Hellenic Plant Protection Journal

Volume 10, Issue 2, July 2017 ISSN 1791-3691 Hellenic Plant Protection Journal

A semiannual scientifi c publication of the BENAKIBEE PHYTOPATHOLOGICAL INSTITUTE EDITORIAL POLICY The Hellenic Plant Protection Journal (HPPJ) (ISSN 1791-3691) is the scientifi c publication of the Benaki Phytopathological Institute (BPI) replacing the Annals of the Benaki Phytopathological Institute (ISSN 1790-1480) which had been published since 1935. Starting from January 2008, the Hellenic Plant Protection Journal is published semiannually, in January and July each year. HPPJ publishes scientifi c work on all aspects of plant health and plant protection referring to plant pathogens, pests, weeds, pesticides and relevant environmental and safety issues. In addition, the topics of the journal extend to aspects related to pests of public health in agricultural and urban areas. Papers submitted for publication can be either in the form of a complete research article or in the form of a suffi ciently documented short communication (including new records). Only original articles which have not been published or submitted for publication elsewhere are considered for publication in the journal. Review articles in related topics, either submitted or invited by the Editorial Board, are also published, normally one article per issue. Upon publication all articles are copyrighted by the BPI. Manuscripts should be prepared according to instructions available to authors and submitted in electronic form on line at http://www.hppj.gr. All submitted manuscripts are considered and

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EDITORIAL BOARD Editor: Dr F. Karamaouna (Pesticides Control & Phytopharmacy Department, BPI) Associate Editors: Dr A.N. Michaelakis (Entomology & Agric. Zoology Department, BPI) Dr K.M. Kasiotis (Pesticides Control & Phytopharmacy Department, BPI) Dr I. Vloutoglou (Phytopathology Department, BPI) Editorial Offi ce: M. Kitsiou (Library Department, BPI) A. Karadima (Information Technology Service, BPI)

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REVIEW ARTICLE

Endophytic fungi residing in medicinal plants have the ability to produce the same or similar pharmacologically active secondary metabolites as their hosts

A. Venieraki, M. Dimou and P. Katinakis*

Summary Medicinal plants have been used for thousands of years in folk medicines and still are used for their health benefi ts. In our days medicinal plants are exploited for the isolation of plant-derived drugs as they are very eff ective and have relatively less or no side eff ects. However, the natural resources of medicinal plants are gradually exhausted and access to plant bioactive compounds is challenged by the low levels at which these products accumulate in native medicinal plants. For instance, to meet the market demands of 3 Kg per year of vinca alkaloids, powerful plant-derived anticancer drugs, 1.5x106 Kg dry leaves are required. In this regard, this review aims to highlight the fact that endophytic fungi residing in medicinal plants are capable to biosynthesize pharmacologically active secondary metabolites similar or identical to those produced by their host medicinal plant. Fur- thermore, the evolutionary origin of the genes involved in these metabolic pathways as well as the approaches designed to enhance the production of these metabolites by the isolated endophytic fungi are also discussed.

Additional key words: metabolites from endophytic bacteria and actinomycetes, chemical ecology

Introduction present in virtually all organs of a given plant host, and some are seed borne. Endophytes Plant endophytes consist of bacterial and often confer considerable benefi ts to the fungal communities that colonize and spend host plant they inhabit, since they can pro- the whole or part of their life cycle inside mote the growth of host plants, enhance re- the plant tissues, without instigating any sistance to biotic and abiotic stresses (Rodri- noticeable symptoms of infection or visible guez et al., 2009; Hardoim et al., 2015), and manifestation of disease to their hosts (Pe- accumulate bioactive secondary metabo- trini and Fisher, 1990). Evidence of plant-as- lites (Kusari et al., 2012). The ecological role sociated microorganisms found in the fossil- of secondary metabolites produced by en- ized tissues of land plants stems and leaves dophytes is not clear. However, recent stud- suggests that endophyte–plant associa- ies have shown that these metabolites are tions may have evolved along with the evo- involved in deterrence of herbivory (Pannac- lution of higher land plants (Krings et al., cione et al., 2014), protection against fungal 2007). Nearly all vascular plant species stud- (Soliman et al., 2015) or bacterial pathogens ied were found to harbor endophytic bac- (Mousa et al., 2017) and amelioration of plant teria and/or fungi (Rodriguez et al., 2009; abiotic stress (Hamayum et al., 2016). Hardoim et al., 2015). They are found to be Bioactive secondary metabolites derived from medicinal plants are Laboratory of General and Agricultural Microbiology gradually decreasing - Alternative Department of Crop Science, Agricultural University approaches for their production of Athens, Iera Odos 75, GR 118 55 Votanikos, Athens, Greece * Corresponding author: [email protected] Medicinal plants, as a rich source of nat-

