Wheat Information Service Electronic Newsletter for Wheat Researchers

Wheat Inf. Serv. 108, 2009. www.shigen.nig.ac.jp/ewis

No. 108

CONTENTS

Research Information

Elena K. Khlestkina, Marion S. Röder, Andreas Börner Identification of glume coloration genes in synthetic hexaploid and common wheats. p1-3.

Bikram K. Das, Suresh G. Bhagwat AP-PCR analysis of Indian wheat genotypes: genetic relationships and association analysis. p5-10.

Dharmendra Singh, S. K. Singh, K. N. Singh AMMI analysis for salt tolerance in bread wheat genotypes. p11-17.

Topics on Wheat Genetic Resources

Takashi R. Endo National BioResource Project Wheat: Overview. p19.

Taihachi Kawahara, Takashi R. Endo, Tomohiro Ban, Masahiro Kishii, Tsuneo Sasanuma The report of National Bioresource Project-Wheat II. Seed resources, 2008. p20.

Miyuki Nitta, Shuhei Nasuda Annual report of the project “Polymorphism survey among hexaploid wheat and its relatives by DNA markers”. p21-22.

Yasunari Ogihara, Kanako Kawaura, Hisako Imamura Annual report of National Bioresource Project-Wheat II. DNA resources, 2008. p23.

Meeting Reports

Taihachi Kawahara The 6th International Triticeae Symposium (6th ITS). p25-66.

Nikolay P. Goncharov Wheat science at the Sixth International Triticeae Symposium. p67-70.

Others

Instructions to Authors. p71-72.

Wheat Inf. Serv. 108: 1-3, 2009. www.shigen.nig.ac.jp/ewis

Research Information

Identification of glume coloration genes in synthetic hexaploid and common wheats

Elena K. Khlestkina1*, Marion S. Röder2, Andreas Börner2 1Institute of Cytology and Genetics, Siberian Branch, Russian Academy of Sciences, Lavrentjeva Ave. 10, Novosibirsk, 630090, Russia 2Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany *Corresponding author: Elena K. Khlestkina (E-mail: [email protected])

Abstract Crosses of hexaploid wheats having phenotype for glume color distinct from that reported in previous studies, were preformed and used to clarify whether these phenotypes are determined by new or earlier identified Rg alleles. It was found that although a common wheat accession ‘TRI 14341’ has not typical black glabrous glume, its glume color is controlled by the known allele Rg-A1c (chromosome 1AS). Dark brown-black glume color of the synthetic wheat line analysed in the current study was controlled by the two alleles Rg-A1c (1AS) and Rg-D1b (1DS). Rg-D1b was mapped 3.9 cM proximal to the microsatellite marker Xgwm1223, which is more precise mapping result than that obtained in the previous studies.

Introduction phenotypes are determined by new or by earlier Three homoeologous loci controlling glume coloration identified Rg alleles. in hexaploid common wheat (Triticum aestivum L.) or synthetic hexaploid wheat were mapped to the short Materials and methods arms of chromosomes 1AS (Rg-A1), 1BS (Rg-B1) and Two crosses between hexaploid wheat lines and 1DS (Rg-D1) (Khlestkina et al. 2006, 2009). Multiple accessions were used to create F2 populations: (1) allelism was shown for Rg-A1 and Rg-D1, since synthetic hexaploid line (McFadden and Sears 1947) besides recessive colorless (wildtype) allele two having dark brown-black glume was crossed with different dominant alleles were found at each of these white-glumed Russian cultivar ‘Ulyanovka’. 146 F2 loci. Alleles Rg-A1b and Rg-A1c controlling red and plants were obtained, genotyped and scored for glume black glume coloration, respectively, whereas Rg-D1b coloration, and (2) accession ‘TRI 14341’ and Rg-D1c control red and smokey-grey color. (IPK-Genebank collection) having black glabrous Rg-A1b was described only in several Russian T. glume was crossed with white-glumed ‘TRI 542’, aestivum cultivars (Efremova et al. 1998). Rg-A1c was which is known to carry dominant Hg allele for hairy described in T. aestivum near-isogenic line glume (Khlestkina et al. 2006). Fifty-one F2 plants ‘i:S29BgHg’, in which black glume color is known to were obtained and scored for glume coloration and be closely linked to glume hairiness (Arbuzova et al. hairiness. 1998). Rg-D1b inherited from the D-genome DNA from frozen leaves of progenitor of hexaploid wheat (Aegilops tauschii) was ‘Synthetic’/‘Ulyanovka’ F2 progeny was extracted described in synthetic hexaploid wheats only (Kerber according to Plaschke et al. (1995). Six chromosome and Dyck 1969, Jones et al. 1990, Börner et al. 2002). 1A and nine chromosome 1D GWM (Gatersleben Rg-D1c was described in single T. aestivum Russian Wheat Microsatellites) markers were found to be cultivar ‘Golubka’ (Pshenichnikova et al. 2005). polymorphic within the ‘Synthetic’/‘Ulyanovka’ In the current study, crosses of hexaploid wheats population and used for mapping. Experimental having phenotypes distinct from that of the previously details regarding microsatellite assays are described in studied accessions (namely, synthetic wheat having Röder et al. 1998. Linkage maps were constructed dark brown-black glume and T. aestivum accession with MAPMAKER 2.0 (Lander et al. 1987). having rare black glabrous glume phenotype) were Centimorgan distances were calculated applying the preformed and used to clarify whether these Kosambi map-unit function (Kosambi 1944).

Results and discussion Segregation ratio for glume coloration in F2 population ‘Synthetic’/‘Ulyanovka’ was 134 colored: 12 non-colored plants, corresponding to digenic 15:1 (χ2=0.966, P>0.30). Therefore, we suggested that the synthetic wheat may carry black glume color allele Rg-A1c inherited from the tetraploid emmer wheat and red glume color allele Rg-D1b inherited from Ae. tauschii. Chromosome 1A and 1D polymorphic microsatellites (6 and 9 markers, respectively) were used for genotyping of the F2 lines. Linkage maps obtained are presented at Fig. 1. Dobrovolskaya et al. (2006) have mapped one of the two complementary Pp (purple pericarp) genes in population with digenic segregation, using microsatellite genotyping data for selection of genotypes recessive for the other gene. Similarly, suggesting that the synthetic parent of the ‘Synthetic’ / ‘Ulyanovka’ F2 population may carry allele Rg-A1c which was previously mapped 0.6 cM proximal to Xgwm1223 on chromosome 1AS, we selected genotypes having ‘Ulyanovka’ microsatellite alleles at the locus Xgwm1223 and also at the locus Xgwm1104 flanking suggested Rg-A1c location site from the other side. Therefore 27 F2 genotypes were selected, from which 22 had colored and 5 non-colored glumes, corresponding to 3:1 (χ2=0.606, Fig. 1. Microsatellite mapping of the genes P>0.30). This genotyping data were used for mapping controlling glume coloration in the hexaploid glume color gene on chromosome 1D. The gene was synthetic wheat analyzed in the current study. mapped 3.9 cM proximal from 1DS locus Xgwm1223 Genetic distances are given in centimorgans (cM). (Fig. 1). Earlier allele Rg-D1b originating from synthetic ‘W7984’ was mapped 11.6 cM proximal to the microsatellite locus Xgwm1223 (Khlestkina et al. to be usually closely linked and often occur in 2009) using ITMI mapping population, whereas T. tetraploid wheat, whereas in common wheat most of aestivum ‘Golubka’ allele Rg-D1c, controlling the accessions and cultivars have white or red glumes, smokey-grey glume color, was mapped 1.5 cM and the former may sometimes have hairy glumes proximal to Xgwm1223 (Khlestkina et al. 2006). We (Philipchenko 1935, Zeven 1983). Therefore, it was suggest the gene mapped in the current study on interesting to identify which gene determine glume chromosome 1DS is Rg-D1b (Fig. 1), however using color in T. aestivum accession ‘TRI 14341’ having ‘Synthetic’ / ‘Ulyanovka’ cross it was mapped more black glabrous glume. We crossed it with precisely than in ITMI mapping population. After white-glumed accession ‘TRI 542’ known to carry establishment of Rg-D1b precise map position, we dominant allele of the gene Hg on 1AS. There were no used the flanking markers Xgwm1223 and Xgwm0106 plants with white glabrous glumes within F2 progeny genotyping data to select F2 plants carrying of this cross, suggesting black glume gene of ‘TRI ‘Ulyanovka’ alleles, in order to map the synthetic 14341’ to be closely linked with recessive allele of glume color gene on chromosome 1A. Thus, 19 locus Hg. Therefore, we concluded that allele genotypes were selected, from which 13 had colored determining black glume color in ‘TRI 14341’ is and 6 non-colored glumes, corresponding to 3:1 identical to Rg-A1c of ‘i:S29BgHg’. (χ2=0.439, P>0.50). This genotyping data were used Thus, it was found that glume color in T. aestivum for mapping glume color gene on chromosome 1A. accession ‘TRI 14341’ having black glabrous glume is The gene was mapped 0.7 cM proximal from 1AS controlled by allele Rg-A1c (chromosome 1AS), locus Xgwm1223 (Fig. 1), in a position highly whereas dark brown-black glume color of the comparable with that determined for allele Rg-A1c synthetic wheat line analyzed in the current study is controlling black glume color in T. aestivum controlled by the two alleles Rg-A1c (1AS) and near-isogenic line ‘i:S29BgHg’ (0.6 cM proximal to Rg-D1b (1DS). Rg-D1b was mapped more precisely Xgwm1223; Khlestkina et al. 2006). In the (Fig. 1) than in the previous study (Khlestkina et al. near-isogenic line ‘i:S29BgHg’ Rg-A1c was also 2009). It also can be concluded that, using shown to be closely linked to dominant allele at the microsatellite markers, it is possible to map major locus Hg (hairy glume; Khlestkina et al. 2006). These genes even if digenic segregation in the mapping two traits (black glume and hairy glume) were known population is observed.

References association with specific microsatellite allele. Arbuzova VS, Maystrenko OI and Popova OM (1998) Cereal Res Com 37: 37-43. Development of near-isogenic lines of the common Kosambi DD (1944) The estimation of map distances wheat cultivar ‘Saratovskaya 29’. Cereal Res Com from recombination values. Ann Eugenet 12: 26: 39-46. 172-175. Dobrovolskaya OB, Arbuzova VS, Lohwasser U, Lander ES, Green P, Abrahamson J, Barlow A, Daly Röder MS and Börner A (2006) Microsatellite MJ, Lincoln SE and Newburg I (1987) mapping of complementary genes for purple grain MAPMAKER: an interactive computer package colour in bread wheat (Triticum aestivum L.). for constructing primary genetic linkage maps of Euphytica 150: 355-364. experimental and natural populations. Genomics 1: Efremova TT, Maystrenko OI, Arbuzova VS, and 174-181. Laikova LI (1998) Genetic analysis of glume McFadden ES and Sears ER (1947) The genome colour in common wheat cultivars from the former approach in radical wheat breeding. J Amer Soc USSR. Euphytica 102: 211-218. Agron 39: 1011-1026. Jones SS, Dvorak J and Qualset CO (1990) Linkage Philipchenko YA (1934) Genetics of soft wheats, relations of Gli-D1, Rg2, and Lr21 on the short Moscow-Leningrad, USSR, pp. 1-262. arm of chromosome 1D in wheat. Genome 33: Plaschke J, Ganal MW and Röder MS (1995) 937-940. Detection of genetic diversity in closely related Kerber ER and Dyck PL (1969) Inheritance in bread wheat using microsatellite markers. Theor hexaploid wheat of leaf rust resistance and other Appl Genet 91: 1001-1007. characters derived from Aegilops squarrosa. Can J Pshenichnikova TA, Bokarev IE and Shchukina LV Genet Cytol 11: 639-647. (2005) Hybrid and monosomic analyses of smoky Khlestkina EK, Pshenichnikova TA, Röder MS, coloration of the ear in common wheat. Rus J Arbuzova VS, Salina EA and Börner A (2006) Genet 41: 1147-1149. Comparative mapping of genes for glume Röder MS, Korzun V, Wendehake K, Plaschke J, colouration and pubescence in hexaploid wheat Tixier M-H, Leroy P and Ganal MW (1998) A (Triticum aestivum L.). Theor Appl Genet 113: microsatellite map of wheat. Genetics 149: 801-807. 2007-2023. Khlestkina EK, Salina EA, Pshenichnikova TA, Röder Zeven AC (1983) The character brown ear of bread MS and Börner A (2009) Glume coloration in wheat: a review. Euphytica 32: 299-310. wheat: allelism test, consensus mapping and its

4 Wheat Inf. Serv. 108: 5-10, 2009. www.shigen.nig.ac.jp/ewis

Research Information

AP-PCR analysis of Indian wheat genotypes: genetic relationships and association analysis

Bikram K. Das and Suresh G. Bhagwat* Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400085, India *Corresponding author: S. G. Bhagwat (E-mail: [email protected] or [email protected])

Abstract Genetic diversity among forty four Indian wheat genotypes and Chinese Spring was analyzed by arbitrary primed polymerase chain reaction (AP-PCR) using long primers. Using eight primers twenty five polymorphic bands were obtained and dendrogram was made. Only a few genotypes were not differentiated, however, others were clearly identified. Association analysis for rust resistance genes was done. It was found out that AP-PCR analysis could be used for detecting polymorphism in wheat. Two AP-PCR markers viz. SS30.2580(H) and SS26.11100 were identified to be associated with rust resistance gene Sr31 in coupling phase and repulsion phase respectively. The similarity matrix revealed that the genotypes had narrow genetic diversity value among the wheat genotypes used in this study.

Key Words: AP-PCR, Association analysis, Genetic diversity, Wheat

Introduction 1992). Usually arbitrary primers having 10 mer Wheat is the world’s leading cereal grain and in base length are used in RAPD (William et al.1990). India it is the most important food crop next to rice. However, investigations in which long primers A large number of accessions (varieties, genetic (>10 bases) have been used in RAPD (called as stocks and experimental lines) are available in AP-PCR, Welsh and Mc Clelland 1990) are national gene bank. The genetic diversity lacking. Saini et al. (2004) compared AP-PCR assessment of these accessions is necessary to help using long primers with RAPD and showed that the breeders in finding the genetic diversity, parent AP-PCR yields more polymorphism per primer selection, germplasm management and protection than RAPD in mung bean. The use of AP-PCR (Lee 1995). analysis for detecting polymorphism in bread Conventionally, genetic diversity is estimated wheat and study genetic diversity among Indian on the basis of morphological and phenotypic wheat varieties/accessions is not yet reported. characters. In recent years a variety of molecular The aims of this study were : (i) to find the markers have been used in plant genetic diversity utility of AP-PCR analysis using long primers for study. A major advantage of these markers is that detecting polymorphism in wheat, (ii) to study unlike the phenotypic markers these are not genetic diversity among Indian bread wheat affected by environmental changes. In crop plants, varieties; and to (iii) analyze association of a large number of molecular markers in addition to polymorphic band(s) with stem rust resistance analyzing genetic diversity are also used in tagging gene Sr31. genes and subsequently for use in marker assisted selection (Mohan et al.1997). Among the various Materials and methods techniques available, RAPD analysis is a simple, rapid, and effective method for detecting Plant materials polymorphism in wheat (Vierling and Nguyen The bread wheat (Triticum aestivum L.) varieties

5 Table 1. List of wheat genotypes used for AP-PCR analysis

Sr. Genotypes/ Year of Stem rust References No. Varieties release resistance genes present

1 WH-542 1994 Sr31 Sawhney (1998) 2 HUW-206 1985 Sr31 Sawhney (1998) 3 Kanchan 1994 Sr31 Sawhney (1998) 4 Unnath Kalyansona 1988 Sr24/Lr24 Kochumadhavan et al. (1988) 5 Vaishali 1993 Sr24/Lr24 Sawhney (1998) 6 Unnath Sonalika 1988 Sr24/Lr24 Kochumadhavan et al. (1988) 7 Vidisha 1996 Sr24/Lr24 Sawhney (1988) 8 HD-2285 1984 - - 9 HD-2385 1987 - - 10 PBW-373 1995 Sr31 Nayar (Personal Comm.) 11 PBW-343 1994 Sr31 Sawhney (1998) 12 PBW-138 1986 - - 13 PBW-154 1986 - - 14 Raj-3765 1994 - - 15 Sonalika 1967 - - 16 Kalyansona 1967 - - 17 Parbhani-51 1992 - - 18 PBW-299 1992 - - 19 PBW-175 1988 - - 20 HD-2745 - Sr31 Nayar (Personal Comm.) 21 HD-2735 - Sr31 Nayar (Personal Comm.) 22 C-306 1965 - - 23 GW-190 1994 Sr31 Sawhney (1998) 24 UP-2338 1995 Sr31 Sawhney (1998) 25 Chinese Spring - - - 26 Sehore - - - 27 MACS-2496 - Sr31 Sawhney (1998) 28 PBNS-3963 - - - 29 Ajantha 1981 - - 30 Lok-1 1982 - - 31 HD-2177 1978 - - 32 HD-2189 1979 - - 33 HD-1925 1974 - - 34 HD-2327 1985 - - 35 HD-2320 - - - 36 HD-1949 1974 - - 37 HD-1941 1970 - - 38 HD-2668 - - - 39 HD-2667 - Sr31 Nayar (Personal Comm.) 40 HD-2643 1995 Sr31 Nayar (Personal Comm.) 41 HD-2428 1988 - - 42 HDR-77 1990 - - 43 HD-2172 - - - 44 HD-2270 1987 - - 45 HD-2135 1975 - -

6 Table 2. The list and sequence of long primers used for AP-PCR analysis

Sr. Primer Sequence Length No. code (bases)

1 SS1.1 5’-CTC GTCTGA GATCGGAGGAG-3’ 20 2 SS1.2 5’-GGCTCCAAGCACCACTATAC-3’ 20 3 SS5.2 5’-TTCTCAGTTCAATGTTGTCC-3’ 20 4 SS13.1 5’-TTGTGACCTCCACCTACTAGCA-3’ 22 5 SS13.2 5’-GAGGATGAGAGTTGGTTGGTAG-3’ 22 6 SS18.1 5’-CACACACATACCATTCAGATAC-3’ 22 7 SS19.1 5’-GACATCTCTAGTGCACACAT-3’ 20 8 SS19.2 5’-TGAGACACAGACACAACTCT-3’ 20 9 SS26.1 5’-GAAGGGTAATTCAGAGCCA-3’ 19 10 SS26.2 5’-CAGGCATAGTGTCACTCTT-3’ 19 11 SS30.1 5’-CTTCTGCCTCCACCTAAACT-3’ 20 12 SS30.2 5’-TAGGTCCGACAATACGAACG-3’ 20 13 NJ-6 5’-TCTGGAGCCGTCCGTGTCCACGAGG-3’ 25 14 NJ-7 5’-GCCGCGTCCAGGGAAAATGTAGGCA-3’ 25 15 NJ-8 5’-AACTGGAAGAATTCGCGGCCGCAGGAA-3’ 27 16 NJ-9 5’-TTGAGGGATCCACACCACAA-3’ 20 17 ML-C1 5’-GCCTAGCAACCTTCACAATC-3’ 20 18 ML-G2 5’-GAAACCTGCTGCGGACAAG-3’ 19 19 MBC1 5’-GCCTAGCAACCTTCACAATC-3’ 20 20 MBG6 5’-GGCTAGCCGACAATGCGTCG-3’ 20

Table 3. Characteristics of the AP-PCR profiles used for genotyping of wheat varieties/accessions

Sr. Primer No. of No. of % No. code bands polymorphic polymorphism bands amplified

1 SS30.1 7 1 14.2 2 SS30.2 6 3 50.0 3 SS26.1 6 3 50.0 4 SS26.2 10 5 50.0 5 SS19.2 6 1 16.6 6 SS13.1 5 2 40.0 7 SS13.2 9 5 55.5 8 SS5.2 12 5 41.6

used in this study are listed in Table 1. AP-PCR analysis AP-PCR analysis was carried out using 20 long DNA isolation primers (Table 2) using the method of Saini et al. DNA was isolated from seeds according to (2004), with some modifications. PCR was Krishna and Jawali (1997) and DNA was further performed in 25 µl volume, containing 10 mM purified and concentration was estimated as Tris-HCl (pH 9.0), 2.0 mM MgCl2, 50 mM KCl, described in Prasad et. al. (1999). 200 µM of each dNTP, 25 pmoles of primer, 1.0 Unit of Taq DNA polymerase (Bangalore Genei

7 Pvt. India Ltd.) and 50 ng of template DNA. The values ranged from 0.13 to 1.0 and a dendrogram amplification reactions were carried out in Hybaid was constructed based on UPGMA analysis (Fig. PCR express gradient thermal cycler (Hybaid, UK) 2). There were three distinct clusters ‘A’, ‘B’, ‘C’. using the following thermal profile: 95 ºC for 5 The cluster ‘A’ consisted of all the genotypes min, 55 ºC for 5 min, 72 ºC for 5 min (one cycle); which were carriers of Sr31 or 1BL/1RS 95 ºC for 1 min, 55 ºC for 1 min, 72 ºC for 1 min translocation. Cluster ‘B’ consisted of Kalyansona, (42 cycles); final extension at 72 ºC for 10 min. Unnath Kalyansona, Sonalika and Unnath The amplified products were analyzed by Sonalika etc. Cluster ‘C’ consisted of mostly HD electrophoresis at 8-10 V/cm in 1X TBE buffer on varieties developed at IARI, New Delhi. 2.5% Agarose gel (Sigma-Aldrich Corporation, USA). Gels were stained with ethidium bromide. The gels were placed on UV transilluminator and the image was grabbed by gel documentation system from Syngene Corporation (U.K.) using Gene Snap software.

Scoring and data analysis AP-PCR bands were scored as present (1) or absent (0). Data analysis was performed using NTSYS-pc (Numerical taxonomy system, version 2.0, Rohlf 1990). Similarity matrices based on Jaccard’s coefficient were calculated and used to construct dendrogram by UPGMA (unweighed Fig. 1. AP-PCR profile of Indian wheat genotypes pair group method with arithmetic average). using the long primer SS13.2. Lane M: Phi Where mentioned association between band and X174/HaeIII digest DNA molecular standards. trait was done manually. Lanes marked 1 to 45 represents the wheat

genotypes as listed in Table 1 in the same

sequence. The arrows indicate some of the Results and discussion polymorphic bands detected. The PCR conditions were standardized at 55 ºC annealing temperature. Out of 20 primers, eight Using these primers most of the cultivars could primers (Table 3) were found to give reproducible be distinguished from each other because of their profile and only eight primers were considered for unique banding patterns; however few cultivars scoring polymorphic bands. The AP-PCR profile could not be distinguished. From this data of 45 wheat genotypes using primer SS13.2 are PBW-373 and PBW-343; GW-190 and shown in Fig.1. The reproducible bands were MACS-2496; Kalyansona and Unnath analyzed subsequently for genetic Kalyansona; C-306 and Sehore pairs could not be similarity/diversity analysis and association distinguished. The pedigrees of these were studies. observed. PBW-373 and PBW-343; GW-190 and AP-PCR markers using long primers were MACS-2496; C-306 and Sehore pairs were found to be suitable for detecting polymorphism derived from same parentage and their HMW and subsequent association studies. All the chosen glutenin sub-unit patterns (Das et al. 2001) and primers amplified fragments/ bands across the 45 other profiles matched exactly. Among Kalyansona wheat genotypes studied, with the number of and Unnath Kalyansona, Unnath Kalyansona is amplified fragments ranging from 5 to 12 (Table 3). near isogenic to Kalyansona with respect to Of the total 61 amplified bands, 25 were Sr24Lr24 (Kochumadhavan et al. 1988). polymorphic, with an average of 3.3 polymorphic Although the number of primers and bands per primer. The number of polymorphic polymorphic bands used in this study were limited, bands ranged from one (SS30.1 and SS19.2) to these were useful for determining the genetic five (SS26.1, SS13.2 and SS5.2) (Table 3). diversity among Indian wheat genotypes. This Percentage polymorphism ranged from 14.2% to showed the utility of AP-PCR markers in detecting 55.5%. From these eight primers, 25 polymorphic polymorphism in wheat. However use of more bands were obtained. Using these 25 bands, the number of primers can overcome the problem of genetic similarity was calculated by Jaccard’s not distinguishing three pairs of varieties similarity coefficient. The similarity coefficient originating from common pedigre

Fig. 2. Dendrogram of 45 wheat genotypes as revealed by AP-PCR markers.

Careful analysis of each primer profile revealed that an ~580 bp AP-PCR marker (SS30.2580(H)) obtained by primer SS30.2 was found to be linked in coupling phase where as another ~1100 bp AP-PCR marker (SS26.11100) obtained by primer SS26.1 was in repulsion phase. The marker SS30.2580(H) was present in all Sr31 carrier genotypes and absent in all other genotypes which were either susceptible or carry some other gene

(Fig. 3). Similarly marker SS26.11100 was absent in Fig. 4. AP-PCR profiles of wheat genotypes all Sr31 carriers and present in all non-carriers obtained using primer SS26.1. The names of the (Fig. 4). Upon linkage analysis in a segregating genotypes are shown on top of the lanes. ‘*’ population, both the markers were found to be indicates genotypes carrying Sr31 gene. Arrow tightly linked to Sr31 gene (Das et al. 2006). This indicates the position of 1100 bp band. Lane ‘M’ demonstrates that genetic similarity and careful indicates molecular size marker (PhiX174 digested association analysis can identify suitable markers. with Hae III).

The results of this study indicate that AP-PCR can be used to study the genetic relationship and diversity in bread wheat genotypes. Using AP-PCR markers the varieties can be distinguished from each other and use of more markers may help in making DNA finger printing of genotypes.

Fig. 3. AP-PCR profiles of wheat genotypes obtained using primer SS30.2. The names of the Acknowledgements genotypes are shown on top of the lanes. ‘*’ Authors are thankful to Dr. S.F. D’ Souza, Head, indicates genotypes carrying Sr31 gene. Arrow NABTD and Dr. N. Jawali and Dr. A. Saini of indicates the position of 580 bp band. Lane ‘M’ MBD for their encouragement and support; to indicates molecular size marker (PhiX 174 PAU, Ludhiana; IARI, New Delhi; DWR, Karnal digested with Hae III). and NBPGR, New Delhi for providing the seeds of wheat genotypes.

9 References Rohlf FJ (1990) NTSYS-pc Numerical Taxonomy Das BK, Bhagwat SG, Saini A and Jawali N and Multivariate Analysis System, Version (2001) Screening of Indian wheat cultivars for 2.20. Applied Biostatistics, New York. Glu-D1d allele (HMW- glutenin subunits Saini A, Reddy KS and Jawali N (2004) 5+10) by PCR. Ann Wheat News Letter 47: Evaluation of Long Primers for AP-PCR 62-63. Analysis of Mungbean (Vigna radiata [L.] Das BK, Saini A, Bhagwat SG and Jawali N Wilczek): Genetic Relationships and (2006) Development of SCAR markers for Fingerprinting of some Genotypes. Indian J identification of stem rust resistance gene Sr31 Biotech 3: 511-518. in the homozygous or heterozygous condition Sawhney RN (1998) Genetic basis of rust in bread wheat. Plant Breed 125: 544-549. resistance in Indian wheats and the need to Kochumadhavan M, Tomar SMS and Nambisan harness alien genes for durability. In: PNN (1988) Transfer of rust resistance genes Nagarajan S, Singh G and Tyagi BS (eds.) into commercial cultivars of wheat. Ann Wheat Wheat Research Needs Beyond 2000 AD. News Letter 34: 54-55. Narosa Publishing House, New Delhi, India, Krishna TG and Jawali N (1997) DNA isolation pp. 161-175. from single or half seeds suitable for random Vierling R and Nguyen HT (1992) Use of RAPD amplified polymorphic DNA analysis. Anna markers to determine the genetic relationship Biochem 250: 125-127. of diploid wheat genotypes. Theor Appl Genet Lee M (1995) DNA marker and plant breeding 84: 835-838. programs. Adv Agro 57: 265-326. Welsh J, Petersen C and McClelland M (1991) Mohan M, Nair S, Bhagwat A, Krishna TG, Yano Polymorphisms generated by arbitrarily M, Bhatia CR and Sasaki T (1997) Genome primed PCR in the mouse: application to strain mapping, molecular markers and identification and genetic mapping. Nucleic marker-assisted selection in crop plants. Mol Acids Research 19: 303-306. Breed 3: 87-103. Williams JGK, Kubelik AR, Livak KJ, Rafalski JA Prasad S, Reddy KS and Jawali N (1999) and Tingey SV (1990) DNA polymorphisms Abundance of randomly amplified hybridizing amplified by arbitrary primers are useful as microsatellites in mungbean Vigna radiata L. genetic markers. Nucleic Acids Research 18: Wilczek. Asia Pacific J Plant Mol Biol 6531-6535. Biotechnol 7: 1-11.

10 Wheat Inf. Serv. 108: 11-17, 2009. www.shigen.nig.ac.jp/ewis

Research Information

AMMI analysis for salt tolerance in bread wheat genotypes

Dharmendra Singh1, 2, S. K. Singh2 and K. N. Singh1 1 Central Soil Salinity Research Institute, Karnal-132001, India 2 Directorate of Wheat Research, Karnal-132001, India * Corresponding author: Dharmendra Singh (E-mail: [email protected])

Abstract Grain yield data of 15 bread wheat genotypes evaluated under four salt stress conditions for two consecutive years was subjected to the ordinary analysis of variance (ANOVA) and the additive main-effect and multiplicative interaction (AMMI) analysis. The ANOVA indicated that the variance due to genotypes (52.26%) was maximum followed by the variance due to environment (36.05%) and genotype x environments interaction (13.88%) was considered as residual. Thus, the ordinary ANOVA model accounted only for 88.31 per cent of the trial sum of squares that concentrated only on the genotype effects and environment effects. The bioplot from AMMI1 parameters provided the comprehensive understanding of the pattern of the data and indicated that the specifically adapted genotypes would away from the line with IPCA1=0 and next to grand mean level. Thus, 6 genotypes viz., KRL 3-4, Kharchia 65, KRL 99, KRL 19, Amery, BT- Schomburgk and PBW 343 showed general adaptability to all location as these were scattered at the right-hand side of the grand mean level and close to IPCA1 score =0 line. However, environments E1, E3, E4, E5, E6, and E7 had smallest (near to zero) IPCA scores and therefore, relative ranking (not absolute yields) of genotype would be fairly stable in these environments. In addition to the smallest interaction effects, the sites E1, E3, E5 and E7 were high yielding (environment mean yield > grand mean) and thus, deemed to be fit for growing wheat crop in general. Key Words: Wheat grain yield, G x E interaction, AMMI analysis, Adaptability.

