ‘Gialla’ 5 4 5 ‘M-5’ 5 1 3 ‘Insolia’ 5 4 5 ‘M-502’ 5 3 3 grammes were started in the world, one at ‘Iraq’ 3 3 7 ‘M-57’ 5 2 5 the UC Davis (USA) (Chao et al., 1998) and ‘Joley’ 3 4 5 ‘M-6’ 3 4 5 the other at IRTA Mas de Bover (Spain) ‘Kastel’ 3 3 5 ‘M-8’ 5 3 5 (Vargas et al., 1996). More recently, a ‘Kerman’ 3 2 5 ‘M-9’ 5 1 7 third programme to breed scion and root- ‘Larnaka’ 5 3 5 ‘M-P3’ 7 4 3 stocks was initiated in Turkey at the Pista- ‘Lassen’ 3 4 3 ‘M-P9’ 7 1 5 chio Research Institute, Gaziantep (H.S. ‘Latwhardy’ 7 2 7 Nazar’ 5 2 5 ‘Marawhy’ 7 2 7 ‘Peters T-41’ 7 2 5 Atli, 1998 personal communication). In Iran, ‘Mateur’ 7 3 3 ‘Peters T-71’ 7 1 5 some breeding work has been carried out ‘Muntaz’ 3 2 5 ‘Túnez’ 7 2 5 over the years at the Pistachio Research ‘Ouleimy’ 7 1 7 ’02-18' 3 2 3 Institute of Rafsanjan and a few unnamed ‘Pignatone’ 3 2 5 ’25A’ 5 3 3 seedlings are under assessment (Sheiba- ‘Red Aleppo’ 7 2 7 ’40A’ 3 2 7 ni, 1995). The Californian breeding pro- ‘Red Jalap’ 5 2 3 gramme has recently released three culti- ‘Safeed’ 5 3 5 vars: two females ‘Lost Hills’, ‘Golden Hi- ‘Sirora’ 3 2 5 lls’ and one male ‘Randy’ (Ferguson 2006). ‘Sfax’ 3 2 3 The Spanish breeding programme has a ‘Silvania’ 7 3 5 few advanced selections under trialling. ‘T. Serra di Falco’ 3 3 3 ‘White Ouleimy’ 3 2 5 IRTA’s programme to breed new pista- (1): According to IPGRI descriptors for pistachio (P. vera L.)(1997) chio cultivars for the Mediterranean in- 1 Scale: 3 = low, 5 = intermediate, 7 = high. dustry was started in 1989. Cross combi- 2 Scale: 1 = erect, 2 = semi-erect, 3 = spreading, 4 = dropping. nations were planned considering diffe- 3 Scale: 3 = sparse, 5 = intermediate, 7 = dense rent parental origin as they could not fre- ely occur in nature. The main female pa- western Asian regions. Seedlings from di- Genetic enhancement rents used were ‘Aegina’ (Greece), ‘Ba- fferent origins are still being introduced. Pistachio production is facing some im- toury’ and ‘Ouleimy’ (Syria), ‘Kerman’ These native and foreign species were in- portant problems, such as late and alter- (USA), ‘Larnaka’ (Cyprus), and ‘Mateur’ troduced to produce seedling rootstocks nate bearing and a few nut traits like size, (Tunisia), and the main males chosen for experimentation (Vargas et al., 1998). blanks, splitting and kernel colour and fla- were: ‘B’ and ‘C’ (Greece), ‘M-36’ and ‘M- A few of these seedling trees are hybrids vour. Although very little is known on pis- 38’ (Syria), ‘M-502’ (Italy) and ‘Nazar’ (Is- instead of pure species. tachio genetics, many of the production rael). The breeding aims were: precocity, problems may have genetic solutions. regular and high productivity, vigour and P. vera cultivars Most cultivars grown in the pistachio pro- nut quality. Data of first flowering, sex, le- In 1976 a collection of P. vera cultivars ducing countries are restricted to them afing date, tree size measured as trunk was started and some 77 genotypes (45 and were developed by chance. Genetic diameter, disease incidence and first nut females and 27 males) have been intro- diversity has been scarcely explored in records were taken. duced over the years (Table 3). Cultivars this crop. Dioecy represents an important were propagated either onto P. palaesti- limitation to pistachio breeding since A total of 31 controlled crosses among 10 na or P. vera rootstocks. This collection male performance is unknown and, in female and 12 male parents were made holds cultivars from the main producing addition, sex expression in seedlings between 1989 and 1990 yielding some countries and, represents also a good takes between 5 and 8 years. These re- 2,000 seedlings (Vargas et al., 1996). sample of the existing genetic diversity duce efficiency of selection. Only a few Among seedlings significant differences within the cultivated pistachio, conside- pistachio cultivars have been obtained were observed in relation to vigour (Var- ring that less than 100 cultivars have from open pollinated known cultivars, gas and Romero, 1988a and 1998b and been described worldwide (Maggs, ‘Pontikis’ (seedling of ‘Aegina’) in Greece Vargas et al., 1995), leafing dates (Var- 1973). Sheibani (1995) however reported (Pontikis, 1986) and ‘Sirora’ (seedling of gas et al., 1995 and 2001), flowering pre- that in Iran more than 60 named cultivars ‘Red Aleppo’) in Australia (Maggs, 1982). cocity (Vargas et al., 2002) and nut cha- have been described. In the late 1980s, two scion breeding pro- racteristics (Vargas and Romero, 2003).

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 49 Table 5. Pistachio cultivar flowering at IRTA Mas de Bover, Tarragona. Average dates 1990-951.

March April May

20-3 25-3 31-3 5-4 10-4 15-4 20-4 25-4 30-4 5-5

Females ‘Batoury’ (6) ======‘Aegina’ (6) ======‘Mateur’ (6) ======‘White Ouleimy’ (6) ======‘Marawhy’ (5) ======‘Ashoury’ (6) ======‘Red Jalap’ (5) ======‘Bronte’ (6) ======‘Iraq’ (6) ======‘Cerasola’ (6) ======Figure 6. 'Mateur' tree. ‘Gialla’ (5) ======‘Sfax’ (6) ======In the programme a few promising fema- ‘Larnaka’ (6) ======‘Ajamy’ (5) ======les and males have been selected. ‘Avidon’ (6) ======‘Latwhardy’ (4) ======CHARACTERIZATION ‘Insolia’ (6) ======Most cultivars of P. vera have been ex- ‘Boundoky’ (5) ======tensively documented following the IPGRI ‘Ghiandolara’ (5) ======descriptors list for pistachio (1997) and ‘Ouleimy’ (6) ======are recorded in an Excel file following a ‘Muntaz’ (4) ======common format. A few other cultivar cha- ‘Silvana’ (5) ======racteristics like: tree vigour, growth and ‘Bianca’ (6) ======branching habit (Table 4), blooming date ‘Joley’ (6) ======(Table 5) and flowering precocity (Table ‘Capuccia’ (4) ======‘Lassen’ (6) ======6) are given. In addition, important nut ‘Kerman’ (6) ======characteristics like nut length, nut width, ‘Bianca Regina’ (5) ======nut shape, nut and kernel weight and, ‘Pignatone’ (4) ======kernel colour are commented in Table 8. Males Tree vigour ‘A’ (5) ======This growth descriptor is very important ‘B’ (6) ======for pistachio performing, mainly under dry ‘Peters’ (6) ======conditions. Vigour observations range from ‘C’ (6) ======3 to 7 (Table 4). Cultivars showed impor- ‘Naz’ (6) ======‘M-502’ (6) ======tant differences in vigour. Some female ‘M-57’ (6) ======cultivars are vigorous, like ‘Ajamy’, ‘Boun- ‘Túnez’ (6) ======doky’, ‘Marawhy’, ‘Mateur’, ‘Ouleimy’, ‘Red ‘M-47’ (6) ======Aleppo’ and ‘Silvana’ and some showed ‘Enk’ (6) ======lack of vigour, like ‘Bronte’, ‘Joley’, ‘Ker- ‘M-25A’ (5) ======man’, ‘Lassen’, ‘Sirora’, ‘Sfax’ and ‘White ‘Ask’ (6) ======Ouleimy’. Within male cultivars, ‘B’, ‘M-P3’, ‘M-36’ (6) ======‘Peters’ and ‘Túnez’ are vigorous, while ‘M-38’ (6) ======‘Enk’, ‘M-47’, ’02-18' and ’40A’ appear to ‘C-Especial’ (6) ======be less vigorous. Female cultivars from ‘M-8’ (4) ======Syria seem to be largely vigorous. Regar- ’02-18' (4) ======‘M-3’ (4) ======ding quantitative trait inheritance, first re- ‘M-11’ (6) ======sults on different segragating progenies, ‘M-P3’ (6) ======showed vigour is related to parents (Vargas ‘M-37’ (6) ======et al., 1995a; Vargas and Romero, 1998). ‘M-2’ (4) ======‘M-P9’ (6) ======Growth and branching habit ‘M-9’ (4) ======These two tree descriptors are presented ‘M-5’ (4) ======together as are related horticultural con- 1: number of observation years for each cultivar cepts. Growth and branching habit of cultivars has important agronomical effects on training, pruning and harvesting. Pis- minal positions and the remaining lateral bit are ‘Ashoury’ and ‘Batoury’ respectively, tachios have a strongly apical dominant buds are reproductive. Table 4 shows that which means diversity from the same ori- growth habit as most vegetative growth the number of cultivars having erect (1) or gin, Syria. Table 4 also shows, regarding occurs on terminal shoots. On adult trees semi-erect (2) growth habit are 19 and, branching habit (observed as intensity of one year old pistachio shoots produce ve- with spreading or dropping habit there are branching) that out of 37 characterized getative buds only at terminal or subter- 18. Examples of erect and spreading ha- cultivars most (22) have an intermediate

50 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Table 6. Juvenile period of female and male pistachio cultivars at IRTA Mas de Bover.

Females Juvenil period1 Males Juvenil period1

‘Bianca’ 1 ‘M-36’ 2 ‘Bianca Regina’ 1 ‘M-37’ 2 ‘Boundoky’ 1 ‘M-47’ 2 ‘Bronte’ 1 ‘M-8’ 2 ‘Cappuccia’ 1 ‘Tunez’ 2 ‘Red Jalap’ 1 ‘C-Especial’ 3 ‘Silvana’ 1 ‘M-11’ 3 ‘Ajamy’ 2 ‘M-2’ 3 ‘Ashoury’ 2 ‘M-3’ 3 ‘Cerasola’ 2 ‘M-5’ 3 ‘Insolia’ 2 ‘M-57’ 3 ‘Lathwardy’ 2 ‘M-9’ 3 ‘Marawhy’ 2 ‘M-P9’ 3 ‘Ouleimy’ 2 ‘’Ask’ 4 ‘Avidon’ 3 ‘A’ 4 Figure 7. 'Kerman' tree. ‘Lassen’ 3 ‘M-38’ 4 ‘Mateur’ 3 ‘M-P3’ 4 tant and dark green coluored pistachios ‘Muntaz’ 3 ‘Nazar’ 4 is a bonus. ‘Pignatone’ 3 ‘B’ 5 ‘Safeed’ 3 ‘C’ 5 ‘Sirora’ 3 ‘Enk’ 5 Regarding nut and kernel weight at ‘White Ouleimy’ 3 ‘M-25A’ 5 IRTA’s collection, ‘Kerman’ produces large ‘Aegina’ 4 ‘M-502’ 5 nuts but also ‘Ajamy’, ‘Batoury’ and ‘Oulei- ‘El Guetar’ 4 ‘Peters’ 5 my’. Instead ‘Avidon’, ‘El Guetar’, ‘Maraw- ‘Iraq’ 4 hy’ and ‘Red Jalap’ have a small kernel size. ‘Joley’ 4 ‘Larnaka’ 4 Split nuts ‘Batoury’ 5 It is an important nut trait affecting marke- ‘Ghiandolara’ 5 table yield as split nuts are preferred by ‘Kerman’ 5 consumers. Indehiscence of pistachios is ‘Sfax’ 5 considered as a physiological ripening re- 1 Scale juvenile period: 1 = long, 5 = short lated factor but also appears to be related to growing conditions. However from differences observed between cultivars it branching habit, 8 showed sparse and 7 important differences between cultivars in has also an important genetic component. dense branching habit respectively. relation to juvenile period. In Table 6, flo- Wide cultivar differences can be obser- wering precocity is not given in years as ved in Table 8 ranging from 5% to 90% of Blooming date recommended in the IPGRI descriptors splitting. Cultivars such as ‘Batoury’, This phenology descriptor, although very but on a scale from 1 (long) to 5 (short). ‘Bianca’, ‘Bronte’, ‘Latwhardy’, ‘Marawhy’, much influenced by local environmental The juvenile period of male cultivars ap- ‘Ouleimy’, ‘Pignatone’ and ‘White Oulei- conditions, is very useful for cultivar peared to be shorter than in females, be- my’ appear to have much lower levels of choice when planning new orchards. Lar- ing early pollen producers ‘B’, ‘C’, ‘Enk’, split nuts than ‘Ashoury’, ‘Avidon’, ‘Lar- gely the cultivar bloom scaling is maintai- ‘M-25A’, ‘M-502’ and ‘Peters’, and late naka’, ‘Lassen’, ‘Sirora’ and ‘Sfax’ in our ned between places. Pistachio cultivar pollen producers ‘M-36’, ‘M-37’, ‘M-47’, collection. ‘Kerman’ shows an intermedia- flowering average dates (1990-95) are ‘M-8’ and ‘Tunez’. Regarding females, te splitting. In general, Italian genotypes presented in Table 5. Average flowering cultivars like ‘Batoury’, ‘Kerman’ and produce low levels of split nuts. period is around two weeks. Early blooming ‘Sfax’ showed short unproductive periods female cultivars are ‘Batoury’, ‘Aegina’, while ‘Bianca’, ‘Boundoky’ and ‘Red Ja- EVALUATION ‘White Ouleimy’, ‘Mateur’, ‘Marawhy’ etc. lap’ showed the opposite. Molecular markers while ‘Joley’, ‘Ouleimy’, ‘Kerman’, ‘Bianca The information provided by morphologi- Regina’, etc. are late flowering. Early Nut length, nut width, nut shape, nut cal characters is limited. These limitations male bloomers are ‘A’, ‘B’ and ‘Peters’ and kernel weight and kernel colour. have resulted in the development of bio- and, several selected clones as ‘M-P9’, In Table 7, nine nut characteristics are chemical and molecular techniques. Mo- ‘M-9’ and ‘M-5’ are late flowering. It can presented. Cultivar choice is related to lecular genetics has an important role to be observed from Table 6 that flowering nut traits and thus breeding aims for play in characterizing plant genetic diver- of ‘Kerman’ and its pollinator ‘Peters’, scion improvement too. Nut shape varies sity for acquisition, maintenance and use. which shows a long flowering period, from long almond (‘Joley’, ‘Red Jalap’ For diversity studies, isoenzymes are re- does not overlap completely resulting in and ‘White Ouleimy’) to round hazelnut latively cheap and informative, RFLP’s partial pollination. In addition, average le- shape (‘Kerman’). Ovoid shape (1,5 > l / are expensive, RAPD’s are not very infor- afing date in the offspring is correlated (r w < 1,8) which is the most commonly mative and microsatellites promise to be = 0.67) with the mean date of full bloom of found in pistachio, are: ‘Aegina’, ‘As- very informative but expensive. Most cul- the parents (Vargas and Romero, 1998). houri’, ‘Bianca’, ‘Larnaka’, ‘Mateur’, ‘Si- tivars held at IRTA have been isoenzyma- rora’ and ‘Sfax’. Kernel colour varies tically analyzed for seven systems (Rovi- Flowering precocity from light yellow (‘Kerman’) to dark ra et al. 1998; Vargas et al., 1995b). This Juvenility is considered as the period green (‘Bianca’). For some industry work resulted in 12 different isoenzymic from graft or bud to first flower. There are applications kernel colour is very impor- phenotypic combinations for female culti-