© Benaki Phytopathological Institute 52 Venieraki et al. ural products, have been used to treat vari- this review, we aim to show that the large ous ailments and have been the foundation number of medicinal plants used for the for discovery and development of modern isolation of medically important bioactive therapeutics (Pan et al., 2013). Up to 80 % of compounds harbor endophytic fungi capa- people in developing countries are totally ble of host-independent biosynthesis of the dependent on herbal drugs for their prima- same or similar bioactive secondary metab- ry healthcare. More than 51% of small mole- olites as their hosts. This review will also dis- cule drugs approved between 1981 and 2014 cuss the evolution and origin of pathways were based on natural products, the rest be- involved in the biosynthesis of these bioac- ing synthetic (Chen et al., 2016). With the in- tive compounds and potential approaches creasing demand for herbal drugs, natural aiming to enhance their production. health products and secondary metabolites, the use of medicinal plants is growing rapid- ly throughout the world (Chen et al., 2016). Medicinal plants harbor endophytic However, we are facing the accelerated loss fungi capable of mimicking their host of wild medicinal plant species; one third plant secondary metabolite profi le- of the estimated 50.000-80.000 medicinal Case studies on medicinal plants plant species are threatened with extinction producing metabolites of known from overharvesting and natural anthropo- medical importance genic habitat destruction (Chen et al., 2016). Furthermore, the feasibility of access to Since the fi rst report of endophyte Tax- plant bioactive compounds is challenged by omyces andreanae that produces the same the low levels at which these products accu- bioactive secondary metabolite taxol (pacli- mulate in native medicinal plants, the long taxel) as its host Taxus brevifolia in 1993 (Sti- growth periods required for plant matura- erle et al., 1993), several studies have shown tion, and the diffi culty in their recovery from that plant-derived secondary metabolites other plant-derived metabolites (Staniek et are produced by endophytes (Zhao et al., al., 2014). For example, the taxol concentra- 2011). In this section, we will present a liter- tion is about 0.001–0.05% in Taxus brevifolia, ature survey aiming to show that medicinal which is the most productive species. Thus, plants used for isolation of medically impor- 15 kg of Taxus bark, three trees, are required tant secondary metabolites usually harbor for production of 1 g, while every cancer pa- endophytic fungi which are capable of host- tient requires about 2.5 g (Malik et al., 2011). independent biosynthesis of these metabo- Therefore, it is important to fi nd alter- lites. In each one of the presented case stud- native approaches to produce the medici- ies, emphasis will be placed on the plant nal plant-derived biologically active com- species, the organ where the bioactive compounds, in particularly those derived from pound is accumulated and the organ from endangered or diffi cult-to-cultivate plant which the active compound-producing fun- species, to meet the medical demand. This gi were isolated. can be achieved by the application of plant cell and tissue culture, heterologous pro- Salvia sp. (Lamiaceae) duction, total chemical synthesis, semi-syn- Salvia species have many important thesis, or by starting with a microbially - pro- medicinal properties with proven pharma- duced or plant-extracted natural product cological potential. Some of these proper- occurring more abundantly in nature (Ata- ties may be mediated by biologically active nasof et al., 2015; Rai et al., 2016; Ramirez-Es- polyphenols or terpenoids (Wu et al., 2012). trada et al., 2016) or by exploiting the abili- Two kinds of bioactive compounds, tanshi- ty of endophytic fungi residing in plants to nones (tanshinone I, tanshinone IIA, tanshi- produce the same or similar bioactive com- none IIB, isotanshinone I, and cryptotanshi- pounds as their hosts (Zhao et al., 2011). In none) and salvianolic acids (salvianolic acid