Introduction be released as varieties, yield trials are conducted Excess amount of salt in the soil adversely affects with a set of genotypes at different salinity stress plant growth and development. Nearly 20% of the environments which is always affected by G x E world’s cultivated area and nearly half of the interactions (Zobel 1990). A significant G x E world’s irrigated lands are affected by salinity interaction for a quantitative traits, such as yield (Ashraf 1994, Bray et al. 2000). Increased salt can seriously limits efforts in selecting superior tolerance in crops is widely recognized as an genotypes for both new crop production and effective way to overcome the limitations of crop improved cultivars development (Kang 1990) and production in a salinized area (Munns and James would reduce the usefulness of subsequent 2003). For improving the salt stress tolerance of analysis of means and inferences that would crop varieties by plant breeding, it is necessary to otherwise be valid. identify donor genotypes that have proven The analysis of variance (ANOVA) is useful tolerance to salt stress during all the growth stages. for identifying the sources of variability but it Genotype x environments (G x E) interaction plays provides no insight into the particular pattern of a major role in evaluation of genotypes under the underlying interaction. On the other hand, the different environment (salinity stress) to identify AMMI analysis model is additive and effectively genotypes suitable to different stresses (Munns and describes the main (additive) effects, while the James 2003). With an aim to make interaction (residual from the additive model) is recommendations about the suitable genotype to non additive and requires other techniques, such as

11 principal component analysis (PCA) to identify interaction patterns. Thus, ANOVA and PCA Which yields the AMMI of model λk is the models combine to constitute the additive eigen value of interaction principal components Main-effect and Multiplicative Interaction axis (IPCA) k, γik and αjk are correspondingly the (AMMI) model (Gauche and Zobel 1988, Zobel et genotype and environment eigenvectors (i.e., al. 1988). The AMMI model is, therefore, a hybrid IPCA scores) for the axis k, N is the number of statistical model incorporating both ANOVA (for axes retained in the model, and εij is the residual. additive component) and PCA (for multiplicative Beginning with the ordinary ANOVA component) for analyzing two-way procedure (Snedecor and Cocharan 1980) for two (genotype-by-environment) data structure. The way analysis of variance, the AMMI analysis first model has, in recent past, been recommended for separates additive variance (µ, gi and ej) from the statistical analysis of yield trials, and was multiplicative variance (interaction), and then preferred over other customary statistical analyses, applies PCA to the interaction, i.e., to the residual such as ordinary ANOVA, principal component portion of the ANOVA model to extract a new set analysis and linear regression analysis (Gauch of coordinate axes which account more effectively 1988, Zobel 1988). An experiment was conducted for the interaction patterns (Gauch 1988, Gauch to evaluate the performance of the genotypes and Zobel 1988, Nachit 1992). Direct estimation under different salt stress environments with the of G x E interaction is obtained by the product of 0.5 following objective (i) to determine the nature and IPCA score (s) (λk γik) times the environment 0.5 magnitude of G x E interaction effect on grain IPCA score(s) (λk γjk). The eigen values in PCA yield in diverse environment, (ii) to determine are equivalent to sum of squares, and the degrees environment where wheat genotypes would be of of freedom for IPCA axes were calculated adapted and produce economically competitive as Gollob (1968): df= G+E-1-2k for axis k. yields. AMMI generates a family of models with different values of N. The simplest model with Materials and methods AMMI0 with N equal to zero considers only the Fifteen genotypes (KRL 99, Schomburg, KRL 105, additive effects,namely genotypes and HD 4530, Perenjori, HD 2009, PBW 343, environments means to explain the data matrix. BT-Schomburg, Amery, KRL 3-4, Ducula 4, The second model AMMI1 considers main effect Cunderdin, KRL 19, Camm, and Kharchia 65) and one interaction principal component axis to were evaluated in a randomised block design with interpret residual matrix. Similarly, AMMI2 three replicates at Central Soil Salinity Research involves main effects and two interaction principal Institute, Karnal in micro plot under four salt component axes for non additive (interaction) stress environments namely Normal (pH2 :8.2), variation, and so on. When one interaction PCA Saline (5.9ds/m), Sodic Low (pH2 :9.2) and Sodic axis account for most G x E, a feature of AMMI High (pH2 :9.4) for two consecutive years 2004-05 model is the biplot procedure in which genotypes and 2005-06. The yield was recorded and the data and environment-taking mean values on abscissa was subjected to ordinary ANOVA and AMMI and IPCA1 scores on ordinate are plotted on the model for analysis. The basic liner model (the same diagram, facilitating inference about specific ANOVA model) used in the analysis of yield trial interactions as indicated by the sign and magnitude is of the from: of IPCA1 values of individual genotypes and environments. The statistical analyses were carried γij = µ+gi+ej+δij out by the software (Gauch 1986).

Where γij is the observed response value (e.g., Results and discussion yield) of genotype (cultivar) i in environment j; µ is the grand mean; gi is the effect for genotype i Partitioning of variance (deviation of g from µ), i=1,…k; ej is the effect for environment j (deviation of e from µ), j =1,…n; Since AMMI model uses additive ANOVA for and δij is the interaction (=γij- γi- γj + γ..). It is partitioning of variance due to genotype and possible to partition the interaction component δij environments and analyses its residual (i.e., G x E into the sum of multiplicative functions of i and j interaction), analysis for AMMI (Table 1) can also (Mandel 1971). be used for a study of the results of ANOVA. It can be seen from this table that the mean squares γij = µ+gi+ej + Σλk γikαjk +εij for genotypes, environments and G x G interaction

12 Table1. Pooled AMMI analysis of variance for grain yield of 15 wheat genotypes under four different environments for two crop seasons

Source d.f. Sum of squares Mean squares R2a

Trails 119 426. 35 3.58** 100.00

Genotypes 14 218. 04 15.57** 52.26

Environments 7 150.40 21.49** 35.05

G x E interaction 98 57.90 0.59 13.88

IPCA 1 20 32.65 1.63** 66.96

Residual 44 9.15 0.21 33.24 were found to be highly significant. This due to the presence of highly levels uncontrolled suggested that broad range of diversity existed variation but not due to the real G x E interaction. among genotypes and among environment and that Above analysis, however, seems to suggest the the performance of genotype was differential over presence of a complex, multidimensional variation environments. Of the total treatment variation in the genotype-by-environment data as the first (trails SS), the proportion of variation of variance seven IPCA axes were demonstrated to be highly due to difference in genotypes was largest (52.26 significant by an F-test (P>0.001). The AMMI per cent) followed by the variance due to G x E models with many IPCA axes are expected to interactions (13.88 %: considered as residual in involve rather more noise than the highly complex case of ANOVA), and variance due to interactions among genotypes and environment. environment (36.05 %) Thus, ordinary Anova Further, if the AMMI model incluses more than model accounted only for 88.31 per cent of the one IPCA axes, assessment and presentation of trail SS concentrating only on the genotype effects genetic stability are not as that from the AMMI and environment effects. Therefore, it could tell model (Gauch 1982, Gauch 1988, Gauch and use (through statistical test ) whether genotypes, Zobel 1988, Gauch and Zobel 1994, Nachit et al. environments and genotype x environment 1992). The second and higher IPCA axes, despite interaction exerted a significant effect, but it did significant in the present study, were polled in to not tell us which genotypes environments and residual. Thus, AMMI model (AMMI model with genotype x environment combinations were first IPCA axis) was accepted for further study. responsible, nor did it tell us how their responses differ. Conclusively, ANOVA provided no insight AMMI1 Biplot: Interpretting specific pattern into the particular patterns of genotypes or environments that gave rise to interactions, but The results of AMMI1 analysis can also be easily described only the main effects effectively. These comprehended with help AMMI1 biplot results were also confirmed to the observations as presented in Fig. 1. The mean performance and made by Snedecor and Cochran 1980. Thus, in the IPCA1 scores for both the genotypes and present investigation, ANOVA model was not environment used to construct the biplot (Fig. 1) found to be adequate for analyzing the bread are presented in Table 2 and Table 3. The biplot - wheat yield data, as G x E interactions were highly a graphical representation - from AMMI1 analysis significant. Therefore, ANOVA model was is a used tool in under standing more combined with PCA model to further analyze the comprehensively the specific pattern of main residuals of the ANOVA model, which infact effect and G x E interactions of both the genotypes contains G x E interaction. Gauch (1988) and environments simultaneously (Crossa et al suggested further analysis of the effects of G x E 1991, Kempton1984, Zobel et al. 1988). The interactions even if they are indicated to be non bioplot of parameters accounted for 88.31 per cent -significant by an F-test in ANOVA. The residual of the trail SS. It is clear from the biplot that the SS which accounted for 33.04 % of the G x E SS points for environment were more scattered than with 13.88 % of G x E df was also found to be the point for genotypes; this indicated that highly significant. This situation seems to arise variability due to environments was higher than

Fig. 1. Biplot for grain yield showing main and interaction effects for both genotypes and environments.

Table 2. Mean yields (g/plant) and IPCA 1 score for 15 wheat genotype (two years data) S. No. Genotypes Mean Yield (g/plant) IPCA 1 scores

1. KRL99 5.141 0.279

2. Schomburgk 2.500 0.386

3. KRL105 5.893 1.664 4. HD4530 1.011 -0.281

5. Perenjori 3.161 -0.465

6. HD2009 2.006 0.023

7. PBW343 3.812 -1.106

8. BT-Schomburgk 3.871 0.252

9. AMERY 4.082 0.123

10. KRL3-4 5.508 -0.681

11. Ducula4 3.037 -0.468

12. Cunderdin 2.996 0.081

13. Camm 2.596 0.347 14. KRL19 (Check) 4.658 0.311

15. Kharchia 65 (Check) 5.188 -0.464

Table 3. Mean yields (g/plant) and IPCA 1 score for eight environments

S. No. Environment Mean Yield (g/plant) IPCA 1 score

1. E1 5.069 1.431

2. E2 2.060 0.329

3. E3 3.473 0.859

4. E4 2.360 0.517 5. E5 5.305 -0.596

6. E6 3.554 -0.913

7. E7 4.555 -0.651 8. E8 3.165 -0.617

15 that due to genotypes difference. This is also interaction and thus will suggest higher yield of evident from the ANOVA. In Fig. 1, displacement genotypes. along the abscissa (horizontal axix) reflects difference in main effects whereas displacement 2. Identifying favorable environments for wheat along the ordinate (vertical axix) exhibits genotypes differences in the interaction effects. When a genotype and an environment fall in the upper or Environment that appears almost in a lower portion from the line indicating IPCA1=0 in perpendicular line have similar means and those the biplot, their interactions is positive. However, that fall almost in a horizontal line have similar the genotypes and environments of opposite interaction pattern. AMMI bioplot (Fig.1) thus portions from the IPCA1=0 line show negative exhibited that environment differed in maim interaction. In other words the genotypes and effects but they exhibited nearly similar environments with similar signs (either positive or interactions. The environment E1 and E5 had negative) of IPCA1 scores exhibit negative similar main effect but differed in interaction with positive and vice versa. Thus, with the help of genotypes. The environment E2, E3, E4, E6 and biplot, the results of present investigation can be E8 differed in both main effect and interactions; interpreted as follow: the ranking in such environment are likely to be quite variable, thus making it complex to produce 1. Identifying high yielding stable genotypes variety recommendations. Further the environment E1, E3, E5 and E7 were highest yielding and According to the AMMI model, the genotypes highly interacting, hence are most suitable only for which are characterized by means greater than the specifically adapted genotypes. However, grand mean and the IPCA score nearly zero are environment E1, E3, E4, E5, E6, E7, and all had considered as generally adaptable to all smallest (near to zero) IPCA scores; relative environment. However, the genotype with high ranking (not absolute yields) of genotype would be mean performance and with large value of IPCA fairly stable in this environment. In addition to the score are consider as having specific adaptability smallest interaction effects, the sites E1, E3, E5 to the environments. AMMI Analysis was also and E7 were high yielding (environment mean conducted and the stability of genotypes was yield > grand mean) and therefore, deemed to be predicted on the basis of mean performance and suitable for growing wheat crop in general. the magnitude of IPCA1 scores in soybean (Zobel Selection of environment and requirement of et al. 1988), maize and wheat (Crossa et al. 1990, environment for wheat crop may, therefore, be 1991), sorghum (Zavala-Garcia et al. 1992) and recommended on the basis of the main features of barley (Ramagossa et al. 1993). On the bioplot, the the respective environments. The results points for the generally adapted genotypes would confirmed that analysis with its biplot is a very be at right hand side of grand mean levels (this useful tool in analyzing yield trial data. It explains suggests high mean performance) and close to the comprehensively both the effects due to genotypes line showing IPCA=0 and (this suggests negligible and environments and also their interaction or no G x E Interaction). However, the points for patterns. ANOVA could explain only the the specifically adapted genotypes would be away genotypes and environments but not interaction from the line with IPCA1=0 and next to grand which is a significant feature yield trial. mean level. Thus it was from Fig 1 that 6 genotypes, viz., KRL 3-4, Kharchia 65, KRL 99, References KRL 19, Amery,BT- Schomburgk and PBW Ashraf M (1994) Breeding for salinity tolerance in 343 which were scattered at the right-hand side plants. Crit Rev Plant Sci 13: 17-42. of the grand mean level and close to IPCA1 score Bray EA, Bailey-Serres J and Weretilnyk E (2000) =0 line, were declared by the AMMI1 model as Responses to abiotic stresses. In: Biochemistry having general adaptability to all location. and Molecular Biology of Plants. Gruissem W, However KRL 105 was equipped with high mean Buchnnan B and Jones R eds. American and large IPCA score, hence specifically suited to Society of Plant Physiologists, Rockviile, MD, the favourable environment. Favourable 1158-1249. environment for these genotypes can be Crossa J, Gauch HG and Zobe RW (1990) characterized as with high mean and large IPCA Additive main effect and multiplicative score with same sign as of genotype IPCA1 score. interaction analysis of two international maize Similar sign of IPCA1 scores implies positive cultivar trails. Crop Science 30: 493-500.

16 Crosa J, Fox PN, Pfeiffer WH, Rajaram S and Mandel J (1971) A new analysis of variance Guch HG (1991) AMMI adjustment for model for nonadditive data. Technometrics 13: statistical analysis of an international yield trial. 1-18. Theor Appl Genet 81 :27-37. Munns R and James RA (2003) Screening method Gauch HG (1982) Noise reduction by eigenvector for salinity tolerance: a case study with ordination. Ecology 63: 1643- 1649. tetraploid wheat. Plant and Soil 253: 201-208. Gauch HG (1986) MATMODEL:A Nachit MM, Nachit G, Ketata H, Gauch HG and FORTRAN-77 programe for AMMI, ANOVA Zobel RW (1992) Use of AMMI and liner and Finaly Wilkinson regression models for regression models to analyze genotype two–way data matrices with or without environment interaction in Durum wheat. replication. Microcomputer Power, Ithaca, Theor Appl Genet 83: 597-601. New York. Romagossa I and Molina Cano JL (1993) Gauch HG (1988) Model selection and validation Intergration of statistical and physiological for yield trials with interaction. Biometrics 34: analyses of adaptation of near isogenic barley 705-715. lines. Appl Genet 86: 822-826. Gauch HG and Zobel RW (1988). Predictive and Snedecor GW and Cochran WG (1980) Statistical postdicctive success of statistical analyses of Methods.7th Edn. Iowa State University Press yield trial. Theor Appl Genet 76:1-10. Ames 1A. pp264-265. Gauch HG and Zobel RW (1994) AMMI analysis Zaval-Garcia F, Barmal-Cox PJ and Estian JD of yield trials. Contribution from Soil, Crop (1992) Potential grain from selection for yield and Atmospheric Science and stability for grain sorghum populations. Appl USDA-ARSNAA, Bradfield Hall, Cornell Genet 85: 112-119. University, Ithaca,NY14853-1901. Zobel RW, Wright MG and Gauch HG (1988) Gollob HF (1968) A ststistical model which Statistical analysis of yield trial. Agron J 80: combines feature of factor analysis and 388-393. analysis of variance techniques. Zobel RW (1990) A powerful statistical model for Psychometrika 33: 73-175. understanding genotype by environment Kang MS (1990) Genotype by Environment interaction. In: ‘Genotype by Environment Interaction in Plant Breeding. Department of Interaction in Plant Breeding’ (Ed., M.S.Kang, Agronomy, Louisiana State University, Baton 1990), pp 126-140. Department of Agronomy, Rouge, Louisiana. Louisiana State University, Baton Rouge, Kempton RA (1984) The use of biplots in Louisiana. interpreting variety by environment interactions. J Agric Sci 103: 123-135.

Wheat Inf. Serv. 108: 19, 2009. www.shigen.nig.ac.jp/ewis

Topics on Wheat Genetic Resources

National BioResource Project Wheat: Overview

Takashi R. Endo* Laboratory of Plant Genetics, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan *Corresponding author: Takashi Endo (E-mail: [email protected])

The National BioResource Project (NBRP)-Wheat, characterization of DNA markers that would be useful which started its second term in 2007-2011, stores and in gene isolation, and obtained data is going to be supplies the wild species, landraces, and experimental released in the database KOMUGI. It is also strains of wheat and related species, and also EST and continuing to collect ESTs from different cDNA TAC clones of wheat that have been collected during libraries. The project is implemented by the Graduate the first term of the NBRP-Wheat (2002-2006). These School of Agriculture, Kyoto University (core facility) resources can be requested online (http://www. and the Kihara Institute for Biological Research, shigen.nig.ac.jp/wheat/komugi/top/top.jsp). Besides, Yokohama City University (sub-facility). the second-term NBRP-Wheat is focusing on the

Wheat Inf. Serv. 108: 20, 2009. www.shigen.nig.ac.jp/ewis

Topics on Wheat Genetic Resources

The report of National Bioresource Project-Wheat II. Seed resources, 2008

Taihachi Kawahara1*, Takashi R. Endo2, Tomohiro Ban3, Masahiro Kishii3, Tsuneo Sasanuma3,4 1: Laboratory of Crop Evolution, Plant Germ-plasm Institute, Graduate School of Agriculture, Kyoto University, Mozume, Muko 617-0001, Japan 2: Laboratory of Plant Genetics, Graduate School of Agriculture, Kyoto University, Kitasirakawa-Oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan 3: Division of Evolutionary Genetics, Kihara Institute for Biological research, Yokohama City University, Maioka, Totuka-ku, Yokohama 244-0813, Japan 4Present address: Faculty of Agriculture, Yamagata University, Wakaba, Tsuruoka 997-8555, Japan *Corresponding author: Taihachi Kawahara (E-mail: [email protected])

Maintenance of seed resources: Regeneration of of Agriculture, Kyoto University. In autumn 2008, total of 1,218 strains was finished early in summer 1,502 strains were sown, 419 at KIBR and 1,083 at 2008. Two hundred and ninety-seven landraces of Graduate School of Agriculture, Kyoto University. bread wheat were grown in Kihara Institute for Biological Research, Yokohama City University Distribution of seed resources: Total of 1,922 strains (KIBR) and 921 strains consisting of several wild have been distributed to various researchers and species and landraces were grown in Graduate School institutions throughout the world (Table 1).

Table 1. Number of seed stocks distributed in 2008 Code (Institution) * No. of strains distributed Domestic Overseas Total KT (KIBR) 168 164 332 KU (MOZUME) 444 213 657 LPGKU 86 716 802 TACBOW 20 111 131 Total 718 1,204 1,922 * KIBR: Kihara Institute for Biological Research, Yokohama City University, LPGKU and MOZUME: Graduate School of Agriculture, Kyoto University, TACBOW: Faculty of Agriculture, Tottori University.

Wheat Inf. Serv. 108: 21-22, 2009. www.shigen.nig.ac.jp/ewis

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Annual report of the project “Polymorphism survey among hexaploid wheat and its relatives by DNA markers”

Miyuki Nitta and Shuhei Nasuda* Laboratory of Plant Genetics, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan *Corresponding author: Shuhei Nasuda (E-mail: [email protected])

We are running the project “Polymorphism survey (http://rcshigen.lab.nig.ac.jp/wheat/komugi/strains/abo among hexaploid wheat and its relatives” granted by utNbrpMarker.jsp) on the KOMUGI database. the second phase of NBRP-Wheat. The detail of the project has been described in elsewhere (Nitta and Other activity: We hold a user-meeting of the wheat Nasuda 2009). So, we here provide a summary of our SSR marker on July 31st, 2008 at Kyoto Univ. We activities in the fiscal year 2008 (from April 2008 to explained recent development of the project and the March 2009). participants gave us valuable comments and requests. Three participants (Drs. Nishida, Hatta, and Objectives: (1) To make the genetic stocks more Matsunaka) gave short talks on their works using valuable for molecular genetic studies by adding DNA markers. genotype information of the SSR (simple sequence repeat) markers. (2) To establish a set of SSR markers Acknowledgements that is suitable for surveying DNA polymorphisms We would like to express our gratitude to Drs. G. among wheat strains. Ishikawa, K. Kato, Y. Matsuoka, and S. Takumi for their valuable advices. This work is supported by the Goal of the FY 2008: (1) To obtain amplification National Bioresorce Project-Wheat, the Ministry of profiles of 48 accessions of wheat and its relatives Education, Culture, Sports, Science and Technology, (Table 1 in Nitta and Nasuda 2009) with respect to Japan. 400 SSR markers. (2) To release the data to users by publically accessible methods. References Polymorphism survey: We could obtain Gupta PK, Balyan HS, Edwards KJ, Isaac P, Korzun V, amplification profiles of more than 500 markers in this Röder M, Gautier MF, Joudrier P, Schlatter AR, year. The cumulative numbers of the markers analyzed Dubcovsky J, De la Pena RC, Khairallah M, since the beginning of the project is 824. Overall, we Penner G, Hayden MJ, Sharp P, Keller B, Wang obtained 39552 amplification profiles (824 markers x RCC, Hardouin JP, Jack P and Leroy P (2002) 48 lines). The numbers of markers analyzed were 233 Genetic mapping of 66 new microsatellite (SSR) gwm (Röder et al. 1995, 1998), 122 barc (Song et al. loci in bread wheat. Theor Appl Genet 105: 2005), 125 cfa & cfd (Guyomarc’h et al. 2002; 413–422. Sourdille et al. 2004), 21 gdm (Pestsove et al. 2000), Guyomarc’h H, Sourdille P, Charmet G, Edwards KJ and 323 wmc (Gupta et al. 2002; Somers et al. 2004) and Bernard M (2002) Characterization of markers, which cover most of the markers genetically polymorphic microsatellite markers from Aegilops mapped in hexaploid wheat by Somers et al. (2004). tauschii and transferability to the D genome of bread wheat. Theor Appl Genet 104: 1164–1172. Data release: Our web-based database is still under Nitta M and Nasuda S (2009) A report of the project construction. However, we started to release “Polymorphism survey among hexaploid wheat amplification profiles to domestic researchers within and its relatives by DNA markers” granted by the Japan upon request. So far, we provided marker National Bioresource Project-Wheat, Japan. eWIS information to seven research groups in Japan. The 107: 33-35. request could be made through the web page Pestsova E, Ganal MW and Röder M (2000) Isolation

and mapping of microsatellite markers specific for wheat (Triticum aestivum L.). Theor Appl Genet the D genome of bread wheat. Genome 43: 109: 1105–1114. 689–697. Song QJ, Shi JR, Singh S, Fickus EW, Costa JM, Röder MS, Plaschke J, Konig SU, Börner A, Sorrells Lewis J, Gill BS, Ward R and Cregan PB (2005) ME, Tanksley SD and Ganal MW (1995) Development and mapping of microsatellite (SSR) Abundance, variability and chromosomal location markers in wheat. Theor Appl Genet 110: of microsatellites in wheat. Mol Genet Genomics 550–560. 246: 327–333. Sourdille P, Singh S, Cadalen T, Brown-Guedira GL, Röder MS, Korzun V, Wendehake K, Plaschke J, Gay G, Qi L, Gill BS, Dufour P, Murigneux A and Tixier MH, Leroy P and Ganal MW (1998) A Bernard M (2004) Microsatellite-based deletion microsatellite map of wheat. Genetics 149: bin system for the establishment of 2007–2023. genetic-physical map relationships in wheat Somers DJ, Isaac P and Edwards K (2004) A high (Triticum aestivum L.). Funct Integr Genomics 4: density microsatellite consensus map for bread 12–25.

Wheat Inf. Serv. 108: 23, 2009. www.shigen.nig.ac.jp/ewis

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Annual report of National Bioresource Project-Wheat II. DNA resources, 2008

Yasunari Ogihara*, Kanako Kawaura, Hisako Imamura Plant Genome Science Division, Kihara Institute for Biological Research, Yokohama City University, Maioka-cho Totsuka-ku, Yokohama 244-0813, Japan *Corresponding author: Yasunari Ogihara (E-mail: [email protected])

In the second year of the NBRP-Wheat II, we are Maintenance of DNA resources: Approximately continuously conducting the systematic management 45,000 cDNA clones were newly stored. EST of cDNA clones and ESTs in common wheat. information was processed to open at KOMUGI. Achievements carried out during April 2008 to March 6,162 full-length cDNA clones which have been 2009 are reported here. already sequenced in 2007 were stored.

Collection of DNA resources: Total of 90,306 ESTs Distribution of DNA resources: 70 cDNA clones from four cDNA libraries were sequenced. Tissues were distributed, of which 35 clones were sent to used for these cDNA libraries are listed as follows; domestic researchers and 35 clones to foreign (1) Roots of boron tolerant wheat, Halberd, grown institutes. hydroponically for 18 days, (2) Roots of boron tolerant wheat, Halberd, grown hydroponically for 17 Related activity of DNA resources: The oligo-DNA days and treated with 10 mM Boric Acid for 24 hours, microarray harboring 38k gene probes of common (3) Roots of boron intolerant wheat, Cranbrook, wheat were developed under the collaboration with grown hydroponically for 18 days, (4) Roots of boron Agilent Technology and distributed to domestic intolerant wheat, Cranbrook, grown hydroponically researchers. for 17 days and treated with 10 mM Boric Acid for 24 hours.

Wheat Inf. Serv. 108: 25-66, 2009. www.shigen.nig.ac.jp/ewis

Meeting Reports

The 6th International Triticeae Symposium (6th ITS) May 31 – June 5, 2009 International Conference Hall II & III, Clock Tower Centennial Hall, Kyoto University, Kyoto, Japan

Taihachi Kawahara* Chair of Local Organizing Committee, Plant Germ-plasm Institute, Graduate School of Agriculture, Kyoto University, Japan *Corresponding author: Taihachi Kawahara (E-mail: [email protected])

The Sixth International Triticeae Symposium (6th University) ITS) was held at the Clock Tower Centennial Hall of Shigeo Takumi (Graduate School of Agricultural Kyoto University in Kyoto, Japan from 31 May to 5 Science, Kobe University) June 2009. The symposium was organized under the Hisashi Tsujimoto (Faculty of Agriculture, Tottori joint auspices of the Local Organizing Committee, University) International Organizing Committee and the National Institute of Agrobiological Sciences (NIAS), Japan. It International Organizing Committee was supported by the Kyoto University Foundation Roland von Bothmer (Chair of IOC, Swedish and the Japanese Society of Breeding. Total of 118 University of Agricultural Science, Sweden) researchers and students, from 20 countries including Mary Barkworth (Vice-chair of IOC, Intermountain Australia, Azerbaijan, Canada, China, Czech Republic, Herbarium, Utah State University, USA) Germany, Georgia, Iran, Italy, Japan, Kazakhstan, Bradley Shaun Bushman (USDA-ARS Forage and Mexico, Poland, Russia, Slovak Republic, Spain, Range Research Lab., USA) Sweden, Turkey, UK, and USA, participated in this Vojtech Holubec (Department of Gene Bank, Reserch symposium (Fig. 1). There were 48 oral presentations Institute of Crop Production, Czech Republic) including 8 plenary lectures and 49 posters (Fig. 2). Taihachi Kawahara (Plant Germ-plasm Institute, Like its predecessors, the Sixth Symposium brought Graduate School of Agriculture, Kyoto University, together researchers and students in many different Japan) disciplines who had common interest in the one group Helmut Knupffer (Leibniz Institute of Plant Genetics of grasses, the Triticeae. and Crop plant Research, Germany) Kazuhiro Sato (Research Institute for Bioresource, Local Organizing Committee Okayama University, Japan) Taihachi Kawahara (Chair of LOC, PGPI, Kyoto University) National Institute of Agrobiological Sciences Kazuhiro Sato (Secretary of LOC, RIB, Okayama (NIAS), Tsukuba, Ibaraki, Japan University) Teruo Ishige (President, NIAS) Tomohiro Ban (Kihara Institute for Biological Takuji Sasaki (Vice-President, NIAS) Research, Yokohama City University) Hirokazu Handa (Head, Research Planning Section, Katsuyuki Kakeda (Graduate School of Bioresource, NIAS) Mie University) Takao Komatsuda (Plant Genome Research Unit, Masahiro Hishii (Kihara Institute for Biological NIAS) Research, Yokohama City University) Takao Komatsuda (National Institute of Agrobiological Science) PROGRAM Hideho Miura (Obihiro University of Agriculture and Veterinary Medicine) Sunday, May 31, 2009 Tsuneo Sasanuma (Faculty of Agriculture, Yamagata 16:00-20:00 Registration

16:00-20:00 Poster mounting 15:30-16:00 Barkworth M. (Utah State University, 18:00-20:00 Welcome Reception USA) Identifying genomic groups in the Monday, June 1, 2009 perennial Triticeae by their morphology 09:30-10:00 Opening Session 16:00-16:30 Coffee/Tea Break Chair: K. Sato 16:30-16:50 Taketa S. (Okayama University, Japan) Kawahara T., Chair, Local Organizing Molecular cytogenetic investigation on Committee (Kyoto University, Japan) the origin of two tetraploid Hordeum Bothmer R. von, Chair, International species, H. secalinum and H. capense Organizing Committee (Swedish 16:50-17:10 Rahiminejad M.R. (University of University of Agricultural Sciences, Isfahan, Iran) Sweden) The relationships among the A genome Ishige, T. (President, National Institute bearing Triticum species as evidenced of Agrobiological Sciences, Japan) by SSRs in Iran 10:00-11:00 Plenary Lectures I 17:10-17:30 Kakeda K. (Mie University, Japan) Chair: R. von Bothmer Molecular phylogeny of the genus 10:00-10:30 Heslop-Harrison J.S. (Leicester Hordeum using thioredoxin-like gene University, UK) sequences Triticeae cytogenomics: results, 17:30-19:30 Poster Session implications and applications 17:30-19:30 Happy Hour 10:30-11:00 Endo T. (Kyoto University, Japan) Dissection of barley genome by the Tuesday, June 2, 2009 gametocidal system 09:30-11:00 Domestication and Evolution I 11:00-11:30 Coffee/Tea Break Chair: V. Holubec 11:30-13:00 Systematics and Phylogeny I 09:30-10:00 Mori N. (Kobe University, Japan) Chair: R. von Bothmer Genetic diversity, evolution and 11:30-12:00 Blattner F.R. (Leibniz Institute of Plant domestication of wheat and barley in the Genetics and Crop Research, Germany) Fertile Crescent Phylogenetic and population level 10:00-10:20 Murai K. (Fukui Prefectural University, analyses to understand evolutionary Japan) processes in Hordeum (Poaceae) Adaptation of flowering-time in 12:00-12:20 Sun G. (Saint Mary's University, tetraploid wheat by selection of Canada) flowering-time genes under Molecular evolution and origin of domestication tetraploid Elymus species 10:20-10:40 Wang N. (National Institute of 12:20-12:40 Goncharov N.P. (Siberian Branch of the Agrobiological Sciences, Japan) Russian Academy of Sciences, Russia) Molecular evolution of cleistogamy in Taxomony and molecular phylogeny of barley natural and artificial wheat species 10:40-11:00 Nishida H. (Okayama University, Japan) 12:40-13:00 Muramatsu M. (Okayama University, Structural variation in 5’ upstream Japan) region of photoperiodic response genes, Wild Triticeae species indigenous to Ppd-A1 and Ppd-B1, in wheat Japan, their features and results of 11:00-11:30 Coffee/Tea Break hybridization studies involving wheat 11:30-13:00 Domestication and Evolution II and barley cultivars. Chair: F.R. Blattner 13:00-14:30 Lunch & Poster View 11:30-12:00 Komatsuda T. (National Institute of 14:30-15:30 Plenary Lectures II Agrobiological Sciences, Japan) Chair: J.S. Heslop-Harrison Domestication and spike architecture of 14:30-15:00 MacKay M. (Bioversity International, barley (Hordeum vulgare) Italy) 12:00-12:20 Ball T.B. (Brigham Young University, A strategy to enhance the effective and USA) efficient conservation and use of ex situ Typologic and morphometric analysis of plant genetic resources phytoliths produced by wheat and 15:00-15:30 Sasaki T. (National Institute of barley Agrobiological Sciences, Japan) 12:20-12:40 Takumi S. (Kobe University, Japan) Rice as a model for Triticeae genome Natural variation in morphological traits analysis in central Eurasian wild wheat 15:30-17:30 Systematics and Phylogeny II progenitor Aegilops tauschii Coss. Chair: J.S. Heslop-Harrison 12:40-13:00 Saisho D. (Okayama University, Japan)