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 51 Table 7. Mean records of nine pistachio nut characteristics(1)

Cultivar Nut length Nut width Nut Nut shape Percentage Nut Kernel Kernel dry Kernel (l) (mm) (w) (mm) thickness (l / w) of split weight weight weight x colour1 (mm) nuts (%) (g) (g) 100 / nut dry weight

Nr. Mean Nr. Mean Nr. Mean Nr. Mean Nr. Mean Nr. Mean Nr. Mean Nr. Mean Nr. Mean ‘Aegina’ 15 19,26 15 10,94 15 9.73 15 1,76 12 35,41 23 0,88 23 0,45 23 50,96 8 2 ‘Ajamy’ 8 19,72 8 11,78 8 10.95 8 1,67 4 55,25 8 1,01 8 0,53 8 54,65 3 2 ‘Ashoury’ 13 18,93 13 10,60 13 10.02 13 1,78 9 64,44 18 0,91 18 0,40 18 45,69 6 2 ‘Avidon’ 4 16,01 4 10,38 4 9.55 4 1,54 4 60,75 8 0,75 8 0,34 8 47,05 3 2 ‘Batoury’ 9 20,50 9 12,02 9 11.31 9 1,70 7 20,14 14 1,12 14 0,52 14 48,36 5 3 ‘Bianca’ 12 19,99 12 11,00 12 9.87 12 1,81 11 15,72 18 0,88 18 0,43 18 50,54 8 2 ‘Bianca Regina’ 8 18,95 8 10,86 8 9.55 8 1,74 5 5,60 5 0,83 5 0,35 5 45,02 3 3 ‘Boundoky’ 9 16,55 9 10,80 9 9.88 9 1,53 5 42,80 10 0,80 10 0,42 10 54,97 3 2 ‘Bronte’ 13 19,05 13 10,69 13 9.58 13 1,78 10 13,40 17 0,89 17 0,41 17 47,03 6 3 ‘Capuccia’ 5 19,66 5 11,28 5 10.12 5 1,74 3 3,33 3 0,75 3 0,38 3 53,15 2 2 ‘Cerasola’ 6 19,95 6 11,20 6 10 6 1,78 6 36,00 10 0,97 10 0,42 10 44,09 4 3 ‘El Guetar’ 1 14,98 1 10,33 1 8.69 1 1,45 2 90,00 3 0,77 3 0,37 3 49,88 1 2 ‘Ghiandolara’ 9 20,49 9 10,77 9 9.94 9 1,90 5 45,00 10 0,93 10 0,44 10 48,36 3 3 ‘Gialla’ 7 19,96 7 11,20 7 10.15 7 1,77 4 24,75 5 0,99 5 0,43 5 46,52 3 2 ‘Insolia’ 5 20,11 5 11,16 5 10.15 5 1,80 3 15,33 5 0,89 5 0,44 5 50,70 2 3 ‘Iraq’ 14 18,73 14 11,62 14 11.07 14 1,61 8 53,37 15 0,98 15 0,48 15 50,91 6 2 ‘Joley’ 7 20,29 7 11,04 7 9.95 7 1,84 5 37,40 9 0,87 9 0,43 9 52,13 3 2 ‘Kerman’ 6 19,31 6 14,12 6 11.96 6 1,36 3 47,66 6 1,15 6 0,57 6 52,57 3 2 ‘Larnaka’ 9 19,87 9 10,98 9 10.45 9 1,81 7 56,42 14 0,94 14 0,45 14 49,26 4 2 ‘Lassen’ 6 17,76 6 11,55 6 11.81 6 1,54 3 68,00 6 0,91 6 0,46 6 53,33 3 2 ‘Lathwardy’ 7 18,86 7 11,16 7 10.09 6 1,54 4 12,00 8 0,90 8 0,45 8 49,63 4 2 ‘Marawhy’ 9 18,50 9 11,13 9 10.03 9 1,66 4 19,50 7 0,80 7 0,37 7 47,54 3 2 ‘Mateur’ 11 19,46 11 11,26 11 10.04 11 1,72 10 43,20 20 0,88 20 0,43 20 50,63 4 2 ‘Ouleimy’ 8 22,19 8 12,10 8 10.92 8 1,83 4 18,75 7 1,22 7 0,58 7 48,17 3 2 ‘Pignatone’ 5 19,12 5 11,09 5 9.50 5 1,72 4 11,75 7 0,85 7 0,37 7 45,07 4 3 ‘Red Jalap’ 7 19,08 7 10,34 7 10.07 7 1,84 6 37,16 10 0,83 10 0,36 10 44,54 4 2 ‘Safeed’ 1 18,54 1 11,25 1 10.06 1 1,65 3 38,66 6 0,88 6 0,43 6 49,60 2 2 ‘Sirora’ 5 20,10 5 11,90 5 10.11 5 1,69 4 67,75 8 0,95 8 0,47 8 50,67 3 1 ‘Sfax’ 9 17,65 9 11,20 9 10.50 9 1,57 7 58,57 14 0,89 14 0,43 14 49,83 5 2 ‘Silvania’ 7 19,91 7 10,99 7 10.05 7 1,81 4 21,75 7 0,89 7 0,43 7 49,96 3 2 ‘T. Serra di Falco’ 2 20,65 2 10,87 2 10.01 2 1,90 2 16,50 3 0,86 3 0,42 3 49,15 1 3 ‘White Ouleimy’ 8 21,08 8 11,17 8 10.44 8 1,88 5 17,60 8 0,93 8 0,44 8 48,71 3 3

(1)According to IPGRI descriptors for pistachio (P vera L.) (1997) Nr. = Number of samples recorded 1 Scale: 1 = yellowish, 2 = yellowish green, 3 = green vars and 2 for males. It allowed also the and Vargas et al. 2005). However this ge- cal characters with respect to true types identification of 2 female cultivars nebank has been extensively characteri- from Syria. (‘Bianca’ and ‘Red Jalap’) and 4 male cul- zed using ten isoenzyme systems (Rovira tivars (‘A’, ‘C-Especial’, ‘M-P3’ and ‘M- et al. 1995; Monastra et al., 1998). Isoen- ACKNOWLEDGEMENTS P9’) as hybrids. Also, the RAPD marker zymic polymorphism revealed the most We are indebted to many researchers linked to sex expression, found using variable species was P. atlantica followed from around the Mediterranean region bulked segregant analysis, can be useful by P. vera, P. palaestina and P. integerri- who have provided seeds, scion-wood for (Hormaza et al., 1994). A joint work to as- ma. P. terebinthus and P. khinjuk were budding or grafting and thus allowing the sess the phenotypic and genetic diversity polymorphic only for two isoenzymes. establisment of these genebanks. Pista- at DNA level within P. vera germplasm Thus the dioecious mating system of Pis- cia genetic resources at IRTA Mas de collected in Mediterranean countries con- tacia which enforces outcrossing and high Bover are maintained and characterized firmed also the existing high degree of levels of heterozygosity has a different with funding from the Spanish Instituto de polymorphism (Caruso et al., 1998). effect depending on the species. In addi- Investigación y Tecnología Agraria y Ali- tion, a few wild accessions were identified mentaria (INIA) of the Ministry of Agricul- The Pistacia species genebank of IRTA as being of interspecific hybrid origin, P. ture, Food and Fisheries (Project RF 01- has not been morphologically characteri- palaestina seedling trees from Greece 036 and RF 04-0011) as the Spanish pista- zed although the interest of some proge- and USA showed distinctive hybrid patter- chio genebank. The breeding work is being nies as rootstocks and for reforestation ns and also clear differences in their leaf granted by the INIA Projects (SC 97-049, have been studied (Vargas et al. 1998 morphology and some other morphologi- RTA01-081, RTA04-030 and RF04-015).