© Benaki Phytopathological Institute Endophytic fungi & secondary metabolites in medicinal plants 53 and rosmarinic acid) have been found in the the endophytes for the production of vinca roots and leaves of S. miltiorrhiza, respec- alkaloids revealed that only endophytic fun- tively. Tanshinones belong to diterpenoid gi residing in the leaves of C. roseus were ca- quinones, and are considered as potent an- pable of producing vinblastine and vincris- ti-carcinogenic, antiatherosclerosis, and an- tine. These endophytic fungi were identifi ed tihypertensive, whereas salvianolic acids are as Fusarium oxysporum, Talaromyces radicus phenolic acids, which are mainly respon- and Eutypella spp. The drugs were purifi ed sible for benefi cial eff ects on cardiovascu- by TLC and HPLC and authenticated using lar and cerebrovascular diseases (Chun-Yan UV-Vis spectroscopy, ESI-MS, MS/MS and et al., 2015). Several Salvia species produce 1H NMR. Culture fi ltrates of the fungi yield- the bioactive phenolic labdane-type diter- ed >55 μg/L of vinblastine or vincristine, re- penes rosemarinic acid, carnosic acid and spectively (Kumar et al., 2013; Palem et al., carnosol. These compounds show distinct 2015; Kuriakose et al., 2016). anti-oxidant activity with carnosic acid carnosol being approved food additives (Wu et Coleus forskohlii (Willd.) Briq. (Lamiace- al., 2012). Salvia divinorum produces a novel ae) diterpenoid, salvinorin A, which is a power- Coleus forskohlii or Indian Coleus is a trop- ful hallucinogen in humans and shows a se- ical perennial shrub of the Lamiaceae fam- lective, high effi cacy agonist activity (Butel- ily and grows in the subtropical temperate man and Kreek, 2015). climates of South-east Asia and India. The Eighteen endophytic fungal strains have plant is extensively cultivated in southern been isolated from the roots of Salvia milti- India and the roots are used in Indian folk orrhizae, the site of tanshinones accumula- medicine for treating a broad range of hu- tion, and 58 fungal strains from the leaves, man health disorders (Kavitha et al., 2010). the main site of salvianolic acid accumula- The roots of the herb contain a pharmaco- tion. Liquid culture extracts of all the fungi logically active compound called forskolin were screened for the presence of tanshino- that accumulates in the root cork (Patera- nes or salvianolic acid, respectively. One fun- ki et al., 2014). The approved and potential gus in each case was proven to produce tan- applications of forskolin range from allevia- shinones or salvianolic acid compared with tion of glaucoma, anti-HIV or antitumor ac- authentic standards. However, the yield was tivities, treatment of hypertension and heart quite low; about 4μg/L for tanshinones and failure to lipolysis and body weight control 47μg/L for salvianolic acid (Ming et al., 2013; (Pateraki et al., 2017). Li et al., 2016). Screening of endophytic fungi isolated from inner tissues of root and stems of Catharanthus roseus (L.) G.Don (Apocyn- C. forskohlii for the production of forskolin aceae) revealed that one of the endophytic fun- Catharanthus roseus is well known for the gi identifi ed as Rhizoctonia bataticola was production of several anticancer vinca alka- able to stably synthesize forskolin and inter- loids such as vincristine, vindesine, vinorel- estingly, release it into the broth (Mir et al., bine, vinblastin and the recently discovered 2015). vinfl unine (Kumar et al., 2014). The two major anticancer vinca alkaloids, vincristine Macleaya cordata (Willd.) R.Br. (Papaver- and vinblastine, used in chemotherapy reg- aceae) imens, have been isolated from leaves (Ku- Sanguinarine (SA) is a benzophenanthri- mar et al., 2014). dine alkaloid isolated from Macleaya cor- The diff erent C. roseus plant organs har- data leaves, and is known to have a wide bour a plethora of endophytic fungi (Khar- spectrum of biological activities, such as war et al., 2008; Kumar et al., 2013; Palem et antibacterial, antihelmintic, antitumor and al., 2015; Kuriakose et al., 2016). Screening all anti-infl ammatory (Wang et al., 2014). SA is