Evolutionary process of six-rowed spike Sciences, Japan) in domesticated barley Biological nitrification inhibition (BNI) 13:00-14:30 Lunch & Poster View potential in Triticeae 14:30-16:00 Biodiversity and Genetic Resources I 10:20-10:40 Schnurbusch T. (Leibniz Institute of Chair: M. Barkworth Plant Genetics and Crop Plant Research, 14:30-15:00 Knüpffer H. (Leibniz Institute of Plant Germany) Genetics and Crop Plant Research, Reduced transcript levels at the Bot3 Germany) locus in barley (Hordeum vulgare L.) Genetic resources of Triticeae - confer increased tolerance to high boron cultivated species and genebank supply collections 10:40-11:00 Chen G. (National Institute of 15:00-15:20 Holubec V. (Crop Research Institute, Agrobiological Sciences, Japan) Czech Republic) Genetic targeting of drought sensitive Annual Triticeae genebank collection gene eibi1 of wild barley (Hordeum 15:20-15:40 Tomita M. (Tottori University, Japan) spontaneum) Quantitative variation of Revolver 11:00-11:30 Coffee/Tea Break transposon-like gene in synthetic wheat 11:30-13:00 Genomics and Breeding II and its structural relationship with Chair: S.R. Larson LARD element 11:30-12:00 Kumlehn J. (Leibniz Institute of Plant 15:40-16:00 Saeidi H. (University of Isfahan, Iran) Genetics and Crop Plant Research, Biodiversity of the D-genome species Germany) Aegilops tauschii in Iran Genetic engineering in cereals: Current 16:00-16:30 Coffee/Tea Break technologies for the elucidation of gene 16:30-18:00 Biodiversity and Genetic Resources II functions Chair: T. Schnurbusch 12:00-12:20 Sato K. (Okayama University, Japan) 16:30-17:00 Yen C. (Sichuan Agricultural University, Map based cloning of dormancy QTL in China) barley The tribe Triticeae Dumort. (Poaceae) 12:20-12:40 Dou Q.W. (North West Plateau Institute 17:00-17:20 Kishii M. (Yokohama City University, of Biology, China) Japan) Diversity of Triticeae as forage crops Synthetic wheat production for wheat 12:40-13:00 Konovalov F. (Vavilov Institute of breeding General Genetics, Russia) 17:20-17:40 Garg M. (Tottori University, Japan) Revealing genetic diversity in closely Exploration of Triticeae resource for related A-genome diploid wheat species wheat end product quality improvement (Triticum boeoticum, T. monococcum, T. 17:40-18:00 Cheng J. (Guizhou University, China) urartu) by retrotransposon display Natural variation in grain selenium 13:00-14:30 Lunch concentration derived from Israeli wild 13:00-14:30 Poster View barley, Hordeum spontaneum 14:30-16:00 Genomics and Breeding III 18:00-18:20 Yanagisawa T. (National Agricultural Chair: T.D. Colmer, Research Center for Western Region, 14:30-15:00 Sreenivasulu N. (Leibniz Institute of Japan) Plant Genetics and Crop Plant Research, Recent breeding objectives of hulled Germany) and hull-less barley for food in Japan Revealing seed storage metabolism from genetical genomic and systems Wednesday, June 3, 2009 biology approaches 09:30-20:30 Symposium Excursion 15:00-15:20 Turuspekov Y. (Montana State University, USA) Thursday, June 4, 2009 Hardness locus sequence variation in 09:30-11:00 Genomics and Breeding I association with grain quality in spring Chair: G. Muehlbauer barley (Hordeum vulgare L.) 09:30-10:00 Colmer T.D. (The University of Western 15:20-15:40 Tsujimoto H. (Tottori University, Japan) Australia, Australia) Mutual interchange of genetic variation Waterlogging and salinity tolerance in and genomic information between wild wild Hordeum species: physiological and cultivated species in Triticeae basis and prospects for use in wheat 15:40-16:00 Prieto P. (Consejo Superior de improvement Investigaciones Científicas, Spain) 10:00-10:20 Subbarao G.V. (Japan International Development and cytogenetic analysis Research Center for Agricultural of Hordeum chilense chromosome 4

introgression lines into durum wheat 7. Rollo J, Jacobs SWL, Rashid A, Barkworth, ME. 16:00-16:30 Coffee/Tea Break Using discriminant analysis to identify genomic 16:30-17:40 Genomics and Breeding IV groups within the perennial Triticeae Chair: J. Kumlehn 8. Knüpffer H. Geographical distribution patterns of 16:30-17:00 Matsumoto T. (National Institute of morphological characters in cultivated barley Agrobiological Sciences, Japan) (Hordeum vulgare L.) inferred from botanical Transcriptional landscape of malting varieties barley, Haruna Nijo - using a 9. Mori N, Watatani H, Ishii T, Kondo Y, Kawahara T, custom-made oligo array from cDNA Nakamura C. Intraspecific variation of chloroplast sequences DNA in Aegilops speltoides 17:00-17:20 Shavrukov Y. (University of Adelaide, 10. Ohta A, Kawahara T, Yamane K. Morphological Australia) variations of spike and the geographical distribution Salinity tolerance and sodium exclusion of subsection Emarginata species, genus in genus Triticum Triticum-Aegilops, close wild relatives of wheat 17:20-17:40 Malik A.I. (The University of Western 11. Pourkheirandish M, Komatsuda T. The regulatory Australia, Australia) network underlying the six-rowed spike in barley Submergence tolerance in Hordeum 12. Sakuma S, Pourkheirandish M, Matsumoto T, marinum Koba T, Komatsuda T. The barley vrs1 gene evolved 18:30-21:00 Symposium Dinner from duplication of a well-conserved HD-Zip I-class homeobox gene in the Poaceae Friday, June 5, 2009 13. Morihiro H, Takumi S. Intraspecific variation in 09:30-12:00 Plenary lectures III leaf shape-related traits in a wild einkorn wheat Chair: H. Knüpffer species Triticum urartu Thum. 09:30-10:00 Larson S.R. (Utah State University, 14. Tanno K, Bothmer von R, Yamane K, Takeda K, USA) Komatsuda T. Allopolyploidy of the Hordeum Gene, genomic, and trait discovery murinum complex indicated by a nucleotide research in perennial Triticeae grasses sequence of cMWG699 10:00-10:30 Tsunewaki K. (Kyoto University, Japan) 15. Turuspekov Y, Abugalieva S. The variation of SSR Plasmon analysis in wheat profiles in wild and cultivated barley 10:30-11:00 Coffee/Tea Break 16. Aliyeva AJ, Aminov NKH. A novel source of 11:00-11:30 Muehlbauer G. (University of germplasm for the development of branched ear Minnesota, USA) wheat The barley coordinated agricultural 17. An X, Wang D, Yan Y. Isolation and molecular project (CAP): integrating genomics characterization of three novel HMW glutenin with breeding subunits from Aegilops tauschii 11:30-12:00 Takeda K. (Okayama University, Japan) 18. Taguchi J, Kiribuchi-Otobe C, Matsunaka H, Ban Features of East Asian barley and their T. Analysis grain characteristics of tetraploid wheat genetic analyses gene pool to diversify genetic background of durum 12:00-12:30 Business session & Closing remark wheat 19. Bordbar F, Rahiminejad MR, Saeidi H, Blattner Poster List FR. Study of diversity and relationships of the D 1. Bushman BS, Barkworth ME. DNA markers: genome species of Aegilops–Triticum from Iran another tool in the toolbox 20. Gregová E, Medvecká E, Šramková Z, Mihálik D. 2. Jakob SS, Blattner FR. Phylogenetic and Estimation of quality of Triticum durum Desf. wheat phylogeographic analyses of Hordeum murinum on the basis of gliadin and glutenin characterization (Poaceae) 21. Mihálik D, Šramková Z, Medvecká E, Horevaj V, 3. Gerus DE, Agafonov AV. Levels of study of Šliková S. Genetic variability in bread wheat StH-genomic Elymus species of Asian Russia and (Triticum aestivum L.) of Slovakia based on North-Eastern Kazakhstan in connection with a polymorphism for high molecular weight glutenin problem of “species-phantoms” subunits 4. Hasheminejad N, Saeidi H, Yoosofi M, 22. Miyazaki T, Ban T. Multiplex Quantitative Rahiminejad MR. Taxonomy and interspecific analysis for trichothecene genes expression of relationships of Agropyron Grant. in Iran Fusarium graminearum causing head blight on 5. Zhang H-Q, Fan X, Huang Y, Sha L-N, Zhou Y-H. wheat spikes Genome constitution of Hystrix komarovii (Poaceae: 23. Igartua E, Molina-Cano JL, Gracia MP, Casas AM, Triticeae) Moralejo M, Ciudad FJ, Lasa JM. New tools for the 6. Ohta S, Fujita Y, Maesaka Y, Hattori M, Iwasaki R. accessibility of the Spanish barley core collection A biosystematic study in Aegilops neglecta – Ae. 24. Mosulishvili M, Maisaia I, Shanshiashvili T, columnaris species complex Akhalkatsi M. Triticum species in Georgia: diversity,

conservation, and taxa of special interest Orczyk W. Posttranscriptional silencing of CKX 25. Geraybeyova N, Sadiqov H, Rahimova O, genes, regulating cytokinin level in barley by RNA Sadiqova S, Karimov A, Mammadova N, Babayeva interference S, Abbasov M. Assessment of genetic diversity 42. Niwa S, Kikuchi R, Handa H, Ban T. Genetic among Azerbaijan barley genotypes (H. vulgare L.) variability of MRP gene constituting ‘Qfhs.kibr- based on Hordein alleles 2DS’ QTL to reduce Fusarium mycotoxin 26. Sharma S, Röder MS. Study of sequence accumulation among hexaploid wheats polymorphism and genetic diversity of sucrose- 43. Tanaka H, Arakawa T, Tsujimoto H. Alien glutenin phosphate synthase genes in bread wheat and its A, subunits expressed in common wheat endosperm B and D genome progenitors affect on the composition 27. Šliková S, Šramková Z, Gregová E, Mihálik D. 44. Tonooka T, Aoki E, Yoshioka T, Taketa S, Composition of high-molecular-weight glutenin Kiribuchi-Otobe C. Characterization of a subunits in European wheats β-glucanless mutant in barley 28. Zaharieva M, Dreisigacker S, Crossa J, Payne T, 45. Kidou S, Yokota S, Yoshida K. Virus-induced gene Misra S, Hanchinal RR, Mujahid MY, Trethowan R. silencing of P23k in barley leaf reveals Genetic diversity within Triticum turgidum L. subsp. morphological changes involved in secondary wall dicoccon (Schrank) Thell. (cultivated emmer) and its formation utilization in wheat breeding 46. Zhang LQ, Liu DC, Zheng YL, Yen Y. 29. Abugalieva S, Abugalieva A, Quarrie S, Spontaneous amphidiploidization via unreduced Turuspekov Y. Identification and mapping of QTLs gametes is a universal phenomenon for Triticum for grain protein content in common wheat turgidum - Aegilops tauschii hybrids 30. Aydin Y, Cabuk E, Mert Z, Akan K, Bolat N, 47. Mizuno N, Yamasaki M, Matsuoka Y, Kawahara T, Cakmak M, Uncuoglu AA. Investigations on yellow Takumi S. Population structure of central Eurasian rust disease resistance by useful genes and markers wild wheat progenitor Aegilops tauschii Coss. in gene-rich regions on wheat chromosomes 48. XF Zhang, DC Liu, WL Yang, JZ Sun, DW Wang, 31. Bińka-Wyrwa A, Orczyk W, Nadolska-Orczyk A. HQ Ling and AM Zhang. Molecular markers for Regulation of transformation efficiency in polyploid systematic characterization of low molecular weight cereals by type and number of selection cassettes glutenin subunits in common wheat (Triticum 32. Buwan R, Takahashi H, Kato K, Sato Y-i, aestivum L.) Komatsuda T, Nakamura I. Sequence variation of 49. Nishinaka M, Okumoto Y, Kato K, Kawahara T, the 20th exon within PolA1 gene among Triticeae Tanisaka T. Genetic diversity of high molecular species weight glutenin subunits in wheat landraces 33. Cagirgan AMI, Ullrich SE, Ozbas MO. High frequency and a wide spectrum of mutations in ‘BARONESSE’ barley fields ABSTRACTS & TITLES 34. Miroshnichenko D, Poroshin G, Dolgov S. Characterization of growth and yield of transgenic Oral Presentation wheat plants overexpressing vacuolar Na+/H+ antiporter genes 1. 35. Kikuchi R, Kawahigashi H, Ando T, Tonooka T, Triticeae cytogenomics: results, implications and Handa H. The flowering pathway under short day in applications barley 36. Martín AC. The potential of Hordeum chilense Heslop-Harrison JS (Pat)1, Contento A1, cytoplasm in the development of CMS systems in Graybosch RA2, Ali N1, Kuhn G CS1, Saeidi H1,3, Triticeae crops Kalpande H1,4, Schwarzacher T1 37. Martinek P, Dobrovolskaya O, Röder MS, Börner 1Department of Biology, University of Leicester, LE1 A. Agronomic traits and genetic determination of 7RH, UK winter wheat lines (Triticum aestivum L.) with 2USDA-ARS, University of Nebraska, Lincoln, NE multirow spike 68583, USA 38. Martinek P. Breeding Triticale (X Triticosecale 3Faculty of Science, University of Isfahan, Isfahan, Wittmack) for improved breadmaking quality Iran 39. Miroshnichenko D, Poroshin G, Dolgov S. Gene 4Marathwada Agriculture University, Maharashtra, flow from genetically modified to cultivated wheat India plants 40. Mishina K, Manickavelu A, Sato H, Katsumata M, The study of the genomics of the Triticeae has been a Sassa H, Koba T. Observation of pollen tube growth story of continuous progress over nearly a century. and molecular mapping of Kr genes in common After the definition of species and polyploids, the wheat-rye hybridization seminal work of Kihara and Sears showed the basic 41. Nadolska-Orczyk A, Zalewski W, Galuszka P, number of x=7 and developed the concepts of the

genome. Following these studies, relationships and the wheat lines carrying segments of barley chromosomes. genetics of characters including disease resistance and Thus, it has become possible to dissect the barley quantitative traits were investigated, where aneuploids genome. In this talk I will describe the progress in the and synthetic hybrids were invaluable, leading up to dissection of the barley genome. our current state of knowledge about genes and DNA sequences. These academic advances have been 3. paralleled by exploitation of the knowledge in plant Phylogenetic and population level analyses to breeding. In this talk, I will discuss our cytogenomics understand evolutionary processes in Hordeum research in the Triticeae, studying genome evolution at (Poaceae) a large scale. Tandemly repeated satellite DNA sequence arrays that make a substantial parts of Blattner FR Triticeae genomes, and include some of the most Leibniz Institute of Plant Genetics and Crop Research conserved sequence motifs as well as abundant (IPK), D-06466 Gatersleben, Germany variable sequences which are not conserved even between closely related genomes. I will report our Hordeum consists of 31 species, distributed in increasing understanding of the chromatin code – the temperate and arid regions of the Northern changes in methylation and histones which are Hemisphere, South Africa and southern South responsible for epigenetic effects, and may play an America. About 50% of the species thrive in this latter important role in nuclear architecture and genome region, making it the diversity center of the genus. To interactions in polyploids. Finally, I will discuss the analyze evolutionary processes in the genus and to applications of this work in the use of hybrids to explain the high species number in the Americas, interchange genetic variation and widen the genepool phylogenetic and phylogeographic analyses together available to plant breeders. Related publications and with molecular dating approaches, reconstruction of information are available from www.molcyt.com. historical biogeography, and ecological niche modeling were conducted for species and species 2. groups of the genus. Dissection of barley genome by the gametocidal The genus originated about 12 million years (My) system ago in Western Asia and colonized its extant distribution area by at least seven intercontinental Endo TR long-distance dispersals. About 4 My ago it reached Laboratory of Plant Genetics, Graduate School of South America, where a rapid radiation took place Agriculture, Kyoto University, Kyoto 606-8502, Japan during the last 2 My. While high speciation rates are characteristic for the Americas, Eurasian Hordeum lost Barley is one of the major cereals in the world. The most of its genetic and species diversity through analysis of genomes is important in modern breeding, extinction during the Pleistocene. Eurasian species but the barley genome is too huge and complicated were mostly restricted to small refugial populations that it is still difficult to arrange sequenced pieces of during ice-age cold cycles, while particularly in the genome in order. It is desirable to have the genome southernmost Patagonia large populations survived the divided into small pieces in separate plant lines. ice-age without spatial or genetic restrictions. Each of the barley chromosomes has been added to Therefore, speciation in Eurasia involved mostly common wheat and such barley chromosome addition severe genetic bottlenecks, while South America was lines are useful in locating the positions of genes and characterized by vicariance-driven speciation. These DNA markers on a chromosome. It has been proved differences are clearly reflected in chloroplast that the added barley chromosomes in common wheat diversity, resulting in specific patterns in the Old and can be fragmented genetically by the gametocidal New World. system. This system involves unique chromosomes, called gametocidal (Gc) chromosomes, that were 4. derived from some wild species of the genus Aegilops Molecular evolution and origin of tetraploid related to common wheat. When the Gc chromosome Elymus species exist in common wheat in monosomic condition, two types of gamete are produced, one carrying the Gc Sun G chromosome, the other without the Gc chromosome, Biology Department, Saint Mary's University, Halifax, and chromosome breakage occurs only in the latter NS, B3H 3C3 Canada gamete. Such Gc-induced chromosomal breakage leads to either the sterility of gametes or the It is well known that Elymus arose through production of fertile gametes carrying chromosomal hybridization between representatives of different mutations. Although no molecular mechanism of the genera and several different polyhaplomic genomes gametocidal system is known, this system has been have been described. Cytogenetically, five basic applied successfully to the generation of common genomes (St, H, Y, P, and W) in different

combinations have been found in the genus. The vast Triticum species except hexaploid Timopheevii group majority of species are tetraploids and they are species and the artificial ones. characterized by having the StY genome or the StH Presence of competing wheat classifications and genome. It is not known where the Y genome using illegitimate species names are causing confusion originated, although it is a common in Elymus from in different groups of research community. The Central and East Asia. Phylogeny and origin of possibility of using different classifications of genus tetraploid Elymus is far from clear. It has been Tritucum L. for molecular-biological, genetic and hypothesized from isozymic and cytological studies of phylogenetic investigations, for collecting and Elymus species that the Old and New World taxa may identifying wheat accessions and breeding practice is be of separate origin of the H genome in the StH discussed. genome species. To test this hypothesis, and estimate the phylogenetic relationship of polyploid Elymus 6. species within the Triticeae, single copy of nuclear Wild Triticeae species indigenous to Japan, their gene, the second largest subunit of RNA polymerase II features and results of hybridization studies (RPB2), was analyzed with Elymus species containing involving wheat and barley cultivars StH or StY genomes and diploid species. Our data indicated that the Eurasian and American StH genome Muramatsu M species have independent alloploid origins with Ezucho 3-6, Okayama 700-0028 Japan different H-genome donors, and provides some insight on the origin of Y genome and its relationship to other Studies on the wild Triticeae species indigenous to genomes in Elymus. Japan have revealed some biological phenomena; the main results are summarized here: (1) Observations 5. made for many years indicate that three species, Taxonomy and molecular phylogeny of natural and Elymus tsukushiensis, E. humidus, and E. ciliaris are artificial wheat species especially tolerant of the humid monsoon climate. (2) From the cross pollination of these three species with Goncharov NP, Golovnina KA, Kondratenko EYA species of Triticum, Aegilops, Secale and Hordeum, it Institute of Cytology and Genetics, Siberian Branch of was possible to obtain F1s by using embryo rescue. In the Russian Academy of Sciences, 10 Lavrentyev crosses of E. tsukushiensis and E. humidus as aven., Novosibirsk 630090, Russia female by Hordeum, hybrid plants had many sectors and polyhaploid plants of the female parents The effective use of wheat genetic resources is often resulted. In the combination involving Triticum, connected with their conservation strategy and an amphiploid line was produced with colchicine mechanism of their utilization. Producing of wheat treatment when E. ciliaris was the parent, whereas amphiploids with genomes of related species is an with E. tsukushiensis it was not possible to obtain effective way for this purpose. The problem of such amphiploids in spite of repeated treatments. The same amphiploid preservation has been under investigation behavior occurred in hybrids E. tsukusiensis x rye. during last hundred years. Artificial amphiploids Attempts to double the chromosome number of E. should be given names and places in the genus tsukushiensis and E. ciliaris have never succeeded. It Triticum taxonomy for preservation in genebanks. In appears that the formation of polyploids may be under this connection the carefulness in working with genus genetic control. The term, “definitomodis genetic Triticum classification is very important not only to conditions” is proposed to designate such control. The find the solution of problems in wheat origin and its term was suggested by the late Dr. G. Redei. (3) For phylogeny, but also to collect and estimate wheat the number of spikelets per spike node, which is 1~2 biodiversity preservation. The inheritance of in Elymus and 1 in Triticum, the genes involved may domesticated and taxonomically important characters be on the chromosome of the homoeologous group 2. has been studied in present work. An attempt to Nullisomics of that group in hexaploid wheat cv. integrate the results of different comparative-genetic Chinese Spring often have two spikelets resembling analyses of wheats and their molecular taxonomy has Elymus, indicating dosage effect of the duplicated been made. The correspondence of earlier genes; that is, the decrease from six to four doses evolutionary specifications to the phylogenetic may induce the Elymus-like spikelet phenotype. relationships within the genus Triticum species has been estimated using chloroplast and nuclear DNA 7. sequence data. The results of the provided molecular A strategy to enhance the effective and efficient analysis indicated close relationship of all hexaploid conservation and use of ex situ plant genetic species. Three different variants of investigated genes resources have been detected in diploid A genome Triticum. The detailed analysis showed that one of these variants Mackay MC1, Guarino L2, Street KA3 was a progenitor for all A genomes of all polyploid 1Bioversity International, Via dei Tre Denari 472/a,

00057 Maccarese, Rome, Italy ancestor about 40 MY ago. Both genomes however 2Global Crop Diversity Trust, C/o FAO, Viale delle retain the same order of genes in corresponding Terme di Caracalla, 00153 Rome, Italy genomic blocks. This synteny between rice and the 3International Centre for Agricultural Research in the Triticeae has been clarified by cross-genetic mapping Dry Areas (ICARDA), P.O. Box 5466, Aleppo, Syrian and further indicates conservation of gene order across Arab Republic regions of the chromosomes. In some cases, rice gene homologs could be identified based on synteny The conservation and use of plant genetic resources irrespective of its function in Triticeae. The syntenic (PGR) has a history dating back to the first relationships in cereal crops extend to the sequence domestication of plants by humans. In the 20th level as revealed by comparative analysis of specific Century significant efforts were made to collect and regions including the regulatory sequence of conserve these resources in ex situ genebanks. This corresponding genes. Although the genome size of was done initially at an institutional level but by the Triticaceae is several times larger than rice due to end of the Century there was a whole range of high-degree of insertion of transposable elements, conservation and use paradigms, including private, macrocolinearity and microcolinearity in both state, national, regional and international collections. genomes must be exploited to accelerate genome Each genebank had its own information management analysis. The high-quality rice genome sequence and system, mandate, standards and modus operandi. subsequent achievements in rice genomics should Despite this diverse array of arrangements for provide a model towards a comprehensive conserving PGR, they still need to be studied and characterization of Triticaceae genome structure and properly documented to ensure their effective use. To function. The availability of Triticaceae genome facilitate this on a global level standards are required: information will provide additional tool for crop standards of data quality and quantity for improvement to ensure a stable food supply. documentation; standards to uniquely identifying genotypes; standards to identify duplication; standards 9. for aggregating information across genebanks. The Identifying genomic groups in the perennial ongoing development and deployment of such Triticeae by their morphology standards is leading PGR towards a workable global structure that will provide access to, and practical Rollo J1, Jacobs SWL2, Rashid A3, Barkworth ME1 ways to identify and use, the millions of accessions 1Intermountain Herbarium, Dept. of Biology, Utah stored in the world’s genebanks. This paper will State University, Logan, Utah, 84322-5305, USA describe how the jigsaw puzzle of this global system 2National Herbarium of New South Wales, Mrs is being put together. Macquaries Road, Sydney, New South Wales, 2000, Australia 8. 3University of Peshawar Botanic Garden, University Rice as a model for Triticeae genome analysis of Peshawar, Peshawar, Northwest Frontier Province, Pakistan Sasaki T, Matsumoto T National Institute of Agrobiological Sciences, 1-2, Our goal was to determine whether morphology could Kannondai 2-chome, Tsukuba, Japan be used to predict the genomic constitution of perennial Triticeae with solitary spikelets. We scored Rice genome research has generated major advances 77 characters (58 quantitative, 19 qualitative) on 219 in plant science including a wide array of genetic specimens. The specimens represented 78 taxa and 13 resources and genomic information. Utilization of the different genomic groups. Discriminant analysis of the rice genome sequence information is now a quantitative characters indicated that it is possible to two-faceted strategy aimed at increasing world food use morphology to place perennial Triticeae with production. The first target focuses on complete solitary spikelets in their genomic group. Which understanding of rice biology based on the genome characters are most important depends on the sample composition and function to elucidate the components size of the groups in the analyses. The qualitative involved in grain yield, resistance to biotic/abiotic characters proved to be of little value. The data have stress, and adaptability to extreme environmental been used to initiate development of an online, conditions. The second target focuses on using the rice multi-access key to the groups and to aid in genome as a model for understanding other cereals development of a dichotomous key to the groups crops through comparative approaches. The based on our findings. These have been tested on completion of the rice genome sequence in 2004 led to specimens that were not included in the analyses. elucidation and isolation of many important genes that support rice molecular breeding and functional 10. characterization of genes from other cereal species. Molecular cytogenetic investigation on the origin of Rice, and the Triticeae diverged from a common two tetraploid Hordeum species, H. secalinum and

H. capense dicocoides and T. aestivum) collected from Iran and some accessions from other areas were examined Taketa S1, Nakauchi Y2, Bothmer von R3 using SSR markers. Thirty-two polymorphic primers 1Research Institute for Bioresources, Okayama were used to detect the biodiversity and relationships University, Kurashiki 710-0046, Japan among the A genome bearing species. A total of 411 2Faculty of Agriculture, Kagawa University, Miki alleles were revealed from which 349 were 761-0795, Japan polymorphic. Among the species studied T. durum 3Department of Crop Science, Swedish University of showed the maximum number of polymorphic loci Agricultural Sciences, SE-230 53 Alnarp, Sweden (186) and T. dicocoides the minimum (31). The observed (HO) and expected heterozygosity (HE) We previously reported that the two tetraploid varied from 0 to 0.98 (average of 0.49) and 0 to 0.92 Hordeum species, H. secalinum and H. capense are (average of 0.79), respectively. UPGMA cluster allotetraploids carrying the Xa genome of H. marinum analysis based on Nei 1972 coefficient classified the and the I genome of an unidentified diploid species genotypes examined into three main groups which (Taketa et al. Hereditas 130: 185-188, 1999). In the were corresponded to three ploidy levels. Analysis of present study, intraspecific variation in each tetraploid Molecular Variance (AMOVA) results indicated that species was investigated with regard to intergenomic most of the genetic variance occurred among translocations and chromosomal distribution of rDNA populations between the assumed groups, although sites. Genomic in situ hybridization revealed that three there was significant variance within the populations H. secalinum accessions examined did not carry (75.5%). A significant correlation was not detected intergenomic translocations, but that two of three H. between the poloidy levels of the species studied (p < capsense accessions carried a pair of intergenomic 0.05). Results of analysis showed that lower genetic translocations. Multicolour fluorescent in situ distance was observed in diploid groups (0.1) hybridization was applied to analyse chromosomal comparing with tetra- and hexaploids populations distribution of rDNA sites. In H. secalinum, two (0.14-0.36). T. urartu and T. turgidum showed the rDNA patterns were found and they differed in the highest genetic distance (0.36) as similar as genetic presence or absence of an additional 5S rDNA site in distance of T. dicoccum and T. aestivum (0.36). The the long arm of a submetacentric chromosome of the results of AMOVA analysis were in accordance with Xa-genome origin. The additional 5S rDNA site was cluster analysis and showed that genotypes were not also found in all H. capense accessions examined. The differentiated based on their geographical regions. additional 5S rDNA site is characteristic of H. marinum subsp. gussoneanum, but this site is absent in 12. H. marinum subsp. marinum. Polymorphisms in 5S Molecular phylogeny of the genus Hordeum using rDNA site infer that H. secalinum has two lineages, thioredoxin-like gene sequences one having subsp. gussonuanum and the other having subsp. marinum, as the Xa-genome donor. We suppose Kakeda K1, Taketa S2, Komatsuda T3 that H. capense is originated from a limited number of 1Graduate School of Bioresources, Mie Univ., H. secalinum accessions that were introduced through Tsu514-8507, Japan migration of European people to South Africa. 2Research Institute for Bioresources, Okayama Univ., Kurashiki 710-0046, Japan 11. 3National Institute for Agrobiological Sciences, The relationships among the A genome bearing Tsukuba 305-8602, Japan Triticum species as evidenced by SSRs in Iran Phylogenetic relationship in the genus Hordeum was Ehtemam MH1, Rahiminejad MR2, Saeidi H2, investigated based on nucleotide sequences of the Sayed Tabatabaei BE1, Krattinger S3, Keller B3 thioredoxin-like (HTL) gene. HTL gene was originally 1Department of Agriculture, Isfahan University of isolated in diploid H. bulbosum as a single copy gene Technology, Isfahan, 84156-83111, Iran closely linked to the self-incompatibility (S) locus. We 2Department of Biology, University of Isfahan, Iran amplified PCR fragments homologous to HTL from 11 3Institute of Plant Biology, University of Zurich, species including 16 taxa (25 accessions), which cover Switzerland mainly diploid accessions together with several tetraploid accessions in H. marinum and H. murinum. Microsatellites are polymorphic, multi-allelic and We determined nucleotide sequences of the variable co-dominantly inherited molecular markers, which region (949 to 1270 bp) that mainly includes 5’-UTR have become the marker of choice in plant genetics and 2 introns. Phylogenetic analysis based on these and breeding studies. Genetic relationships of 55 sequence data clearly demonstrated a divergence of 4 genotypes belonging to 8 Triticum of A genome basic genomes H (H. vulgare and H. bulbosum), X (H. bearing species (T. monococcum, T. boeoticum, T. marinum), Y (H. murinum) and I (other species) in the urartu, T. durum, T. turgidum, T. dicoccum, T. genus Hordeum. Phylogenetic clustering also inferred