52 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 BIBLIOGRAPHY Maggs, D.H., 1973. Genetic resources Vargas, F.J.; Romero, M.A. 2003. Fruit in pistachio. FAO Plant Genetic Resour- characters in pistachio progenies. Op- ces Newsletter, 29: 7-15. tions Méditerranéennes, Série A XIII Batlle, I.; F. J. Vargas and M.A. Rome- GREMPA Meeting, 63: 49-56. ro. 1996. Natural occurrence, conservation and uses of Pistacia species in Maggs, D.H. 1990. The austalian pista- Spain. (Padulosi, S., T. Caruso and E. chio ‘Sirora’. Fruit Varieties Journal, 44 Vargas, F.J.; Romero, M.A.; Clavé, J.; Barone, editors). Taxonomy, distribu- (4): 178-179. Batlle, I. 1995. First results of vigour tion, conservation and uses of Pistacia and leafing in pistachio progenies. Acta genetic resources. Report of a Works- Maggs, D.H., 1982. An introduction to Horticulturae, 419: 273-278. hop, June 1995, Palermo, Italy. IPGRI, pistachio growing in Australia. Ed. CSI- Rome, Italy: 42-45. RO. Australia, 37 p. Vargas, F.J.; Romero, M.A.; Monastra, F.; Mendes Gaspar, A.; Rouskas, D., 1997. Sélection de variétés de pista- Batlle, I.; M.A. Romero and F. J. Var- Monastra, F.; M. Rovira; F.J. Vargas; chier adaptées à l’aire nord méditerra- gas. 2002. Posibilidades del cultivo del M.A Romero; I. Batlle; D. Rouskas; A. néenne. Options Méditerranéennes, pistachero en España. Fruticultura Pro- Mendes Gaspar. 1997. Caractérisation Serie B: Etudes et Recherches, (16): fesional, 130: 17-24. isoenzymatique de diverses espèces du genre Pistacia et leurs hybrides. Etude 93-119. Caruso, T.;C. Iannini; E. Barone; F.P. de leur comportement comme porte- Vargas, F.J.; Romero, M.A.; Vargas, I. Marra; F. Sottile; C.I. Greco; M.R. Sabi- greffe du pistachier Pistacia vera L. Op- 2001. Blooming and leafing time in pis- na; G. Martelli; F. Monastra; I. Batlle; tions Méditerranéennes, Serie B: Etu- tachio progenies. Cahiers Options mé- F.J. Vargas; M.A. Romero; B.E. Ak; G. des et Recherches 16: 133-142. diterranéennes, XI GREMPA Seminar Zakynthinos; D. Rouskas; S. Padulosi 56: 41-46. and M. Laghezali. 1998. Genetic and Pontikis, C.A., 1986. ‘Pontikis’ pista- phenotypic diversity in pistachio (P. chio. HortScience, 21 (4): 1.074. vera L) germplasm collected in medite- Vargas, F.J.; Romero, M.A.; Vargas, I. rranean countries. Acta Horticulturae, Rovira, M.; I. Batlle; M. Romero and 2002. Flowering precocity in pistachio 470: 168-178. F.J. Vargas, 1998. Characterization of progenies. Acta Horticulturae, 591: 297- pistachio cultivars using isozymes. Op- 303. Chao, C.T., D.E. Parfitt; L. Ferguson; C. tions Méditerranéennes. X GREMPA Kallsen and J. Maranto. 1998. Breeding Seminar, 33: 113-121. Vargas, F.J.; Romero, M.; Clavé, J.; and genetics of pistachio: the Califor- Aletà, N. 2005. Pistacia progeny trial for nian program. Acta Horticulturae 470: Rovira, M.; I. Batlle; M.A. Romero and woodland use. IV ISHS International 152-161. F.J. Vargas. 1995. Isoenzymic identifi- Symposium on pistachios and almonds. cation of Pistacia species. Acta Horti- Book of Abstracts, 39-40. Couceiro, J.F.; J.M. Coronado and M.A. culturae, 419: 265-272. Mendiola. 1998. Current situation of research into the pistachio tree (Pistacia Sheibani, A. 1995. Pistachio production I. Batlle, M.A. Romero, M. Rovira and F.J. vera L.) in the region of Castilla-La in Iran. Acta Horticulturae, 419: 165- Vargas Mancha (Spain). Options Méditerra- 168. IRTA - Mas de Bover, Mediterranean Fruit néennes. X GREMPA Seminar, 33: 225- Trees 233. Vargas, F.J. and M.A. Romero. 1998a. Ctra. Reus-El Morell, km 3.8, Vigour and juvenile stage in pistachio E-43120 Constantí (Tarragona) Cruceiro, J.F., Coronado, J.M. Men- progenies. Options Méditerranéennes, [email protected] chén, M.; Mendiola, M. A.. 2000. El cul- X GREMPA Seminar, 33: 105-111. tivo del pistachero. Ed. Agrolatino: 112 pp. Vargas, F.J.; Romero, M.A. 1998b. Vi- gour in pistachio progenies. Acta Horti- Ferguson, L. 2006. Californian pistachio culturae, 470: 162-167. industry. NUCIS Newsletter 13: 37-40. DOMESTICATION Vargas, F.J.; M.A. Romero and J. Cla- OF THE PISTACHIO NUT Guerrero, J.; Cruceiro, J. F.; A. Moria- vé. 1998. Nursery behaviour of pista- na. 2002. Selection of terebinth (P. tere- chio rootstocks. Acta Horticulturae, 470: (PISTACIA VERA) binthus) trees as seed producers for 231-236. pistachio (P. vera L.) rootstocks in the Castilla-La Mancha region (Spain). NU- Vargas, F.J.; M.A. Romero; R. Rovira CIS Newsletter 11: 25-29. and I. Batlle. 1996. Pistachio cultivar improvement at IRTA-Mas Bové. Pro- INTRODUCTION In the last five decades several hundred Guerrero, J., Gijón, M. C., Moriana, A., ceedings of the IX GREMPA Meeting on Cruceiro, J. F. 2006. Pistachio cultiva- Pistachio, Bronte-Sciacca, Italy, May prehistoric and early historic sites have tion in Castilla-La Mancha: twenty years 1993. been excavated by archaeologists in later. NUCIS Newsletter 13: - . South-West Asia, temperate Europe and Vargas, F.J.; Romero, M. and Batlle, I. 1999. Aspectos básicos del cultivo del in the Mediterranean Basin. Large Hormaza, J. I.; L. Dollo and V. S. Polito. amounts of well-preserved (mostly char- 1994. Identification of a RAPD marker pistachero: situación, problemática y to sex determination in Pistacia vera perspectivas. Fruticultura Profesional, red) plant remains were retrieved from using bulked segregant analysis. Theo- Especial Frutos Secos II, 104: 98-105. these digs. They were carefully exami- retical and Applied Genetics 89: 9-13. Vargas, F.J.; Romero, M.; Clavé, J.; ned, expertly identified, and radiocarbon 14C dated. Central Asia was explored ar- IPGRI. 1997. Descriptors for Pistachio Batlle, I., 1995a. First results of vigour (Pistacia vera L.). International Plant and leafing in pistachio progenies. Acta chaeologically as well, but on a much lo- Genetic Resources Institute, Rome, Horticulturae, 419: 273-277. wer scale. In the same time span, the bo- Italy. tanical and the genetic evidence extrac- Vargas, F.J.; Romero, M.; Plana, J. and Batlle, I. 2001. Variedades de pistache- ted from the living plants (the cultigens IPGRI. 1998. Descriptors for Pistacia and their wild relatives) also increased spp. (excluding Pistacia vera L.). Inter- ro de interés agronómico. Actas de Hor- national Plant Genetic Resources Insti- ticultura 29: Volum 2: 649-656. explosively. Consequently, (in terms of tute, Rome, Italy. crop domestication and the initiation of Vargas, F.J.; Romero, M.; Plana, J.; agriculture) South-West Asia, temperate Rovira, M.; Batlle, I., 1995b. Characteri- Joley, LL. E. 1969. Pistachio. In: Hand- Europe and the Mediterranean Basin pre- book of North American Nut Tress, R.A. zation and behaviour of pistachio culti- Taynes (ed), 348-361 pp. W. F. Hum- vars in Catalonia (Spain). Acta Horticul- sently rank among the most intensively phrey Press Inc., Geneva, New York. turae, 419: 181-188. explored parts of the World (Zohary and Hopf 2000).

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 53 This paper aims at a short review of the archaeological, botanical and genetic evidence already available on the domestication of the pistachio nut Pistacia vera. However it is not easy to reach a summary. On the one hand, there is reliable evidence, obtained from the living plants, indicating that the wild progenitor of this nut crop is geographically confined to Central Asia. In addition, as far as archaeo-botanical information, Central Asia is still po- orly explored. Consequently, the archaeological information about the domestication of the pistachio nut is still grossly insufficient. However, some relevant pieces of archaeo-botanical evidence do exist, mostly in South-West Asia. These provide some clues for answering the questions: when, where and how the domestication of the pistachio nut could have taken place. In this note, an attempt Figure 1. Savannah-like park forest of Pistacia vera in the Badkhyz, South Turkmenia. is made to combine (i) the relevant Çfossil evidenceÈ already discovered at archaeological digs, with (ii) the information extracted from the living plants.

BOTANICAL AND HORTICULTURAL BACKGROUND The following four deciduous tree-size Pistacia species grow wild in the semiarid steppes, steppe-forests and Savan- nah-like vegetation of Central and South- West Asia, as well as in the Mediterra- nean basin (Zohary, M. 1952; Browicz 1984, 1988; Zohary, D. 1996). Only one of these four wild Pistacia species, namely the pistachio nut, P. vera, was domesticated, and gave rise also to vegetatively propagated, female clonal cultivars. The three other wild species, namely P. atlantica, P. khinjuk and P. terebinthus (including subsp. palaestina) stand taxonomi- cally somewhat more remote from the Map 1. Distribution range of wild forms of Pistacia vera. Based mainly on K. Browicz (1988). crop. However, they are closely associated with pistachio horticulture, being widely used as stock material for scion RANGES OF DISTRIBUTION the Canary Islands. P. khinjuk spreads grafting (budding) of domestic P. vera AND THEIR INDICATIONS from Central Asia to Sinai. P. terebin- clones. In some places these three spe- As already noted, Pistacia vera is a Cen- thus (including subsp. palaestina) is a cies also replace P. vera as pollen do- tral Asian element. Wild forms of this spe- characteristic arboreal element of the nors, and are used for pollinating the do- cies are fully inter-fertile with the crop’s maquis vegetation all over the Mediter- mestic female clones. In addition the cultivars. They are widely accepted as ranean basin. fruits borne by all these wild growing pis- the wild progenitor from which the domes- tacia taxa have a long tradition of being tic clones have evolved. The distribution The fact that the wild forms of P. vera collected from the wild, and used for con- area of the wild populations of P. vera is are geographically confined to some sumption as nuts, or for oil extraction. confined (Map 1) to parts of the Middle parts of Central Asia (Map 1) is a real This practice still survives in some areas Asian republics (South-East Uzbekistan, asset, since it delimits, rather specifical- with traditional ÇprimitiveÈ farming. In Tadzhikistan, Kirgizia) and North Afgha- ly, the geographic area in which this Central Asia, the local wild stands of P. nistan, and it extends westward to the Ko- crop could have originated. In other vera, with their relatively larger nuts (10- pet Dagh range in most southern Turkme- words, it suggests that this nut crop was 20 mm in length) seem to be the most at- nistan and North-East Iran (Browicz introduced into horticulture roughly in tractive source for collection from the wild 1988; Kamelin 1990). Within their distri- the same general territory in which the (Browicz 1988). However even the smal- bution range, wild forms of P. vera fre- wild populations of P. vera thrive today. ler fruits of the three other Pistacia spe- quently appear as a dominant arboreal This is especially true, if the biomes in cies are still being collected by local inha- component of Savannah-like park forests, this part of the world had not changed bitants all over their much wider distribu- steppe forests and similar semi-arid envi- drastically since the domestication of tion ranges. In the last few decades one ronments (Fig.1). In contrast, distribution P.vera (that I suspect, occurred only is also faced with an accelerated activity, ranges of the three other wild Pistacia some 2500-3000 years ago). Further by using trees of the four wild species for species are much wider: P. atlantica ex- support for Central Asian domestication in situ grafting of domestic clones. tends from Central Asia to Morocco and is given in the next section.

54 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 NUT REMAINS AND THEIR usually with the olive tree (Olea euro- CONTRIBUTION: REFERENCES paea), lentisco (Pistacia lentiscus) and Remains of the nutshells of the four Pis- myrtle (Myrtus communis); it is located tacia species constitute the main Çfossil Browicz, K., 1984. Chorology of trees mainly in coastal and inland areas of Mo- and shrubs in south-west Asia and adja- evidenceÈ retrieved from archaeological cent regions. Institute of Dendrology, rocco, Algeria and Tunisia, this area re- excavations, and they were soundly iden- Polish Academy of Science. Polish presenting 18% of the world carob pod tified by archaeo-botanists. However, a scientific Publishers, Warszawa-Poz- and 30% of total seed (garrofín). In Tuni- large part of the reported archaeo-botani- nan. Vol. 3: 88 pp. sia, carob trees are distributed in different cal information appeared in reports, sup- ecological areas, and usually they are Browicz, K., 1988. Chorology of trees plements in books, articles in local jour- and shrubs in south-west Asia and adja- found in groups of isolated trees and the- nals and newsletters, in many languages. cent Regions. Institute of Dendrology. re are very few commercial orchards. The All this is difficult to collate, particularly Polish Academy of Science. Polish potential production of carob pods is for people working with the living plants. scientific Publishers, Warszawa-Poz- around 2.500 t, and used mainly for do- na_. Vol. 6: 86 pp. Altogether Pistacia remains have been mestic-consumption and animal feeding "in situ". At industrial level, only one kib- uncovered from about 35-40 archaeologi- Kamelin, R. W. (ed.), 1990. Pistachio- cal sites, most of them in South-West tree in Bbadkhyz [in Rusian]. Komarov ble processing company exports some Asia and in the Mediterranean basin. Few Botanical Institute, USSR Academy of 60-70 t of seed to the EU. sites are from Central Asia, East Iran, Science. Leningrad. 230 pp. Afghanistan and Pakistan. They repre- The traditional use of the carob bean until Zohary D., 1996. Taxonomy. in: Padulo- sent a variety of prehistoric and early his- si, S., Caruso, T. and Barone, E. (eds.) the 1960s was its pulp (90% of the pod torical ages - from epi-palaeolithic to Taxonomy, distribution, conservation weight approximately) for animal feeding. Greek and Roman times. Fortunately, in and uses of Pistacia genetic resources. In Europe, currently, the most valued part these remains, one is able to distinguish International Plant Genetic Resources of the fruit is the kernel for gum extraction between nutshells coming from (i) wild Institute, Rome - Italy: 1-11 pp. (E-410), which is used as stabilizer and and cultivated forms of P. vera, and (ii) Zohary, D. and Hopf, M., 2000. Domes- thickener for the food industry. During the those produced by the three other Pista- tication of crops in the Old World (3rd last decades this use has recovered inter- cia species. The fruits of P. vera (both edition). Oxford University Press. est for its cultivation (Batlle and Tous, wild and domestic forms) are twice as big Oxford, UK. XI + 316 pp. 1997). In traditional carob producing (or even more) than those produced by P. Zohary, M., 1952. A monographic study countries (Spain, Italy, Portugal and atlantica, P. khinjuk and P. terebinthus of the genus Pistacia. Palestine Journal Cyprus) the carob tree is growing with (Browicz 1988). In addition, P. vera is uni- of Botany (Jerusalem series), 5: 187- grafted varieties producing fruits with high que in the hardness of its nutshells, that 228. pulp content (90%), while wild carobs are commonly split longitudinally at maturation. normally found in the North of Africa, pro- *) For survey of the literature published on plant remains retrieved from ar- ducing pods of high seed yield (18-20%). Definite signs of P. vera presence were chaeological excavations (including Pis- The genetic variability of predominantly not discovered in South-West Asia and tacia species) consult the following an- seedling trees is very high, which justifies the Mediterranean Basin before Greek nual reviews, prepared (from 1989/1990 the interest of surveying the native mate- onwards); first by J. Schulze-Motel and and Roman times. This in spite of the fact later by H. Kroll: ÇLiterature on ar- rial in this region. Therefore, in the year that both these territories are archaeo- chaeological remains cultivated plantsÈ 2002 a joint project AECI started between boanically well explored, and South-West published in Vegetation History and Ar- Spain (IRTA-Mas de Bover) and Tunisia Asia is geographically a neighbour of chaeobotany, Vols. 1-11. (INRGREF and INSAT) entitled ÇSelec- Central Asia. Such absence strongly sug- tion, conservation and valuation of the ca- gests late domestication of the pistachio robÈ. In this article we present the results nut (in Central Asia) and even more so - D. Zohary of the native carob tree survey made in Department of Evolution, Systematics and late arrival of P. vera culture (to South- Ecology Tunisia, in order to know the areas of West Asia). A possible explanation for The Hebrew University, Jerusalem 91904, highest density and diversity of this spe- this delay is the way this nut crop is vege- Israel cies. Also, we identified the most promi- tatively propagated. The pistachio nut E-mail: [email protected] sing commercial trees or seedlings. does not lend itself to rooting. Instead, it requires grafting, a technique that appea- MATERIAL AND METHODS red in South-West Asia and in the Mediter- The methodology followed was similar to ranean Basin only in the first millennium that used in early carob surveys made in BC (Zohary and Hopf 2000: 143). Theo- CAROB TREE GERMPLASM Spain (Tous et al., 1995 and 2005). We phrastus II.v.3 in the 4th Century AD is IN TUNISIA first identified the main carob production the earliest literary source mentioning areas of Tunisia (Map 1) with the help of grafting in the Mediterranean Basin.] technicians of the INRGREF and INSAT. The exploration was carried out in July of Further east, few sites excavated in Cen- INTRODUCTION 2003 and 2004 which is the period when tral Asia (even somewhat South-East of The carob tree (Ceratonia siliqua L.) is a carob ripening begins and flowering over- the present distribution area of wild P. polygamous, thermophilous and typical laps in this country. We travelled by car, vera) yielded older nutshells remains. evergreen species of the Mediterranean some 2.000 km on five different itinera- They were dated as belonging to Chalco- basin. The European Union (Spain, Italy, ries, covering the main carob density lithic and/or Bronze Age times. However, Portugal, Greece and Cyprus) produce areas: I) Ariana-Tunis, II) Cap Bon (Soli- as nutshells coming from wild populations more than 70% of the world crop, estima- man-Korbous), III) Zaghouan-Jradou, IV) and those produced by domesticated clo- ted currently around 300.000 t of pods, Bargou-Oueslatia and V) Monastir-Khe- nes overlap to some extent in their sizes and it is mainly concentrated in Spain nis-Sayada. (Browicz 1988), it is yet hard to decide (100.000 t). In North African countries ca- whether these remains represent collec- rob trees are usually found as wild, in iso- During the visits information was taken tion or cultivation. lated tree populations, and associated relating to growing areas, agronomic as-