© Benaki Phytopathological Institute 54 Venieraki et al. used in feed additives for livestock produc- leaves of Cephalotaxus have been used in tion (Kantas et al., 2014). Most of the SA cur- Chinese folk medicine as anticancer agents, rently used in herbal supplements and med- and its biological active constituents were icines is extracted from M. cordata. Recently, proved to be alkaloids. Among these alka- SA has gained increasing attention as a po- loids, homoharringtonine (HHT) was shown tential agent in the treatment of cancer (Yu eff ective against acute myeloid leukemia et al., 2014). and has recently been approved by the Screening of endophytic fungi isolated Food and Drug Administration for the treat- from leaves of M. cordata revealed that one ment of chronic myeloid leukemia (Hu et al., of 55 isolates has the capacity to produce SA 2016). (Wang et al., 2014). A large number of endophytic fungi have been obtained from Cephalotaxus Cajanus cajan (L.) Millsp. (Fabaceae) phloem. The bioactive compounds isolated Cajanus cajan (pigeon pea) is a grain le- from their culture extracts were character- gume crop in semitropical and tropical ar- ized as sesquiterpenoids, anthraquinones eas of the world. The extract of pigeon pea and aromatic compounds, which exhibited leaves exhibit therapeutic eff ects on sickle cytotoxic and antibacterial activities (Lu et cell anemia, plasmodiosis, and hepatic dis- al., 2012; Xue et al. 2012; Zheng et al., 2011). orders. Moreover, pigeon pea roots are used The hunt for an HHT-producing endophytic as a sedative, a vulnerary preparation. The fungus was eventually successful following active constituents of pigeon pea are fl a- the screening of 213 strains isolated from vonoids and stilbenes. Cajaninstilbene acid the bark of Cephalotaxus trees grown in Chi- (CSA) is one of the major stilbenes found in na and Thailand. The fungus was identifi ed pigeon pea. Pharmacological studies have as Alternaria tenuissima and stably produced revealed that CSA exhibited anti-infl amma- 100 μg/L HHT (Hu et al., 2016). tory and analgesic eff ects. In addition, CSA has an antioxidant activity similar to that of Cinchona spp. (Rubiaceae) the natural antioxidant resveratrol (Liang The bark of the stem and roots of variet al., 2013). Cajanol is a isofl avone isolat- ous trees of the genus Cinchora produce ed from pigeon pea roots. Pharmacological quinine alkaloids (quinine, quinidine, cin- studies have shown that cajanol has anti- chonidine and cinchonine), which were the plasmodial, antifungal and antimicrobial only eff ective treatment of malaria for more activities. In addition, cajanol has been de- than four centuries. Cinchona bark and its scribed as a novel anticancer agent, which alkaloids remained the most effi cient treat- induced apoptosis in human breast cancer ment of malaria until the 1940s when chlo- cells (Luo et al., 2011). roquine and other synthetic antimalarial A total of 245 endophytic fungi isolated compounds were developed (Kaufman and from roots, stems and leaves of pigeon pea Ruveda, 2005). With the development of re- plants were screened for the production of sistant malaria strains, the quest for new an- cajaninstilbene acid or cajanol. Three fungal timalarial compounds was successful with strains isolated from leaves were capable of the discovery of artemisinin from a Chinese producing CSA and one strain isolated from herbal medicine based on Artemisia annua roots stably produced cajanol at a concen- L. (Tu, 2011). tration of 500μg/L (Zhao et al., 2012; Zhao et Twenty-one endophytic fungi have been al., 2013). isolated from Cinchona ledgeriana young plant stems and screened for the presence Cephalotaxus hainanensis H.L.Li (Cepha- of Cinchora alkaloids. These fungi com- lotaxaceae) prised of Phomopsis, Diaporthe, Schizophyl- Cephalotaxus hainanensis H. L. Li is an in- lum, Penicillium, Fomitopsis and Arthrinium digenous conifer tree of China. The bark and species while fermentation studies demon-