2 clades separating one containing H and Y, and the activate transition from vegetative to reproductive other containing X and I genomes, although the phase (termed flowering). VRN2 is a repressor of bootstrap value of the latter was lower than that of the flowering, and down-regulated by vernalization. former. In the I genome, 4 American species (H. VRN1 is a homolog of Arabidopsis brachyantherum, H. chilense, H. pubiflorum, H. APETALA1/FRUITFULL MADS-box genes, and pusillum) were confirmed to be closely related each VRN2 encodes a putative zinc finger and a CCT other and divergent from Asian species (H. domain transcription factor. Genetic and molecular brevisubulatum, H. bogdanii, H. roshevitzii). Two studies suggested that the VRN2 protein suppresses diploid cytotypes of H. marinum (ssp. marinum and VRN1 expression. VRN3 is a homolog of Arabidopsis ssp. gussoneanum) were suggested to be involved in FLOWERING LOCUS T, which proteins function as the formation of the tetraploid cytotype (ssp. the florigen that moves from the leaves into the shoot gussoneanum). Two tetraploid cytotypes of H. apex to determine floral meristem identity, leading to murinum (ssp. murinum and ssp. leporinum) shared 2 the floral organ formation. We performed expression, distinct sequences, one related to that of the diploid mutant and transgenic studies to clarify the genetic cytotype (ssp. glaucum) and the other unique to them. network of these flowering-time genes, VRN1, VRN2, and VRN3. Expression analysis of a VRN1 deletion 13. mutant suggested that VRN3 is up-regulated by VRN1. Allelic diversity at chloroplast microsatellite loci Furthermore, transgenic analysis indicated that VRN3 among polyploid wheat species suppresses VRN2 expression. Based on these results, we recently present the VRN1-VRN3-VRN2 triangle Mori N model for the regulation of floral transition in wheat, Laboratory of Plant Genetics, Graduate School of in which VRN1 is upstream of VRN3 with a positive Agricultural Science, Kobe University, Kobe, Japan feedback loop through VRN2. In this paper, I discuss the molecular mechanism of adaptation of The domestication of wheat and barley was the most flowering-time in tetraploid wheats on the basis of the important step in the emergence of farming VRN1-VRN3-VRN2 triangle model. Especially, I communities that later led to the ancient civilisations would like to focus on selection of flowering-time of Mesopotamia. Several lines of evidence indicate genes under domestication. that emmer wheat (Triticum turgidum subsp. dicoccum, genome: AABB, 2n = 28) was the earliest 15. domesticated wheat derived from wild emmer (T. Molecular control of cleistogamy in barley turgidum subsp. dicoccoides, genome: AABB) and that the domestication occurred within the Fertile Wang N1, Nair S1, Turuspekov Y1, Pourkheirandish Crescent of southwest Asia. Chloroplast DNA M1, Sinsuwongwat S1, Sameri M1, Kanamori H2, fingerprinting of wild and domesticated emmer wheat Honda I3, Watanabe Y3, Stein N4, Wicker T5, Tagiri revealed that two distinct maternal lineages were A1, Nagamura Y1, Matsumoto T1, Komatsuda T1 involved in their domestication and thus suggested 1National Institute of Agrobiological Sciences, Plant that domestication of emmer wheat occurred Genome Research Unit, 1-2-1 Kannondai, Tsukuba, independently at least two times. Further survey in Ibaraki 305 8602, Japan common wheat revealed that only one of the two 2Institute of the Society for Techno-innovation of maternal lineages of emmer wheat might have Agriculture, Forestry and Fisheries (STAFF), 446-1 transmitted to common wheat through allopolyploidy Ippaizuka, Kamiyokoba, Tsukuba, Ibaraki, 305-0854, evolution. Based of these results the process and Japan geography of wheat domestication will be discussed. 3National Institute of Crop Science (NICS) Tsukuba, Kannondai 2-1-18, Ibaraki 305-8518, Japan 14. 4Group Genome Diversity, Dep. Genebank Leibniz Adaptation of flowering-time in tetraploid wheat Institute of Plant Genetics and Crop Plant Research by selection of flowering-time genes under (IPK) Corrensstr. 3, D-06466 Gatersleben, Germany domestication 5Institute of Plant Biology, University of Zürich, Zürich CH8008, Switzerland Murai K Fukui Prefectural University, 4-1-1 Open/close flowering is considered to be one of the Matsuoka-kenjojima, Eiheiji-cho, Fukui 910-1195, most important traits for alternative breeding Japan programs in both barley and wheat, such as for increase outcrossing rate or conversely restrict gene There are three major gens responsible for variation in flow of GM. In open flowers lodicules force lemma vernalization requirement in temperate cereals such as and palea apart by the swelling itself, however, some wheat and barley, VRN1, VRN2, and VRN3. natural variants that tightly closed throughout the Vernalization up-regulates VRN1 and VRN3, which anthesis exist in cultivated barley. In this report, the

closed flowering is defined cleistogamy in the strict involved in the control of Ppd-1 expression. In sense and the isolation of Cly1, which lies in a addition, no important allelic variations were detected genomic region of chromosome 2HL, showing some in the coding region of Ppd-A1 and Ppd-B1. Therefore, colinearity with a part of rice chromosome 4. The it was strongly suggested that these structural colinearity is interrupted by a micro-inversion. A fine variations explain the difference between mapping allowed Cly1 to be located within a 0.7cM photoperiod-sensitive and -insensitive alleles. interval defined by two ESTs, and the locus co-segregated with an ortholog of a rice transcription 17. factor. A barley EST homologous with this ortholog Domestication and spike architecture of barley was used to screen a barley BAC library. The (Hordeum vulgare) development of additional markers narrowed the location of Cly1 to a 7kbp region which contained Komatsuda T1, Sakuma S1, 2, Koba T2, only a single ORF. The expression of this sequence Pourkheirandish M1, Matsumoto T1, Gottwald S3, (presumed to be Cly1) was specific to the lodicule, Hensel G3, Kumlehn J3, Stein N3 and transcripts were abundant in the immature spikes. 1National Institute of Agrobiological Sciences (NIAS), The Cly1 coding sequence of a core collection of 274 Plant Genome Research Unit, Kan-non-dai 2-1-2, cultivars was resequenced, and this produced a clear Tsukuba, Ibaraki 305 8602, Japan correlation between cleistogamy and the presence of a 2Graduate School of Horticulture, Chiba University, synonymous SNP located in a conserved domain Matsudo, Chiba 271-8510, Japan within the C-terminal region. In the presence of the 3Leibniz Institute of Plant Genetics and Crop Plant recessive allele, transcription was suppressed, Research, D-06466 Gatersleben, Germany suggesting that the development of the lodicule fails when the Cly1 protein accumulates. The barley spike is composed of triplets (each with one central and two lateral spikelets) arranged 16. alternately at the rachis nodes. This arrangement of Structural variation in 5’ upstream region of spikelets is common in Hordeum species but unique in photoperiodic response genes, Ppd-A1 and Ppd-B1, Triticeae. All three spikelets of the six-rowed barley in wheat cultivars are fully fertile and able to develop into grains, but the lateral spikelets of two-rowed barley Nishida H, Yoshida T, Akashi Y, Kato K are reduced in size and sterile. Wild barley (H. vulgare Graduate School of Natural Science & Technology, ssp. spontaneum) is two-rowed and its arrow-like Okayama University, 1-1-1, Tsushima-Naka, Kita-ku, triple spikelets, a product of disarticulation of the Okayama 700-8530, Japan rachis, are result of an adaptive specialization that took place under native conditions. Some 10,000 years Three dominant alleles Ppd-A1, Ppd-B1, and Ppd-D1 ago, early farmers generated prototypes of cultivated confer photoperiod-insensitivity, while their recessive barley with non-brittle rachis. Later, six-rowed spikes alleles confer photoperiod-sensitivity in wheat. To that stably produced three times the usual grain date, no allelic variation have been found to explain number were produced during domestication. The the difference between photoperiod-insensitive and six-rowed spike 1 (vrs1) gene was isolated by a -sensitive alleles in Ppd-A1 and Ppd-B1, whereas the map-based approach. The vrs1 locus included HvHox1 large deletion in 5’ upstream region of Ppd-D1 allele, encoding a homeodomain-leucine zipper I–class that does not exist in ppd-D1 allele, is considered to protein, a potential transcription factor common in be an causal structural variation. In this study, Ppd-1 plants. Analysis of a plenty of Swedish mutant lines homoeoalleles from two Japanese cultivars suggested mutational events at regulatory regions of “Chihokukomugi” (genotype: Ppd-A1 ppd-B1 Vrs1 in addition to a variety of structural changes of ppd-D1) and “Abukumawase” (genotype: ppd-A1 encoded polypeptides by codon changes, altered Ppd-B1 Ppd-D1) were analyzed to identify such the splicing, new stop codons and frame shift. Reverse allelic variation. As for Ppd-A1, a 1085 bp deletion genetics using TILLING population was successful in was detected in the 5’ upstream region of Ppd-A1 the detection of new mutant alleles. In six-rowed allele, while it was not in ppd-A1 allele. This deletion barley cultivars, analogous mutational events were covers most part of the deletions in Ppd-D1 allele and detected, which indicate a multiple origin of six-rowed in tetraploid’s Ppd-A1 allele reported before. As for barley. Vrs1 is strongly transcribed in lateral spikelet Ppd-B1, a 308 bp insertion was detected in the 5’ primordia in the triple mound stage as shown by in upstream region of Ppd-B1 allele, while it was not in situ hybridization analysis. Down-regulation of Vrs1 ppd-B1 allele. This insertion site exists within the gene expression by inverted-repeat RNAi in deletion region of Ppd-A1 and Ppd-D1. Furthermore, transgenic barley lines provided evidence of the the approx. 100 bp region around the insertion site is biological function of Vrs1 on the determination of conserved among species, barley, rice, and spike row number. Evolutionary patterns of Vrs1 in Brachypodium, suggesting that this sequence is cereals will be presented.

examined traits in the Ae. tauschii accessions. 18. Geographically, significant latitudinal clines were Typologic and morphometric analysis of phytoliths detected for anther size, internode length and spike produced by wheat and barley length. Anther tended to be small in the eastern region. Internode also tended to be short, whereas spike to be Ball TB long in the eastern region. Based on these results, we 370 A JSB, Brigham Young University, Provo, UT discuss the patterns of intraspecific divergence and 84602, USA morphological diversification in the course of Ae. tauschii’s long dispersal from Transcaucasus to China. Solid deposits of amorphous hydrated silica are formed at specific intracellular and extracellular 20. locations in many plant taxa, including all taxa in Evolutionary process of six-rowed spike in Triticeae. These deposits of silica are called phytoliths, domesticated barley literally meaning "plant-rocks." Many plants produce phytoliths with morphological characteristics unique Saisho D1, Pourkheirandish M2, Komatsuda T2 to the taxon. When plant tissue decomposes, any 1Research Institute for Bioresources, Okayama phytoliths formed are released into the surrounding University, 2-20-1 Chuo, Kurashiki, 710-0046 Japan environment thus becoming microfossils of the plants 2National Institute of Agrobiological Science, 2-1-2 that produced them. Analysis of microfossil phytoliths Kannondai, Tsukuba 305-8602, Japan can provide information to researchers in a wide variety of disciplines, including, archaeobotany, The origin of six-rowed spike was one of the most paleoecology, phytogeography and systematics. This seminal evolutionary events in domesticated barley. It paper reviews current methodologies and results of has revealed that six-rowed spike morphology was typologic and morphometric analysis of wheat and caused by recessive mutation in Vrs1 locus, which barley phytoliths. It further presents paradigms for encoded homeodomain-lecine zipper I homeobox distinguishing between phytoliths produced by these gene (HvHox1). In order to investigate the taxa at the genus and species level. evolutionary process of the six-rowed barley, we performed comprehensive molecular polymorphic 19. analysis using wild and domesticated barley Natural variation in morphological traits in central accessions collected from all over the world. Eurasian wild wheat progenitor Aegilops tauschii Approximately 2 kb spanning the entire region of Coss. HvHox1 gene was sequenced in 136 wild barley core-collection distributed from ICARDA, 267 world Takumi S1, Morihiro H1, Nishioka E1, Kawahara T2, representative collection of domesticated barley from Matsuoka Y3 RIB, Okayama University, 41 North African 1Graduate School of Agricultural Science, Kobe accessions from NIAS and 13 Western Asian University, Rokkodai-cho 1-1, Nada-ku, Kobe accessions from USDA. Of the 224 six-rowed types 657-8501, Japan found in our sample panel, we identified four types of 2Graduate School of Agriculture, Kyoto University, vrs1 alleles, designed to vrs1.a1, vrs1.a2, vrs1.a3 and Muko 617-0001, Japan vrs1.a5. The frequencies of each allele in this panel 3Fukui Prefectural University, Matsuoka, Eiheiji, were 72 %, 10 %, 8 % and 9 %, respectively. A Yoshida, Fukui 910-1195, Japan haplotype genealogy was constructed using the diallelic substitution nucleotide polymorphisms Aegilops tauschii Coss. (syn Ae. squarrosa L.) is a observed in HvHox1 gene. Each of the four types of wild autogamous diploid wheat species. It has a wide vrs1 alleles was assigned to significantly separate natural species range in central Eurasia, spreading lineages on this phylogenic tree. The assessments of from northern Syria and Turkey to western China. Ae. geographic distribution of each vrs1 alleles showed tauschii is also known as the D genome progenitor of the contrasting patterns among the alleles. The two hexaploid bread wheat. The genealogical and alleles (vrs1.a1 and vrs1.a3) were spread to the geographic structure of variation in morphological worldwide range from Europe to East Asia. In contrast, traits was analysed for Ae. tauschii using a diverse the residual two alleles were locally distributed to array of 203 sample accessions that represented the either ‘Occidental’ regions (vrs1.a2) or ‘Oriental’ entire species range. Our previous studies reported regions (vrs1.a5). These evidence indicated that the that significant longitudinal and latitudinal clines were recessive vrs1 mutation events responsible for detected for spikelet size, and that there exist six-rowed barley were repeatedly occurred in process significant longitudinal and latitudinal clines in of the barley domestication and that the repetition of flowering time. Totally 12 traits including anther- and the migrations to westward and / or to eastward in the pistil-shape and internode length were used in this Old World could generate the geographic distribution study. Large natural variation was found for the all patterns of vrs1 alleles revealed in this study.

conservation, must serve a broad spectrum of users. A 21. proper determination of material is a prerequisite and Genetic Resources of Triticeae - cultivated species any evaluation represents added value to the collection. and genebank collections The Prague Gene Bank wild Triticeae collection has been gathered since 1985. There are nearly 1800 Knüpffer H accessions, belonging to 23 genera, and 133 species, Leibniz Institute of Plant Genetics and Crop Plant of which the most important annuals represent 9 Research (IPK), Corrensstrasse 3, D-06466 genera, 46 species and 1300 accessions. Infestation Gatersleben, Germany by pests and diseases was observed while collecting in wild populations. Nursery evaluation was directed Of the ca. 350 species and ca. 30 genera estimated for mainly to powdery mildew and rusts, cereal aphids the Triticeae, 111 species of 19 genera are either and viral infection. Infestation by aphids in field cultivated or useful wild species. An overview is given conditions was very selective towards accessions but of these species and their main uses. More than also species and genomes. Aegilops S-genome species 1,250,000 accessions of Triticeae are maintained in were the most infested by all observed aphids the world’s genebanks, thus comprising one-fifth of (Metopolophium dirhodum, Rhopalosiphum padi and the estimated world germplasm holdings. Based on a Sitobion avenae). Triticum boeoticum, T. monococcum survey of online information resources, Triticeae and T. timopheevii showed a high level of field accessions belonging to 35 genera (among them 12 resistance to viral diseases (WDV and BYDV) hybrid genera) and almost 300 species are compared with T. urartu and T. dicoccoides. When the documented to occur in almost 300 genebank plants were infested by leafhopper (Psammotettix collections. Summaries of the world holdings per alienus) artificially, all plants were eventually diseased. genus, species, regions where the genebanks are The vectors showed a high level of non-preference to located, and the largest collections of the major genera certain accessions. The accessions of Crithopsis are provided. For the larger genera (in terms of total delilaena and Heteranthelium piliferum showed the number of genebank accessions), i.e., Triticum, highest level of resistance to rusts, aphids and viral Hordeum, ×Triticosecale, Aegilops, Secale, Elymus infection. and Agropyron, the worldwide germplasm collections are surveyed with more detail. Existing international 23. or regional cooperation programmes, germplasm Quantitative variation of Revolver transposon-like databases and cultivar registers with pedigree gene in synthetic wheat and its structural information for these genera are briefly described. For relationship with LARD element the major cereal crops, core collections and genetic stocks collections are also mentioned. Due to its Tomita M1, Noguchi T1, Kawahara T2 growing importance as a model plant in genomics 1Molecular Genetics Laboratory, Faculty of research, the genus Brachypodium closely related to Agriculture, Tottori University, Tottori, Japan the Triticeae is also included in the surveys. The 2Plant Germ-plasm Institute, Graduate School of presentation aims at providing background Agriculture, Kyoto University, Muko, Japan information for plant breeders and crop plant researchers about the germplasm available in ex situ Revolver is a new multi-gene family dispersed like genebank collections, to make this wealth of material transposons in the Triticeae genomes. Revolver more easily accessible. encompasses 3041 bp and has 20 bp of terminal-inverted repeated sequences at both ends and 22. contains a transcriptionally active gene encoding a Annual wild Triticeae Gene Bank collection DNA binding-like protein. It is like Class II transposable elements. Revolver showed considerable Holubec V quantitative variation through the evolution of the Dept. of Gene Bank, Crop Research Institute, wheat-related species. The highest copy number of Drnovska 507, 16106 Praha 6 Ruzyne, Czech Revolver was 19,000 contained in Secale cereale (RR) Republic and the lowest was 2,000 in hexaploid wheat Triticum aestivum (AABBDD). Next, copy numbers were Gene bankers have a greater responsibility for determined in artificially synthetic hexaploid wheat material than bankers have for money. No crisis may lines between Aegilops squarrosa (DD) and tetraploid decrease the value of PGR except for improper wheat species, T. dicoccoides, T. dicoccum, T. maintenance. PGR must be saved in perpetuity and, ex carthlicum, T. turgidum, and T. durum (AABB). situ should back up possible, simultaneous in-situ Eleven lines out of 23 synthetic wheat lines showed conservation. The collection must be built with the significantly lower copies than the sum of their aim of gathering the largest genetic diversity possible. parental plants and 10 lines were equal to the sum, The Gene Bank collection, with the exception of suggesting that the polyploidy was negative stress

causing loss of Revolver. The members of Revolver be of high importance for more investigation in the family showed also structural variation especially in future. length. Revolver did not share any similarity with autonomous transposable elements. On the other hand, 25. the LARD LTRs, which were regarded as solo LTRs The tribe Triticeae Dumort. (Poaceae) of the non-autonomous retrotransposon LARD in barley, showed 60% homology to both 5' and 3' ends Yen C, Yang JL of some variants of Revolver. As to the region of 2 kb Triticeae Research Institute, Sichuan Agricultural between the ends, LARD LTRs lacked the region of University, Wenjiang 611130, Sichuan, China Revolver from the first exon to the middle of the first intron and resulted in non-coding sequences. LARD The tribe Triticeae is a group of taxa of Poaceae, LTR was situated as a structural part of a LTR which including very important cereal crops and very retrotransposon, while Revolver is a multi-gene family important forage grasses. Many useful and special consisting of the exon-intron structure. Evolutionally genes are kept in the gene pool of these grasses, which relationship between Revolver and LARD was are good for crop and forage improvement. So, these discussed. taxa have very high economic value. The taxonomy of Triticeae is a tool for organism recognition, an 24. understanding of phylogenetic relationships among Biodiversity of the D-genome species Aegilops organisms, and also a guide for germplasm utilization. tauschii in Iran The traditional morphological taxonomy was basically based on the comparative studies of morphological Saeidi H1,2, Rahiminejad MR1, Heslop-Harrison characters and geographic distribution. Morphological JS2 characters are phenotypes of an organism, which are 1Department of Biology, Faculty of Science, produced by functional reaction(s) between or among University of Isfahan, Isfahan, Iran dominant gene(s) and environmental condition(s). 2Department of Biology, University of Leicester, External morphological characters cannot reflect Leicester, LE1 7RH, UK internal recessive inheritance. Similar environmental conditions produce convergence natural selection and The biodiversity and phylogeography of Ae. tauschii different environmental conditions produced in Iran was assessed using morphological, molecular divergence natural selection. Therefore, traditional cytogenetic, SSR, inter-retroelement insertional morphological taxonomy cannot avoid some mistakes polymorphism (IRAP) and AFLP markers. From the in its determinations. Only cytogentic or molecular taxonomic point of view, all of the infraspecific taxa genomic analysis can avoid these mistakes. Genomic of the species were found in Iran. A high level of analysis can exactly determine genomic relationships genetic diversity between populations was revealed by and differences of species. According to recent all molecular markers, but different markers revealed investigations of genomic constitutions of tribe different level of diversity. SSR markers showed high Tritceae, we recognized 30 genera in this tribe. The levels of diversity without significant correlation with taxonomical changes and genomic constitution of the taxonomic groupings. These markers are suitable these genera are listed in this paper. for studying diversity within or between close populations. In addition to the high genetic diversity, 26. IRAPs showed a phylogeographic pattern within the Synthetic wheat production for wheat breeding subspecies and varieties. The genetic diversity of the species significantly decreases from north to east and Kishii M1, Mujeeb-Kazi A2 west of the Country, suggesting the patterns of spread 1Yokohama City University, Kihara Biological and centre of diversity of the subspecies. AFLPs Research Institute, Maioka-cho, Totsuka-ku, indicated the presence of two subgene-pools of the Ae. Yokohama, 244- Japan tauschii in Iran which was expected by other 2National Institute of Biotechnology & Genetic molecular markers but it was not clearly demonstrated. Engineering (NIBGE), Faisalabad, Pakistan There was no notable difference between diversity within the subspecies (subsp. strangulata and subsp. Synthetic hexaploid wheats (SH) are a new diversity tauschii) based on all molecular data, suggesting source for wheat improvement. Since 1980s, more occurrence of high gene flow between the two than 1,200 have been produced by randomly crossing subspecies in this region. In molecular cytogenetic the wild D genome diploid donor species (Aegilops studies, the in situ hybridization of tandem repeats tauschii, 2n=2x=14) of various ecological origins and (dpTa1) showed their distribution follows subspecific 51 durum wheat lines (Triticum turgidum; 2n=4x=28, taxonomy. Considering the needs for introducing new AABB). This huge genetic resource captures genetic characteristics and alleles for wheat improvement diversity of both parents and addresses a maxima of purposes, Ae. tauschii Iranian gene-pool is assumed to biotic and abiotic stresses. From the large number of

synthetic wheats produced suitable combinations have homoeologous group 1 chromosomes of wild species been identified for all three rusts, spot blotch, karnal of wheat. Several new alleles of HMW-GSs were bunt, Septoria tritici, barley yellow dwarf virus, identified and named. Dough strength of addition powdery mildew, drought, salinity, waterlogging, heat lines was evaluated for 3 consecutive years and 11 tolerance, improved bread making quality and some addition lines with strong dough were selected. micronutrients. Few targeted synthetics were also Rheological parameters of 6 addition lines revealed produced by utilizing T. dicoccum and T. dicoccoides better quality for bread-making. Among these selected accessions. Consistent with the contributions of the ‘D addition lines, Agropyron intermedium proteins genome’ exploiting the diversity of the ‘A and B showed best rheological characteristics followed by genomes’ also received attention generating Ag. elongatum and Ae. searsii. Cloning and AAAABB and AABBBB hexaploids. The first SH sequencing of HMW-GS genes of wild species from production phase (upto 2004) ended by generating selected addition lines showed great diversity among winter habit synthetics. Distribution of the above SH these genes. Wild species whose HMW-GSs gene germplasm to other researchers led to the development aligned with those of D-genome of wheat showed of the ITMI (International Triticeae Mapping much better quality characteristics. Substitution lines Initiative) population and the wheat microsatellite of chromosome 1D that eventually appeared from (SSR) map. addition lines showed very bad characteristic for From a global genebank survey conducted after bread-making quality. Preferential elimination of 2004, it turned out that 1/3 to 1/2 of the Ae. tauschii chromosome 1D was also observed during transfer of accessional holdings had been utilized for synthetic these alien chromosomes to selected regional cultivars wheat production. The survey generated information and thus needs considerable attention. on some missing geographical origin areas of Ae. tauschii. Thus in order to produce further new 28. synthetic wheats (post 2004) and fully exploit the Ae. Natural variation in grain selenium concentration tauschii diversity range in a trait targeted manner, derived from Israeli wild barley, Hordeum parental phenotypic and molecular characterization spontaneum studies were initiated to further increase the SH number. In this later targeted approach, emmer wheat Cheng J1, Wang F1, Yan J1, 2, Xiao T3, Ning Z3, (T. dicoccum) accessions were screened for heat and Chen G4, Nevo E2 drought tolerance for transfers to bread wheat. Using 1Institute of Triticeae Crops, Guizhou University, identified character positive accessions for these Guiyang, 550025, China complex traits in bread wheat improvement poses the 2Institute of Evolution, University of Haifa, Haifa, question whether genetic expression at the hexaploid 31905, Israel level could occur. Suppression of genetic expressivity 3State Key Laboratory of Environmental is a recognized constraint around practical utilization Geochemistry, Institute of Geochemistry, Chinese of SH wheats and warrants attention. Academy of Sciences, Guiyang, 550002, China 4Cold and Arid Regions Environmental and 27. Engineering Research Institute, Chnese Academy of Exploration of Triticeae resource for wheat end Sciences, Lanzhou 730000, China product quality improvement As the progenitor of cultivated barley, wild barley, Garg M, Tanaka H, Tsujimoto H Hordeum spontaneum is widespread in the Near East Laboratory of Plant Genetics and Breeding Sciences, Fertile Crescent. In Israel, wild barley ranges from Tottori University, Japan mesic Mediterranean areas to xeric northern and central areas of the Negev desert. Due to its rich Wild species of wheat are useful source of genetic adaptive diversity, H. spontaneum has proven to be an variation for crop improvement. They have been important genetic resource for crop improvement. In utilized for improving the tolerance of wheat to the current study, the grain selenium concentration different biotic and abiotic stresses. However, their (GSeC) originating from Israeli wild barley, Hordeum potential for wheat quality has not been much spontaneum was investigated. Ninety-four genotypes investigated. In this study, we used 177 disomic of wild barley from 9 populations were grown in the addition lines belonging to 17 wild species of wheat. central area of Guizhou province, China. Detection of These lines were screened initially by poly-acrylamide GSeC by hydride generation atomic fluorescence gel electrophoresis for identification of addition lines spectrometry (HG-AFS) method was carried out. The carrying seed storage proteins like obtained results showed that there are remarkable high-molecular-weight glutenin subunits (HMW-GSs), differences of GSeC between and within populations. low-molecular-weight glutenin subunits (LMW-GSs) GSeC among the 94 H. spontaneum genotypes ranged and gliadins from wild species. The loci of HMW-GSs, from 0 to 0.387 mg.kg- 1 , with an average of 0.047 LMW-GSs and gliadins were observed on mg.kg- 1 . The highest genotype of GSeC was No. 7

originated in Sede Boqer population, while the lowest so barley has the possibility of the functional foods. one was 25_1 originated from Atlit population. The mean value of GSeC of each population varied from 30. 0.010 to 0.105 mg.kg- 1 .The coefficient of variation Waterlogging and salinity tolerance in wild (CV) of each population ranged from 28 ％ (Mt. Hordeum species: physiological basis and prospects Hermon population) to 163％ (Caesarea population). for use in wheat improvement Spearman’s Rho correlations between GSeC and 1,2 1,3 ecogeographical data from 9 local populations in Colmer TD , Islam AKMR 1 Israel were tested; and there were significant Future Farm Industries CRC, The University of correlations between the GSeC and 12 of all 14 Western Australia, 35 Stirling Highway, Crawley, WA, 6009, Australia ecogeographic indexes. One way ANOVA indicates 2 significant difference of GSeC among five habitat soil School of Plant Biology, The University of Western types. These results displayed that there are obvious Australia, 35 Stirling Highway, Crawley, WA, 6009, Australia genotypic differences of H. spontaneum in selenium 3 uptake and accumulation ability. Therefore, wild School of Agriculture and Wine, The University of barley, H. spontaneum harbors considerable Adelaide, Waite Campus, Glen Osmond, SA, 5064, differences in GSeC, and those can be used for genetic Australia studies of barley selenium nutrition and further for crop improvement. The genus Hordeum contains many wild species that grow in diverse habitats (non-saline to saline, dryland 29. to wetland). We evaluated waterlogging and salinity Recent breeding objectives of hulled and hull-less tolerances, and selected physiological traits, in a range barley for food in Japan of Hordeum taxa. Species (and sub-species) with the X genome and several with the H genome were Yanagisawa T waterlogging tolerant, whereas those with the I National Agricultural Researcerh Center of Western genome, Y genome and the remaining H genome Region (WeNarc), Barley Research Team, Senyu species, were much less tolerant. Waterlogging 1-3-1, Zentsuji, Kagawa 765-8508, Japan tolerant species displayed traits for improved internal aeration of roots. H. marinum (X genome) was not In Japan, barley is used not only for alcohol beverages only one of the most waterlogging tolerant species, but food such as miso, rolled barley and barley tea. but was also salt tolerant. H. marinum maintains low + - Breeding efforts for genetic improvement for seed leaf Na and Cl concentrations, even when exposed to composition and grain quality, as well as increasing levels of salinity that can kill wheat. H. yield and introducing disease resistance, is important marinum-wheat amphiploids have been produced, breeding objectives of barley for food in Japan. firstly with Chinese Spring and recently with several Last year, two-rowed hull-less barley Australian varieties. The amphiploids show improved salinity and waterlogging tolerances, with better root “Yumesaki-boshi” was released. This cultivar is + - resistant to barley yellow mosaic virus and powdery aeration and regulation of leaf Na and Cl mildew, and is moderately resistant to scab. Since concentrations. The current amphiploids are in H. grain size is large, pearled grain is high whiteness; this marinum cytoplasm, and suffer some cytoplasmic cultivar is tentatively used for rolled barley. male sterility. Selected amphiploids will be Barley is susceptible to a browning reaction after transferred to wheat cytoplasm to restore normal heating and browning reaction is correlated to its fertility. The aim initially is to develop feed wheat polyphenol content. Proanthocyanidin is a kind of for moderately affected salt land. Furthermore, in a polyphenol, so proanthocyanidin-free mutants (ant 28) parallel program, 6 out of the 7 possible disomic were useful to reduce the browning reaction in boiled additions lines (1Hm, 2Hm, 4Hm, 5Hm, 6Hm and pearled barley. “Tochinoibuki” and “Shirotae-Nijo” 7Hm) of individual H. marinum chromosomes to (two-rowed hulled) were released last year. These Chinese Spring wheat have been produced. These cultivars will be used for retort-packed food. addition lines will be useful in determining the (1,3) (1,4)-beta-D-glucan (beta-glucan) is major chromosomal controls of salt- and waterlogging- components of polysaccharides in cell walls of barley tolerance in H. marinum, as expressed in a wheat endosperm. Beta-glucan is dietary fiber, is favorable background. for human foods because it lowers cholesterol. High beta-glucan contents (>10%) hull-less barley is 31. recently tested in the performance test for Biological Nitrification Inhibition (BNI) potential recommended variety. in Triticeae Recently whole barley grain barley and certain dry milled barley grain products are appropriate sources of Subbarao GV, Ishikawa T, Ito O beta-glucan soluble fiber for the health claim in USA, JIRCAS (Japan International Research Center for

Agricultural Sciences), Ibaraki 305-8686, Japan Barley (Hordeum vulgare L.) is an annual cereal grain and has been ranked number four among the staple Nitrification is the key process in the global nitrogen foods of the world. Today’s cultivated barleys are not cycle that results in the formation of nitrate through well adapted to high soil boron (B) although toxicity microbial activity, negatively affecting the availability to B has been known for some time. The Algerian of nitrogen to plants, causing low nitrogen-use landrace Sahara 3771 proved of being highly tolerant efficiency in agricultural systems. The ability of to B and thus, represents one of the most B-tolerant certain plants to suppress soil nitrifier activity by barleys currently known. It carries four quantitative releasing inhibitors from roots, a plant function, trait loci (QTL) conferring tolerance to toxic B termed “biological nitrification inhibition (BNI)”. conditions. One gene (Bot1) underlying the tolerance Using recombinant luminescent Nitrosomonas QTL on chromosome 4H of barley has recently been europaea to quantify BNI release, we found that identified and is a putative membrane-bound B wheat wild relative Leymus racemosus releases about transporter with similarity to bicarbonate transporters 20 times more BNIs than cultivated wheat and in animals; it functions as an efflux transporter to - effectively suppresses NO3 formation in soil. The move B out of the plant (Sutton et al. 2007; Science high BNI-capacity in L. racemosus is controlled by 318:1446 ff.). Bot1 was the first B tolerance gene to chromosome Lr+n, which was introduced into be identified in plants. In this work, we describe the Leymus-wheat chromosome addition lines via cloning of a second high B tolerance QTL in barley, inter-specific crosses. The high BNI release capacity mapping to the 6H B tolerance locus, Bot3. Using from Leymus was successfully expressed in wheat heterologous expression systems in yeast and Xenopus lines. Our recent results from barley indicate oocytes, we show here that this gene can facilitate the substantial genetic variability for BNI-capacity. transport of B across membranes. Higher tolerance to Unlike synthetic nitrification inhibitors that block only B in Sahara 3771 is mediated through lower transcript the AMO pathway, BNIs block both AMO and HAO levels of Bot3 in root tips of barley plants possibly enzymatic pathways of Nitrosomonas, making the owing to a repeat insertion into the promoter region of inhibitory effect more stable. Recent field evaluations Bot3 approximately 2 kb upstream of the start codon. of high-BNI capacity tropical pasture grasses Moreover, we observed lower shoot B accumulation in (Brachiaria spp.) demonstrated substantial reductions a rice (Oryza sativa L.) mutant possessing a point (>90%) in soil nitrification rates and nitrous oxide mutation in the orthologous rice transporter gene. emissions. Given the wide-spectrum of genomic Based upon our results we conclude that under high structure, the unique capacity for inter-generic gene soil B Bot3 entails lower shoot B accumulation and flow and the ability to form allopolyploid genomes, thus, effectually aids to higher B tolerance in Sahara Triticeae could offer a range of genetic tools/options 3771. to exploit and introduce sufficient BNI-capacity into wheat, barley and rye. Potential implications from 33. such an approach in facilitating a shift towards Genetic targeting of drought sensitive gene eibi1 of + sustainable NH4 -dominated cereal production wild barley (Hordeum spontaneum) systems are the subject of discussion. Chen G1,2, Pourkheirandish M1, Sameri M1, Wang 32. N1, Duan Z2, Shi Y2, Li C2, Nevo E3, Komatsuda T1 Reduced transcript levels at the Bot3 locus in 1Plant Genome Research Unit, National Institute of barley (Hordeum vulgare L.) confer increased Agrobiological Sciences, Tsukuba 305-8603, Japan tolerance to high boron supply 2Ecology and Agriculture Department，Cold and Arid Regions Environmental and Engineering Institute, 1,3 1 2 Schnurbusch T , Hayes J , Tyerman SD , Chinese Academy of Sciences, Lanzhou 730000, 1 1 2 Baumann U , Pallotta M , Ramesh S , Langridge China 1 1 P , Sutton T 3Institute of Evolution, University of Haifa, Mount 1 Australian Centre for Plant Functional Genomics, Carmel, Haifa 32905, Israel The University of Adelaide, Waite Campus, PMB 1 Glen Osmond, SA 5064, Australia The plant cuticle plays multiple roles in the protection 2 The University of Adelaide, School of Agriculture, of plant from various environmental stresses, Food & Wine, Australian Centre for Plant Functional especially the drought stress. The spontaneous cuticle Genomics, Waite Campus, Urrbrae, SA 5064, mutant eibi1 derived from Israeli wild barley was Australia characterized and fine mapped in the present study. 3 Present address: Leibniz-Institute of Plant Genetics eibi1 showed the highest relative water loss rate and Crop Plant Research (IPK), Genebank among the known wilty mutants, which indicates that Department, Corrensstr. 3, D-06466 Gatersleben, eibi1 is one of the most drought-sensitive mutants. Germany eibi1 was neither an ABA-deficient nor an ABA-insensitive mutant. The eibi1 leaves had a larger

chlorophyll efflux rate in 80% ethanol than the permits a convenient integration of any gene or gene wild-type leaves. The stomatal movement of eibi1 was fragment by GATEWAY-based recombination and normal. eibi1 had a more than 10 times larger comprises derivatives with various promoters. As a transpiration rate than the wild type in the dark. These result, constitutive as well as endosperm-, epidermis- lines of evidences indicated that eibi1 was defective in and egg cell-specific expression systems have been the cuticle. The scanning electronic microscope established. Further promoters and selectable markers (SEM) analysis revealed that the cuticular layer of of choice can be readily integrated due to the vectors' eibi1 leaf was broken, not continuously covering the modular configuration. Through coupling genetic leaf surface, which caused endless water loss. This transformation with haploid technology, we have been SEM result confirmed the conclusion that eibi1 was able to rapidly produce homozygous transgenic barley defective in the cuticle. The eibi1 caused pleiotropic lines. Based upon the enabling technology established, effects including semi-dwarf, low fertility, kinked we have embarked on numerous projects aiming to peduncle, drought sensitive, and hulless seed. eibi1, a functionally analyse candidate genes, and to generate monogenic and recessive mutant, was mapped to the transgenic barley lines with improved performance. pericentromeric region of chromosome 3H. To facilitate map-based cloning of EIBI1, we conducted a 35. high resolution mapping of eibi1 with 1682 F2 Map based cloning of dormancy QTL in barley individuals of Morex × eibi1. Barley-rice synteny was employed to search for ESTs for eibi1 mapping and to Sato K1, Matsumoto T2, Ooe N1, Takeda K1 identify candidate genes for eibi1. Comparison of the 1Research Institute for Bioresources, Okayama barley high-resolution genetic map and rice physical University, 2-20-1 Chuo, Kurashiki, 710-0046 Japan map revealed the inversion of the eibi1 region in 2National Institute of Agrobiological Science, 2-1-2 barley against its corresponding region in rice. Kannondai, Tsukuba 305-8602, Japan However, the collinearity within this inverted region was well conserved. eibi1 was located in a region of Seed dormancy is a trait of wild barley to escape 0.11 cM in barley genetic map which corresponding drought in the summer of arid condition. Dormancy in region in rice chromosome 1 physical map was 112.8 cultivated barley has different crop functions e.g. kb. delays malting process or prevents pre-harvest sprouting. Thus, the cloning of dormancy gene in 34. barley contributes to understand domestication Genetic engineering in cereals: Current process and optimize the trait for efficient uses. Rates technologies for the elucidation of gene functions of seed germination were used to evaluate dormancy on physiologically matured grain samples after dried Kumlehn J and stored frozen until use. Many genetic factors Institute of Plant Genetics and Crop Plant Research controlling seed dormancy has been reported as (IPK) Gatersleben, Germany quantitative trait loci. Among these loci, a locus on the centrometic region of chromosome 5H (Qsd1) has The development of reliable Agrobacterium-mediated been most frequently identified and showed largest transformation technologies for Triticeae cereals has effect among mapping populations. This QTL was greatly stimulated a variety of approaches to the identified on the EST map of Haruna Nijo (H. vulgare functional analysis of genes and the generation of ssp. vulgare) and wild barley H602 (H. vulgare ssp. genetically engineered breeding lines. In the Triticeae spontaneum) by both doubled haploid population and model species barley, three types of regenerable recombinant chromosome substitution lines (RCSLs). recipient cells or tissues have been successfully At least four QTLs are segregating in this cross. employed for the stable Agrobacterium-based RCSLs having only Qsd1 in the segment of wild DNA-transfer, each having specific advantages. While barley on the genetic background of Haruna Nijo were the use of immature zygotic embryos results in selected and a total of 919 B3F2 plant was used to exceedingly high efficiency of transgenic plant develop a high-resolution map of the QTL. By using formation, embryogenic pollen cultures enable us to rice genome and barley cDNA sequences, possible instantly produce true-breeding transgenic lines, barley transcript based markers were generated to map whereas isolated ovules facilitate the generation of the locus. By selecting heterozygous plants of B3F2, transgenic plants without use of a selectable marker. 4,792 B3F3 plants were genotyped to evaluate In addition to the barley platform established, recombinants between QTL and transcript based Agrobacterium-mediated transformation is also markers. After sequencing BAC clones of Haruna Nijo perfomed in winter and spring wheat as well as in rye. and H602, the possible mutations were identified To make the creation of plasmids for the between two gene sequences. Functional analyses of transformation of cereals more efficient, a set of these genes are underway. generic binary vectors for over-expression and RNAi-approaches has been generated. The vector set 36.