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 55 Map 1. Native carob tree distribution in Tunisia, marking the five Figure 1. Carob pods showing size shape diversity from the are IV surveyed populations (I a V). (Bargou-Oueslatia). pects (potential carob yield, easy pod dropping, Oidium tolerance, etc.) and commercial characteristics (kernel yield) of each variety and/or most interesting ecotypes, as well as fruit samples for their later description, weight, measures and kibbling. In the laboratory several morphological parameters of the fruit were studied (length, weight and kernel yield) and their seed (weight, thickness and gum content). Scion woods from the selected types will be collected and introduced by grafting on seed rootstocks at the IRTA’s carob gene bank in spring 2005, for their conservation and assessment.

RESULTS AND DISCUSSION Current situation and growing areas In Tunisia carob trees are usually found as wild types, on coastal and inland areas, its presence being outstanding from sea level to 950 m above it. Carob populations also have been observed as Figure 2. Research group that surveyed the native carob trees in Tunisia. far as 120 km from the coast in area III (Bargou-Oueslatia), contrarily to what appears in the traditional Spanish regions racterized by hot dry summers with high dividual trees which show a complex flo- (Catalonia, Valencia and Balearic is- temperatures. Carob tree growth takes ral biology, of which trees with female flo- lands), where they usually grow well until place in marginal land, normally in calca- wers (55% of the total) and male flowers 500 m altitude and 40-50 km from the reous and basic pH soils. (37 %) are outstanding. Some scattered coast (Tous and Batlle, 1990). The obser- hermaphrodite types (8%) are found, vations carried out in Tunisia show the There are few regular orchards, with wide mainly, in the Bizerte and Cap Bon areas presence of carob populations adapted to plant densities (80-100 trees/ha), and (Afif et al., 2003). Also we observed that different ecological conditions, with rain- they were established during the decades as the altitude increases the proportion of fall habitats ranging from 1000 mm in Ta- of the 1950s and 1960s of the last cen- male trees rises in relation to those of fe- barka (north), lowering to 460 mm in Tu- tury, mainly growing the ‘Sfax’ cultivar male flowers (Area III). The bloom period nis, 300 mm in Souassi-Monastir and re- (Crossa-Raynaud, 1960). The native ca- takes place between August and Septem- aching down to 215 mm in Sfax, at the rob has not generally been propagated by ber, being necessary male and/or herma- eastern part of the country (Rejeb et al., farmers: it was spread by natural dissemi- phrodite trees to pollinate the female 1991). Overall, the climate is arid, cha- nation. This distribution has originated in- trees.

56 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Figure 3. Native carob tree usually grow in groups of isolated trees Figure 4. Traditional image of Tunisia: Carob tree associated with olive (area III: Zaghouan). and agave.

Table 1. Main carob tree populations surveyed in Tunisia.

Population Preselected trees code Surveyed area (altitude, m) Bioclimatic characterization (rainfall/year, mm)

Ariana-Tunis (A) A1*, A2, A3 I (20-50 m) Humid lower (460 mm) Cap Bon (CB) - II (100-300 m) Semiarid superior-medium (425 mm) Zaghouan (Z) Z6 III (200-800 m) Semiarid superior-medium (400 mm) Jradou (J) J3, J4 III (400-500 m) Semiarid superior-medium (400 mm) Bargou (B) B8, B9, B10, B11, B12, B15, B19, B20 IV (500-700 m) Semiarid medium-lower (350 mm) Sayada-Monastir (S) S4, ‘Sfax’** V (50-200 m) Semiarid lower (300 mm)

* Hermaphrodite ecotype ** Cultivar

Table 2. Carob pod morphological and commercial traits of some ecotypes preselected in Tunisia, compared with other Spanish growing cultivars. Period 2003-2004.

Code Pod length Pod weight Seed yield Seed weight Seed thickness Gum content (cm) (g) (%) (g) (cm) (% endosperm)

‘A-1’ (H) 20,3 16,8 14,8 0,21 0,41 49,77 Z-6’ 18,3 8,5 18,4 0,18 0,36 53,45 ‘J-3’’ 19,3 14,4 16,7 0,18 0,37 - ‘J 4’ 19,7 14,5 18,4 0,17 0,34 56,01 ‘B-8’ 17,3 12,3 14,3 0,18 0,38 52,56 ‘B-10’ 15,2 8,9 24,1 0,20 0,42 55,79 ‘B-19’ 12,5 8,7 24,7 0,16 0,47 57,66 ‘S-4’ 21,0 16,6 23,2 0,21 0,44 53,07 ‘Sfax’* 17,7 33,2 7,21 0,22 0,45 53,28 ‘Rojal’** 17,2 19,12 11,44 0,18 0,39 55,11 ‘Duraió’** 16,0 15,93 17,57 0,24 0,45 57,76

*Main cultivar in Tunisia; ** Spanish grown cultivars

Characteristics of the plant material are usually medium and with a high seed Important pod trait differences between Sixteen promising trees were identified, yield) and ÇGraftedÈ (the smaller number, wild populations and cultivated trees were one of them being a hermaphrodite type, the fruits being outstanding for their large observed, which agree with parallel work and another tree is the main cultivated size and average-high pulp content and about characterization of the native carob variety (‘Sfax’). This cultivar was introdu- mainly based on the ‘Sfax’ type). germplasm of Tunisia (Naghmouchi et al., ced to overseas regions, as California 2004). (Coit, 1967) and Australia (Tous, 1995). Table 1 shows a relationship of the stu- Selected trees produced carob with high died populations, with the most promising We observed a wide range of pod size, yield in kernels and showed some out- trees, with indication of their range area. weight, shape and kernel yield. Pod size standing agronomic traits. In general, trees The grafted population, where the female varies from 12,5 cm, for the ‘B-19’ type, appear to be healthy although no sprays variety ‘Sfax’ is outstanding, is located in to 20 cm for ‘S-4’ and ‘A-1H’ types. The are used. In a few trees leaf symptoms of the coastal areas of Tabarka-Bizerte, Cap average weight of the fruit oscillates mildew (Oidium ceratoniae) were found. Bon and Monastir-Sayada (Map 1). The between 8,7 g (‘B-19’ type) and 33,2 g Two main carob populations were distin- results obtained, regarding the morpholo- (‘Sfax’ variety). Pod shape varies from guished: "ungrafted" or "wild" (the greater gical and commercial traits of the most in- straight to curved. Kernel yield ranges number all wild seedling trees; the fruits teresting ecotypes, are shown on Table 2. from 14-25% for wild types to 7-8% for

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 57 Figure 5. In some Tunisian inland zones, it is common to find sheep and goats feeding directly on the carob groves.

the ‘Sfax’ grafted population but, in gene- CONCLUSIONS ACKNOWLEDGEMENTS ral, it is higher that in other Spanish From the results of the survey and fruit This work has been granted by the growing areas (see Table 1). The highest characterization of the carob germplasm "Agencia Española de Cooperación seed percentages were found in the ‘B- in Tunisia the following aspects are outs- Científica" (AECI) through a joint resear- 19’ (24,7%), ‘B-10’ (24,1%), ‘S-4’ (23,2%) tanding: ch project of PCI Hispano-Tunecina and ‘Z-6’ (18,4%) types. The size of the (ref:78/03/R) on the selection of native seed is also very variable, from 0,16 g - In the surveyed area, several carob pro- carob germplasm in Tunisia. We are gra- (‘B-19’) to 0,22 g (‘Sfax’), the same as the ducing zones (coastal and inland) were teful to Tunisian institutions (INRGREF gum content, with variations from 49,7% established. Also a large microclimate and INSAT) for their helpful indications (‘A-1H’) to 57,6% (‘B-19’). It is necessary and a high range of elevation diversity during this survey. to consider that currently kernel yield is has been observed in the areas where an important commercial trait for carob the native material survey has been loca- material preselection but, also that the ted. REFERENCES main feature to carry out its final selection should be the total production of Afif, M., Soyah, N., Khouja, M.L., Bous- seed per tree. - A high genetic variability within the Tuni- said, M., 2003. Le caroubier (Ceratonia sian carob germplasm has been obser- siliqua, L): Analyse preliminaire de la ved. The fruits of the studied populations diversite genetique des populations na- The criteria for fruit evaluation, relating to turelles en Tunisie (typeset copy). commercial (fruit weight and seed yield) have a wide variability, especially in the and agronomic traits (large fruits and aspects related to fruit weight and kernel Batlle, I., Tous, J., 1997. Carob tree. easy pod dropping to improve the har- yield. Ceratonia siliqua, L. Promoting the conservation and use of underutilized and vesting) are focussed on making a first neglected crops. 17. Institute of Plant tree preselection in the field. According - The selected genotypes gave pods of Genetics and Crop Plant Research, to these results, 6 outstanding indivi- high kernel yield in wild ecotypes, and Gatersleben/International Plant Genetic duals have been selected, belonging to thus it is necessary that they perform a Resources Inst., Rome, Italy. 92 pp. the populations Ariana-Tunis (A-1H), good agronomic and productive beha- Coit, J.E., 1967. Carob varieties for the Zaghouan-Jradou (Z-6 and J-4), Saya- viour in collections and trials for their semi-arid southwest. Fruit Varieties da-Monastir (S-4) and Bargou (B-10 and possible use in future commercial or- and Hort. Digest, 21: 5-9. B-19). chards.

58 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Naghmouchi, S., Khouja, M.L.,Tous, J., Rejeb, M.N., Boussaid, M., 2005. Le caroubier (Ceratonia siliqua, L): Une ri- chessse genétique et un patrimoine bio- logique d’une grande importance pour la Tunisie (typeset copy).

Rejeb, M.N., Laffray, D., Louguet, P., 1991. Physiologie du Caroubier (Cera- tonia siliqua, L) en Tunisie. En: Physio- logie des Arbres et Arbustes en zones arides et semi-arides: 417-426. Ed. Groupe d’Etude de l’Arbre. París.

Tous, J., 1995. Situación del algarrobo en Australia. Boletín Agropecuario ‘La Caixa’, 35: 43-49.