© Benaki Phytopathological Institute Endophytic fungi & secondary metabolites in medicinal plants 55 strated that all produce quinine and quini- 3β-D-glucoside whereas a second endo- dine, as well as cinchonidine and cinchonine phytic fungus was found to secrete pei- (Maehara et al., 2011; Maehara et al., 2013). misine and peiminine. Interestingly, a large number of the remaining endophytes were Passifl ora incarnata (Passifl oraceae) able to produce large amounts of antioxi- Passifl ora consists of 500 species that dants, such rosemarinic acid (Pan et al., 2014; are found mostly in warm and tropical re- Pan et al., 2015; Pan et al., 2017). gions. Passifl ora incarnata leaves were found to contain several active compounds, in- Huperzia serrata (Thunb. ex Murray) Tre- cluding alkaloids, phenols, glycosyl fl a- vis (Huperziaceae) vonoids, and cyanogenic compounds. The Huperzia serrata is a traditional Chinese major compounds present in P. incarnata herb medicine and has been extensively are C-glycosyl fl avonoids (vitexin, isovitex- used for the treatment of a number of ail- in, orientin and chrysin) and b-carbolinic al- ments, including contusions, strains, swell- kaloids (harman, harmin, harmalin, harmol, ings, schizophrenia, myasthenia gavis and and harmalol). Among these natural prod- organophosphate poisoning. These phar- ucts, chrysin has shown interesting biolog- maceutical applications of H. serrata are ical activities, including antibacterial, an- mainly due to its biologically active lycopo- ti-infl ammatory, anti-diabetic, anxiolytic, dium alkaloids. Among the lycopodium al- hepatoprotective, anti-aging, anticonvul- kaloids, huperzine A (HupA) was found to sant and anticancer eff ects (Seetharaman et possess potent acetylcholine esterase inhi- al., 2017). bition (AChEI) and is clinically used for the Three endophytic fungi identifi ed as Al- treatment of Alzheimer’s disease (Zhao et tenaria alternata, Colletotrichum capsici, and al., 2013). The content of HupA in the leaf is C. taiwanense were isolated from leaves of P. richer than that in the stem and root of H. incarnata and production of fungal chrysin serrata (Gu et al., 2005). was confi rmed through UV-vis spectrosco- Several groups have isolated endophyt- 1 py, FT-IR, LC-ESI-MS, and H1 NMR analysis of ic fungi from leaves, stems and roots of H. their extracts. The quantitative HPLC analy- serrata. Screening culture extracts of these sis revealed that the yield of chrysin from A. fungi for HupA production revealed that alternata was higher when compared with most of HupA-producing fungi were iso- previously reported bioresources (Seethara- lated from leaf tissues (Su et al., 2017). The man et al., 2017). HupA-producing endophytic fungi belong to Penicillium griseofulvum, Penicillium sp., Fritillaria cirrhosa D.Don (Liliaceae) Aspergillus fl avus, Mycoleptodiscus terrestris, Bulbus Fritillaria have been used in tra- Trichoderma sp., Colletotrichum gloeospori- ditional Chinese medicine for more than oides strain ES026 and Shiraia sp.. The pro- 2000 years, and at present, they are among ductivity of these strains is less than 60-90 the most widely used antitussive and ex- μg/L, with Shiraia sp. Slf14 being the best pectorant drugs. The major biological ac- producer (327.8 μg/L) (Su et al., 2017). Inter- tive ingredients of Bulbus Fritillaria cirrho- estingly, many H. serrata endophytic fungi sa are steroidal alkaloids, such as peimisine, with AChE inhibitory activity did not contain imperialine-3β-D-glucoside, and peimine HupA in their extracts (Su et al., 2017; Wang (Wang et al., 2011). et al., 2016) suggesting that some endophyt- Several dosens of endophytic fungi were ic fungi produce new compounds with ac- isolated from fresh bulbus of Fritillaria uni- tivity against AChE. bracteata var. wabensis. One of these fungal endophytes, Fusarium redolens 6WBY3 was Rhodiola spp. (Crassulaceae) capable of producing and secreting in the Rhodiola rosea is a perennial herbaceous culture medium peimisine and imperialine- plant that belongs to the family Crassu-

© Benaki Phytopathological Institute 56 Venieraki et al. laceae. This species is mainly distributed in is possible, however it does not appear to be high altitudes of >2,000 m in the Arctic and practical as the overall yield was only 10.5%, mountainous regions throughout Asia and requiring 13 steps (Wei et al., 2011). Europe. This typical alpine plant has been Three fungal endophytes have been iso- widely used as an important food crop and lated from S. nigrum stems, leaves and fruits. folk medicine since ancient times by many Their culture extracts were screened for the countries, such as Sweden, Russia, India, potential production of steroidal alkaloids. and China (Chiang et al., 2015). Rhodiola rhi- The stem derived endophytic fungal strain zome, as a traditional folk medicine, stimu- A. fl avus was able to steadily produce sola- lates mental and physical endurance, coun- margine with a titer of about 250–300 μg/ L teracts depression, improves sleep quality, which is higher than the plant callus culture and prevents high-altitude sickness. Mod- method (El-Hawary et al., 2016). ern pharmacology research suggests that Rhodiola rhizome has received consider- Piper longum L. and Piper nigrum L. (Pip- able attention because of its biological be- eraceae) havior, including antioxidant and anti-aging Piperine is a major alkaloid present in the properties, anti-microwave radiation, anti- fruit of Piper longum and Piper nigrum and it hypoxia and adaptogenic activities. Most of is known to have a wide range of pharma- these eff ects are ascribed to phenolics, such ceutical properties including antibacterial, as salidrosides and p-tyrosol, and glycosides antifungal, hepato-protective, antipyretic, like rosavins (Chiang et al., 2015). anti-infl ammatory, anti-convulsant, insec- Screening of 347 endophytic fungal ticidal and antioxidant. The amount of pip- strains isolated from rhizomes of R. crenula- erine varies in plants belonging to the Pip- ta, R. angusta and R. sachalinensis revealed eraceae family; it constitutes 2% to 7.4% of that four endophytic fungi were capable of both black pepper and white pepper (Cor- producing salidrosides and p-tyrosol (Cui gani et al., 2017). Screening of endophyt- et al., 2015). One of these endophytic fungi ic fungi isolated from both plant species re- identifi ed as Phialocephala fortinii was able vealed the presence of piperine in culture to stably produce large amounts of salidro- extracts of endophytic Periconia strains iso- sides and p-tyrosol, 2.3 and 2 mg/ml of cul- lated from leaves of P. longum (Verma et al., ture medium, respectively (Cui et al., 2016). 2011) and Colletotrichum gloeosporioides from the stems of P. nigrum (Chithra et al., Solanum nigrum L. (Solanaceae) 2014). Solanum nigrum L., family Solanaceae is a well-known medicinal plant which possess- Digitalis lanata Ehrh. (Plantaginaceae) es several biological activities such as anti- Glycosides from plants of the genus Dig- oxidant, hepato-protective, antiinfl amma- italis have been reported to be cardiotonic tory, antipyretic, diuretic, antimicrobial and and are widely used in the treatment of var- anticancer activities due to its fl avonoid and ious heart conditions namely atrial fi brilla- steroidal alkaloids content (Jain et al., 2011). tion, atrial fl utter and heart failure. The bio- Solamargine, one of the major steroidal al- active glycosides accumulate in the leaves kaloids in S. nigrum has been demonstrated and to a less extent in other organs of the to exhibit potent anticancer activity against plant (Alonso et al., 2009). colon, prostate, breast, hepatic and lung A total of 35 fungal endophytes were iso- cancer cell lines (Jain et al., 2011). Solamar- lated from stems and leaves, and screened gine is always found in a complex mixture for the production of secondary metabo- with other glycoalkaloids such as solasonine lites. Crude extracts of fungal cultures re- and solanine, which makes solamargine iso- vealed the production of glycoside digox- lation from the plant quite diffi cult (Milner et in from cultures of fi ve endophytic strains al., 2011). Chemical synthesis of solamargine (Kaul et al., 2013).