High frequency of karyotype variation revealed by We have explored the usability of retrotransposon sequential FISH and GISH in breeding population display (also known as SSAP method, of plateau perennial grass forage Elymus nutans Sequence-Specific Amplification Polymorphism; Waugh et al., 1997) to infer the relationships within a Dou QW1, Chen ZG1, Liu YA1, Tsujimoto H2 group of A-genome diploid wheat species Triticum 1Key Laboratory of Adaptation and Evolution of boeoticum, T. urartu, T. monococcum and T. sinskajae. Plateau Biota, Northwest Plateau Institute of Biology, The presence/absence data for 200 polymorphic SSAP the Chinese Academy of Sciences, Xining, 810008, bands representing the insertion sites of Sasanda and China BARE-1/WIS retrotransposon families were obtained 2Faculty of Agriculture, Tottori University, Tottori, for 49 accessions. For a given accession set the 680-8550, Japan majority (>90%) of bands that separated in a sequencing gel were polymorphic. The marker system Karyotyping was conducted to 12 random selected was also successfully tested on several representatives samples of grass forage Elymus nutans (StHY, 2n=42) of Triticeae and closely related taxa, including wheat which are widely planted in QingHai-Tibet plateau in species of different ploidy, Aegilops, Hordeum, Secale China by sequential FISH and GISH. The results and Avena species. Our modified SSAP procedure is showed that genome constitutions of 12 samples were relatively tolerant to DNA amount, and the bands can different from each other and nearly each chromosome be visualized by silver staining. Various approaches to included high frequent variants. The St genome shared retrotransposon display data analysis have been the highest number of variants of 42, 38.2% of total compared, including Dollo parsimony and several genome variants; while the Y genome shared the distance-based methods. Diploid wheat phylogeny lowest number of variants of 33, 30.0% of total. High issues are discussed. frequency of translocation was revealed that 7 of 12 samples, 58.3% of total, carried non-Robertsonian 38. translocation chromosomes between different Revealing seed storage metabolism from genetical genomes. 4 types of different translocations were genomic and systems biology approaches demonstrated. 4 samples carried the type 1 and 3 samples carried each of the other 3 types respectively. Sreenivasulu N, Pietsch C, Radchuk V, Röder M, High frequency of heterogeneous karyotype was Weschke W, Wobus U revealed that 7 samples, 58.3% of the total, showed Leibniz-Institut für Pflanzengenetik und that one or more than one chromosome lacked the Kulturpflanzenforschung (IPK), Corrensstraße 3, homologous chromosomes which shared the same or D-06466 Gatersleben, Germany similar repetitive sequence FISH patterns. In our ongoing efforts we developed large scale EST 37. resources (Zhang et al. 2004) and throughput gene Revealing genetic diversity in closely related expression profiling platforms in barley (Sreenivasulu A-genome diploid wheat species (Triticum et al. 2002, 2004, 2006), which have brought boeoticum, T. monococcum, T. urartu) by substantial progress in elucidating biochemical retrotransposon display pathways of barley seed metabolism in barley (Sreenivasulu et al. 2008a, 2008b; Neuberger et al. Konovalov FA1, Goryunova SV1, Shaturova AS1, 2008; Strickert et al. 2007). Very recent findings shed Fisenko AV1, Melnikova NV1, Kudryavtsev AM1, light on the interplay of many cellular and metabolic Goncharov NP2 events that are coordinated by a complex regulatory 1Vavilov Institute of General Genetics, GSP-1, network during barley seed development. Studying Gubkina 3, 119991 Moscow, Russia expression data of nearly 12,000 seed-expressed genes 2Institute of Cytology and Genetics, Lavrentiev ave. revealed, for instance, the participation of tissue- 10, 630090 Novosibirsk, Russia specific signaling networks controlling ABA-mediated starch accumulation (via SNF1 kinase and a set of LTR retrotransposons constitute a rapidly evolving transcription factors) in the endosperm and part of plant genomes due to their transpositional participation of ABA-responsive genes in establishing activity and susceptibility to deletions. embryo desiccation tolerance. In an attempt to gain Retrotransposon-based DNA markers are known to insight into the underlying genetic factors that govern provide a high level of polymorphism which can be differences in storage product accumulation we used for discriminating closely related accessions. If a compared changes in gene expression patterns during particular retrotransposon family does not have a seed development among 30 lines carrying defined strong target site preference, it can be assumed that an wild barley introgressions and dissected the candidate ancestral state (absence at a given site) is known for regulatory genes involved in altering the process of each inserted copy and that independent insertion storage product accumulation through genetical events into the same site are unlikely to occur. genomics approach. With the focus of using the

‘developing seed’ as model for systems biology Faculty of Agriculture, Tottori University, Tottori studies we further investigated transcriptional and 680-8553, Japan metabolic networks during grain development, developed models of the developing barley grain and The increase of wheat yield by breeding has slowed implemented magnetic resonance-based techniques. down in the last decade. It will not be easy to solve the These tools will be used to explore transgenic model food crisis which we are facing today without another systems for identifying key regulatory networks green revolution. Wild species of the tribe Triticeae related to seed quality traits. grow in a wide range of environments around the world and have large genetic variation. This variation 39. should have been efficiently used for wheat Hardness locus sequence variation in association improvement. But the method to introduce wild with grain quality in spring barley (Hordeum chromosome segments to wheat chromosomes has got vulgare L.) behind without using recent genomic information. New molecular breeding system needs to be Turuspekov Y1,4, Beecher B2, Darlington Y1, developed to introduce only small chromosome Bowman J3, Blake TK1, Giroux MJ1 regions to wheat using accumulated information of 1Dept. of Plant Sciences and Plant Pathology, 119 wheat and barley genomes and that of meiosis studied Plant BioSciences, Montana State University, in model organisms. The problem that occurs in Bozeman, MT 59717-3150, USA transferring genes to different genomes is the 2E.202, FSHN Facility East, USDA-ARS, Washington co-transfer of undesired deleterious genes located near State University, Pullman, WA 99163, USA the aimed gene. Application of fine molecular maps of 3Dept. of Animal and Range Sciences, 230C Linfield wheat and barley will make it possible to pin point the Hall, Montana State University, Bozeman, MT 59717, location of the desired gene and to transfer only a USA smallest portion to a wheat chromosome. 4current address: Inst. of Plant Biology and Homoeologous pairing may be induced by ph1b or PhI Biotechnology, NCB RK, Almaty 050040, Kazakhstan genes, but crossing and chromosome observation is necessary. Isn’t it possible to induce homoeologous Grain hardness has an important impact on the pairing by chemical and physical stimulation? In this end-use quality of cereal grains. The Hardness (Ha) presentation, I will discuss the problems and possible locus in barley contains the Hina, Hinb-1, Hinb-2, and solutions to these problems occurring when the Gsp genes and was shown to be associated with grain genetic variation of wild Triticeae species is used for hardness and dry matter digestibility (DMD) variation. wheat improvement. In this work, 73 spring barleys accessions selected for DMD variation were assessed for variation in DMD, 41. seed quality, and Hardness (Ha) locus component Development and cytogenetic analysis of Hordeum gene alleles. In order to determine whether Ha is chilense chromosome 4 introgression lines into associated with grain quality traits, we assessed the durum wheat relationship of the Ha locus in the presence or absence of head type (2 or 6-row) variation. To accomplish this, Prieto P, Ramírez MC, Martín A the barley Ha locus component genes (Hina, Hinb-1, Consejo Superior de Investigaciones Científicas Hinb-2, and Gsp) were sequenced from each accession (CSIC), Apartado 4084, 14080 Córdoba, Spain and sorted by prevalence. The most common Ha haplotype (HINA, HINB-1, HINB-2 alleles) was Wheat is one of the most important food crops in the present in 42 accessions with the remaining 39 world and the wild specie Hordeum chilense Roem. et dispersed over 24 haplotypes. Seeds from two rowed Schult is a valuable genetic resource for wheat accessions with the most common Ha haplotype were breeding. In fact H. chilense carries interesting significantly softer in grain texture (P<0.001) and had agronomic genes like resistance to the root-knot increased starch content (P<0.001) and DMD nematode Meloidogyne naasi on chromosome 1HchS; (P<0.05). The results indicate that selection for tolerance to salt on chromosomes 1Hch, 4Hch and 5Hch; individual Ha locus haplotypes may be useful in resistance to Septoria on chromosome 4Hch; and high modifying seed size, DMD, and starch content in carotenoid pigment content and resistance to common 2-rowed barley. bunt, both located on chromosome 7Hch. In breeding programs addition lines are useful as starting point for 40. the transfer of agronomic traits to wheat, but the Mutual interchange of genetic variation and introgression of specific characters requires the genomic information between wild and cultivated identification of recombinants that remove unwanted species in Triticeae regions of the H. chilense chromosome. With the aim of introgress resistance to Septoria in durum wheat we Tsujimoto H have developed inter-specific crosses between a

4Hch(4B) substitution line in bread wheat (T. aestivum, 43. 2n = 6x = 42, AABBDD) and the 4D(4B) substitution Salinity tolerance and sodium exclusion in genus line in durum wheat cv. Langdon (T. turgidum, 2n = Triticum 4x = 28, AABB). Backcrosses by durum wheat cv. Yavaros, which is a better adapted cultivar to our Shavrukov Y, Langridge P, Tester M agro-climatic region, have been also developed. A set Australian Centre for Plant Functional Genomics, of monosomic and disomic 4Hch(4B) chromosome University of Adelaide, Urrbrae, SA 5064, Australia substitutions and additions have been generated in durum wheat. Alien chromosome segments have been The ability of plants to exclude sodium from the shoot reduced in size by further spontaneous translocations is one of the major components of salinity tolerance. between chromosome 4A wheat and chromosome The mechanism of sodium exclusion is presented in 4Hch of H. chilense. We present the identification and genus Triticum and the considerable variability in characterisation of all these introgression lines in sodium exclusion within different species is durum wheat by multicolour fluorescent in situ demonstrated. The diploid species T. monococcum hybridisation. revealed a large (50-fold) variability in sodium exclusion in contrast to T. urartu, which was 42. significantly less variable (10-fold). These species Transcriptional landscape of malting barley, with the A genome are known to be salt sensitive, Haruna Nijo - using a custom-made oligoarray whilst T. (Aegilops) tauschii, a diploid species with the from cDNA sequences D genome, was very salt tolerant, but had only moderate variability in sodium exclusion (10-fold). Matsumoto T1, Ikawa H2, Fujii N1, Tanaka T1, The tetraploid species T. turgidum ssp. durum (both Sakai H1, Itoh T1, Nakamura S3, Sato K4 cultivated and landraces) and wild emmer T. 1National Institute of Agrobiological Sciences (NIAS), dicoccoides (all with the AB genome) showed a range Kan-non-dai 2-1-2, Tsukuba, Ibaraki 305 8602, Japan of variability in both salinity tolerance and sodium 2STAFF-Institute, Ippaizuka 446-1, Kamiyokoba, exclusion. The general pattern (from most sensitive Tsukuba, Ibaraki 305-0854, Japan and with highest Na+ accumulation) was as follows: 3National Institute of Crop Sciences (NICS), Research durum (cultivated) < durum (landraces) < wild emmer. Team for Barley and Wheat Biotechnology, Cultivated durum wheats had minimal or no variability, Kan-non-dai 2-1-18, Tsukuba, Ibaraki 305 8518, Japan whereas landraces of durum wheats had greater 4Research Institute for Bioresources Okayama variability, with two excellent genotypes having been University, Chuo 2-20-1, Kurashiki, Okayama identified which combine very low sodium 710-0046, Japan accumulation with very high salinity tolerance. Wild emmer was extremely variable. Hexaploid bread In order to understand barley transcriptome under the wheat, T. aestivum with the ABD genome, is known to normal and the stressed conditions, we have designed be more salt tolerant, having an effective mechanism 60-nucleotide oligomers from 3' end sequences from for sodium exclusion but only low variability. As a the ca. 36 000 full-length cDNA clones of two-rowed result of spontaneous hybridisation during their origin, malting barley, Haruna Nijo, and these were used for cultivated durum and bread wheats have a limited 36K Agilent-type oligoarray construction. Microarray gene pool and low variability in salinity tolerance. experiments indicated that about 1/8 of the genes Introgression of new genes from different progenitors tested were drastically influenced under either of the and relatives of Triticum that cope better with salt 42 stress conditions. Using clustering function of the stress can significantly improve salinity tolerance in GeneSpring analysis software, these affected genes are both cultivated durum and bread wheats. classified into several categories, such as co-expressed genes in root with salt (NaCl), dehydration, and ABA 44. stresses (such as ABA-inducible protein PHV A1 and Submergence tolerance in Hordeum marinum ACC oxidase), indicating there are common responsive pathway to these diverse stimuli. While we Malik AI1,2,3, Pedersen O1,4, Colmer TD1,2 found known annotation in the stress responsive genes, 1School of Plant Biology, The University of Western many other genes had no known functions in barley, Australia, 35 Stirling Highway, Crawley, WA 6009, rice, and Arabidopsis. Functional information revealed Australia in this study will not only endow novel understanding 2Future Farm Industries CRC, The University of to barley gene function, but also give some hints to Western Australia, 35 Stirling Highway, Crawley, elucidate biological functions of the orthologous genes WA 6009, Australia in grasses. This work is supported by the grant from 3Graduate School of Agricultural and Life Sciences, the Ministry of Agriculture, Forestry and Fisheries, The University of Tokyo, Yayoi 1-1-1, Bunkyo, Tokyo Japan (MAFF: TRC1008). 113-8657, Japan 4Freshwater Biological Laboratory, University of

Copenhagen, Helsingørsgade 51, 3400 Hillerød, Moreover, 1375 Pseudoroegneria, 442 Elymus, and Denmark 1798 Leymus EST simple sequence repeat (SSR) have been identified. PCR primers flanking these In the tribe Triticeae, the genus Hordeum contains EST-SSRs have been tested for amplification and many wild species that grow in a range of dryland to polymorphism on parental genotypes of existing or wetland habitats, and those growing in wet areas could proposed new mapping populations. A total of 375 naturally experience flash flooding. Submergence Leymus EST-SSR markers amplified one or more tolerance in three H. marinum accessions (H21, H90 segregating markers in the Leymus mapping and H546) was tested for 7 days during the vegetative populations. These Leymus EST markers were used to stage. Waterlogging alone (i.e. O2-deficient root-zone) align the Leymus linkage groups to the rice genome had no effect on whole plant growth of H21 and H90, sequence and identify homoeologous groups relative however, H546 was reduced to 82% of the aerated to wheat, barley, and other Triticeae genomes. control. Complete submergence of the shoots ([CO2] Bacterial artificial chromosome (BAC) genomic DNA of 200 µM in floodwater) reduced whole plant growth libraries representing approximately 6.1 haploid to 61%, 31% and 50% of the aerated controls of H21, genome equivalents of tetraploid Leymus have also H90 and H546, respectively. Underwater net developed and used to compare homoeologous photosynthesis by submerged leaves was about genomic DNA sequences containing the rice lax and one-third of that in air. Tissue sugars were reduced in maize barrenstalk1 orthogene. Likewise, genetic maps submerged plants, but not in waterlogged plants. are being constructed from interspecific hybrids of Porosity (% gas volume per unit tissue volume) in leaf caespitose Elymus lanceolatus and rhizomatous sheaths and roots enabled diffusive O2 transport from Elymus wawawaiensis. Other traits of interest in the floodwaters during darkness, and of O2 produced Elymus mapping populations include resistance to during photosynthesis during light periods, to the roots billbug (Parvulus parvula) feeding. in a hypoxic root medium. The adverse effects of submergence were reduced when dissolved CO2 in the 46. floodwater was increased. In summary, the study Plasmon analysis in the Triticum-Aegilops complex shows that some accessions of H. marinum are highly tolerant of waterlogging (i.e. shoots in air, roots in Tsunewaki K O2-deficient medium), and there is also diversity in 6-14-10 Kasagadai, Nishi-ku, Kobe, Hyogo 651-2276, tolerance to complete submergence. Japan

45. This paper reviews past results of our group on USDA-ARS gene, genomic, and trait discovery plasmon analysis of the Triticum-Aegilops complex. research in perennial Triticeae grasses Planned content of the talk is as follows: (1) Classification and characterization of the plasmon; Larson SR, Bushman BS, Mott IW, Wang RR-C, genetic effects to 47 plasmons on 21 wheat characters Jensen KB, Robins J, Jones TA were analyzed using 563 eu- and alloplasmic lines, in USDA-ARS Forage and Range Research Laboratory, which 12 common wheat genotypes and 47 Utah State University, Logan, UT, 84322-6300, USA Triticum-Aegilops plasmons were combined in all possible combinations, except one lethal combination, Long-term grass breeding efforts at the USDA-ARS and characteristics of the individual plasmons were Forage and Range Research Laboratory, traditionally clarified, based on which they were classified into 16 focused on perennial Triticeae range grasses, have groups. (2) Classification of the plastomes and been expanded to include the development of chondriomes; their genetic diversities were studied by molecular markers and genetic maps that can be used RFLP analyses of chloroplast and mitochondrial to investigate the genetic control of functionally DNAs, respectively. Based on the data, genetic important perennial grass traits. Genetic maps distances between all pairs of plasmons were constructed from tetraploid interspecific hybrids of tall estimated for both the plastomes and chondriomes, caespitose Leymus cinereus and relatively short based on which their phylogenetic trees were rhizomatous Leymus triticoides have been used to constructed. Sites of some plasmagene mutations were identify chromosome regions associated with plant speculated in the chondriome phylogeny. (3) Genetic height, growth habit, flowering, seasonal biomass diversity of the plasmon; combining the results of the accumulation, seed shattering, seed germination, above works, 48 plasmons were classified into 19 biotic and abiotic stress resistance, inflorescence traits types, to which plasmon symbols were designated. related to seed production, and forage quality (fiber, Characteristics of individual plasmon types will be protein, and mineral content). A total of 27,273 described. Differentiation of the plasmon at the expressed gene sequence tag (EST) unigenes have diploid level and maternal lineages of polyploid been isolated and sequenced from Pseudoroegneria species became evident. Origin of some polyploid (8780), Elymus (7212), and Leymus (11281). species were assumed to be more recent than some

other relatives, three examples being Timopheevi vs. Emmer in the tetraploid wheat, Ae. ventricosa vs. Ae. Sato K, Takeda K crassa of the Vertebrata, and Ae. triaristata vs. Ae. Research Institute for Bioresources, Okayama ovata of the Polyeides section, of the genus Aegilops. University, 2-20-1 Chuo, Kurashiki, 710-0046 Japan Referring to the genome symbols designated by previous workers, the genome-plasmon constitutions Cultivated barley was considered domesticated in of all Triticum-Aegilops species became elucidated, Middle East and distributed to East Asia. Abundant owing to which species relationships of this complex morphological variation, which was exotic to became clear in most part for both genome and Occidental barley germplasm, has been found in East plasmon, the first case at generic level throughout the Asian barley landraces. The cause of this plant and animal kingdoms. At the end, some morphological variation is still not understood. plasmon-related problems remained unsolved will be However, the uses of barley flour in Tibetan people pointed out. and steamed pearled grain in East Asian area might keep these variation unselected. The growing 47. condition of East Asian barley is also very different The barley coordinated agricultural project (CAP): from the one in Occidental barleys. An obvious integrating genomics with breeding different growing condition in East Asian barley is the cultivation in the paddy field after rice. The habit is Muehlbauer GJ still common in Japan, Korea and southern part of Barley CAP consortium and AGOUEB consortium China. These areas have rainy season in early summer caused by the Asian monsoon climate. There are some The barley coordinated agricultural project (CAP) is typical abiotic and biotic stresses due to these special an integrated project that leverages community growing conditions. Since barley cultivars are grown strength in genomics, statistics, computer science and in the paddy field where soil moisture is constantly breeding to enhance the efficiency of barley breeding. high, plants are continuously exposed to excessive The basic idea is to genotype and phenotype advanced water. There might be repeated severe selections in breeding lines from 10 U.S. barley breeding programs this condition for barley and we have some promising and to utilize the combined datasets to conduct tolerant accessions from East Asia. There is a special association-based analysis to identify quantitative trait semi-drawf barley genotype uzu which has been loci (QTL). During the first phase of the project we mainly distributed in western part of Japan. This group worked with our international collaborators and of barley shows tolerance to lodging in wet field mapped 2,943 single nucleotide polymorphisms (SNP). condition. Higher precipitation after flowering also From these mapped SNPs, an international SNP promotes fungal disease development in barley. genotyping platform was developed. To date, 2880 Fusarium head scab attacks barley spikes and damages breeding lines have been genotyped with over 3,000 grains by mycotoxin contamination. The promising SNPs and phenotyped for over 40 traits including source of resistance to this disease has been also found agronomic, disease resistance, and food and malting in East Asian barley germplasm. Some results of quality. The SNP genotyping has provided the genetic analysis of these stress tolerances will be opportunity to understand linkage disequilibrium (LD) presented. in U.S. barley breeding germplasm, and to understand the genetic relationships between breeding programs. Poster Presentation The model that fits the 10 U.S. barley breeding programs is seven populations with LD ranging from 1. 20-30 cM. All data have or will be deposited in a DNA markers: another tool in the toolbox centralized database called The Hordeum Toolbox. We used the combined genetic and phenotypic Bushman BS1, Barkworth ME2 datasets to detect QTL for a variety of traits including 1USDA-ARS Forage and Range Research Lab, Logan, malting quality, winterhardiness, dormancy, preharvest UT, USA sprouting, drought tolerance and resistance to 2Intermountain Herbarium, Utah State University, Fusarium head blight, spot blotch, net blotch, African Logan, UT, USA stem rust (Ug99), and common root rot. The success of identifying marker-trait associations has lead to the A scientific name has several meanings, and is often first stages of implementing marker-assisted selection. taken for granted by the larger scientific community. An example of the breeding utility of mapping However, if ambiguity exists for taxa, the scientific resistance to Fusarium head blight will be described. names may be incorrect and lead to erroneous conclusions with lasting impact. Nowhere is this a 48. greater problem than within the grasses, and Triticeae Features of East Asian barley and their genetic is not exempt. In this presentation we discuss analyses instances of taxonomic confusion, situations that give

rise to taxonomic confusion, and how the use of molecular markers can add another valuable tool to Gerus DE, Agafonov AV the toolbox of a taxonomist. We highlight situations Central Siberian Botanical Garden, SB RAS, where one or few genes are responsible for traits with Zolotodolinskaya st., 101, Novosibirsk, 630090, tremendous phenotypic differences, where lasting Russia hybrid populations give rise to new species names, where overlapping characteristics preclude diagnostic There are 72 species of the genus Elymus L. in the morphological characters but can be resolved with former Soviet Union according to S. Czerepanov DNA sequences, and where DNA sequences and (1995).It is difficult to determine an exact number of morphology suggest different taxonomic species for the vast area of due to contradictory data of interpretations. Our conclusion is that a molecular the authors. North-Eastern Kazakhstan is the area biologist working with grasses that have taxonomic closely related in respect of floristics with the ambiguity should have a plant taxonomist as a friend, Altai-Sayan region of Russia. Fourteen new Elymus and vice-versa. species were described from North-Eastern Kazakhstan over the last 17 years. Species from Asian 2. Russia and North-Eastern Kazakhstan can be Phylogenetic and phylogeographic analyses of characterized by the following levels of study: a) Hordeum murinum (Poaceae) species well studied by the methods of biosystematics with data on the genetic analyses of some Jakob SS, Blattner FR morphological characters; b) species studied by means Leibniz Institute of Plant Genetics and Crop Research of genetic markers (DNA, grain proteins, histone H1); (IPK), D-06466 Gatersleben, Germany c) species widely represented by herbaria; d) “species-phantoms” not studied, known only by The Hordeum murinum taxon complex consists of type-specimens, without mature seeds. The greatest diploid H. murinum subsp. glaucum and the two number of “species-phantoms” was described from polyploid subspecies murinum (4x) and leporinum (4x, North-Eastern Kazakhstan. Only 44 species of the 6x). The taxa are native in the Mediterranean and genus Elymus have StH constitution or unknown one. adjacent areas of Eurasia and northern Africa, and Among them, 20 species belong to level a, 22 species were introduced worldwide as weeds in man-made – b, 22 species – c, 18 species – d. Taxonomic habitats. For the polyploids it is unclear if they problems connected with study of “species-phantoms” originated via (segmental) allopolyploidization or are will be discussed in the poster. autopolyploids. AFLP analysis of individuals covering all taxa, 4. ploidy levels, and the entire native distribution Taxonomy and interspecific relationships of resulted in two clades, separating diploid subsp. Agropyron Grant. in Iran glaucum from all polyploids. Also extensive sequencing of cloned PCR products of the nrDNA ITS Hasheminejad N1, Saeidi H1, Yoosofi M2, region showed a clear separation of diploid from Rahiminejad MR1 polyploid cytotypes with only few diploid alleles 1Dept. of Biology, Faculty of Sciences, University of found within single polyploid individuals. As this Isfahan, Isfahan, Iran could be a result of recent hybridization, we 2Dept. of Biology, University of Payame Noor sequenced also a single-copy nuclear gene (TOP6). University, Najafabad, Isfahan This locus clearly showed that subsp. glaucum contributed to the polyploids. Phylogenetic analysis The genus Agropyron has been restricted to: A. revealed that the second parent belongs to the H. cristatum (L.) Gaertn. and A. desertrum (Fisch.) murinum genome group (Xu genome) but is nowadays Schultes with seven infra-specific taxa in the former in extinct. This latter species was most probably the Iran. Taxonomic status and inter taxa relationships of maternal parent of the polyploids, as their chloroplast the genus in Iran were examined using taxonomic, type is clearly different from the types occurring in cytotaxonomic and molecular (SSR markers) studies. extant diploid individuals. None of the markers used Based on x=7, with no populational variability the was able to separate subsp. murinum from leporinum Iranian materials of A. desertrum belongs to only although these taxa are morphologically and tetra- and that of A. cristatum to tetra- and hexa-ploid geographically clearly separated, which might indicate levels. Regarding the karyotyping features these two a young age of these taxa and cytotypes. species are very similar. Our SSR results showed that while the highest genetic diversity appeared among 3. the populations belonging to northern parts and the Levels of study of StH-genomic Elymus species of lowest within the eastern populations there no clear Asian Russia and North-Eastern Kazakhstan in distinction between the two species. The northern, connection with a problem of “species-phantoms” north-western and western populations showed very