Tous, J., Batlle, I., 1990. El algarrobo. Ed. Mundi-Prensa. Madrid, 102 pp. Walnut, orchard in Tarragona, Spain. Tous, J., Batlle, I., Romero, A., 1995. Prospección de variedades de algarrobo en Andalucía. Información Técnica Since the mid 1990s, East Malling Re- culture, Seed and Plant Improvement Re- Económica Agraria (ITEA) 91 V (3): search, Kent has been establishing a col- search Institute (SPII), Faculty of Agricul- 164-174. lection of walnut varieties and timber se- ture / University of Tehran, Agricultural Tous, J., Batlle, I., Rallo, J., Romero, lections with the aim of selecting genoty- Biotechnology Research Institute of Iran, A., 2001. Prospección de variedades de pes that are well adapted to growing in Farmer’s House, Iranian Society for Horti- algarrobo en las islas Baleares. Investi- the UK. Graftwood of genotypes reported cultural Science and sponsored by Agri- gación Agraria (Producción y Protec- to have characters useful for future bree- cultural Bank and Rafsanjan Pistachio ción Vegetales). Vol 16 (2): 187-203. ding was supplied by colleagues from a Producer’s Co-operative (RPPC). range of European countries and North America, who are involved in walnut im- J.Tous1, M. Rovira1, A. Romero1, M. Afif3, The opening ceremony started with a wel- M. L. Khouja2, S. Naghmouchi2, provement programmes. In total, some 60 come lecture by Dr. Amanollah Javansh- M. Boussaid3 J. regia varieties and selections are being ah, convener of the Symposium and head 1IRTA Mas de Bover. grown at EMR and are being recorded for of the Iran’s Pistachio Research Institute Ctra. Reus-El Morell, km 3,8 budburst, disease resistance, vigour and (IPRI), in which he extended his thanks E-43120 Constantí, Spain 2Institut Nacional des Recherches en Génie fruit production. It is hoped that promising and gratitude to the distinguished partici- Rural, Eaux et Forêts (INRGREF). varieties can be identified and propaga- pants, particularly those from other coun- Ariana (Tunisia) ted for multisite trials in the near future. tries, and also very cordially thanked the 3 Institut National des Sciences Appliquées members of scientific and executive com- et de Technologie (INSAT). Tunis (Tunisia). E-mail: [email protected] K. Russell mittees and editorial committee and his East Malling Research colleagues of the Iran’s Pistachio Re- New Road, East Malling search Institute. Kent ME 19 6BJ, England E-mail: [email protected] The second presentation was delivered by H.E. Dr. Ali Ahoonmanesh, Hon, De- NOTES AND NEWS puty Minister and head of AREO. At first he extended his warmest welcome to the CONGRESSES prominent participants and the esteemed WALNUT RESEARCH AT EAST AND MEETINGS scientists, participating in this Sympo- MALLING RESEARCH sium. He stated that Iran is the oldest and biggest producer and exporter of pistachio and the fourth producer of almond in IV INTERNATIONAL the world and Iran demands for higher Interest in growing walnuts for fruit and SYMPOSIUM ON PISTACHIO contribution regarding policy making of timber in the UK has undergone a revival AND ALMOND these crops at international level. Then in recent years. This was clearly demons- he gave a brief description of AREO’s or- trated when in response to requests for ganizational chart, mandate, responsibili- information regarding the distribution of ty and research activities of its affiliated walnut trees in the UK, over 10,000 indi- The 4th International Symposium on Pis- national research institutes and centers. viduals reported their walnut trees to tachio and Almond under the auspices of EMR. The most northern fruiting trees in the International Society for Horticultural The following speech was presented by the UK were located in Ullapool, on the Science (ISHS) was very successfully the Iranian Pistachio Producers Coopera- western coast of Scotland - over 600 mi- held in Tehran-Iran from 22 to 25 May tive representative. He propounded for les north of London. However, the area 2005. The number of participants from solving most causes of damages in agri- of walnut grown commercially is still very Iran and all continents of the world was cultural production that farmers should small and restricted largely to southern high. The symposium was organized by: have access to advanced technologies England. Research is required to identify Agricultural Research and Education Or- and research highlights for enhancing the varieties and selections best suited to the ganization (AREO), Iran’s Pistachio Re- quality and quantity of their products and UK conditions; often late spring frosts search Institute (IPRI), Deputy of Horti- in this regard making a close celebration and cool, wet summers. cultural Affairs, Ministry of Jihad-e-Agri- among national research institutes.

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 59 IV International Symposium on Pistachio and Almonds. 22-25 May - 2005 - Tehran - Iran.

The first key note lecture was presented The output of the Symposium was satis- sits. The conference was preceded by a by Prof. Talaei, the dean of Agriculture factory especially the message of Dr. technical tour in Veneto. Held in Sorrento, Faculty in Tehran University. He gave Kaiser from the United States of America the town which gave its name to one of some general information about the stra- who, in Persian language, evoked the the oldest Italian walnut varieties, the tegic and geographical position of Iran, its emotional feelings of all participants pre- Symposium was attended by a large population and other relevant information. sent in the meeting. number of participants coming from 28 The Second key note lecture was given countries, among them various underde- by Dr. Alavi in the field of Aflatoxin and Finally the whole executive and scientific velopped countries were represented. Mycotoxin. The last key note lecture was committee members wished all deared delivered by Dr. S. Chaichi in the field of participants good health, success and Several papers debated the Economic analysis on Almond breeding in Iran. prosperity. and commercial interest for walnut cultivation, including the lecture of the repre- Then the total of 141 Oral presentations sentative of the International Nut Council and 175 Posters, divided into eight ses- which detailed the worldwide situation of sions, were presented in three halls. A. Javanshah walnut production and consumption. The H. Hokmabadi Iran’s Pistachio Research Institute synthesized results showed that the Eu- The Symposium’s exhibition contained 14 P.O. Box 77175.435, Rafsanjan, Iran ropean countries, with France as leader, stands on different pistachio and almond Telephone: 0098 391 4225201 are at the moment developing a modern industries. At the end of the second day a Fax: 0098 391 4225208 walnut production system, by applying or- splendid dinner party hosted by Dr. Ka- E-mail: [email protected] ganic farming methods. Another fact E-mail: [email protected] lantari, director of Khaneh Keshavarz, brought to our attention by the Spanish was served at the diplomatic club, enter- presentation is the demand from consu- tained with delicious traditional Iranian mers for specific quality standards. The dishes. perspectives offered by the walnut market have stimulated interest from Northern On the third day, in a special ceremony, SUMMARY OF THE European countries like UK and the Nether- the ISHS medal was awarded to Dr. Ama- V INTERNATIONAL WALNUT lands as well as Southern American and nollah Javanshah by Dr. R. Socias i Com- CONGRESS non-traditional producing Asian countries. pany, ISHS representative, and all those researchers already retired in the field of In Genetics and Breeding an overview of Pistachio and Almond were appreciated the different lines of research carried out, with special gifts for their valuable and The V Symposium was held in Sorrento, ranging from taxonomical classification of effective efforts. Italy from the 5th to the 9th of November Juglandaceae to breeding programmes, 2004. The Symposium presented the cur- were presented. An interesting contribu- At the end of the afternoon session, the rent status of the Juglans regia species in tion was a study carried out by cpDNA in- closing ceremony was held and there Tur- all its different aspects (fruit production, tergenetic spacer markers which demons- key was selected as the host for the next timber production, liquor and relative ‘fi- trated that the origin of the section Ju- Symposium. lieres’). This approach stems from the in- glans is ancient in contrast to the fossil discriminate nature of the various biologi- evidence, and that the evolution of Per- The day after the Symposium closing ses- cal research aspects, regardless of the fi- sian walnuts and butternuts happened at sion, the three days tour organized by nal product. Organised by two conveners, a different rate than black walnuts. In the Symposium organizing committee, star- D. Avanzato and M.E. Malvolti with diffe- UK, renewed interest for walnut led to the ted from Isfahan and progressed down rent backgrounds, the conference aimed establishment of research activities devo- the south of Iran to Kerman and Rafsan- to present an integrated view through ted to early growth of walnut in the UK jan. scientific sections as well as technical vi- and the behaviour of progenies/prove-

60 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 nances from Kyrgyzstan, as well as pro- plant agamic regeneration, which in wal- ning trees, and classified on the basis of grammes aimed at genetic improvement. nut seems not to be very common, morphological characters and molecular The University of California, famous for embryogenesis of different tissues was markers. Phytophtora spp. causes similar its breeding programme since 1948, are examined. In the absence of normal root symptoms to drought or nutritional defi- developing controlled pollination crosses and shoot production from walnut soma- ciency. The decline and death can be due in young orchards, for increasing the tic embryos, the effect of abscissic acid to a combination of Phytophtora spp. Clo- breeding efficiency, by traditional and ad- on embryos maturation was also investi- nes of different walnut hybrids were as- vanced techniques. Backcross breeding gated, with early stage development as sessed for resistance to P. citricola, and aims at introducing the gene for the cher- best suited to ABA treatment. the tolerant clones can be used for impro- ry roll virus resistance. The genetic varia- ving rootstocks. Phytophtora spp. and tion among and within populations is un- Mechanisms controlling plant growth and Brenaria nigrifluence cause serious da- dertaken to verify the interaction of gene- production in different conditions were mage in Iran. Pathogens were isolated to tic and environmental characteristics. The discussed in the Biology and Physiology study the reaction of different cultivars preliminary analysis of leave images by session. One important aspect taken into and Çfalse woodÈ plants. Anthracnose is an optical scanner provides a new me- consideration was adaptation to low tem- one of the most important diseases and thod for the characterisation and assess- peratures. To understand the processes Italian provenances identified as resistant ment of genetic variability. Developing involved, attention was focused on two can be used as source germplasm for countries are carrying out a traditional proteins related to sugar transport. For breeding and production of improved ma- broad spectrum of research activities on walnut starch degradation and the subse- terial. In Northern California, walnut blight J. regia. They still contain a wide genetic quent release of sucrose it is of major im- was controlled using cupper compounds and morphologic variability, interesting portance to repair metabolism. The im- augmented with Manex. An early season for breeders and scientists alike. Appro- portance of winter biology as preparation bactericide spray programme seems to priate programmes have been started to for the next vegetative season was de- be promising. validate the selection and testing of local monstrated. The utilisation of walnut in varieties and increase the growing effi- mixed plantations was studied using oxy- Problems concerning the hedging training ciency and production quality. Several gen stable isotopes, as tracers for the system and their suitable cultivation me- presentations showed worldwide efforts water usage by the different species. It thods were discussed in Orchard and Fo- for collection, evaluation and characteri- showed the potential benefit from asso- restry Management. Results suggest that sation of material for different producti- ciation with some woody species or less frequent pruning is likely to increase ons. Discussion on genetic resources shrubs. Close relationships exist betw- productivity and that irrigation and nu- conservation was carried out, concluding een photosynthesis efficiency of intercro- trient management in the year after the that germplasm should be protected on pped plants. Thus, it can be assumed hedging have the highest potential impact global level as richness for humanity. that a relation exists between leaflet area on canopy development. Also some hed- and photosynthetic efficiency. Also the ging strategies are more advantageous In Biotechnology great attention was gi- interaction between phenolic compounds than others and more advantageous than ven to inter-specific hybrids due to the and different cultivar stage developments non-pruned treatment by taking care of possibility of concentrating more advanta- was demonstrated. Moreover some pheno- the solar light management. Another as- geous characteristics of parents, such as lic compounds seem to be involved in the pect related to the orchard management timber and nut quality, fast growth, right defence mechanisms of the walnut plant. is the water use. Research carried out by stem and disease resistance. There were Bongi et al. showed that J. regia seems to presentations about the application of The difficulty of rooting and grafting of be a rootstock adapted to slower and molecular markers as well as advanced walnut species remains under investiga- wider water supplies than riparian root- biotechnologies, both used for improving tion. Focus lies on seed treatment for stocks in Argentina. For intensive cultiva- selected plants, with economical and na- seedling production before the grafting tion the necessity for systematic manu- tural values. Currently, the most useful step and the influence of genetic prove- rings and irrigation as well as soil structu- markers seem to be microsatellites nance on quality. Problems of low plant re adaptation in poor areas clearly emer- (SSR). Walnut nuclear SSR have been growth due to coercion of root apparate ged. developed in the USA by Woeste and were encountered, but overcome by hor- applied in Italy to discriminate J. regia mone treatment of the roots. Still under In Nut and Wood Processing research ai- and J.nigra genotypes and their hybrids, investigation is the propagation of root- med at the improvement of moisture con- and to characterise cultivars. Modification stocks and hybrid resistance to disease tent after nut drying, a factor which can of native germplasm for commercial rea- and nematodes as well as concern over compromise shelf-life. The interest in sons is necessary when certain desired the production of ownrooted cloned va- shelf-life has stimulated the research into traits are missing. A genetic improvement rieties. Also under research are some the role of lipoxygenases (LOX) on walnut programme on J. nigra tested the effect of physiological aspects to clarify mecha- quality and the presence in purified lipid genotype and kanamycin selection on nisms of Çin vitroÈ rooting or embryo re- bodies can indicate a role of this enzyme agrobacterium mediated transformation generation. Due to their practical impor- in nut quality. The nutritional value of the efficiency, yielding stable transgenic li- tance for plant nurseries, data on walnut nut was also investigated and emphasis nes. It needs reminding that Agrobacte- grafting were reported using both traditio- was placed on the content of Taurina, po- rium tumefaciens is however the cause of nal and new methods, like hypocotil graf- orly present in vegetables, but well pre- crown gall disease. This infection, wide- ting and hot callusing. sent in animal tissues as a metabolite of spread in Californian orchards and quan- sulphured amino acids, in the nut. The tifiable by the pathogen presence in the Selection of disease resistant varieties is chemical composition and health property soil, was studied by soil DNA extraction, of interest to both scientists and produ- of the walnut brandy was also studied. PCR amplification and electrophoresis. A cers alike, reducing the amount of chemi- The presence of phenolic compounds positive correlation between observed di- cal compounds needed and thus preser- deemed to have anti-cancerogenic pro- sease incidence in the field and the pre- ving the environment. Various Phyto- perties, as discussed. An exposition of sence of A. tumefaciens gives a useful phthora species were isolated from diffe- different walnut products, coming from all tool. Finally, due to the importance of rent plant parts and from soil under decli- regions of the globe, was displayed.