Capsicum annuum L. (Solanaceae) seven fl avolignans (silybin A, silybin B, isosi- Capsaicin, the pungent alkaloid of red lybin A, isosilybin B, silychristin, isosilychris- pepper (Capsicum annuum), is present in tin, and silydianin) with reported chemopre- large quantities in the placental tissue, the vention and hepatoprotective properties internal membranes and, to a lesser extent, (Feher and Lengyel, 2012). the other fl eshy parts of the fruits of Cap- Twenty one endophytic fungi were iso- sicum. The pharmacological properties of lated from stems, leaves, roots, and seeds of capsaicin include cardio protective infl u- S. marianum and were examined for produc- ence, anti-lithogenic eff ect, anti-infl ammation of fl avolignans (El-Elimat et al., 2014). tory and analgesia, thermogenic infl uence, Two of these compounds, silybin A and si- and benefi cial eff ects on gastrointestinal lybin B, have been extracted as fermenta- system (Srinivasan et al., 2016). tion products of two strains of Aspergillus ii- An endophytic fungal strain identifi ed as zukae isolated from the leaves and stems of Alternaria alternata has been isolated from S. marianum, respectively. Subcultur of one fruits of C. annuum and has been found to fl avonolignan-producing strain revealed an produce and secrete capsaicin up to three attenuation of the production of fl avono- generations (Devari et al., 2014). lignans. However, when autoclaved leaves of the host plant were added to the growth Ginkgo biloba L. (Ginkoaceae) medium, the production of fl avonolignans Ginkgo tree contains in bark and leaves could be resumed (El-Elimat et al., 2014). fl avones and terpenoide lactones, among which, bilobalide and ginkgolides (terpe- Vinca minor L. (Apocynaceae) and Neri- noide lactones) have been shown to be um indicum Mill. (Apocynaceae) benefi cial to human health (Usai et al., 2011). Vincamine indole alkaloids (vincamine, Ginkgolide B has revealed potent antagonis- tabersonine and catharanthine) are widely tic eff ects on platelet activating factors in- found in plants of the Apocynaceae family volved in the development of a number of and show benefi cial properties for human, renal cardiovascular, respiratory and central such as prevention of cerebrovascular, pre- nervous system disorders (Usai et al., 2011) caution of chronic ischemic stroke, and re- while bilobalide was found to exert neuro- duction of vascular dementia or memory protective eff ects (Kiewert et al., 2008). impairment (Saurabh and Kishor, 2013). Vin- Screening of 27 endophytic fungal camine is a precursor compound for other strains isolated from the bark of G. biloba medicinal alkaloids such as 11-bromovin- trees revealed that only one isolate F. ox- camine, ethyl-vincamine and vinpocetine, ysporum SY0056, was capable of produc- which have shown potential clinical thera- ing Ginkgolide B (Cui et al., 2012). The search peutic eff ect (Manda et al., 2015). Vincamine for bilobalide -producing endophytic fun- is accumulated in the leaves and stems of gi was far more copious; a total of 57 fungal Vinca minor and Nerium indicum. Though strains were isolated from stem, root, leaf, abundant chemical synthesis and semi-syn- and bark of the plant G. biloba and their ex- thesis research results have been reported, tracts were evaluated for the presence of bi- the main sources of vincamine indole alka- lobalide. Only the isolate Pestalotiopsis uvi- loids are stems and leaves of Vinca minor L. cola GZUYX13 residing in leaves was proven Eleven fungal strains have been isolated to be a bilobalide-producing fungus (Qian from the stems and roots of Nerium indicum et al., 2016). and fungal culture extracts were screened for the presence of indole alkaloids (Yin and Silybum marianum (L.) Gaertn. (Aster- Sun, 2011). One fungal strain, CH1, produces aceae) vincamine alkaloids as its host plant as de- Silymarin is a bioactive extract of the termined by TLC, HPLC and LC–MS analy- fruits of Silybum marianum and contains sis. The yield of vincamine, ethyl-vincamine,