similar genetic diversity. Based on the results of this rachis internodes had elliptic glumes. Ae. columnaris study it is suggested that A. desertrum to be lumped as was the extreme of the latter, while Ae. neglecta var. a subspecies into A. cristatum; the only Agropyron contorta was the extreme of the former. The variation species in Iran with eight infra-specific taxa in. showed a clear geographical cline from west to east, or coast to inland. Furthermore, 32 accessions of 5. tetraploid Ae. neglecta (9 of var. contorta and 23 of Genome constitution of Hystrix komarovii the other varieties) and 8 of Ae. columnaris were (Poaceae: Triticeae) artificially crossed with each other within and among taxa and geographical regions to clarify the degree of Zhang H-Q, Fan X, Huang Y, Sha L-N, Zhou Y-H reproductive isolation barriers. Intraspecific F1 hybrids, Triticeae Research Institute, Sichuan Agricultural especially those in Ae. neglecta, showed wide-ranged University, Wenjiang 611130, Sichuan, China fertility. The fertility in interspecific Key Laboratory of Crop Genetic Resources and neglecta-columnaris F1 hybrids was not lower than Improvement, Ministry of Education, Sichuan that in intervarietal hybrids in Ae. neglecta. From Agricultural University, Yaan 625014, Sichuan, China these results, we conclude that tetraploid Ae. neglecta and Ae. columnaris make a species complex in a The genome constitution of Hystrix komarovii was biological species with a wide genetic variation. examined by genomic in situ hybridization (GISH) and genome-specific RAPD assay. GISH of 7. Pseudoroegneria spicata (St genome) and Hordeum Using discriminant analysis to identify genomic bogdanii (H genome) probes confirmed the presence groups within the perennial Triticeae of the StH genomes in H. patula, but did not identified its presence in H. komarovii. The Ns and Ee genomic Rollo J1, Jacobs SWL2, Rashid A3, Barkworth, ME1 probes from diploids Psathyrostachys juncea and 1Intermountain Herbarium, Dept. of Biology, Utah Lophopyrum elongatum did not discriminate the State University, Logan, Utah, 84322-5305, USA genomes of H. patula, whereas they produced strong 2National Herbarium of New South Wales, Mrs hybridization signals on the two genomes of H. Macquaries Road, Sydney, New South Wales, 2000 komarovii. Results of genome-specific RAPD assay Australia were comparable with those of GISH, except that the 3University of Peshawar Botanic Garden, University Ee- and Eb-genome-specific RAPD bands were absent of Peshawar, Peshawar, Northwest Frontier Province, in H. komarovii. The results indicated that H. Pakistan komarovii might contain the Ns and Ee genomes, but lack the StH genomes. Further cytological data are As part of an investigation into the diagnosability of required for verifying the genome constitution of H. genomic groups, we scored 58 quantitative characters komarovii. on 219 specimens of perennial Triticeae with solitary spikelets in their genomic group. The specimens 6. represented 78 taxa and 13 different genomic groups. A biosystematic study in Aegilops neglecta – Ae. When all the specimens were used in the analyses, columnaris species complex 98% of the specimens were placed in the correct genomic group. Jackknife analysis was less successful. Ohta S, Fujita Y, Maesaka Y, Hattori M, Iwasaki R In Jack knife analysis, one specimen is excluded from Department of Bioscience, Fukui Prefectural the analytical phase and used as test case. Using this University, Fukui, Japan procedure, only 80% of the specimens were placed in the correct genomic group. There were some subsets Aegilops neglecta Req. ex Bertol. consists of that for which identification was reasonably successful. tetraploid and hexaploid cytotypes. Ae. columnaris For instance, the Australasian species, all of which Zhuk. is a tetraploid species closely related to the contain the W genome, were successfully tetraploid cytotype of Ae. neglecta. The aim of the distinguished from the non-Australasian species 94% present study is to clarify the morphological variation of the time. When the analysis was restricted to the and cytogenetic relationship in tetraploid Ae. neglecta StH and StY genomic groups (Elymus sensu stricto and Ae. columnaris. Spike characters of 131 and Roegneria, respectively), the jack knifed success accessions of tetraploid Ae. neglecta and 12 of Ae. rate was 73%. It dropped to 69% if the StYP columnaris collected from the whole distribution area (Kengyilia) and P (Agropyron) groups were included. were measured. The morphological variation was continuous but showed a significant correlation 8. between the two characters, the length of rachis Geographical distribution patterns of internodes at the base of spike and the shape of empty morphological characters in cultivated barley glumes: accessions with longer basal rachis internodes (Hordeum vulgare L.) inferred from botanical had circular glumes while those with shorter basal varieties

turgidum ssp. dicoccoides), six accessions of Knüpffer H domesticated emmer wheat (T. turgidum ssp. Leibniz Institute of Plant Genetics and Crop Plant dicoccum), and T. aestivum cv. Chinese Spring were Research (IPK), Corrensstrasse 3, D-06466 used as the references. Average number of alleles in Gatersleben, Germany Ae. speltoides (4.00) was larger than that in the wild emmer wheat (3.26). Estimated diversity index (H) The German Genebank at IPK has a collection of the was also slightly larger in Ae. speltoides (0.37) than genus Hordeum with 23,000 accessions, it is the 6th that in the wild emmer wheat (0.33). All 24 largest barley collection worldwide. Most of these microsatellite loci except for WCt20/21 were highly belong to cultivated barley, Hordeum vulgare. Many polymorphic and 92 accessions of Ae. speltoides were of the barley accessions represent landraces or classified into 80 plastotypes. Forty-eight and two traditional cultivars. Based on a number of plastotypes were identified in the wild and morphological spike characters, the accessions are domesticated emmer wheat, respectively. Phylogenetic being grouped into botanical varieties using the analysis revealed that all the plastotypes found in infraspecific classifications of Mansfeld (1950) and emmer wheat and those found in Ae. speltoides were Lukyanova et al. (1990) distinguishing 192 and 218 clustered into two separate plastogroups. This result varieties, respectively. Such a classification of indicated the clear differentiation of plastom between genebank material is very useful for purposes of the the wild emmer wheat and Ae. speltoides. The 80 maintenance of the collection. The characters used are, plastotypes found in Ae. speltoides were further among others, kernel row number (2 or 6), karyopsis grouped into two subgroups, suggesting an type (covered, naked), spike density (lax, dense, very intraspecific differentiation within Ae. speltoides. dense), glume width (narrow, broad), awn length and hoodedness (long, short, hooded, awnless), spike 10. colour (yellow, various colours), awn roughness Morphological variations of spike and the (rough, smooth), kernel colour (amber, various colours geographical distribution of subsection Emarginata – considered for naked barleys only). In addition, species, genus Triticum-Aegilops, close wild growth habit (spring, winter, intermediate) is also relatives of wheat considered. Using various external data sources with pedigree Ohta A1, Kawahara T2, Yamane K1 and breeding information, named cultivars were 1Grad. Sch. Bio. Env. Sci., Osaka Pref. U., Japan classified into “recent” and “traditional” (released till 2Grad. Sch. Agr., Kyoto U., Japan 1940). Forty-seven countries, or groups of smaller countries, were found to be represented by at least 15 To enhance use of wild relatives of cultivated plants as accessions of landraces or traditional cultivars with a genetic resource, it is important to understand the known botanical variety. The names of botanical genetic diversity and its genetic background. We varieties were translated into the corresponding investigated morphological variations of spike of four character combinations. Thus it is possible to show the subsection Emarginata species, Aegilops searsii, Ae. distribution of some morphological characters by bicornis, Ae. longissima and Ae. sharonensis. A total countries (or groups of countries). of 102 accessions from the collection maintained at the Laboratory of Crop Evolution, Plant Germ-plasm 9. Institute, Graduate School of Agriculture, Kyoto Intraspecific variation of chloroplast DNA in University were grown at the experiment field of Aegilops speltoides Kyoto University, Japan in 2007-2008. The result showed a correlation between the morphological Mori N1, Watatani H1, Ishii T2, Kondo Y1, variations of spike (spike length, awn length, spikelet Kawahara T3, Nakamura C1 density) and the geographical distribution 1Lab. Plant Genetics, Graduate School of Agricultural (Mediterranean site or inland site). Our result showed Science, Kobe University, Kobe, Japan the contradiction between the morphological 2Lab. Plant Breeding, Graduate School of Agricultural similarity of spike observed in this study and previous Science, Kobe University, Kobe, Japan studies of the molecular phylogenetic relationships 3Plant Germplasm Institute, Graduate School of among 4 species. It suggested the possibility of Agriculture, Kyoto University, Kyoto, Japan convergent evolution of spike morphologies in subgenus Emarginata. In order to clarify the genetic Chloroplast DNA polymorphism was investigated to factor that caused the differences in the spike clarify the intraspecific variation in Aegilops morphologies, we examined the relationships between speltoides Tausch. Allelic diversity at 24 microsatellite the spike morphologies and climate of collection sites loci was surveyed using 92 accessions of Ae. of the Emarginata accessions. Climate data was speltoides collected across its natural distribution area. obtained from Worldclim database. As a result, a cline Sixty-three accessions of wild emmer wheat (Triticum of spike morphology was found along geographical

gradient of climate change. Our results suggested that 648 Matsudo, Chiba 271 8510, Japan winter temperature affects spike morphologies. Spike of cultivated barley (Hordeum vulgare ssp. 11. vulgare) is composed of three spikelets at each rachis The regulatory network underlying the six-rowed node. In two-rowed barley, the central one is fertile spike in barley and the two lateral ones are sterile, whereas in the six-rowed type, all three are fertile. This characteristic Pourkheirandish M, Komatsuda T is determined by the allelic constitution at the National Institute of Agrobiological Sciences (NIAS), six-rowed spike 1 (vrs1) locus on the long arm of Plant Genome Research Unit, Kan-non-dai 2-1-2, chromosome 2H, with the recessive allele (vrs1) being Tsukuba, Ibaraki 305 8602, Japan responsible for the six-rowed phenotype. The Vrs1 (HvHox1) gene encodes a homeodomain-leucine Many important phenotypic traits are controlled by zipper (HD-Zip) transcription factor which is common networks of genes. The architecture of the Hordeum in plant kingdom. Here we show that the Vrs1 gene spike, which is characterized by the presence of three evolved in the Poaceae via a duplication, with a spikelets at each rachis node, is unique among the second copy of the gene, HvHox2, present on the short Triticeae. Because the two-rowed spike is universal in arm of chromosome 2H. Micro-collinearity and wild barley, it is likely to be the ancestral form, and polypeptide sequences were both well conserved that the six-rowed spike arose by natural mutation between HvHox2 and its Poaceae orthologues, but during the domestication process. Recessive alleles Vrs1 is unique to the barley tribe. The Vrs1 gene present at five independent genes are capable of product lacks a motif which is conserved among the producing a six-rowed spike. The six-rowed spike 1 HvHox2 orthologues. A phylogenetic analysis locus (vrs1 or Vrs1) is located on chromosome arm demonstrated that Vrs1 and HvHox2 must have 2HL. Wild barleys and two-rowed cultivars carry the diverged after the separation of Brachypodium dominant Vrs1 allele, while all six-rowed cultivars distachyon from the Pooideae. The loss of Vrs1 carry the recessive vrs1 allele. Over 90 induced vrs1 function both in natural variants, and in many induced mutants have been generated from two-rowed barley, mutants (including the total deletion of the gene) supporting the notion that cultivated six-rowed barley indicates that Vrs1 is non-essential for plant growth was derived from a two-rowed progenitor. Recessive and development. From these analyses it is suggested mutations (vrs2, vrs3, vrs4 and vrs5, which is that Vrs1 arose following the duplication of synonymous with int-c) have been induced at the four indispensable gene HvHox2, and acquired its new other loci, located on, respectively, chromosome arms function during the evolution of the barley tribe. 5HL, 1HL, 3HL and 4HS, but no cultivated six-rowed type is determined by a variant at any of these genes. 13. The recessive alleles at these four loci all act to Intraspecific variation in leaf shape-related traits enhance the development of the lateral spikelets, in a wild einkorn wheat species Triticum urartu although the degree of this enhancement depends Thum. upon the spikelet's position along the spike. Vrs1 encodes a member of the homeodomain-leucine zipper Morihiro H, Takumi S (HD-ZIP) I class of transcription factors, which binds Graduate School of Agricultural Science, Kobe to its gene target as a homo- or a heterodimer. The University, Rokkoda-cho 1-1, Nada-ku, Kobe six-rowed spike genes may form a gene network 657-8501, Japan involved in the control of lateral spikelet development. The expression of Vrs1 in vrs2, vrs3, vrs4 and vrs5 Einkorn wheat contains three species; cultivated backgrounds should be informative as to the identity Triticum monococcum and wild T. boeoticum and T. of some of the genes acting upstream of Vrs1 in this urartu. Triticum monococcum was domesticated from network. T. boeoticum. Triticum urartu is an A genome donor for the polyploid species of Triticum. Our previous 12. study showed that both nuclear and chloroplast The barley vrs1 gene evolved from duplication of a genomes of T. urartu were clearly differentiated from well-conserved HD-Zip I-class homeobox gene in those of T. monococcum and T. boeoticum, and that the the Poaceae T. urartu accessions were classified into two major haplogroups based on their chloroplast DNA Sakuma S1,2, Pourkheirandish M1, Matsumoto T1, variations. In this empirical study, we analyzed Koba T2, Komatsuda T1 intraspecific variation of eight leaf shape-related traits 1National Institute of Agrobiological Sciences (NIAS), using 30 accessions of T. urartu, 2 T. boeoticum Plant Genome Research Unit, Kan-non-dai 2-1-2, accessions and 2 T. monococcum accessions. Principal Tsukuba, Ibaraki 305 8602, Japan component analysis of the 8 leaf shape traits indicated 2Graduate school of Horticulture, Chiba University, that two subgroups were diverged in the T. urartu

population and that this intraspecific differentiation donors were not identified for their original diploid was corresponding to the two haplogroups based on species nevertheless they were at least within the the chloroplast DNA variations. The subgroup diversity of the genus Hordeum. Regarding the two diversification of T. urartu was largely caused by leaf subspecies of tetraploid, nucleotide sequences of 4x length. Significant difference of the flag and its lower ssp. murinum and 4x ssp. leporinum were highly leaf length was observed between the two subgroups. similar, further study using the different loci is One subgroup with the short leaf length contained necessary for the classification of the two subspecies. accessions mainly collected in Armenia and Lebanon, and another with the long leaf length included a lot of 15. accessions in Iran and Turkey. On the other hand, four The variation of SSR profiles in wild and cultivated or five different clusters could be divided in the T. barley urartu accessions revealed by the AFLP analysis of total DNA. Interestingly the subgroup diversification Turuspekov Y, Abugalieva S was not consistent with the clusters based on nuclear Institute of Plant Biology and Biotechnology, Almaty DNA variations. 050040, Kazakhstan

14. Nineteen SSR primers were analyzed to order to Allopolyploidy of the Hordeum murinum complex assess the genetic diversity of 68 barley cvs. and 13 indicated by a nucleotide sequence of cMWG699 wild populations of H. spontaneum K. from Israel, Turkmenistan, and Kazakhstan. In total 254 alleles Tanno K1,2,3, Bothmer von R4, Yamane K5, Takeda from 22 SSR loci were revealed using 6% K1, Komatsuda T2 polyacrilamide gel electrophoresis. The results are 1Research Institute for Bioresources, Okayama following: a) amount of polymorphism for wild barley University, Japan (He=0.71) was higher than for cultivated (He=0.63); 2National Institute of Agrobiological Sciences, Japan b) higher amount of genetic variation of cultivated cvs. 3Yamaguchi University, Japan from Kazakhstan in compare with European samples, 4Swedish University of Agricultural Sciences, Sweden and it was within a range of genetic diversity for wild 5Osaka Prefectural University, Japan barley; c) of the total genetic diversity of Hordeum, 69.83% was within populations, 9.28% between Hordeum murinum is one of the most widely populations within a species, and 20.89 between distributed plants in the genus Hordeum and is well species; c) the structure of genetic diversity for H. known as a nuisance weed in the world temperate spontaneum was 36.10% within populations, 50.16% zoon. This species is composed of three subspecies between populations of a region, and 13.74% between with three ploidy levels, namely ssp. glaucum (2x=14), regions; e) the level of polymorphism did not ssp. murinum (4x=28) and ssp. leporinum (4x=28 and positively correlated with sample size of wild 6x=42). Their morphological features, however, populations; and f) the genetic distance of wild resemble each other, they are frequently collectively populations did not relate with their geographic referred to as the “murinum complex”. distance. Results confirmed high potential of SSR Many cytological studies suggest allopolyploidy markers for genetic diversity analysis and efficient nature of the murinum complex while autopolyploidy identification of barley genotypes. is also suggested by an inter-specific study. The present study is aim to clarify the allo- vs. auto- 16. polyploidy status of the murinum complex based on A novel source of germplasm for the development molecular phylogenetic analysis, especially focusing of branched ear wheat on nucleotide variations and divergence of the polyploid genomes. Aliyeva AJ, Aminov NKH A single nuclear DNA locus, cMWG699, was Department Cytogenetics, Institute of Genetic analyzed, this nucleotide sequence has been used in a Resources, 155, Azadlig Ave., AZ 1106, Baku, series of phylogenetic studies of the Hordeum species. Azerbaijan PCR-RFLP analysis with HhaI and SspI for 80 H. murinum accessions revealed polymorphism between A novel source to produce branched-ear trait in durum diploid and polyploids, and double-digestion with the wheat was developed from the complex hybridization two enzymes showed polymorphism between tetra- among Triticeae species. After a synthetic hexaploid and hexa-ploids. Nucleotide sequence of sub-clones wheat (T. durum x Ae. squarrosa, 2n=42) was crossed clearly showed allopolyploid nature of the polyploid with rye (S. segetale Roshev, 2n=14), hybridization murinums. between the three generic incomplete amphidiploid The sequence study clearly indicated that one (Aegilotriticale, 2n=42) and common wheat (T. donor of the hexaploid was ssp. glaucum (2x) as has aestivum cv. Chinese Spring, 2n=42) was made. A line been suggested by cytological studies. The other ‘171ACS’ obtained from the latter hybridization was

further crossed with durum and common wheat gene pool to diversify genetic background of accessions. All the resulted F1 plants had normal durum wheat spikes. In F2 generation, all populations derived from the crosses between durum wheats and a line Taguchi J1, Kiribuchi-Otobe C2, Matsunaka H2, ‘171ACS’ segregated for normal and branched ears. Ban T1 The branched ear was not observed from the crosses 1Kihara Inst. Biol. Res., Yokohama City U., between ‘171ACS’ and common wheat. The Yokohama, Kanagawa 244-0813, Japan phenotype of branched ears differed from known 2NICS, Tsukuba, Ibaraki 305-8518, Japan branched ear wheat i.e. T. turgidum L. or T. vavilovii Jakubz, but closer to the morphology of T. vavilovii. It has been not cleared what kind of grain Each spikelet showed hierarchical non-terminated characteristics are related to quality of tetraploid structure. The branched-ear trait is controlled by a wheat gene pool. This study we examined genetic single recessive gene estimated by the above diversity for several traits associated with pasta Mendelian segregation of the F2 population and the making qualities among tetraploid wheat germplasms backcross population of the same cross. The result conserved in KIBR collection. indicated that branched-ear development was We analysed 96 acessions of tetraploid wheat suppressed by the factor derived from the D-genome including Triticum timopheevi, T. turgidum (convs. of common wheat. To confirm the existence of this durum, turgidum, dicoccum, orientale, pyramidale and factor in D-genome, crossings between '171ACS' and carthlicum) and D genome chromosome substitution synthetic tetraploid wheat (T. urartu x Ae. tauschii, lines of Langdon durum for each homoeologous A and AADD) are underway. B chromosomes. We identified genotype of HMW-gulutenin subunits by SDS-PAGE, genotype of 17. NAC gene which is considered to improve protein and Isolation and molecular characterization of three Fe, Zn content in wheat grains (Uauy et al. 2006) by novel HMW glutenin subunits from Aegilops PCR, measured seed protein content by Dumas tauschii combustion analysis (rapid-N®, elementar) for N/Protein, amylase content with AutoAnalyser An X1,2, Wang D2, Yan Y1 (Technicon), and content of grain yellow pigment 1Key Laboratory of Genetics and Biotechnology, (GYPC) with absorption spectrophotometer. College of Life Science, Capital Normal University, Regarding with genotype of HMW-gulutenin subunits, Beijing 100037, China T. timopheevi and T. turgidum conv. turgidum had 2The State Key Laboratory of Plant Cell and specially Glu-B1j, Glu-B1h allele, respectively. T. Chromosome Engineering, Institute of Genetics and turgidum conv. turgidum had lower protein content Developmental Biology, Chinese Academy of and T. timopheevi had higher protein content relative Sciences, Beijing 100101, China to T. turgidum conv. durum. Amylose content was about 27 % in almost of all line, but only T. High molecular weight glutenin subunits (HMW-GSs) timopheevi showed 22%. Regarding with NAC gene, T. exhibit abundant allelic variations in wheat related turgidum conv. turgidum and T. timopheevi carried species. Three novel HMW-GSs from Aegilops NAM-B2 addition to NAM-A1 and NAM-B1 allele tauschii were identified by SDS-PAGE, RP-HPLC and which T. durum carried. About GYPC, T. timopheevi MALDI-TOF-MS. Their complete coding genes were contain 1.4 times content relative to T. turgidum conv. amplified and cloned with AS-PCR primers, named as durum. The D genome chromosome substitution lines Glu-1Dx1.6t Glu-1Dx3t and Glu-1Dx5.2t, respectively. of Langdon durum showed broad range of variability The primary structure of the three subunits was highly on the traits. This study showed valuable diversity of similar to that of the previously reported Glu-1Dx the pasta making traits to improve durum wheat with subunits in wheat, but also displayed unique features. tolerance to biotic and abiotic stresses from the Particularly, Glu-1Dx5.2t subunit contained an extra tetraploid wheat gene pool. This work was supported cysteine residue in the repetitive domain in addition to to use KIBR germplasms contributed by NBRP the four conserved cysteine residues commonly found project. in the N-terminal domain of homoeologous x type subunits. The structural features of Glu-1Dx5.2t 19. resemble closely those of the good quality subunit Study of diversity and relationships of the D 1Dx5 that is frequently used in breeding wheat genome species of Aegilops–Triticum from Iran varieties with superior bread making properties. Based on this and previous studies, the origin and evolution Bordbar F1,2, Rahiminejad MR2, Saeidi H2, of 1Dx5 subunit in common wheat are discussed. Blattner FR1 1Leibniz Institute of Plant Genetics and Crop Research 18. (IPK), D-06466 Gatersleben, Germany Analysis grain characteristics of tetraploid wheat 2Department of Biology, University of Isfahan,

Isfahan, Iran weight glutenin subunit (HMW-GS) and low molecular weight glutenin subunit (LMW) As hexaploid bread wheat (Triticum aestivum) composition using SDS-PAGE and A-PAGE. The data contains a D genome, Aegilops species bearing the D indicated the prevalence of the null allele (95 %) and 1 genome can be used as potential sources of useful subunit (5 %) at the Glu-1A and four alleles, namely alleles to be incorporated into cultivated wheat. These 6+8 (33%), 7+8 (32 %), 13+16 (18%) and 20(9 %) Aegilops species posses a D genome component represented at the Glu-1B. Protein subunit Glu-1A1 derived from their diploid progenitor A. tauschii. We was correlated positively with improved dough studied 76 accessions of D genome species of the strength as compared to subunit null. On the Aegilops-Triticum complex from different parts of Iran chromosome Glu-1B subunit 6+8 was associated with together with 39 accessions of A. ventricosa, A. slightly stronger gluten type than 7+8 and 13+16, vavilovii, T. aestivum and A. juvenalis from other while subunit 20 was associated with weak gluten geographic regions. For estimation of genetic diversity properties. On the basis of electrophoretic separation within and between species, 24 highly informative D of gliadin fraction it was found that 49 genotypes genome-specific microsatellite markers were used, contained γ-45, 1 genotype γ-42 and 6 genotypes together with sequences from nuclear rDNA spacers another. Cultivars having the low molecular weight and a chloroplast intergenic spacer region. (LMW) glutenin allele LMW-2 (or gliadin band γ-45) Analyses of SSR diversity showed A. tauschii to generally gave stronger gluten than lines with allele possess a wide range of alleles, as is evident from high LMW-1 (or gliadin band γ-42). The combined better PIC values. Phylogenetic trees indicate A. tauschii to alleles at Glu-B1 (coded bands 6+8, 13 + 16, 7 + 8) be an old lineage with high genetic differentiation of and Glu-3 (patterns LMW- 2) showed linear the populations found in Iran. All our results revealed cumulative effects for dough strength. These results A. tauschii to consist of two different gene pools. could provide a more complete understanding of the Some of the accessions are more closely related to A. studied collections diversity on high molecular cylindrica, while other accessions of A. tauschii subunits and it will be useful to breeders who now clustered with the accessions of T. aestivum. The possess a tool to formulate crosses by choosing analyzed accessions of A. cylindrica showed low PIC varieties with appropriate characters. values and very low genetic distances compared to the other species. The results of this study confirmed that 21. the D genome of A. tauschii, A. cylindrica and T. Genetic variability in bread wheat (Triticum aestivum are different from the D genomes found in A. aestivum L.) of Slovakia based on polymorphism crassa, A. juvenalis, A. vavilovii and A. ventricosa, for high molecular weight glutenin subunits and indicate a cryptic taxon within A. tauschii, as the two groups defined by molecular methods are not in Mihálik D1, Šramková Z2, Medvecká E3, Horevaj accord with the already described subspecies of this V4, Šliková S1 taxon. 1Research Institute of Plant Production, Bratislavská cesta 122, 921 68 Piešťany, Slovak Republic 20. 2Slovak University of Technology in Bratislava, Estimation of quality of Triticum durum Desf. Faculty of Chemical and Food Technology- Institute wheat on the basis of gliadin and glutenin of Biochemistry, Nutrition and Health Protection, characterization Radlinského 9, 812 37 Bratislava, Slovak Republic 3Constantine the Philosopher University, Faculty of Gregová E1, Medvecká E2, Šramková Z3, Mihálik Natural Sciences, Department of Chemistry, Tr. A. D1 Hlinku 1, 949 74 Nitra, Slovak Republic 1Research Institute of Plant Production, Bratislavská 4Hordeum, s.r.o., Nový Dvor 1052, SK-92521 cesta 122, 921 68 Piešťany, Slovak Republic Sládkovičovo, Slovak Republic 2Constantine the Philosopher University, Faculty of Natural Sciences, Department of Chemistry, Tr. A. The genetics and biochemistry of high molecular Hlinku 1, 949 74 Nitra, Slovak Republic weight glutenin subunits (HMW GS) in wheat is very 3Slovak University of Technology in Bratislava, well known now. The results have shown that three Faculty of Chemical and Food Technology- Institute loci coding for HMW GS are highly polymorphic in of Biochemistry, Nutrition and Health Protection, nature without being influenced by the environment. Radlinského 9, 812 37 Bratislava, Slovak Republic Forty-six samples of common wheat varieties registered in the period 1976 – 2008 in Slovakia, kept The aim of the studies was the electrophoretic in the collection of the Genebank Piešťany, were characterization gliadin and glutenin proteins and surveyed to determine their high molecular weight evaluation of in kernels of Triticum durum Desf.. All glutenin subunits as separated by polyacrylamide gel 56 accessions originating from different geographical electrophoresis (SDS-PAGE). The high-molecular- areas of Europe were evaluated for high molecular weight (HMW) glutenin subunits, group of storage

proteins of wheat, are encoded by genes at three infected by F. graminearum at flowering time. All of complex loci (Glu-A1, Glu-B1 and Glu-D1) located on the Tri genes expression was promoted in a FHB the long arms of the homoeologous chromosomes 1A, susceptible wheat (Gamenya) more than resistance 1B and 1D respectively. Total proteins where extracted one (Sumai 3). We chose wheat genes associated with from crushed half grains and separated according to FHB resistance to introduce the multiplex PCR to the method of Wrigley (1992). The HMW glutenin examine interaction between wheat and F. subunits were classified according to the numbering graminearum gene expression. This method is system of Payne and Lawrence (1993). A total number cost-effective to the microarray analysis and high of 10 alleles were detected at all Glu-1 loci, 3 throughput then the real time PCR method. belonging to Glu-A1, 5 to Glu-B1 and 2 to Glu-D1 locus, respectively. The allele most strongly associated 23. with good quality, Glu-D1 subunits 5+10, was present New tools for the accessibility of the Spanish barley in 82,6 % of the studied wheat genotypes. The most core collection common banding patterns were Null (on chromosome 1A), 7+9 (on chromosome 1B) and 5+10 (on Igartua E2, Molina-Cano JL1, Gracia MP2, Casas chromosome 1D), respectively. The results of the AM2, Moralejo M1, Ciudad FJ3, Lasa JM2 study showed that 44 of the analysed accessions were 1IRTA, Av. Rovira Roure 191, E-25198 Lérida, Spain found to be homogenous. The other 2 consisted of at 2EEAD-CSIC, Avda Montañana 1005, E-50059 least 2 protein lines (phenotypes). Zaragoza, Spain 3ITACyL, P.O. Box 172, E-47071 Valladolid, Spain 22. Multiplex quantitative analysis for trichothecene The Spanish Barley Core Collection (SBCC) was genes expression of Fusarium graminearum causing conceive as a resource for research, and was head blight on wheat spikes assembled as a representative sample of the genetic variability present in the collection of over 2000 Miyazaki T, Ban T accessions held at the National repository for plant Kihara Inst. Biol. Res., Yokohama City U., Yokohama, genetic resources (CRF-INIA). The SBCC consists of Kanagawa 244-0813, Japan mostly inbred lines derived from landraces, adapted to Southern European conditions. It is a unique Fusarium graminearum attacks spike of Triticeae germplasm resource, holding a great amount of species and causes head blight (FHB) disease. It diversity, originated in an abiotically stressed results in contamination of trichothecene mycotoxins environment, isolated from mainstream gene pools, in the graitns as a global threat of food production and already studied to an unparalleled level among hygiene. Wheat resistance level to mycotoxin European landraces. It has been thoroughly studied for contamination varies among genotypes, however it over 30 agronomic, disease resistance, morphological, remains incompletely understood whether resistance and quality traits, revealing great potential to become to fungal invasion produces secondary effect or a source of novel alleles for adaptive traits and disease specific genes works on low level accumulation of the resistance. Its adaptation to Mediterranean climates mycotoxins. We applied multiplex quantitative gene also anticipates the existence of drought tolerance expression analysis for the trichotecene production traits, as a result of adaptation to the prevailing (Tri) genes of F. graminearum to reveal effective semi-arid conditions. Most information has been made wheat genotypes and germplasms to reduce mycotoxin available to the scientific community through the web production. We designed reverse chimera primers, site http://www.eead.csic.es/EEAD/barley/index.php, which consisted of 3’ side specific sequence of Tri which includes passport data, evaluation data for genes and universal primer sequence tail, to synthesize morphological, agronomical and disease resistance single strand cDNA from total RNA. Forward chimera traits. These data have been also summarized in a primers with a universal tail designed to amplify recent book published by the Spanish Institute for different size of each Tri gene added for 1st step PCR Agri-Food Research and Technology (in English): up to 3cycles to quantify the expression. Then, Spanish Barley Core Collection, JM Lasa universal primer sets, which forward primer was (coordinator), Monografías INIA: Serie Agrícola, labeled with fluorescent at 5’ end, amplified fragments 25-2008 (ISBN 84-7498-526-9). around 100 to 600bp length to be detected by the capillary sequencer (Genome Lab GeXP, BECKMAN 24. COULTER). As a result, Tri genes (Tri5, Tri6, Tri8, Triticum species in Georgia: diversity, conservation, Tri10, Tri11) expression was detected from 1.0 mg and taxa of special interest lyophilized hypae cultivated in a liquid medium and correlated with final myctoxin production level by Mosulishvili M1, Maisaia I2, Shanshiashvili T2, means of ELISA. Likewise, we could evaluate of Tri Akhalkatsi M1 genes expression level from a wheat spike which 1Ilia Chavchavadze State University. 32