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 61 A special half-day workshop was held on During the first day, information was gi- ting gathered a particularly impressive walnut cultivation in China. The recent re- ven on walnut for fruit, analysing from the participation: 160 delegates representing organisation of its agriculture meant that current crop situation until the potentiality research and industry of over 24 countries walnut became also important for its timber of new walnut orchard planning. Also, se- attending the congress. About 220 papers production, and not only for its traditional veral technical contributions were sche- on many aspects of chestnut research fruit production. The Italian National Re- duled on pests and diseases affecting were presented and discussed. The mee- search Council (CNR) has been particularly this nut tree and about the most suitable ting was organised under the auspices of active in this area, with complementary Chi- training systems for planting new or- the International Society for Horticultural nese partners (Chinese academy for Agri- chards. Finally an overview was given on Science, in cooperation with the Universi- cultural Sciences, CAAS, and the Chinese the possitive effects of nut consumption ty of Tras-os-Montes, Regional Agricultu- Academy of Forestry, CAF) for the pro- on human health. The first day ended ral Service, Agricultural College of Bra- motion and introduction of innovative with a very lively round table where the gança and the Portuguese Society of Hor- agricultural models aimed at improving the walnut market, quality requirements and fi- ticulture. agricultural production, and create new pro- nancial support for orchards were debated. ducts responding to the recent evolution of There were two invited speakers in the Chinese production systems to industriali- The second day, a field visit to the IRTA’s Opening Ceremony: Prof. Giancarlo sation. Some results of the project have Estació Experimental of Lleida and two Bounous, Chairman of the ISHS Working been presented together with those obtai- young orchards allowed the numerous at- Group on Chestnut presented a keynote ned by Chinese researchers. Factors affec- tendance to observe Çin situÈ several of paper entitled ÇThe Chestnut, a Multipur- ting "in vitro" micro-shoot rooting of pre- the walnut planting developments com- pose Resource for the New MillenniumÈ cious Chinese cultivars were detailed as mented the day before. and Dr. Sandra Anagnostakis, plant pa- well as the application of parthenocarpy in thologist, CAES, Connecticut, USA, pre- genetic purification of walnut variety. The third day was focussed on walnut sented "Chestnuts in the US for Food and timber production, its technical and com- Timber". Dr. Stephanos Diamandis, NA- The symposium included a pre-sympo- mercial aspects and the evolution of the GREF, Thessaloniki, Greece, presided sium tour in Veneto region, where a mo- silviculture of this species in France and the Closing Ceremony. dern orchard farm cooperative (Nogalba) Spain. The workshop was ended by a was visited. The technical tour included round table on reforestation of agrarian Five thematic sessions, each highlighted the visit of Petillegno company in order to land, the environmental impact of the by an invited speaker, formed the main see a wood processing industry, which agroforestal orchards and timber value. part of the meeting: uses wood coming from Italy and the rest Chestnut Growing, Economy, Marketing, of the world. The second part was a visit Among others, thanks to the financial Harvest and Fruit Processing; of "Besana" enterprise that is specialised support of the European project Interreg Biology, Physiology and Ecology; in processing, packaging and marketing of III PORTA it was possible to organise this Genetics, Breeding, Biotechnology and Italian and foreign nuts and other dry fruits. Workshop, between regions of the south Plant Development; of France and Catalonia. The participa- Plant Protection; At the end of the symposium Damiano tion was considerable, with about 200 Orchard and Forest Management. Avanzato was elected Chair of the ISHS people attending the Workshop on walnut Walnut Group and Australia was chosen as for fruit and with about 160 people the A technical tour was arranged on 21st Oc- the seat of the next Walnut Symposium. last day, in the frame of walnut for timber. tober. The participants had the opportuni- The French party was notorious, with 10 ty to visit the Padrela Mountain region and contributions in French, and with an at- the Terra Fria Chestnut area. D. Avanzato Consiglio per la Ricerca e la Sperimentazione tendance of 40 people the first two days in Agricoltura and about 30 people the last day. The conclusions of the congress were Istituto Sperimentale per la Frutticoltura, also presented by Dr. Diamandis with em- Roma, Italy IRTA, Mas de Bover - Mediterranean Fruit phasis laid on the chestnut gall wasp, Dr- E-mail: [email protected] Trees was in charge of the organization yocosmus kuriphilus, as a major threat for M. E. Malvolti of the Workshop. The Workshop’s asses- orchards and forests of Europe, ink disease Consiglio Nazionale della Ricerca sment forms filled by the participants re- and blight which seem to be spreading in Istituto di Biologia Agro-Ambientale e flected that most of their expectations Portugal and other countries as well, chest- Forestale di Porano, Italy were fulfilled, as to the quality of the oral nut wood technology, which should be en- E-mail: [email protected] contributions and the possibility of ex- couraged, as well as more efforts on the im- changing experiences among producers provement of the propagation systems and of neighbouring countries. research on chestnut marketing on national SPANISH-FRENCH WORKSHOP and international level. ON WALNUT N. Aletà The IV ISHS International Chestnut Con- IRTA Mas de Bover, Constantí, Spain E-mail: [email protected] gress will be held in Beijing, China, Septem- ber 2008. The manuscripts presented at this From the 22nd to the 24th of November meeting were reviewed by the members of 2005 a ÇSpanish-French Workshop on the Editorial Board and appear in the volu- walnut: fruit and timber productionÈ III INTERNATIONAL CHESTNUT me 693 of Acta Horticulturae. (ÇJornades hispano-franceses sobre el CONGRESS noguer: la producció de fruita i fusta») was held. The three days Workshop took C. Gomes de Abreu Convener 3rd International Chestnut Congress place in the frame of a suitable and newly UTAD refurbished old building of the village Po- The III International ISHS Chestnut Con- Vila Real, Portugal ble de Mafumet, Tarragona, Spain, kindly gress was held in Chaves, Northeast Por- E-mail: [email protected] provided by the townhall. tugal, on 20-23 October 2004. The mee-

62 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 TO BE HELD: BIBLIOGRAPHY

Almond and Pistachio

XIV GREMPA Meeting on Almond and Pistachio ALMOND Date: September, 2007 Place: Saronis, Attica, Greece Ahrens, S., Venkatachalam, M., Mistry, A. Convener: G. Zakynthinos M., Lapsley, K. and Sathe, S. K., 2005. Al- Address: Agricultural Products Technology Department mond (Prunus dulcis L.) protein quality. Technological Educational Institute of Kalamata Plant Foods for Human Nutrition 60(3): Antikalamos 24100 Kalamata GREECE 123-128. Tel.: 30 27210 45274 / Fax: 30 27210 45234 E-mail: [email protected] Alonso, J. M. and Company, R. S. I., 2005. Homage to Francesco Monastra Differential pollen tube growth in inbred Date: November, 2007 self-compatible almond genotypes. Euphy- tica 144(1-2): 207-213. Place: Messina, Sicily, Italy Conveners: F. Sottile and F. Marra Address: Dipartimento di Colture Arboree Alonso, J. M. and Company, R. S. I., 2005. Università di Palermo - Facoltà di Agraria Self-incompatibility expression in self-com- Viale delle Scienze, 11 patible almond genotypes may be due to in- 90128 - Palermo, Italy breeding. Journal of the American Society Tel: 39 09 170 49 000 / Fax: 39 09 170 49 025 for Horticultural Science 130(6): 865-869. E-mail: [email protected] Blooming almonds in Teruel, Spain. Amarowicz, R., Troszynska, A. and Shahi- V International Symposium on Pistachios and Almonds di, F., 2005. Antioxidant activity of almond Date: 2009 seed extract and its fractions. Journal of Place: Sanliurfa, Turkey Food Lipids 12(4): 344-358. Convener: B. E. Ak Address: University of Harran, Faculty of Agriculture, Department of Horticulture, Baiano, A. and Del Nobile, M. A., 2005. 63200 Sanliurfa, Turkey. Shelf life extension of almond paste pastries. Tel.: (90)4142472697 / Fax: (90)4142474480 Journal of Food Engineering 66(4): 487-495. E-mail: [email protected] Bi, G., Scagel, C. F., Cheng, L. and Fuchi- Hazelnut gami, L. H., 2005. Effects of copper, zinc and urea on defoliation and nitrogen reser- VII International Congress on Hazelnut ves in nursery plants of almond. Journal of Date: 23th - 27th June Horticultural Science & Biotechnology Place: Viterbo, Italy 80(6): 746-750. Convener: L. Varvaro Address: Hazelnut Research Center, CeFAS, viale Trieste 127, 01100 Viterbo, Italy. Browne, G. T. and Viveros, M. A., 2005. Tel.: (39)0761357461 / Fax: (39)0761357473 Effects of phosphonate and mefenoxam E-mail: [email protected]; [email protected] treatments on development of perennial http://www.hazelnut2008.it cankers caused by two Phytophthora spp. on almond. Plant Disease 89(3): 241-249. Chestnut Chalak, L., Elbitar, A., Rizk, R., Choueiri, E., IV International Chestnut Symposium Salar, P. and Bove, J. M., 2005. Attempts to Date: September 9-12, 2008 eliminate Candidatus phytoplasma phoeni- Place: Beijing, China cium from infected Lebanese almond varie- Convener: Dr. Ling Qin ties by tissue culture techniques combined Address: Beijing Agricultural College, No. 7 Beinong Road, Changpin District, Bejing or not with thermotherapy. European Jour- 102206, China. nal of Plant Pathology 112(1): 85-89. Tel.: (86)1080799136 or (86)1080799126 / Fax: (86)1080799004 E-mail: [email protected] Chen, J., Groves, R., Civerolo, E. L., Vive- ros, A., Freeman, A. and Zheng, Y., 2005. Walnut Two Xylella fastidiosa genotypes associated with almond leaf scorch disease on the same location in California. Phytopathology VI ISHS International Symposium on Walnut 95(6): 708-714. Date: January 2009 Place: Victoria, Australia Conveners: H. Adem Cherif, A., Sebei, K., Boukhchina, S., Kallel, E-mail: [email protected] H., Belkacemi, K. and Arul, J., 2004. Kernel fatty acid and triacylglycerol composition for three almond cultivars during matura-

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Groves, R. L., Chen, J., Civerolo, E. L., Reidel, E. J., Brown, P. H., Duncan, R. A., He- Matos, M. C., Rebelo, E., Lauriano, J., Se- Freeman, M. W. and Viveros, M. A., 2005. erema, R. J. and Weinbaum, S. A., 2004. medo, J., Marques, N., Campos, P. S., Ma- Spatial analysis of almond leaf scorch di- Sensitivity of yield determinants to potassium tos, A. and Vieira-Da-Silva, J., 2004. CO2 sease in the San Joaquin Valley of Califor- deficiency in ‘Nonpareil’ almond (Prunus dul- assimilation and water relations of almond nia: Factors affecting pathogen distribution cis (Mill.) DA Webb). Journal of Horticultural tree (Prunus amygdalus Batsch) cultivars and spread. Plant Disease 89(6): 581-589. Science & Biotechnology 79(6): 906-910. grown under field conditions. Photosynthe- tica 42(3): 473-476. Guldur, M. E., Caglar, B. K., Castellano, M. Ribo, J. M., Crusats, J., El-Hachemi, Z., Fe- A., Unlu, L., Guran, S., Yilmaz, M. A. and liz, M., Sanchez-Bel, P. and Romojaro, F., Martelli, G. P., 2005. First report of almond Miranda, C., Santesteban, L. G. and Royo, 2004. High-resolution NMR of irradiated al- leaf scorch in Turkey. Journal of Plant Pa- J. B., 2005. Variability in the relationship monds. Journal of the American Oil Che- thology 87(3): 246-246. between frost temperature and injury level mists Society 81(11): 1029-1033.

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FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 65 nut blight on a national scale. Adv. Hort. se in the chestnut weevil Curculio sikkimen- of shake in sweet chestnut (Castanea sati- Sci., 20 (1): 50-54. sis (Coleoptera: Curculionidae). Journal of va Mill.). Forestry 78(2): 175-186. Insect Physiology 51(12): 1352-1358. Eriksson, G., 2006. Forest tree gene con- Myers, B. R., Walck, J. L. and Blum, K. E., servation. Theory and application for Cas- Hillman, B. I. and Suzuki, N., 2004. Viruses 2004. Vegetation change in a former chest- tanea sativa. Adv. Hort. Sci., 20 (1): 101- of the chestnut blight fungus, Cryphonectria nut stand on the Cumberland Plateau of 106. parasitica. Advances in Virus Research, Tennessee during an 80-year period (1921- Vol. 63. San Diego, Elsevier Academic 2000). Castanea 69(2): 81-91. Fernandez, J. L., Ballester, A. and Riguei- Press Inc. 63: 423-472. ro, A., 2005. Phenolic content of microcut- Ovesna, J., Kucera, L., Jiang, L. J. and tings of adult chestnut along rooting induc- Igual, J.M.; Peix, A.; Valverde, A.; Salazar, Vagnerova, D., 2005. Characterisation of tion. Plant Cell Tissue and Organ Culture S.; Alvarez, J.; Rodríguez, C.; Santa, I., Chinese elite cultivars and genetic resour- 83(2): 153-159. 2006. Soil biochemical properties and di- ces of chestnut by AFLP. Biologia Planta- versity of culturable phyllosphere bacteria rum 49(1): 125-127. Fernando, D., Richards, J. and Kikkert, J., in chestnut and oak forests: three study ca- 2005. ‘In vitro’ germination and transient ses at the Sierra de Francia (Salamanca, Panagou, E.Z.; Vekiari, S.A.; Mallidis, C., GFP expression of American chestnut Spain). Adv. Hort. Sci., 20 (1): 21-27. 2006. The effect of modified atmosphere (Castanea dentata) pollen. Plant Cell Re- packaging of chestnuts in suppressing fun- ports. Juhasova, G., Adamcikova, K. and Robin, gal growth and related physiochemical C., 2005. Results of biological control of changes during storage in retail packages Gavaland, A.; Pelletier, S., 2006. Multi-pur- chestnut blight in Slovakia. Phytoprotection at 0 and 8¼C. Adv. Hort. Sci., 20 (1): 82-89. pose management of chestnut: the French 86(1): 19-23. situation. Adv. Hort. Sci., 20 (1): 70-81. Pannatier, E. G., Luster, J., Zimmermann, Kazmierczak, P., Kim, D. H., Turina, M. and S. and Blaser, P., 2005. Acidification of soil Giudici, F. and Zingg, A., 2005. Sprouting Van Alfen, N. K., 2005. A hydrophobin of solution in a chestnut forest stand in ability and mortality of chestnut (Castanea the chestnut blight fungus, Cryphonectria southern Switzerland: Are there signs of re- sativa Mill.) after coppicing. A case study. parasitica, is required for stromal pustule covery? Environmental Science & Techno- Annals of Forest Science 62(6): 513-523. eruption. Eukaryotic Cell 4(5): 931-936. logy 39(20): 7761-7767.