© Benaki Phytopathological Institute 58 Venieraki et al. and tabersonine was 1.279, 1.279 mg/L, 0.102 strain Colletotrichum gloeosporioides isolat- mg/L, respectively (Na et al., 2016). In a simi- ed from the fruits was found to produce the lar study, 10 endophytic fungal strains were active constituent phillyrin as was judged by isolated from the roots, stems and leaves of TLC, HPLC and HPLC-MS analysis (Zhang et the plant V. minor. One fungal strain isolat- al., 2012). ed from the stems was found to produce vincamine although with a relatively lower Miquelia dentata Bedd. (Icacinaceae), yield as compared to that of another fungal Camptotheca acuminata Decne. (Nys- strain isolated from N. indicum (Yin and Sun, saceae) and Nothapodytes nimmoniana 2011). (Graham) Mabb. (Icacinaceae) Camptothecine (CPT), a quinoline indole Rheum palmatum L. (Polygonaceae) alkaloid and its analog, 10-hydroxy camp- Rheum palmatum is a medicinal plant tothecine (10-OH-CPT) are potent inhibitors and its air-dried roots have been used in of the eukaryotic topoisomerase I and are the traditional medicine. R. palmatum pres- currently used as effi cient anticancer drugs ents cathartic eff ect on the digestive move- against a broad band of tumor types such ment of the colon, protects the damaged as small lung and refractory ovarian can- liver, and has antibacterial, anti-infl amma- cers. (Kai et al., 2015). CPT and 10-OH-CPT tion, and anti-aging properties. The most ef- are naturally produced by several plant spe- fective biologically active compounds in the cies of the Asterid clade. Among them how- roots of the genus Rheum are anthraquino- ever, the major sources of commercial CPT nes including emodin, rhein, physcion, al- in the world market are Camptotheca acum- oe-emodin. Pharmacological tests revealed inata and Nothapodytes nimmoniana (Uma that rhein can alleviate pain and fever and Shaanker et al., 2008). Exceptional high lev- inhibits infl ammation (You et al., 2013). els of CPT and 10-OH-CPT are also found Fourteen endophytic fungal strains have in the fruits and seeds of Miquelia dentata been isolated from R. palmatum: 12 strains (Ramesha et al., 2013). were isolated from the root, 2 strains from Twenty-three fungal isolates were ob- the stem. The strain R13, isolated from the tained from diff erent fruit parts of M. denta- roots, was capable to produce the bioactive ta. All fungal isolates produced CPT though compounds rhein and emodin. The yield of in varying quantities (Shweta et al., 2013). rhein in R13 can reach 5.67 mg/L (You et al., Three fungal species, A. alternata, Phomop- 2013). sis sp. and Fomitopsis sp., were identifi ed as CPT-producers with the highest yield of CPT Forsythia suspensa (Thunb.) Vahl. being obtained from A. alternata (73.9 μg/g (Oleaceae) DW) (Shweta et al., 2013). CPT-producing en- The main chemical constituents of F. sus- dophytic fungi have also been isolated from pensa are composed of lignans including C. acuminata (Pu et al., 2013) and N. nimmo- phillyrin and forsythiaside, triterpenic ac- niana (Bhalkar et al., 2016). ids including oleanolic acid and ursolic acid. Phillyrin was reported to have various biological activities such as antioxidant, anti-in- Biochemical convergence or horizontal fl ammatory, anti-hyperlipidemia and anti- gene transfer confer the ability to the pyretic activities (Qu et al., 2008). Studies on endophytic fungi to produce the same phillyrin have shown its presence mainly in bioactive compounds as their host the leaves and fruits of the plant F. suspensa (Piao et al., 2008). The discovery of endophytic fungi pro- A total of 24 fungal strains were isolated ducing the same or similar bioactive com- from stems, leaves and fruits of F. suspensa pounds as their hosts raises the question as and screened for phylirin production. One to whether parallel pathways evolved sim-