Chavchavadze Ave., 0179 Tbilisi, Georgia information on genetic polymorphism degree within H. 2Tbilisi Botanical Garden & Institute of Botany. 1 spontaneum populations, as well as within revealed by Botanikuri str., 0105 Tbilisi, Georgia us H. ishnaterum Coss. variety and to study differences between spontaneum and vulgare varieties Georgia is one of the centers of evolution for many based on hordein electrophoretic components. The cereal crops and is rich with genetic resources of crop electrophoretic analysis of 61 barley genotypes was wild relatives. Earlier investigators of Georgian crop carried out according to modified method of F.A. plants include N. Vavilov, who gave great attention to Poperelya et.al. (2001). In an electrophoretic survey of the Transcaucasus in his research and visited Georgia eight H. spontaneum and one H. ishnaterum Coss. only 16 times. Georgian scientists V. Menabde and A. genotypes high polymorphism degree was observed Dekaprelevich studied phylogenetics of Georgian for hordein coding locus – HRD A and low endemic wheat species. The first evidence of farming polymorphism for HRD B and HRD F loci. civilization discovered in Georgia dates back to the Comparison of electrophoregram for hordein coding Mesolithic period. Wheat, barley and millet were loci in spontaneum and 52 vulgare (50 local and 2 already cultivated in Georgia in the Eneolithic and introduced) genotypes revealed 100 % polymorphism Early Bronze periods. Anterior Asia is the native place degree within HRD A -5, 6, 8 and from 6.2 to 8.8% of 12 wheat endemic (s.str.) species, 8 of them polymorphism within HRD A 1, 2 loci. The originate from the Transcaucasus, and among these polymorphism percentage for HRD B 1, 2, 3, 4 was endemics of the Transcaucasus, 5 species (s.str.) within the range of 6.2-62.5%. Considerable genetic originate from Georgia. Georgian endemic taxa are variation was noted for HRD F1 ranging from 6.2 valuable genetic material for modern selection. to100%, whereas for HRD F9 and HRD F 4, 6, 7 it Particularly, tetraploid (Triticum timopheevii) and varied from 69.2 to 81.8% and from 6.2 to 15.3% hexaploid (T. zhukovskyi) wheats are characterized by respectively. The results proved that in studied high resistance to diseases. T. carthlicum is genotypes the polymorphism was observed mainly in characterized by immunity to diseases, short growing HRD A and partly in HRD B hordein blocks. period and is cold-resistant. In the past this species was widely spread in Georgia in the Javakheti (Upper 26. Kartli) region. Georgia is very rich in agricultural Study of sequence polymorphism and genetic plants and it is a country of unique diversity of old diversity of sucrose-phosphate synthase genes in varieties and landraces of wheat and other cereals. bread wheat and its A, B and D genome Many varieties were lost during the last 20 years but progenitors some were saved in seed banks and farms; 14 species of wheat, 144 varieties and 150 forms were registered Sharma S, Röder MS till 1940-50s. The number of varieties and forms has Leibniz Institute of Plant Genetics and Crop Plant been catastrophically decreased and suffered rapid Research (IPK), Corrensstr. 3, 06466 Gatersleben, genetic erosion. These taxa must be rediscovered and Germany conserved both ex situ in seed and living plant collection and in situ in their original habitat in Sucrose phosphate synthase (SPS) is a main Georgia. regulatory enzyme involved in sucrose biosynthesis pathway in many crop plants, including wheat. Present 25. study was designed to study the structure of SPS Assessment of genetic diversity among Azerbaijan genes in wheat. SPS is organized in several gene barley genotypes (H. vulgare L.) based on Hordein families in the wheat genome. Genome specific alleles primers for SPS gene family II were designed for SPS genes using exon anchored primers that could amplify Geraybeyova N, Sadiqov H, Rahimova O, one or more introns, which are supposed to be more Sadiqova S, Karimov A, Mammadova N, Babayeva polymorphic than the exons. Each of the individual S, Abbasov M SPS loci of the three wheat A, B and D genomes was Genetic Resources Institute of ANAS, Baku, studied. Approximately, 4kb of each genome could be Azerbaijan compared and analysed. Detailed investigation of introns and exons was conducted. Sequences Although H. vulgare ssp. spontaneum (C.Koch) Thell alignment was conducted to search for the SNPs is prevalent plant in Azerbaijan widely distributed present in each of the three genomes. So developed almost all over the country, precise and detailed SPS genome specific primers are being used for the information on genetic diversity is lacking and little is PCR amplification in multiple accessions of T. known regarding population structure and geographic aestivum, T. turgidum subsp. durum, T. urartu, Ae. differentiation of this plant. Therefore, We analyzed H. speltoides and Ae. Searsi. Sequence polymorphism, spontaneum populations collected from different separately for each wheat species, will be grouped into regions of Azerbaijan with the objectives: to get distinct haplotypes. Genetic mapping and genetic

diversity studies in common wheat, for SNP sites will Karnataka, India also be conducted using either pyrosequencing or 4PARC, Pakistan direct sequencing. Linkage disequilibrium analysis 5Sydney University, Australia will also be performed. Synthetic hexaploid wheat (SHW) obtained by 27. crossing durum wheat (AB- genomes) and the wild Composition of high-molecular-weight glutenin relative Aegilops tauschii Coss. (D- genome) allowed subunits in European wheats a significant increase of cultivated wheat genetic diversity. Backcross derived lines (SBL), obtained by Šliková S1, Šramková Z2, Gregová E1, Mihálik D1 crossing SHW to elite bread wheat cultivars, have 1Research Institute of Plant Production, Bratislavská been already extensively used in wheat breeding. cesta 122, 921 68 Piešťany, Slovak Republic Higher genetic diversity was identified in cultivated 2Slovak University of Technology in Bratislava, emmer, Triticum turgidum L. subsp. dicoccon Faculty of Chemical and Food Technology- Institute (Schrank) Thell., compared to durum wheat, of Biochemistry, Nutrition and Health Protection, suggesting that using emmer wheat to develop new Radlinského 9, 812 37 Bratislava, Slovak Republic SHW could provide access to new alleles and traits. Moreover, cultivated emmer has proven to be a Seed storage proteins are considered to be usable valuable source of drought and heat tolerance and the markers for the studies of wheat genetic resources first SBL generated using emmer wheat showed High-molecular-weight glutenin subunits (HMW-GS), higher yield under drought-prone conditions in present in the endosperm of hexaploid wheat (Triticum Mexico, Pakistan and India compared to those using aestivum L.), are the main components of gluten, durum wheat. These lines were found to be genetically which is the main contributor to the rheological and diverse and distant from drought tolerant durum wheat bread-making properties of wheat flour. The objective based SBL and traditional bread wheat cultivars and of our study was to determine the HMW-GS breeding lines, confirming their interest as new composition of 57 wheat cultivars originated from six sources of diversity. A research project has European countries, kept in the collection of the consequently been initiated to i) discover novel Genebank Piešťany. Protein profiles were examined genetic diversity from emmer wheat, ii) develop new by sodium dodecyl sulphate polyacrylamide gel SHW from diverse emmer wheat accessions, iii) electrophoresis (SDS-PAGE). Fourteen different describe and compare diversity within emmer and alleles/allelic pairs were identified for the three durum based SHW to pyramid useful diversity and iv) glutenin loci studied, Glu-A1 (3), Glu-B1 (9) and develop new SBL by crossing the most diverse Glu-D1 (2). All together, twenty-five glutenin patterns primary emmer based SHW to local elite bread wheats were detected for HMW-GS, occurring at various from different origins and breeding programs frequencies depending on country of origin. The most (CIMMYT, India, Pakistan, Australia). common allelic composition was 0 (Glu-A1), 7+9 As part of this project a collection of 300 (Glu-B1) and 5+10 (Glu-D1). Allele “null” was the accessions from different geographical regions most frequent (50-100%) subunit at Glu-A1 locus in covering the emmer wheat area of distribution was wheat cultivars from all of the six countries. established at CIMMYT. Molecular characterization Consequently, the results also suggested that higher was carried out to understand the genetic structure of variability occurred at Glu-B1 locus compared to the species and identify accessions of interest. A Glu-A1 and Glu-D1. Rare alleles such as 7, 18, 20, 22 subset of genetically diverse accessions was created to and 13+16 were found at Glu-B1 locus. The most be further used for development of new pre-breeding frequent glutenin subunits encoded by Glu-D1 were wheat germplasm with enchanced drought and heat 5+10, except cultivars originated from Italy and Great tolerance. Britain, where subunits 2+12 were predominant (60-76, 9%). 29. Identification and mapping of QTLs for grain 28. protein content in common wheat Genetic diversity within Triticum turgidum L. subsp. dicoccon (Schrank) Thell. (cultivated Abugalieva S1, Abugalieva A1, Quarrie S2*, emmer) and its utilization in wheat breeding Turuspekov Y1 1Institute of Plant Biology and Biotechnology, NCB Zaharieva M1, Dreisigacker S1, Crossa J1, Payne T1, RK, Almaty, Kazakhstan Misra S2, Hanchinal RR3, Mujahid MY4, 2John Innes Centre, Norwich NR47UH, UK Trethowan R5 *Current address: Kraljice Natalije 39, 11000 1CIMMYT, Mexico Belgrade, Serbia 2Agharkar Institute, Pune, Maharashtra, India 3University of Agriculture Sciences, Dharwad, In this work we have identified quantitative trait loci

(QTL) for grain protein content (GPC) in 95 doubled screen F2 populations from the cross of İzgi2001 x haploid (DH) lines of a mapping population derived ES14, PI178383 X Harmankaya99 and Sönmez2001 x from a cross between the common wheat genotypes Aytın98 for bulk segregant analysis to find out Chinese Spring and SQ1 under the conditions of molecular markers linked to yellow rust resistance. Southeast Kazakhstan. The GPC of DH lines was The presence of polymorphic markers that is significantly different between rainfed and irrigated associated with yellow rust resistance may sites (P<0.05). In total, ten QTLs for GPC were found significantly enhance the success of selection for under the two treatments for moisture availability. yellow rust resistant genotypes in wheat breeding Two QTLs for GPC under rainfed conditions were programs. predicted to be novel in comparison with previous reports. The novel QTLs were mapped onto 31. chromosomes 2BS and 5DL in the population grown Regulation of transformation efficiency in under rainfed conditions. Closely-linked DNA polyploid cereals by type and number of selection markers were identified for the majority of mapped cassettes QTLs. The results could be implemented in a local breeding program for the improvement of wheat grain Bińka-Wyrwa A, Orczyk W, Nadolska-Orczyk A quality using marker-assisted selection. The study is Plant Transformation and Cell Engineering Lab, Plant the further contribution to our understanding of Breeding and Acclimatization Institute, Radzikow, regions of the wheat genome that contribute to the 05-870 Blonie, Poland control of GPC in common wheat. The aim of this study was to optimize the process of 30. selection and expression of transgenes in wheat and Investigations on yellow rust disease resistance by triticale plants obtained as the result of useful genes and markers in gene-rich regions on Agrobacterium-mediated transformation. We have had wheat chromosomes developed this transformation method for cultivars of both species. Currently we would like to explain the Aydin Y1, Cabuk E1, Mert Z2, Akan K2, Bolat N3, relationships occurring between the type and the Cakmak M3, Uncuoglu AA4 number of expression cassettes represented by two 1Marmara University, Faculty of Science and Letters, selection genes: nptII and bar. Ten pGREEN Department of Biology,34722, Istanbul, Turkey (www.pgreen.ac.uk) vectors contained single or two in 2Field Crop Research Institute, P.O Box: 226, tandem arranged selection cassette(s) were Lodumlu, Ankara, Turkey constructed. The cassettes were driven by the same or 3Anatolian Agricultural Research Institute, P.O Box: different promoters: nos, 35S or Ubi1. All vectors 17, 26001, Eskişehir, Turkey were electroporated to A. tumefaciens, strain Agl1 and 4The Scientific and Technological Research Council of used for transformation of two cultivars of wheat Turkey (TUBITAK), Marmara Research Center (Kontesa, Torka) and one cultivar of triticale (Wanad). (MRC), Genetic Engineering and Biotechnology Selection efficiency in these cultivars transformed Institute (GEBI), P.O Box: 21, 41470, Gebze, Kocaeli, with 35S::nptII, nos::nptII, 35S::nptII/35S::nptII and Turkey nos::nptII /35S:: nptII ranged from 0 to 5%. It was higher in combinations of Kontesa and Wanad High density genetic linkage and physical maps are transformed with one copy of nos::nptII or 35S::nptII available for wheat and there are many markers and than with two copies of the same selection cassette. important genes, including Yr which controlling The second cultivar of wheat, Torka could not be yellow rust disease resistance. One of the important transformed by any of three combinations containing gene-rich regions is present on the short arm of wheat nptII. The single plant was selected after homoeologous group 1 chromosomes. Resistant transformation with doubled cassette of 35S::nptII. (İzgi01, Sönmez2001, PI178383) and susceptible The same cultivars of wheat and triticale transformed (Aytın98, ES14, Harmankaya99) bread wheat with one or two cassettes of bar under nos, 35S or genotypes were screened by 43 molecular markers Ubi1 promoters gave the opposite results. Selection located at gene rich regions on physical maps of wheat efficiency of Kontesa, Torka and Wanad transformed 1B chromosome. Polymorphic band patterns were with 35S::bar, nos::bar, 35S::bar/35S::bar, nos::bar/ obtained with 17 out of 43 markers in wheat 35S::bar and 35S::bar/Ubi1::bar, nos::bar/Ubi1::bar genotypes. Furthermore, 7 Yr genes (Yr9-Yr15-Yr26- ranged from 0 to 6.2%. It was higher in combinations YrH52 on 1B, Yr7 on 2B, Yr17 on 2A, Yr18 on 7D) containing two copies of the bar cassette comparing were used to investigate the DNA sequence with the combinations containing one copy. The differences between wheat genotypes regarding to highest selection efficiency was observed in yellow rust resistance. Based on the obtained results, combinations of Kontesa, Torka and Wanad polymorphic markers and Yr genes sequence transformed by two in tandem arranged bar cassettes differences between wheat genotypes were used to driven by 35S and Ubi1 (35S::bar/Ubi1::bar) and/or

nos and Ubi1 (35S::bar/Ubi1::bar). assuming that the genetic background, Baronesse Acknowledgements: This work was financed by the barley, is sensitive to mutate easily by any Polish Ministry of Science and Higher Education, environmental/agrochemical force, we irradiated the grant PBZ-MNiSW-2/3/2006/31. cultivar twice by gamma rays of 100-300Gy, but almost nothing was selected at the station except some 32. chlorophyll mutations in M2 generations of the two Sequence variation of the 20th exon within PolA1 different irradiated materials although few mutants gene among Triticeae species were selected by re-screening of the on-farm grown M3 populations yielding still low frequency and very Buwan R1, Takahashi H1, Kato K2, Sato Y-i3, narrow spectrum of mutations. Intensive herbicide use Komatsuda T4, Nakamura I1 in the area may be the reason for the high frequency & 1Graduate School of Horticulture, Chiba University, wide spectrum of the mutants observed in Baronesse Matsudo, Chiba 271-8510, Japan barley fields at Washington State. 2Faculty of Agriculture, Okayama University, Okayama 700- 8530, Japan 34. 3Research Institute of Human and Nature, Characterization of growth and yield of transgenic Kamigyo-ku, Kyoto 602-0878, Japan wheat plants overexpressing vacuolar Na+/H+ 4National Institute of Agrobiological Sciences, antiporter genes Tsukuba 305-8602, Japan Miroshnichenko D, Poroshin G, Dolgov S PolA1 gene encodes for the largest subunit of RNA ‘Biotron’, Branch of Institute of Bioorganic Chemistry polymease I complex and is present as a single copy RAS, Pushchino, Moscow Region, Russia per plant genome. Because sequences of the 20th exons were highly polymorphic in Oryza and Petunia, Soil salinity is one of the environmental abiotic stress nucleotide sequences of PolA1 20th exons were factors limiting agricultural productivity in many analyzed for 13 Triticum, 14 Hordeum and 3 related region of the world. Soil salinity severely affects the species. Phylogenetic analysis of the sequences productivity of cereals, including wheat. One possible showed that Triticum and Hordeum species were mechanism by which plants could survive salt stress is distinctly separated into two major clades, except that to compartmentalize sodium ions away from cytosol. SS genome species of Triticum were clustered with To improve the plant growth and yield of wheat in Hordeum species. Although Secale cereale was saline soils, we have generated transgenic wheat grouped into Triticum clade, Dasypyrum villosum plants overexpressing vacuolar Na+/H+ antiporter belonged to Hordeum clade. genes using the biolistic-mediated transformation method. More than 50 independent transgenic lines 33. with expression of hvnhx2 gene isolated from barley High frequency and a wide spectrum of mutations (Hordeum vulgare L.) and agnhx gene isolated from in ‘BARONESSE’ barley fields salt-brush (Atriplex gmelini) were produced. Second generation of homozygous transgenic wheat plants Cagirgan AMI1, Ullrich SE2, Ozbas MO1 showed increased tolerance to salinity (100-200 mM 1Department of Field Crops, Faculty of Agriculture., NaCl). Several transgenic wheat plants with higher Akdeniz University, Antalya,Turkey levels of vacuolar Na+/H+ antiporter transcripts 2Department of Crop and Soil Sciences, Washington exhibited better biomass production at the vegetative State University, Pullman, WA, USA growth stage in saline condition These results demonstrate the feasibility of engineering salt Intensive agricultural systems increase agrochemicals' tolerance in plant. use such as fertilizer and pesticides. Lowering inputs such as no-till or reduced tillage is proposed as a way 35. to sustainable agriculture by lowering costs and so The flowering pathway under short day in barley increasing income from per unit area. However, the new production system generates problems of weeds Kikuchi R1, Kawahigashi H1, Ando T2, Tonooka T3, and hence increases use of amount of herbicides to Handa H1 control weeds. In 1997, Cagirgan and Ullrich observed 1Plant Genome Research Unit, National Institute of a high frequency and wide spectrum of mutations such Agrobiological Sciences, Japan as genetic male sterility, dense/lax spike, smooth awn, 2Institute of the Society for Techno-innovation of anthocyanin-less/more, eceriferum, etc in farmers' Agriculture, Forestry and Fisheries, Japan fields at Washington State, USA. Such huge 3Barley Research Subteam, National Institute of Crop variability would only be observable in a genetic Science, Japan background responding very well to a very efficient mutagen treatment. Considering this situation and Flowering is the essential events for reproductive

success in higher plants, and it is significantly affected has been used as cytoplasms donor to produce by photoperiod. Plants classified into three types, long alloplasmic lines in several crops: Triticum aestivum, day (LD), short day (SD) or day natural (DN) plants, Triticum durum, Secale cereale and Triticale. In all of according to photoperiod response. Barley (Hordeum them, alloplasmic lines display the positive vulgare) is a facultative LD plant, and its flowering is characteristics of stable sterility under different not only promoted under LD conditions but can also environmental conditions and no important occur even under SD conditions, although it is delayed. side-effects of the cytoplasm as developmental or FLOWERING LOCUS T (FT), which encodes a floral abnormalities, showing only slightly reduced protein with a PEBP domain, plays a central role in height and some delay in heading. Restorer of pollen flowering of Arabidopsis, and the FT homologues of fertility appear to be located at least in two H. chilense many species have been isolated and analysed. In this chromosomes, 1Hch and 6Hch. Considering the features study, five barley PEBP genes, HvFT1-3, HvTFL1 and displayed by this cytoplasm, it offers a real potential HvMFT1, were analysed to clarify their functional for the development of viable technology for hybrid roles in flowering. HvFT1 was expressed at phase cereal production. transition, and its transgenic rice showed most robust flowering initiation, suggesting that HvFT1 is the key 37. gene for flowering in barley. HvFT2 transgenic rice Agronomic traits and genetic determination of also showed early heading, but it was expressed only winter wheat lines (Triticum aestivum L.) with under SD conditions in barley. These results indicate multirow spike that the role of HvFT2 is limited to SD conditions, unlike HvFT1. HvFT3 was mapped to chromosome Martinek P1, Dobrovolskaya O2,3, Röder MS2, 1HL, the same chromosome of Ppd-H2, a major QTL Börner A2 for flowering under SD conditions. HvFT3 was 1Agrotest Fyto, Ltd., Havlíčkova 2787, 767 01 expressed in Morex carrying Ppd-H2, but not in Kroměříž, Czech Republic Steptoe carrying ppd-H2. Gene structural analysis 2Leibniz Institute of Plant Genetics and Crop Plant revealed that Morex has an intact HvFT3, whereas Research, Corrensstrasse 3, 06466, Gatersleben, most of this gene has been lost in Steptoe. These data Germany strongly suggest that HvFT3 is candidate for Ppd-H2, 3Institute of Cytology and Genetics, SB RAS, and barley has an adaptive mechanism to adjust Lavrentieva ave 10, 630090 Novosibirsk, Russia flowering even under unfavorable SD conditions using a combination of different FT-like genes. Two groups of spike morphology in wheat (Triticum aestivum L) are distinguished based on the number of 36. spikelets rising from individual nodes of the spike The potential of Hordeum chilense cytoplasm in the rachis: 1) normal spikelets (NS), where one spikelet development of CMS systems in Triticeae crops rises from one spike rachis node and 2) supernumerary spikelets (SS), where more than one fertile spikelet Martín AC rises from one rachis node. Attention is given to newly Departamento de Mejora Genética Vegetal, Instituto developed winter wheat lines with multirow spike de Agricultura Sostenible (C.S.I.C.), Apdo. 4084, (MRS) which is characterised by a higher number of E-14080 Córdoba, Spain spikelets with a few florets that rise from individual nodes of spike rachis. The yields of five different lines Cytoplasmic male sterility (CMS) is a maternally with MRS were compared with current standard inherited trait resulting from incompatibility between varieties (Akteur, Eurofit, Simila and Rapsodia) with nuclear and cytoplasmic genomes characterized by an NS on three different levels of nitrogen doses gradated inability to produce viable pollen but without effects by 30 kg.ha-1 from 90 to 150 kg.ha-1 and at the same on female fertility. CMS has been generated in several level of fungicide treatment; the previous crop was crop species by substituting their normal cytoplasm oilseed rape. Nitrogen was applied in the form of with the one from closely-related species, while limestone ammonium nitrate or urea at the three basic restoration of fertility is generally produced by the timings: regeneration, production and late introgression of nuclear genes from the cytoplasm topdressings. Each variant of treatment was three donor species. One of the limitations of this system is times replicated in 10-m2 plots. The average yield of the deleterious side-effects from the alien cytoplasm MRS lines in 2007 and 2008 was 96% at low, 99% at as kernel shrivelling, preharvest sprouting and a large medium and 101% at high intensity. The good yield reduction in seed germinability amongst others, reaction of lines with MRS to nitrogen dose could be together with male sterility instability. Hordeum explained by higher spike sink capacity of MRS, chilense, a diploid wild barley native to Chile and where higher number of kernels per spike can be Argentina which posses some traits potentially useful developed under higher growing intensities without for wheat breeding and which exhibits high disease infection. The MRS is determined by one crossability with others members of the Triticeae tribe, recessive gene which was designated mrs1. The MRS

trait was mapped by genotyping F2 populations using associated with the use of transgenic plants is the flow microsatellite markers. It was found that Mrs1 locus is of genes to plants in the environment. Potential risks located on the chromosome 2DS, about 10cM from of gene escape from transgenic crops through pollen the centromere. It will be necessary to improve the and seed dispersal have slowed down full utilization genetic background of the MRS lines by hybridisation of gene technology in crop improvement. Although no with important present wheat varieties. transgenic wheat varieties have yet been officially approved for extensive commercial cultivation in the 38. world, it is apparent that, as an important world cereal Breeding Triticale (X Triticosecale Wittmack) for crop, transgenic wheat varieties could be released into improved breadmaking quality the environment for commercial production, and probably within the near future. Martinek P In 2004-2008 crop-to-crop gene flow in spring Agrotest Fyto, Ltd., Havlíčkova 2787, 767 01 wheat was investigated. As the pollen source, a Kroměříž, Czech Republic transgenic homozygous line expressing recombinant DNA encoding two marker genes, such as bar and gfp Outcomes of breeding triticale (X Triticosecale genes was used. Among the marker genes available, Wittmack) for the improvement of breadmaking the bar gene, encoding phosphinothricin acetyl characteristics of grain for making food products from transferase and conferring resistance to herbicide yeast-leavened dough are presented. The breeding ammonium glufosinate, are particularly suitable for programme is based on the employment of triticale investigating gene flow in controlled experimental donors with translocations of chromosome 1R, to field trials. Green fluorescent protein marker gene gfp which the segments of wheat chromosome 1D were was used to monitor in vivo foreign gene expression at transferred. These translocations carry the glutenin different stages of experiments and for segregation allele Glu-D1d encoding HMW subunits 5+10. This study. Analyses of phenotypic and molecular data allele positively affects breadmaking quality. The showed that gene flow was greatly affected by the donors of chromosome 1R translocations were direction of the dominant wind and the distance provided by A. J. Lukaszewski (Univ. of California, between the targets. The overall hybridization rate of USA). Two-year data on grain quality (baking and transgenic seeds collected from non-transgenic rheological tests) in selected triticale lines from 2007 receptor plants growing at one meter from transgenic and 2008 harvests are given. Most grain quality plants varied year after year from 0,15% (2004) to parameters in these lines are close to those of winter 0,41% (2005). When non-transgenic receptor plants wheat classified into B category (bread quality). were grown in plots at two or three meter from Though the breadmaking quality of triticale has been transgenic plants, only a few seeds were produced partly improved, there has still been a problem of from fertilization with transgenic donor (less than higher α-amylase activity that causes a low falling 15% from total amount of transgenic hybrids). A number, which is usually markedly lower than in strong asymmetric distribution of the gene flow was wheat. We suppose that the triticale developed for detected in different parts of plots and the highest baking purposes will retain most properties typical for values (0.90%) were recorded following the direction this crop: better amino acid composition of storage of the dominant wind. protein in grain, better ability to use nutrients from soil, generally higher level of health status as 40. compared to wheat, resistance to some unfavourable Observation of pollen tube growth and molecular abiotic factors (particularly tolerance to aluminium mapping of Kr genes in common wheat-rye ions) and ability to produce good yields in regions hybridization where wheat growing is not profitable. It will lead toward extending the use of triticale in the field in Mishina K1, Manickavelu A2, Sato H1, Katsumata which wheat has been dominating until now. The M1, Sassa H1, Koba T1 perspective of newly developed translocations in 1Graduate School of Horticulture, Chiba U., 648 practical breeding is discussed. Matsudo, Chiba 271-8510, Japan 2Laboratory of Plant Genome Science, Kihara Institute 39. for Biological Research, Yokohama City U., Maioka Gene flow from genetically modified to cultivated 641-12, Totsuka-ku, Yokohama 244-0813, Japan wheat plants Crossability of common wheat with alien species, e.g., Miroshnichenko D, Poroshin G, Dolgov S rye, wild and cultivated barley, is known to be ‘Biotron’, Branch of Institute of Bioorganic Chemistry controlled by Kr gene family. Reproduction barrier RAS, Pushchino, Moscow Region, Russia caused by Kr genes decrease hybrid seed set, but molecular mechanisms are still unclear. In this study, One of the most discussed environmental effects we attempted to clarify the pollen tube behaviour of

rye on the wheat stigma by using confocal microscope, cassette and 5 for Golden Promise and 3 for Scarlett and to localize the QTLs controlling the crossability in with pMCG/CKX2 silencing cassette. Relative wheat-rye crosses by using molecular markers. In high activity of CKX enzyme, measured in roots of T1 crossability cultivar Chinese Spring (CS), pollen tubes seedlings (three repeats of bulk samples) from T0 of rye reached near the micropyle at one hour after plants transformed with pMCG/CKX1 silencing pollination, while in low crossability cultivar Hope, cassette ranged from 0.38 to 1.23 of the activity of the Mara and Cheyenne (Cnn), pollen tube growth was wild in vitro plants. Forty of 52 lines of Golden inhibited, and pollen tubes seldom reached to the Promise showed significantly lower enzyme activity. upper part of ovary. These results show that There was a positive correlation between enzyme reproduction barrier caused by Kr inhibits alien pollen activity and T0 plant productivity (the number of seeds tube growth. Also we conducted QTL mapping using per plant and the mass of thousand kernels) as well as F2 population consisted of 130 individuals derived the mass of the roots. Lower CKX activity influenced from a cross between CS and the chromosome higher plant productivity and bigger mass of the roots. substitution line of CS having chromosome 5B of Cnn, The relative enzyme activity measured in the roots of with 58 microsatellite makers. We found two QTL the lines transformed with the pMCG/CKX2 silencing regions controlling the crossability with rye on the cassette was similar to control and ranged from 0.88 to chromosome arms of 5BS and 5BL, between the 1.21 in all lines but one - this line showed a markers WMC47 and BARC142, and between significantly higher enzyme activity (2.37 ± 0.02). The BARC140 and GWM554, respectively. The effect of productivity of these T0 plants and the root mass from QTL region on 5BS on crossability with rye was their T1 seedlings were lower comparing with the greater than that of 5BL. This result coincident with control. Semiquantitative and quantitative analysis of the report by Tixier et al. (1998), who localized Kr1 expression of both genes subjected to silencing - and SKr genes on the long and short arms of HvCKX1 and HvCKX2 in different tissues of Golden chromosome 5B, respectively. We are now going to Promise and Scarlett plants have been done. Obtained test F6 RIL population derived from the same cross data were the basis for choosing the most appropriate with 91 molecular makers to identify the precise tissues to study expression/silencing of HvCKX in all locations of Kr1 and SKr genes on chromosome 5B. transgenic lines. This research is supported by Ministry of Science 41. and Higher Education; Research Project nr Posttranscriptional silencing of CKX genes, CZECHY/259/2006. regulating cytokinin level in barley by RNA interference 42. Genetic variability of MRP gene constituting Nadolska-Orczyk A1, Zalewski W1, Galuszka P2, ‘Qfhs.kibr-2DS’ QTL to reduce Fusarium Orczyk W1 mycotoxin accumulation among hexaploid wheats 1Plant Breeding and Acclimatization Institute, Radzikow, 05-870 Blonie, Poland Niwa S1, Kikuchi R2, Handa H2, Ban T1 2Departament of Biochemistry, Palacky University, 1Kihara Inst. Biol. Res., Yokohama City U.,Yokohama, Olomouc, Czech Republic Kanagawa 244-0813, Japan 2NIAS, Tsukuba, Ibaraki 305-8602, Japan Cytokinins are important plant hormones, which play an essential role in plant growth and development. The QTL ‘Qfhs.kibr-2DS’, which controls Fusarium Their level in different tissues is regulated by mycotoxin accumuration in wheat grains has been cytokinin oxidase/dehydrogenase enzymes. The reported on 2DS chromosome of hexaploid wheat cv. enzymes are coded by a family of CKX genes. Two of Gamenya and Nobeokabouzu-komugi. The gene for them, HvCKX1 and HvCKX2 were isolated in barley multi drug resistance-associated protein (MRP) was (Galuszka et al. 2004). We studied the effect of tracked down as a candidate gene constituting HvCKX silencing in plants transformed with hpRNAi Qfhs.kibr-2DS (Handa et al. 2008). In this report, we binary vector pMCG161 (http://www.chromdb.org/ demonstrated genetic variability of the MRP gene mcg161.html). Two vectors containing the fragments structure among various hexaploid wheat germplasms. of HvCKX1 and the conservative fragments of The MRP genes consist of 11 exons on wheat HvCKX2 in sens and antisens orientation were homoeologous chromosome 2A, 2B and 2D were constructed. The silencing cassettes were introduced sequenced fully or partially from the BAC library of into two barley cultivars: Scarlett and Golden Promise hexaploid wheat cv. Chinese Spring (CS). To examine by Agrobacterium-mediated (pMCG/CKX1 silencing genetic variability among wheat germplasms with cassette) and biolistic (pMCG/CKX2 silencing different level of mycotoxin accumulation, eight sets fragment) transformation. The numbers of selected, of genome specific primers for chromosome 2D to putative transgenic lines obtained were: 52 for Golden amplify several genomic regions of the MRP gene on Promise, 1 for Scarlett with pMCG/CKX1 silencing chromosome 2DS were designed based on the

sequence of introns, which connected to exsons. Two University of Tsukuba, Tsukuba, Ibaraki 305-8577, types of the MRP gene structure were detected; Japan Chinese wheat type (ex. Sumai 3 and CS) and the 3Research Institute for Bioresoueces, Okayama others (ex. Gamenya and Nobeokabouzu-komugi) University, Kurashiki, Okayama 710-0046, Japan with SNPs and in/del polymorphism. One SNPs marker for the second exon region was developed to We reported a novel mutant gene for β-glucanless identify the MRP genotype for various wheat grain (bgl) in barley (Breeding Science 59:47-54, germplasms collected from the world showing various 2009). In this study, we analysed the effects of the bgl resistance level to Fusarium head blight, and we will gene on growing properties and quality characteristics discuss association between MRP genotype and using near-isogenic line (NIL). We developed a NIL phenotype response on mycotoxin accumulation. This with bgl using a Japanese two-rowed cultivar work was supported by a grant from the Ministry of ‘Nishinohoshi’ as a recurrent parent. The NIL grew Agriculture, Forestry and Fisheries of Japan normally in the field and showed normal seed fertility, (Genomics for Agricultural Innovation, TRC-1005). but had shorter culms and awns than the recurrent parent. The NIL had significantly longer spikes than 43. the recurrent parent; however, the number of spikes Alien glutenin subunits expressed in common was much reduced. Low yield in the NIL was wheat endosperm affect on the composition probably caused by the reduced number of spikes. The NIL showed partial chlorosis in the leaves and awns, Tanaka H, Arakawa T, Tsujimoto H probably due to a pleiotropic effect of the bgl gene. Laboratory of Plant Genetics and Breeding Science, Heading date was 9 days later in the NIL than in the Faculty of Agriculture, Tottori University, Japan recurrent parent. β-glucan content in vegetative organs was much reduced in the NIL. The reduction of culm The seed storage proteins of common wheat (Triticum and awn lengths in the NIL might be attributed to the aestivum L.) are responsible for the ability of flour to reduction of β-glucan in the cell walls. The content of form cohesive dough, required to make strong dough arabinoxylan in the grains of the NIL was significantly such as bread. However, the narrow genetic base of higher than the recurrent parent. The NIL showed hexaploid wheat has limited the allelic combinations completely floury texture in the endosperm. that are available for the improvement of Microscopic observation revealed that the NIL had bread-making quality. Screening for good-quality thinner cell walls in the endosperm. The NIL had soft subunits in wheat related wild species of wheat is and friable grain texture, probably due to the thin important for improving the bread-making quality of endosperm cell walls. wheat. Changes of glutenin composition of wheat endosperm were investigated in 6 alien chromosome 45. addition lines. Each line has group-1 chromosome Virus-induced gene silencing of P23k in barley leaf from wheat related wild species expected to carry reveals morphological changes involved in glutenin genes. SDS-PAGE and two-dimensional gel secondary wall formation electrophoresis have resolved a total of 15 protein bands and 39 protein spots from alien chromosomes, Kidou S1, Yokota S2, Yoshida K3 respectively. Agropyron elongatum disomic addition 1Cryobiosystem Research Center, Faculty of line (DAL) had 11 protein spots with the greatest Agriculture, Iwate University, Morioka 020-8550, numbers among them. We also found changes of the Japan spot density from common wheat in DALs. One spot 2Faculty of Pharmaceutical Science, Nagasaki changed to higher density in Aegilops umbellulata International University, Sasebo, Nagasaki 859-3298, DAL, whereas two spots changed to lower density in Japan Hordeum chilense DAL. These observations might 3Taisei Corporation Technology Center, Nase344-1 show post-translational control and/or changes of gene Totuska Yokohama, 245-0051, Japan expression due to presence of additional alien chromosome in common wheat. P23k is a monocot-unique protein that is highly expressed in the scutellum of germinating barley seed. 44. Previous expression analyses suggested that P23k is Characterization of a β-glucanless mutant in involved in sugar translocation and/or sugar barley metabolism. However, the role of P23k in barley physiology remains unclear. Here, to elucidate its Tonooka T1,2, Aoki E1, Yoshioka T1, Taketa S3, physiological function, BSMV-based virus-induced Kiribuchi-Otobe C1,2 gene silencing (VIGS) of P23k in barley leaves was 1National Institute of Crop Science, Tsukuba, Ibaraki performed. Expression and localization analyses of 305-8518, Japan P23k mRNA in barley leaves showed up-regulation of 2Graduate school of Life and Environmental Sciences, P23k transcript with increased photosynthetic activity