Gold, M., Cernusca, M. M. and Godsey, L., Kehrli, P. and Bacher, S., 2004. How to Pereira, S., Ramos, A. M., Diaz, M. B., 2004. Consumer preferences for chestnuts, safely compost Cameraria ohridella-infes- Ciordia, M. and Rios, D., 2006. Chemical eastern black walnuts, and pecans. Hortte- ted horse chestnut leaf litter on private composition of chestnut cultivars from chnology 14(4): 583-589. compost heaps. Journal of Applied Ento- Spain. Scientia Horticulturae 107(3): 306-314. mology 128(9-10): 707-709. Gold, M., Cernusca, M. M. and Godsey, L., Privitera, M.; Puglisi, M.; Cenci, R.M.; Beo- 2005. Update on Consumer’s Preferences Kwon, J. H., 2005. Effects of gamma irra- ne, G.M., 2006. Bryophyte biodiversity for for Chestnuts. Horttechnology, 15 (4). 904- diation and methyl bromide fumigation on monitoring chestnut ecosystems on Mount 906. the qualities of fresh chestnuts during sto- Etna (Eastern Sicily). Adv. Hort. Sci., 20 rage. Food Science and Biotechnology (1): 28-32. 14(2): 181-184. Gomes, J., Salgado, P., Sang, H., Kraayen- hof, R. and Torres, J., 2005. Isolation of Queijeiro, J. M., De la Montana, J. and Mi- chestnut chloroplasts: Membrane potentials Lopez, C., Torrado, A., Guerra, N. P. and guez, M., 2006. Identification and morpho- of chestnut and spinach thylakoids. Pho- Pastrana, L., 2005. Optimization of solid- logical description of cultivars of chestnut tosynthetica 43(2): 237-246. state enzymatic hydrolysis of chestnut (Castanea sativa Mill) of the region of Verin- using mixtures of alpha-amylase and glu- Monterrei (Ourense, Spain). Journal of the coamylase. Journal of Agricultural and Gomez, J., Pazos, M., Couto, S. R. and American Pomological Society 60(1): 37-45. Food Chemistry 53(4): 989-995. Sanroman, M. A., 2005. Chestnut shell and barley bran as potential substrates for lac- Romane, F., Aronson, J., Gondard, H., case production by Coriolopsis rigida under Lozovitskii, P. S., 2005. Changes in the Grandjanny, M., Grossmann, A., Le Floc’h, solid-state conditions. Journal of Food En- properties of dark chestnut soils under the E., Renaux, A. and Shater, Z., 2005. Dyna- gineering 68(3): 315-319. impact of long-term irrigation (the Kakhovka mics of chesnut plantations in Cevennes irrigation system). Eurasian Soil Science (France) and consequences for plant diver- 38(5): 551-562. Gondard, H. and Romane, F., 2005. Long- sity. Acta Botanica Gallica 152(4): 431-442. term evolution of understorey plant species composition after logging in chestnut coppi- Lugli, S., 2003. Castanicoltura, non solo Ramos, A. M. and Pereira, S., 2005. Gene- ce stands - (Cevennes Mountains, southern per terreni declivi e marginali. Frutticoltura, tic relationship between Castanea sativa France). Annals of Forest Science 62(4): LXV (12): 81-83. Mill. trees from north-western to south 333-342. Spain based on morphological traits and Manetti, M.C.; Amorini, E.; Becagli, C., isoenzymes. Genetic Resources and Crop Haltofová, P., 2006. Vegetative compatibili- 2006. New silvicultural models to improve Evolution 52(7): 879-890. ty groups of Cryphonectria parasitica (Mu- functionality of chestnut stands. Adv. Hort. rrill) M.E. Barr in the Czech Republic. Adv. Sci., 20 (1): 65-69. Ramos, A.M.; Caruncho, L; Díaz, M.B.; Hort. Sci., 20 (1): 55-58. Ciordia, M.; Ríos, D.; González, J.; Pereira, Mutabaruka, C., Woodgate, G. R. and Buc- S., 2006. Study of Spanish chestnut culti- Higaki, M., 2005. Effect of temperature on, kley, G. P., 2005. External and internal vars using SSR markers. Adv. Hort. Sci., 20 the termination of prolonged larval diapau- growth parameters as potential indicators (1): 113-116.

66 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Sauer, U. and Wilhelm, E., 2005. Somatic Vettraino, A. M., Morel, O., Perlerou, C., Amaral, J. S., Casal, S., Citova, I., Santos, embryogenesis from ovaries, developing Robin, C., Diamandis, S. and Vannini, A., A., Seabra, R. M. and Oliveira, B. P. P., ovules and immature zygotic embryos, and 2005. Occurrence and distribution of Phyto- 2006. Characterization of several hazelnut improved embryo development of Casta- phthora species in European chestnut (Corylus avellana L.) cultivars based in nea sativa. Biologia Plantarum 49(1): 1-6. stands, and their association with Ink Di- chemical, fatty acid and sterol composition. sease and crown decline. European Jour- European Food Research and Technology Sizemskaya, M. L. and Bychkov, N. N., nal of Plant Pathology 111(2): 169-180. 222(3-4): 274-280. 2005. The salt status of meadow-chestnut soils in the northern caspian region upon a Vettraino, A. 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FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 67 Ozkal, S. G., Salgin, U. and Yener, M. E., pium hirsutum L.) alley cropping system in Smith, M. W., Cheary, B. S. and Carroll, B. 2005. Supercritical carbon dioxide extrac- the southern United States. Plant and Soil L., 2005. Temporal weed interference with tion of hazelnut oil. Journal of Food Engi- 263(1-2): 151-164. young pecan trees. Hortscience 40(6): neering 69(2): 217-223. 1723-1725. Allen, S. C., Jose, S. B., Nair, P. K. R., Ozkal, S. G., Yener, M. E., Salgin, U. and Brecke, B. J., Nair, V. D., Graetz, D. A. and Smith, M.W.; Wood, B.W., 2006. Pecan Mehmetoglu, U., 2005. Response surfaces Ramsey, C. L., 2005. Nitrogen mineraliza- tree biomass estimates. HortScience, 41 of hazelnut oil yield in supercritical carbon tion in a pecan (Carya illinoensis K. Koch)- (5): 1286-1291. dioxide. 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68 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Guerrero, J., Moriana, A., Perez-Lopez, D., applied potassium. Journal of Plant Nutri- Cortizo, M., Alonso, P., Fernandez, B., Ro- Couceiro, J. E., Olmedilla, N. and Gijon, M. tion 28(8): 1413-1425. driguez, A., Centeno, M. L. and Ordas, R. C., 2006. Regulated deficit irrigation and J., 2004. Micrografting of mature stone pine the recovery of water relations in pistachio Sahin, O., Demirel, S. and Dilekoglu, M. F., (Pinus pinea L.) trees. Annals of Forest trees. Tree Physiology 26(1): 87-92. 2005. Removal of Pb(II) from aqueous so- Science 61(8): 843-845. lution by antep pistachio shells. Fresenius Hadavi, E., 2005. Several physical proper- Environmental Bulletin 14(11): 986-992. Martinez, F. and Montero, G., 2004. The Pi- ties of aflatoxin-contaminated pistachio nus pinea L. woodlands along the coast of nuts: Application of BGY fluorescence for Sepaskhah, A. R. and Karimi-Goghari, S., South-western Spain: data for a new geo- separation of aflatoxin-contaminated nuts. 2005. Shallow groundwater contribution to botanical interpretation. Plant Ecology Food Additives and Contaminants 22(11): pistachio water use. Agricultural Water Ma- 175(1): 1-18. 1144-1153. nagement 72(1): 69-80. Motta, R. and Lingua, E., 2005. Human im- Hokmabadi, H., Arzani, K. and Grierson, P. Tilkat, E., Isikalan, C. and Onay, A., 2005. pact on size, age, and spatial structure in a F., 2005. Growth, chemical composition, In vitro propagation of khinjuk pistachio mixed European larch and Swiss stone and carbon isotope discrimination of pista- (Pistacia khinjuk stocks) through seedling pine forest in the Western Italian Alps. Ca- chio (Pistacia vera L.) rootstock seedlings apical shoot tip culture. Propagation of Or- nadian Journal of Forest Research-Revue in response to salinity. Australian Journal of namental Plants 5(3): 124-128. 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Effects of shoot gir- riability of Mediterranean Stone pine cone real-time PCR assay for the detection of dling on bud abscission, carbohydrate and production: Yield loss as response to clima- azoxystrobin-resistant Alternaria populatio- nutrient concentrations in pistachio (Pista- te change. Agricultural and Forest Meteoro- ns from pistachio orchards in California. cia vera L.). Journal of Horticultural Science logy 132(3-4): 263-272. Crop Protection 23(12): 1259-1263. & Biotechnology 80(5): 529-536. Mutke, S., Sievanen, R., Nikinmaa, E., Per- Mehrnejad, M. R. and Copland, M. J. W., Yakubov, B., Barazani, O., Shachack, A., ttunen, J. and Gil, L., 2005. Crown architec- 2006. Host-stage selection and oviposition Rowland, L., Shoseyov, O. and Golan-Gol- ture of grafted Stone pine (Pinus pinea L.): behaviour of Psyllaephagus pistaciae, pa- dhirsh, A., 2005. Cloning and expression of shoot growth and bud differentiation. Trees- rasitoid of the common pistachio psylla a dehydrin-like protein from Pistacia vera L. Structure and Function 19(1): 15-25. Agonoscena pistaciae. Biological Control Trees-Structure and Function 19(2): 224- 36(2): 139-146. 230. Ozguven, F. and Vursavus, K., 2005. Some physical, mechanical and aerodynamic pro- Mehrnejad, M. R. and Emami, S. Y., 2005. Yakubov, B., Barazani, O. and Golan-Gold- perties of pine (Pinus pinea) nuts. Journal Parasitoids associated with the common hirsh, A. 2005. Combination of SCAR pri- of Food Engineering 68(2): 191-196. pistachio psylla, Agonoscena pistaciae, in mers and Touchdown-PCR for sex identifi- Iran. Biological Control 32(3): 385-390. cation in Pistacia vera L. Scientia Hort. 103, Rettori, A., Paoletti, E., Nicolotti, G. and 473-478. Gullino, M. L., 2005. Ecophysiological Mila, A. L., Driever, G. F., Morgan, D. P. responses of Mediterranean pines to si- and Michailides, T. J., 2005. Effects of la- Yazdanpanah, H., Mohammadi, T., Abou- mulated sea aerosol polluted with an tent infection, temperature, precipitation, hossain, G. and Cheraghali, A. M., 2005. anionic surfactant: prospects for biomoni- and irrigation on panicle and shoot blight of Effect of roasting on degradation of Afla- toring. Annals of Forest Science 62(4): pistachio in California. Phytopathology toxins in contaminated pistachio nuts. Food 351-360. 95(8): 926-932. and Chemical Toxicology 43(7): 1135- 1139. Onay, A., Pirinc, V., Tilkat, E., Akturk, Z. WALNUT and Yildirim, H., 2004. Somatic embryogenesis of pistachio from female flowers. STONE PINE Akca, Y. and Ozongun, S., 2004. Selection Journal of Horticultural Science & Biotech- of late leafing, late flowering, laterally fruit- nology 79(6): 960-964. Bravo-Oviedo, A. and Montero, G., 2005. ful walnut (Juglans regia) types in Turkey. Site index in relation to edaphic variables in New Zealand Journal of Crop and Horticul- Ozden-Tokatli, Y., Ozudogru, E. A. and Ak- stone pine (Pinus pinea L.) stands in south tural Science 32(4): 337-342. cin, A., 2005. ‘In vitro’ response of pistachio west Spain. Annals of Forest Science nodal explants to silver nitrate. Scientia 62(1): 61-72. Amaral, J. S., Cunha, S. C., Alves, M. R., Horticulturae 106(3): 415-426. Pereira, J. A., Seabra, R. M. and Oliveira, Calama, R. and Montero, G., 2005. Multile- B. P. P., 2004. Triacylglycerol composition Pour, A. T., Sepaskhah, A. R. and Maftoun, vel linear mixed model for tree diameter in- of walnut (Juglans regia L.) cultivars: Cha- M., 2005. Plant water relations and se- crement in stone pine (Pinus pinea): a cali- racterization by HPLC-ELSD and chemo- edling growth of three pistachio cultivars as brating approach. Silva Fennica 39(1): 37- metrics. Journal of Agricultural and Food influenced by irrigation frequency and 54. 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FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 69 Ares, A. and Brauer, D., 2005. Growth and nut production of black walnut in relation to site, tree type and stand conditions in south-central United States. Agroforestry Systems 63(1): 83-90.