and the localization of these transcripts to the vascular bundles and sclerenchyma, where secondary wall 47. formation is most active. VIGS of the P23k gene led Population structure of central Eurasian wild to abnormal leaf development, asymmetric orientation wheat progenitor Aegilops tauschii Coss. of main veins, and cracked leaf edges caused by mechanical weakness. In addition, histochemical Mizuno N1, Yamasaki M2, Matsuoka Y3, Kawahara analyses indicated that the distribution of P23k in T4, Takumi S1 leaves coincides with the distribution of cell wall 1Graduate School of Agricultural Science, Kobe polysaccharides. Considering these results together, it University, Rokkodai-cho 1-1, Nada-ku, Kobe is proposed that P23k is involved in the synthesis of 657-8501, Japan cell wall polysaccharides and contributes to secondary 2Food Resources Education and Research Center, wall formation in barley leaves. This work was Kobe University, Kasai, Hyogo 675-2103, Japan supported partly by the Genomic Agricultural 3Fukui Prefectural University, Matsuoka, Eiheiji, Innovation Program of MAFF (TRC1006). Yoshida, Fukui 910-1195, Japan 4Graduate School of Agriculture, Kyoto University, 46. Muko 617-0001, Japan Spontaneous amphidiploidization via unreduced gametes is a universal phenomenon for Triticum Aegilops tauschii Coss. (syn Ae. squarrosa L.) is well turgidum - Aegilops tauschii hybrids known as the D genome progenitor of hexaploid bread wheat. Ae. tauschii, a wild diploid wheat species, has Zhang LQ1,2, Liu DC1,2,3, Zheng YL1,2, Yen Y4 a wide natural species range in central Eurasia, 1Triticeae Research Institute, Sichuan Agricultural spreading from Turkey to western China. AFLP University, Dujiangyan city, 611830, China analysis using total 122 accessions of Ae. tauschii was 2Key Laboratory of Crop Genetic Resources and conducted to clarify the population structure of this Improvement, Ministry of Education, Sichuan wide spreading wild wheat species. Phylogenetic Agricultural University, Yaan 625014, China analysis revealed that there were two major lineages, 3Northwest Plateau Institute of Biology, Chinese L1 and L2, in the Ae. tauschii population. Bayesian Academy of Science, Qinghai 810001, China population structure analysis based on the AFLP data 4Department of Biology and Microbiology, South showed that L1 and L2 were significantly divided into Dakota State University, Brookings, SD 57007, USA six and three subgroups, respectively. Only four out of the six L1 subgroups were diverged from western Polyploidy has been found to be very common in habitats, Transcaucasus and Azerbaijan-Iran Caspian plants. Polyploidy can be formed via the duplication region, to eastern habitats such as Pakistan and of genomes, either of the same genomes Afghanistan. Other subgroups including L2 were (autopolyploid) or of diverged genomes with limitedly distributed to the western region. Three homoeologous relationships (allopolyploid). The major haplogroups (HG7, HG9 and HG16) were formation and union of unreduced gametes in triploid identified in the Ae. tauschii population based on the F1 hybrid between Triticum turgidum L. (2n=4x=28, chloroplast genome diversity (Matsuoka et al. PLoS AABB) and Aegilops tauschii Coss. (2n=2x=14, DD) ONE 3: e3138, 2008). HG7 accessions were widely might have played an important role in the distributed to both L1 and L2. HG9 accessions were spontaneous origin of hexaploid wheat (T. aestivum L., restricted only to L2, whereas HG16 accessions 2n=6x=42, AABBDD). Experimental studies have belonged to L1, suggesting that HG9 and HG16 were indicated that unreduced gametes by meiotic formed from HG7 after the two-lineage differentiation restitution lead to the fertility of F1 hybrids of T. of the nuclear genome. turgidum and Ae. tauschii and then resulted in spontaneous production of hexaploid wheat. However, 48. previous studies on unreduced gametes were only Molecular markers for systematic characterization involved a few T. turgidum genotypes or their of low molecular weight glutenin subunits in combinations with Ae. tauschii. In the present study, common wheat (Triticum aestivum L.) triploid F1 plants from 115 wide hybridization combinations were obtained by crossing 76 T. XF Zhang1,2, DC Liu1, WL Yang1, JZ Sun1, DW turgidum lines with 24 Ae. tauschii accessions without Wang1, HQ Ling1 and AM Zhang1 embryo rescue. It was indicated that a very high 1State Key Lab of Plant cell and Chromosome frequency (88.70%) of combinations was partial engineering, Institute of Genetics and Developmental fertile and spontaneously produced hexaploid wheats. Biology, Chinese Academy of Sciences, Chaoyang The wide spread of spontaneous amphidiploidization District, Beijing 100101, China 2 in the T. turgidum - Ae. tauschii F1 hybrids found in Graduate School of the Chinese Academy of Sciences, our study supported the multi-origin hypothesis for the Beijing 100049, China hexaploid wheat by some wheat scientists.

Wheat is the staple food for human beings. Wheat flour can be processed into bread, other baked goods Landraces are useful genetic resources for wheat (cakes, cookies, crackers etc.), pasta and noodles, and (Triticum aestivum L.) improvement. However, their a wide range of other products. Low-molecular-weight potentials have not yet been fully investigated. glutenin subunits (LMW-GS) are among the major High-molecular-weight glutenine subunits (HMW-GS) polymeric protein components of wheat flour. Their encoded by Glu-A1, Glu-B1, and Glu-D1 loci are ability to form intermolecular disulphide bonds with major components of wheat seed storage protein, and each other and/or with high-molecular-weight glutenin are greatly related to rheological and bread-making subunits (HMW-GS), is crucial for the formation of properties. In this study, we aimed to disclose the glutenin polymers, which determine the technological geographical distributions of HMW-GS alleles using properties of wheat flour (Weegels et al., 1996; Gupta 360 wheat landraces collected from Asia, Europe, and Shepherd, 1988). LMW-GS is encoded by a Russia, and Africa. HMW-GS composition in each multigene family, displaying high polymorphic protein landrace was determined by sodium dodecyl complexes. Recently, some locus-specific primers of sulphate-polyacrylamide gel electrophoresis LMW-GS genes have been developed (Wang., 2009; (SDS-PAGE) based on the method by Payne and Ikeda et al., 2006; Kawaura et al., 2005; Long et al., Lawrence (1983). Consequently, three, ten and 13 2005; Van Campenhout et al., 1995). Systematic alleles were identified for Glu-A1, Glu-B1, Glu-D1, molecular markers specific for each LMW-GS allele respectively. In all the regions, the major genotype for would have importance and practical applications in HMW-GS was “null (Glu-A1), 7+8 (Glu-B1), 2+12 wheat breeding. In this study, we developed (Glu-D1)”. The ratios of these three alleles, however, systematic molecular markers, referring to the differed among regions. For Glu-A1 locus, the LMW-GS genes isolated from Xiaoyan 54, Glenlea frequency of 2* allele was larger than 1 allele in the and Norin61 and the sequences published in Genbank. eastern side of the Caspian Sea, South Asia, and The specificity of these markers, validated in a large Russia, but opposite relationship was found in other collection of wheat varieties from China, was regions. Glu-B1 locus exhibited great geographical discussed. variations in frequencies of 7+9 and 17+18: the frequencies of these two alleles in South Asia and 49. Russia were quite different from those in their Genetic diversity of high molecular weight respective neighboring regions. The allelic variation of glutenine subunits in wheat landraces Glu-D1 locus was quite large in the west coast of the Caspian Sea, especially in Azerbaijan, although Nishinaka M1, Okumoto Y1, Kato K2, Kawahara T3, extremely small in Iraq, Afghanistan and Iran. A novel Tanisaka T1 allele at Glu-D1 locus was identified from one of the 1Graduate school of Agriculture, Kyoto University, Azerbaijan landraces. To clarify the distribution of this Kyoto 606-8502, Japan novel allele, additional 297 landraces originated in the 2Faculty of Agriculture, Okayama University, regions near the Caspian Sea were investigated, but Okayama 700-8530, Japan we found no landraces with this allele. 3Graduate school of Agriculture, Kyoto University, Muko 617-0001, Japan

Fig. 1. 6th ITS participants in front of the Clock Tower Centennial Hall, Kyoto University.

Fig. 2. Oral presentation in International Conference Hall of the Clock Tower Centennial Hall, Kyoto University.

66 Wheat Inf. Serv. 108: 67-70, 2009. www.shigen.nig.ac.jp/ewis

Meeting Reports

Wheat science at the Sixth International Triticeae Symposium

Nikolay P. Goncharov* Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, 10 Lavrentyev ave., Novosibirsk 630090, Russia *Corresponding author: N. P. Goncharov (E-mail: [email protected])

The Sixth International Triticeae Symposium was wheat’s studies are connected with its agronomic held at the Kyoto University (Kyoto, Japan) from characters. Despite this research there has been May 31 to June 5, 2009. Earlier, Helsinborg, little commercialisation. Logan, Aleppo, Cordoba, Prague hosted the A key challenge facing biologists in the Symposium. The main goal of International twenty-first century is the preservation of Triticeae Symposiums is the tribe Triticeae Dum. biodiversity. Review of K. Tsunewaki “Plasmon It includes the most important cereal crops (wheat, analysis in the Triticum-Aegilops complex” [1-46, barley, rye) and some fodder grasses (Dasypyrum, presentation numbers, hereafter, see Kawahara Elymus, Agropyron and so on). A wild annual grass (2009)] deal with classification and Brachypodium distachyon (L.) Beauv. is a good characterization of the wheat and goat grass model for molecular-genetic (genomic) plasmon, which they classified into 16 groups. investigation in the tribe. Mary Barkworth (1992) Author analyzed genetic effects of 47 believes that the history of botanic studies of the Triticum-Aegilops plasmons on wheat alloplasmic Triticeae is repleted with changes in generic line traits, in which they were combined. concepts, caused by taxonomists' struggle with the Chloroplast DNA polymorphism was investigated task of representing its highly reticulated history to clarify the intraspecific variation in plasmon within the hierarchical classification system donor of B and G wheat genomes Aegilops demanded by scientific nomenclature. speltoides Tausch by N. Mori, H. Watatani, T. Ishii, The 118 participants from 20 countries took Y. Kondo, T. Kawahara, C. Nakamura (2-9). The part in the symposium. There were near 50 oral 80 plastotypes found in Ae. speltoides were further presentations and 49 posters. They were grouped grouped into two subgroups, suggesting an in 4 sections, namely Systematics and Phylogeny, intraspecific differentiation. Domestication and Evolution, Biodiversity and M. Garg, H. Tanaka, H. Tsujimoto (1-27) Genetic Resources, Genomics and Breeding. The believe that wild species of wheat are useful presented reports allowed to receive more source of genetic variability for crop improvement. information about the researches in systematics, Their gene pool has been utilized for improving genetics, plant physiology and breeding and the tolerance of wheat to different biotic and preservation of wheat’s biodiversity in different abiotic stresses. M. Muramatsu (1-6) reviewed gene banks. Like its predecessors, the 6th studies of wild Triticeae species indigenous to Symposium raised great interest in wheats and Japan, their features and results of hybridization their relatives. The 20 oral presentations and 30 with wheat and barley cultivars. Much attention posters were connected with studies of them. was given to introgression from three species, Wheat species are not good models for Elymus tsukushiensis, E. humidus, and E. ciliaris. molecular-genetic investigations. Common wheat Aegilops species bearing the D genome can be (Triticum aestivum L.) has long genome near used as sources of alleles to be incorporated into 17,000 Mbp, which makes its molecular and cultivated hexaploid wheats. Biodiversity and genetic study rather difficult, because most of population structure of the D-genome species

67 Aegilops tauschii Coss. (syn. Ae. squarrosa L.) in Mishina, A. Manickavelu, H. Sato, M. Katsumata, Iran and central Eurasian was studied by H. Saeidi, H. Sassa, T. Koba (2-40) produced molecular M. R. Rahiminejad, J. S. Heslop-Harrison (1-24) mapping of these genes. and N. Mizuno, M. Yamasaki, Y. Matsuoka, T. Mutual interchange of genetic variation and Kawahara, S. Takumi (2-47), respectively. Ae. genomic information between wild and cultivated tauschii is the D genome donor of hexaploid species in Triticeae was viewed by H. Tsujimoto wheats and has high importance for introducing (1-40). The increase of wheat yield by breeding new characteristics and alleles from its gene-pool will help to solve the food crisis which we are into common wheat. S. Takumi, H. Morihiro, E. facing today without another green revolution. Nishioka, T. Kawahara, Y. Matsuoka (1-19) also Wild species of the tribe Triticeae grow in a wide checked natural variation in morphological traits range of environments around the world and have in central Eurasian wild wheat progenitor Ae. large genetic variation. tauschii. Diversity and relationships of the D Breeding properties of a single artificial genome species of Aegilops-Triticum from Iran cultivated species Triticale (× Triticosecale also was studied by F. Bordbar, M.R. Rahiminejad, Wittmack) for improved bread-making quality was H. Saeidi, F.R. Blattner (2-19). observed by P. Martinek (2-38). The perspective of Artificial man-made species are very suitable newly developed translocations in practical for introgression of useful genes and their alleles breeding is discussed. into cultivated wheat species. The synthetic M. Tomita, T. Noguchi, T. Kawahara (1-23) hexaploids obtained by crossing durum wheat discussed evolutional relationships between (BA-genomes) and the wild relative Ae. tauschii Revolver and LARD element in synthetic and allowed a significant increase of cultivated wheat natural wheat species and Aegilops squarrosa. D-genome genetic diversity. M. Kishii, A. Three presentations were connected with Mujeeb-Kazi (1-26) and M. Zaharieva, S. genetic engineering and producing transgenic Dreisigacker, J. Crossa, T. Payne, S. Misra, R. R. plants in cereals. They were made by D. Hanchinal, M. Y. Mujahid, R. Trethowan (2-28) Miroshnichenko, G. Poroshin, S. Dolgov (2-39), carried out the investigation for understanding the by A. Bińka-Wyrwa, W. Orczyk, A. genetic structure of the synthetic species. A subset Nadolska-Orczyk (2-31) and by J. Kumlehn (1-34). of genetically diverse accessions was created to be Basing upon these technologies authors further used for development of new pre-breeding established a numerous transgenic lines with wheat germplasm with enhanced drought and heat improved performance. tolerance. In CIMMYT since 1980s, more than A lot of presentations were connected with 1,200 have been produced by randomly crossing wheat protein quality. The seed storage proteins of the wild D genome diploid donor species (Ae. common wheat, responsible for the ability of flour tauschii) of various ecological origins and 51 to form cohesive dough, are required to make durum wheat lines. strong dough such as bread. However, the narrow Spontaneous amphidiploidization via unreduced genetic base of hexaploid wheat has limited the gametes is a universal phenomenon for Triticum allelic combinations that are available for the turgidum - Aegilops tauschii hybrids, which was improvement of bread-making quality. The described by L. Q. Zhang, D. C. Liu, Y. L. Zheng, presented results could be implemented in a local Y. Yen (2-46). The wide spread of spontaneous breeding program for the improvement of wheat amphidiploidization in the T. turgidum - Ae. grain quality using marker-assisted selection. The tauschii F1 hybrids was found in hexaploid wheat. study is the further contribution to our N. P. Goncharov, K. A. Golovnina, E. Y. understanding of regions of the wheat genome that Kondratenko (1-5) reviewed taxonomy and contribute to the control of GPC in common wheat. molecular phylogeny of natural and artificial J. Taguchi, C. Kiribuchi-Otobe, H. Matsunaka, T. (synthetic) wheat species. The using of Ban (2-18) analysed 96 accessions of tetraploid classifications of genus Triticum L. including wheat including Triticum timopheevii, T. durum, T. synthetic wheats is important for molecular- turgidum, T. dicoccum, T. orientale, T. pyramidale biological, genetic and phylogenetic investigations, and T. carthlicum and D genome chromosome for collecting and identifying wheat accessions substitution lines of Langdon durum. They and breeding practice. examined genetic diversity for several traits Crossability of common wheat with alien associated with pasta making qualities among species, e. g., rye, wild and cultivated barley, is tetraploid wheat germplasms conserved in KIBR known to be controlled by Kr gene family. K. collection. This study showed valuable diversity of

68 the pasta making traits to improve durum wheat by R. Buwan, H. Takahashi, K. Kato, Y-I. Sato, T. with tolerance to biotic and abiotic stresses from Komatsuda, I. Nakamura (2-32). Phylogenetic the tetraploid wheat gene pool. D. Mihálik, Z. analysis of the sequences showed that Triticum and Šramková, E. Medvecká, V. Horevaj, S. Šliková Hordeum species were distinctly separated into (2-21) investigated a genetic variability in Triticum two major clades, except those SS genome species aestivum of Slovakia based on polymorphism for of Triticum clustered with Hordeum species. high molecular weight glutenin subunits. The Diploid wheat (Triticum boeoticum, T. HMW glutenin subunits were classified in 44 monococcum, T. urartu) phylogeny issues are cultivars. E. Gregová, E. Medvecká, Z. Šramková, discussed by F. A. Konovalov, S. V. Goryunova, A. D. Mihálik (2-20) and S. Šliková, Z. Šramková, E. S. Shaturova, A. V. Fisenko, N. V. Melnikova, A. Gregová, D. Mihálik (2-27) estimated quality of M. Kudryavtsev, N. P. Goncharov (1-37). Triticum durum on the basis of gliadin and Adaptation of flowering-time in tetra- and glutenin characterization and high-molecular- hexaploid wheat was viewed in two presentation. weight glutenin subunits in European wheats K. Murai (1-14) discussed the molecular respectively. mechanism of adaptation of flowering-time in X. F. Zhang, D. C. Liu, W. L. Yang, J. Z. Sun, D. tetraploid wheats on the basis of the W. Wang, H. Q. Ling and A. M. Zhang VRN1-VRN3-VRN2 triangle model and focused on (2-48) investigated molecular markers for selection of flowering-time genes under systematic characterization of low molecular domestication. H. Nishida, T. Yoshida, Y. Akashi, weight glutenin subunits in common wheat. The K. Kato (1-16) studied structural variation in 5’ specificity of these markers, validated in a large upstream region of photoperiodic response genes, collection of wheat varieties from China, was Ppd-A1 and Ppd-B1 in Japanese cultivars of discussed. M. Nishinaka, Y .Okumoto, K. Kato, T. common wheat. Therefore, it was strongly Kawahara, T. Tanisaka (2-49) studied genetic suggested that these structural variations explain diversity of high molecular weight glutenines in the difference between day length-sensitive and 297 wheat landraces originated from the Caspian day length-insensitive alleles. Sea regions. Disease resistance also was popular subject of Alien glutenin subunits expressed in common investigation. Fusarium graminearum attacks wheat endosperm effect on the composition was spike of Triticeae species. QTL to reduce studied by H. Tanaka, T. Arakawa, H. Tsujimoto Fusarium mycotoxin accumulation among (2-43). Identification and mapping of QTLs for hexaploid wheats and multiplex quantitative grain protein content in common wheat was analysis for trichothecene genes expression of produced by S. Abugalieva, A. Abugalieva, S. Fusarium graminearum presented by S. Niwa, R. Quarrie, V. Turuspekov (2-29). Based on isolation Kikuchi, H. Handa, T. Ban (2-42) and by T. and molecular characterization of three novel Miyazaki, T. Ban (2-22) respectively. The potential HMW glutenin subunits from Aegilops tauschii, of Hordeum chilense cytoplasm in the the origin and evolution of 1Dx5 subunit in development of CMS systems in Triticeae crops common wheat were discussed by X. An, D. Wang, was discussed by A. C. Martín (2-36). Y. Yan (2-17). Investigations on yellow rust disease resistance by Four presentations were connected with origin useful genes and markers in gene-rich regions on of elemental genomes and related species. Based wheat chromosomes was produced by Y. Aydin, E. on the allelic diversity at chloroplast microsatellite Cabuk, Z. Mert, K. Akan, N. Bolat, M. Cakmak, A. loci among polyploid wheat species the process A. Uncuoglu (2-30). The presence of polymorphic and geography of wheat domestication was markers that is associated with yellow rust discussed by N. Mori (1-13). resistance may significantly enhance the success Genetic relationships of 55 genotypes of selection for yellow rust resistant genotypes in belonging to 8 Triticum of A genome bearing wheat breeding programs. species (T. monococcum, T. boeoticum, T. urartu, T. One of reports “Development and cytogenetic durum, T. turgidum, T. dicoccum, T. dicocoides analysis of Hordeum chilense chromosome 4 and T. aestivum) collected from Iran and some introgression lines into durum wheat” presented by accessions from other areas were studied by M. H. P. Prieto, M. C. Ramírez, A. Martín (1-41) was Ehtemam, M. R. Rahiminejad, H. Saeidi, B. E. connected with pest resistance, namely the Sayed Tabatabaei, S. Krattinger, B. Keller (1-11). resistance to the root-knot nematode Meloidogyne Sequence variation of the 20th exon within naas, just like to Septoria. PolA1 gene among Triticeae species was produced Three presentations were connected with

69 studies of spike morphology and one with leaf H. Knüpffer (1-21). An overview of these species shape-related traits. First of them “Morphological and their main uses is given. The presentation aims variations of spike and the geographical at providing background information for plant distribution of subsection Emarginata species of breeders and crop plant researchers about the Aegilops” was presented by A. Ohta, T. Kawahara, germplasm available in ex situ genebank K. Yamane (2-10). Second (2-16) was connected collections, tending to make this wealth of with novel source of germplasm for the material more easily accessible. development of branched ear wheat and third titled Annual wild Triticeae Gene Bank collection “Agronomic traits and genetic determination of was demonstrated by V. Holubec (1-22). The common winter wheat lines with multirow spike” Prague Gene Bank wild Triticeae collection was presented by P. Martinek, O. Dobrovolskaya, consist of 1800 accessions, belonging to 23 genera, M. S. Röder, A. Börner (2-37). and 133 species including genus Aegilops. H. Morihiro, S. Takumi (2-13) presented results Biodiversity of Georgian wheat species was of studies of intraspecific variation in leaf presented by M. Mosulishvili, I. Maisaia, T. shape-related traits in a wild einkorn wheat species Shanshiashvili, M. Akhalkatsi (2-24). Georgia is Triticum urartu. one of the centers of evolution for many cereal The mechanism of abiotic stress resistance was crops such as T. timopheevii, T. zhukovskyi and T. demonstrated in two presentations. Soil salinity is carthlicum, which were characterized according to one of the environmental abiotic stress factors their reaction to diseases, growing period and limiting agricultural productivity in many regions resistance to abiotic stress. Moreover, existing of the world. Salinity tolerance and sodium germplasm collections are not being effectively exclusion in genus Triticum was presented by Y. used in agricultural science and development Shavrukov, P. Langridge, M. Tester (1-43). programs. Introgression of new genes from different relatives Nowadays progress of wheat studies is of Triticum that cope better with salt stress can connected not only with molecular and genetic significantly improve salinity tolerance in both investigation, but also with good modern cultivated durum and bread wheats. The producing taxonomy that will be suitable for use by transgenic crops with salt tolerance was discussed geneticists, botanists and breeders. New molecular in presentation of D. Miroshnichenko, G. Poroshin, breeding system and MAS need to be developed to S. Dolgov (2-34) titled “Characterization of introduce chromosomes, chromosome segments, growth and yield of transgenic wheat plants genes and alleles into wheat from related species. overexpressing vacuolar Na+/H+ antiporter genes”. This investigations should have been efficiently Conservation ex situ and in situ of wheat used for wheat improvement. collections, their study, replenishment and The symposium was nice organized under the maintenance is still in existence as a source of joint auspices of the Local Organizing Committee, pre-breeding material. It is of fundamental International Organizing Committee and the importance for preserving our food resources and National Institute of Agrobiological Sciences security therefore. The conservation and use of (Tsukuba, Japan). It was supported by the Kyoto plant genetic resources has a history dating back to University Foundation and the Japanese Society of the first domestication of plants by humans. A Breeding. strategy to enhance the effective and efficient conservation and use of ex situ plant genetic resources was presented by M. C. Mackay, L. References Guarino, K. A. Street (1-7). This presentation Barkworth ME (1992) Taxonomy of the Triticeae: showed how the jigsaw puzzle of this global a historical perspective. Hereditas 116: 1-14. system is being put together. Kawahara T (2009) The 6th International Triticeae Genetic resources of Triticeae - cultivated Symposium (6th ITS). Wheat Inf. Serv. 108: species and genebank collections was reviewed by 25-66.

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Instructions to Authors

eWIS welcomes manuscripts that provide test results, but a concise presentation is encouraged. technical tips, protocols, mutant and germplasm (2) Research Opinion & Topics: Reviews, mini- descriptions, map information, and any other reviews, trends and topics in wheat research. information that may be useful in the lab and field. Authors who wish to submit a (mini-)review The articles are informal, non-peer-reviewed, thus do should contact the Editorial Office prior to not constitute formal publications. Only manuscripts submission. that require minimal editing will be considered for (3) Meeting Reports: Announcement of forthcoming publication. meeting and reports on the meeting attended Note well that the data and ideas published in eWIS (4) Others: Any other information useful for wheat are made available by their authors with the researchers understanding that they will not be used in publications without the authors’ specific consent. Title, Affiliation and Abstract This means that the eWIS articles may not be cited In the title page(s), the manuscript category (as without permission from the authors. Copyright is mentioned above), a title, the names of the author(s), retained by the author. affiliation(s) and address(es) of the authors, and the e-mail address, telephone, and fax numbers of the Manuscript Submission corresponding author must be clearly indicated. Only manuscripts written in English will be The Abstract (100-250 words) may not contain considered for publication. Follow the Instructions references. strictly. Refer to the latest eWIS articles for the format. References Electronic submission is mandatory. All manu- References should be cited in the text by the author(s) scripts should be submitted using the online and year, and listed at the end of the text with the manuscript submission system in the eWIS page names of authors arranged alphabetically. When an (http://shigen.lab.nig.ac.jp/ewis/) that is linked from article has more than two authors, only the first the “KOMUGI” page (http://shigen.lab.nig.ac.jp/ author's name should appear, followed by "et al.", in wheat/komugi/top/top.jsp/). the text. The references should be formatted as Editorial Office will inform authors of the status of follows. their manuscript via e-mail as quickly as possible. Journal articles: The “eWIS online submission system” offers easy and Payne PI, Holt LM, Law CN (1981) Structural and straightforward web-based submission procedures. genetical studies on the high molecular weight For text writing, Microsoft Word is recommended. subunits of wheat glutenin. Theor Appl Genet Manuscripts should be double-spaced and 60:229-236. page-numbered starting from the title page. Do not Book chapters: use line numbers. Figures including illustrations, Peacock WJ, Dennis ES, Gerlach WJ (1981) photographs and color plates should be submitted as Molecular aspects of wheat evolution: repeated JPEG files. PDF is not an acceptable file format. DNA sequences. In: Evans LT and Peacock WJ (eds.) Wheat Science - Today and Tomorrow. Manuscript Categories Cambridge Univ. Press, Cambridge, UK, pp. eWIS accepts the following categories of papers: 41-60. (1) Research information: Original research articles Books: in the field of wheat sciences Knott DR (1989) The Wheat Rusts - Breeding for The manuscript should start with a title, the Rust Resistance. Springer-Verlag, New York, names of author(s), affiliation(s), abstract, followed USA. by the text. Abstract may be omitted if not Articles in preparation or articles submitted for necessary. There is no fixed limit on the length publication, unpublished observations, personal

communications, etc. should not be included in the separate file for each table. Refer to the latest eWIS reference list but should only be mentioned in the articles for format. article text (e.g., K. Tsunewaki personal communi- cation). Figures Figures must be numbered consecutively. Prepare a Abbreviations separate file for each figure. Abbreviations should be explained at first occurrence. Outline of the publication process Symbols and Units Authors of accepted manuscripts are informed by Gene names and protein names must carefully be e-mail that a temporary URL has been created from discriminated. Gene names and loci should be which they can obtain their proof. Proofreading is italicized; protein should be upright. The SI units the responsibility of the author. Authors should make (http:// physics.nist.gov/Pubs/SP330/contents.html) proof corrections and send them to Editorial Office by should be used throughout. e-mail. After online publication, corrections can only be made in exceptional cases when Editorial Office Nomenclature permits the necessity. Nomenclature of genes and chromosomes should The final version of accepted manuscripts will be follow the ‘Catalogue of gene symbols for wheat’ published in the ‘Online First’ section of the eWIS (McIntosh et al.: 10th Int. Wheat Genet. Symp. 2003). web page upon receipt of proof corrections. Editorial Office biannually gathers the accepted manuscripts Nucleotide sequences published in the ‘Online First’ into a volume. In The DDBJ/EMBL/GenBank accession numbers must ‘Archive’ of eWIS, all manuscripts are collected as be provided for newly reported nucleotide sequences. PDF format, and open to all wheat researchers. No hard-copy edition will be supplied. For each Tables volume, a PDF edition will be available for Tables must be numbered consecutively. For Table downloading. writing, Microsoft Word is recommended. Prepare a

Wheat Information Service No. 108

Editorial Office

Dr. Yoshihiro Matsuoka (Editor-in-Chief) Department of Bioresources, Fukui Prefectural University, JAPAN

Dr. Tsuneo Sasanuma (Editor, Vice) Department of Bioresource Engineering, Faculty of Agriculture, Yamagata University, JAPAN

Dr. Goro Ishikawa (Editor, Associate) Department of Crop Breeding, Tohoku National Agriculture Research Center, JAPAN

Dr. Shuhei Nasuda (Editor, Associate) Laboratory of Plant Genetics, Graduate School of Agriculture, Kyoto University, JAPAN

Dr. Shigeo Takumi (Editor, Associate) Laboratory of Plant Genetics, Graduate School of Agricultural Science, Kobe University, JAPAN

Advisory Board

Dr. Tomohiro Ban Kihara Institute for Biological Research, Yokohama City University, JAPAN

Dr. Justin D. Faris USDA-ARS Cereal Crops Research Unit, Northern Crop Science Laboratory, North Dakota, USA

Dr. Andreas Houben Institute of Plant Genetics and Crop Plant Research (IPK), GERMANY

Dr. Shahryar F. Kianian Department of Plant Sciences, North Dakota State University, USA

Dr. Evans Lagudah CSIRO Plant Industry, AUSTRALIA

Dr. Hakan Ozkan Department of Field Crops, Faculty of Agriculture, University of Cukurova, TURKEY

Dr. Hisashi Tsujimoto Laboratory of Plant Genetics and Breeding Science, Faculty of Agriculture, Tottori University, JAPAN

Dr. Xueyong Zhang Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, CHINA

1 Sept 2009 www.shigen.nig.ac.jp/ewis