Benito, A., 2005. Nogal - Experimentación y cultivo en Navarra. Navarra Agraria, Sep- tiembre-Octubre: 1-8.

Bohanek, J. R. and Groninger, J. W., 2005. Productivity of European black alder (Alnus glutinosa) interplanted with black walnut (Juglans nigra) in illinois, USA. Agroforestry Systems 64(2): 99-106.

Brezna, B., Hudecova, L. and Kuchta, T., 2006. A novel real-time polymerase chain reaction (PCR) method for the detection of walnuts in food. European Food Research and Technology.

Carsten, L. D. and Papaj, D. R., 2005. Effects of reproductive state and host re- Delaire, M., Frak, E., Sigogne, M., Adam, typical distances based on discriminatory source experience on mating decisions in a B., Beaujard, F. and Le Roux, X., 2005. fruit traits in walnut (Juglans regia). Indian walnut fly. Behavioral Ecology 16(3): 528- Sudden increase in atmospheric CO2 con- Journal of Agricultural Sciences 75(4): 225- 533. centration reveals strong coupling between 227. shoot carbon uptake and root nutrient uptake in young walnut trees. Tree Physiology Koyuncu, M. A., Ekinci, K. and Gun, A., Chifflot, V., Bertoni, G., Cabanettes, A. and 25(2): 229-235. Gavaland, A., 2006. Beneficial effects of in- 2004. The effects of altitude on fruit quality tercropping on the growth and nitrogen sta- and compression load for cracking of wal- Diaz, R. and Fernandez-Lopez, J., 2005. tus of young wild cherry and hybrid walnut nuts (Juglans regia L.). Journal of Food Genetic variation at early ages for several trees. Agroforestry Systems 66(1): 13-21. Quality 27(6): 407-417. traits of interest for timber-production breeding of Juglans regia. Canadian Journal of Colaric, M., Veberic, R., Solar, A., Hudina, Martinez, M. L., Torres, M. M., Guzman, C. Forest Research-Revue Canadienne De A. and Maestri, D. M., 2006. Preparation M. and Stampar, F., 2005. Phenolic acids, Recherche Forestiere 35(2): 235-243. syringaldehyde, and juglone in fruits of dif- and characteristics of activated carbon ferent cultivars of Juglans regia L. Journal from olive stones and walnut shells. Indus- Elmore, J. S., Nisyrios, I. and Mottram, D. of Agricultural and Food Chemistry 53(16): trial Crops and Products 23(1): 23-28. S., 2005. Analysis of the headspace aroma 6390-6396. Juglans regia L.). compounds of walnuts ( McGranahan, G. and Leslie, C., 2004. ‘Ro- Flavour and Fragrance Journal 20(5): 501- bert Livermore’, a Persian walnut cultivar Crews, C., Hough, P., Godward, J., Brere- 506. with a red seedcoat. Hortscience 39(7): ton, P., Lees, M., Guiet, S. and Winkel- 1772-1772. mann, W., 2005. Study of the main consti- Foroni, I., Rao, R., Woeste, K. and Gallitelli, tuents of some authentic walnut oils. Jour- M., 2005. Characterisation of Juglans regia Ozkan, G. and Koyuncu, M. A., 2005. Phy- nal of Agricultural and Food Chemistry L. with SSR markers and evaluation of ge- sical and chemical composition of some 53(12): 4853-4860. netic relationships among cultivars and the walnut (Juglans regia L) genotypes-grown ‘Sorrento’ landrace. Journal of Horticultural in Turkey. Grasas Y Aceites 56(2): 141- Dangl, G. S., Woeste, K., Aradhya, M. K., Science & Biotechnology 80(1): 49-53. 146. Koehmstedt, A., Simon, C., Potter, D., Les- lie, C. A. and McGranahan, G., 2005. Cha- Frak, E., Le Roux, X., Millard, P., Guillau- Paris, P., Pisanelli, A., Todaro, L., Olimpie- racterization of 14 microsatellite markers me, S. and Wendler, R., 2006. Nitrogen ri, G. and Cannata, F., 2005. Growth and for genetic analysis and cultivar identifica- availability, local light regime and leaf rank water relations of walnut trees (Juglans re- tion of walnut. Journal of the American So- effects on the amount and sources of N gia L.) on a mesic site in central Italy: ciety for Horticultural Science 130(3): 348- allocated within the foliage of young walnut effects of understorey herbs and polye- 354. (Juglans nigra x regia) trees. Tree Physio- thylene mulching. Agroforestry Systems logy 26(1): 43-49. 65(2): 113-121. Decourteix, M., Alves, G., Brunel, N., Ame- glio, T., Guilliot, A., Lemoine, R., Petel, G. Hemery, G. E., Savill, P. S. and Thakur, A., Ponder, F., Jones, J. E. and Mueller, R., and Sakr, S., 2006. JrSUT1, a putative 2005. Height growth and flushing in com- 2005. Using poultry litter in black walnut xylem sucrose transporter, could mediate mon walnut (Juglans regia L.): 5-year resul- nutrient management. Journal of Plant Nu- sucrose influx into xylem parenchyma cells ts from provenance trials in Great Britain. trition 28(8): 1355-1364. and be up-regulated by freeze-thaw cycles Forestry 78(2): 121-133. over the autumn-winter period in walnut tree (Juglans regia L.). Plant Cell and Envi- Kara, K., Akca, Y., Balta, M. F., Yarilgac, T. Sabate, J., Cordero-MacIntyre, Z., Siapco, ronment 29(1): 36-47. and Balta, F., 2005. An estimation of geno- G., Torabian, S. and Haddad, E., 2005.

70 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 Does regular walnut consumption lead to Woeste, K., 2004. An on-line database of weight gain? British Journal of Nutrition Juglans cultivar names and origins. Horts- Gregory, D. 2004. The development of a 94(5): 859-864. cience 39(7): 1771-1771. genetic linkage map for almond based on molecular and agronomic markers (in Spa- Schilling, G., 2005. Have we cracked Saturn’s Wu, G. L., Zhang, X. Y., Zhang, L. Y., Pan, nish). PhD thesis. University of Adelaide. walnut? New Scientist 186(2499): 16-17. Q. H., Shen, Y. Y. and Zhang, D. P., 2004. Australia. 263 pages. Phloem unloading in developing walnut fruit Stampar, F., Solar, A., Hudina, M., Veberic, is symplasmic in the seed pericarp, and Kodad, O. 2006. Criterios de selección y de R. and Colaric, M., 2006. Traditional walnut apoplasmic in the fleshy pericarp. Plant and evaluación de nuevas obtenciones autocom- liqueur - cocktail of phenolics. Food Che- Cell Physiology 45(10): 1461-1470. patibles en un programa de mejora genéti- mistry 95(4): 627-631. ca del almendro (Prunus amygdalus Batsch) (in Spanish). PhD Thesis. Lleida University. Zeneli, G., Kola, H. and Dida, M., 2005. Torres, M. M., Martinez, M. L. and Maestri, Phenotypic variation in native walnut popu- D. M., 2005. A multivariate study of the relations of Northern Albania. Scientia Horti- Sánchez, R., 2005. Desarrollo y aplicación lationship between fatty acids and volatile culturae 105(1): 91-100. de marcadores moleculares a la mejora ge- flavor components in olive and walnut oils. nética del almendro Prunus dulcis (Miller) Journal of the American Oil Chemists So- D.A. Webb (in Spanish). Universidad de ciety 82(2): 105-110. Murcia, 222 pages. THESES

Vahdati, K; Leslie, C; Zamani, Z; et al., 2004. Rooting and acclimatization of ‘in vi- Afif, M., 2006. Diversité génétique chez les MASTER THESIS tro’ grown shoots from mature trees of populations nuturelles du caroubier (Cera- tonia siliqua L.) en Tunisie: polymorphisme three Persian walnut cultivars. Hortscience, Catalán, U., 2005. Evaluación de indicado- isoenzymatique et moleculaire (in French). 39 (2): 324-327. res de estrés hídrico en almendro (Prunas University of Tunis, El Manar: 136 pages. amygdalus Batsch) y olivo (Olea europaea Victory, E. R., Glaubitz, J. C., Rhodes, O. L.) (in Spanish). Universitat de Lleida, Es- E. and Woeste, K. E., 2006. Genetic homo- Arrázola, G., 2002. Análisis de glucósidos cola Tècnica Superior d’Enginyeria Agrària: geneity in Juglans nigra (Juglandaceae) at cianogénicos en variedades de almendro: 180 p. nuclear microsatellites. American Journal Implicaciones en la mejora genética. Uni- of Botany 93(1): 118-126. versidad de Alicante: 194 pp. Vargas, I., 2005. Análisis de descenden- cias de cruzamientos controlados en pista- Wang, S., Yue, J., Tang, J. and Chen, B., Buck, E., 2006. Genetic Variation of Casta- chero (Pistacia vera L.) (in Spanish). Uni- 2005. Mathematical modelling of heating nea sativa Mill. University of Wales, Ban- versitat de Lleida, Escola Tècnica Superior uniformity for in-shell walnuts subjected to gor, UK: 195 pages. d’Enginyeria Agrària: 144 p. radio frequency treatments with intermittent stirrings. Postharvest Biology and Techno- Fidan, M. 2005. Determination of Nutrient Zribi, F., 2004. Identification et description logy 35(1): 97-107. Elements Contents For Different Parts of pomologique et phénologique des variétés Some Pistachio Cultivars. (In Turkish with mâles et femelles de pistachier dans deux Weckerle, C., Huber, F. K., Yang, Y. P. and summary in English) Master of Science. sites différents du sud (in French). Mémoire Sun, W. B., 2005. Walnuts among the Shu- University of Harran, Graduate School of de Master. Institut National Agronomique hi in Shuiluo, eastern Himalayas. Walnut Natural and Applied Sciences. Department de Tunisie, L’Institut de l’Olivier and l’Ins- (Juglans regia L.). Economic Botany 59(3): of Horticulture, 68 p. (Supervisor: Prof. Dr. titut des Régions Arides. 287-290. Bekir Erol AK)

FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006 71 THE FAO-CIHEAM INTER-REGIONAL COOPERATIVE RESEARCH NETWORK ON NUTS

Network Coordination Centre Network Coordinator

Nut tree crops IRTA - Mas de Bover. Mediterrean Fruit Trees M. Rovira Stone Pine Crta. Reus - El Morell, km 3.8, E-43120 Constantí (Spain) Tel: 34- 977 328424 Fax: 34- 977 344055 E-mail: [email protected]

Subnetworks Liaison Centre Liaison Officer

Almond IRTA - Mas de Bover. Mediterrean Fruit Trees F.J. Vargas Crta. Reus - El Morell, km 3.8, E-43120 Constantí (Spain) Tel: 34- 977 328424 Fax: 34- 977 344055 E-mail: [email protected]

Haselnut Ankara University. Faculty of Agriculture. A.I.Köksal Department of Horticulture. 06110 - Ankara (Turkey). Tel: 90 - 312 3170550. Fax: 90 - 312 3179119 E-mail: [email protected]

Walnut and Pecan Institut National de la Recherche Agronomique INRA. M. Lafargue Unité de Recherches sur les Espèces Fruitières et la Vigne. B.P. 81 - 33883 - Villenave d’Ornon (France) Tel: 33 - 556 843277. Fax: 33 - 556 843274 E-mail: [email protected]

Pistachio University of Harran, Faculty of Agriculture B. E. Ak Departement of Horticulture 63200 - Sanliurfa (Turkey) Tel: 90 - 414 2472697. Fax: 90 - 414 2474480 E-mail: [email protected]

Chestnut Università degli Studi di Torino. G. Bounous Dipartamento di Colture Arboree. Cattedra di Arboricultura Via Leonardo Da Vinci, 44. 10095 - Grugliasco (TO) (Italy) Tel. 39 - 011 6708653. Fax: 39 - 011 6708658. E-mail: [email protected]

Genetic Resources IRTA - Mas de Bover. Mediterrean Fruit Trees I. Batlle Crta. Reus - El Morell, km 3.8, E-43120 Constantí (Spain) Tel: 34- 977 328424 Fax: 34- 977 344055 E-mail: [email protected]

Economics Servicio de Investigación Agraria. L.M. Albisu Diputación General de Aragón. Apartado 727 50080 - Zaragoza (Spain) Tel. 34 - 976 576361. Fax: 34- 976 575501 E-mail: [email protected]

Regional Office for Europe REU: Ms. Jutta Krause. Viale delle Terme di Caracalla. FAO 00100 Roma (Italy). Jutta Krause Tel: 39 - 06 57054405 Fax: 39 - 06 57055634. E-mail: [email protected]

Instituto Agronómico Mediterráneo de Zaragoza IAMZ. Apartado 202, 50080 Zaragoza. CIHEAM Tel: 34 - 976 71 60 00 Fax: 34 - 976 71 60 01 Dunixi Gabiña E-mail: [email protected]

IRTA Mas de Bover Network Coordinator: M. Rovira Mediterranean Fruit Trees Editor: I. Batlle Ctra. Reus-El Morell, km 3,8 Editorial staff: M. Lannoye E- 43120 Constantí, Tarragona, Spain Tel.: +34-977 32 84 24 Typeset by: Carácter Gráfico, S.L. Fax: +34-977 34 40 55 E-mail: [email protected] E-mail: [email protected] ISSN 1020-0797

72 FAO-CIHEAM - Nucis-Newsletter, Number 13 December 2006