© Benaki Phytopathological Institute Endophytic fungi & secondary metabolites in medicinal plants 59 ply because each lineage has benefi tted as PCR analysis did not reveal the presence from making a given compound completely of Taxus ts gene sequences in SSM001 (Soli- independently of the other or whether hori- man et al., 2013). Hence, the divergence of zontal gene transfer (HGT) events took place the two biosynthetic pathways is support- between the fungi and the plant. ed with conservation only in specifi c en- There is precedent for the indepen- zyme sites to be important for the activi- dent development of the same biosynthet- ty rather than the whole protein structure. ic pathway (biochemical convergence) in Similar fi ndings have been reported in the fungi or plants and other organisms. For in- case of huperzine A producing endophytes. stance, although higher plants and endo- Their fungal amine oxidase genes have phytic fungi produce structurally identical been found to present low similarities to the GAs, profound diff erences have been found corresponding plant genes, and only con- in the GA pathways and enzymes of plants served consensus sequences were present and fungi (Hamayum et al., 2016), e.g. 7-m- by the fungal and plant functional amine ethyl-cyercene-1 found in both the fungus oxidase proteins (Yang et al., 2014; Yang et Leptosphaeria maculans (anamorph Pho- al., 2016; Zhang et al., 2015), which supports ma lingam) and the marine mollusk Ercol- the co-evolution theory rather than the HGT ania funereal is produced by distinct en- theory. This has been well established in the zymes (Cutignano et al., 2012). Cyanogenic case of gibberellin biosynthetic pathways in glucosides linamarin and lotaustralin found fungi and higher plants where diff erences in both the moth Zygaena fi lipendulae and in genes and enzymes indicated converged their food plant Lotus japonicus are biosyn- evolution of GA metabolic pathways (Böm- thesized by distinct enzyme systems(Jensen ke and Tudzynski, 2009). et al., 2010). However, a horizontal gene The list of taxol producing endophyt- transfer event between plants and fungi, al- ic fungi is large and encompasses numer- though rare, should not be excluded (Rich- ous fungi belonging to diverse genera (Sti- ards et al., 2009). erle and Stierle, 2015). A similar situation Several studies have reported the pres- appears to hold for CPT-producing fungi (Pu ence of Taxus tree key genes (ts,dbat and et al., 2013) and HupA-producing endophytic bapt) which are involved in plant paclitaxel fungi (Su et al., 2017) suggesting a horizontal biosynthesis in taxol-producing endophytic transfer of large secondary metabolism gene fungi. These results stimulated the conjec- clusters between fungi. Several studies off er ture that the origin of this pathway in these support to this idea; the complete sterigma- two physically associated groups could have tocystin gene cluster in Podospora anserine been facilitated by horizontal gene transfer was horizontally transferred from Aspergil- (Kusari et al., 2014). Other studies, howev- lus (Slot and Rokas, 2012). Furthermore, it has er, provided evidence that microbial taxol been shown that CTP is also produced by a genes exist independent of the plant genes diverse group of endophytic bacteria (Shwe- (Xiong et al., 2013). Recent data support the ta et al., 2013; Pu et al., 2015) suggesting that latter proposal; genome sequencing and bacterial CPT biosynthesis may represent analysis of the taxol-producing endophyt- an independently assembled pathway from ic fungus Penicillium aurantiogriseum NRRL that in fungi or plants. This may be surpris- 62431 revealed that out of 13 known plant ing since converged evolution of the diter- Taxol biosynthetic genes, only 7 showed low pene GA metabolic pathway in plants, fungi homology(>30%) with genes identifi ed in P. and bacteria is well established (Tudzyns- aurantiogriseum (Yang et al., 2014). Further- ki et al., 2016). Therefore, extensive genome more, polyclonal antibodies against Yaxus sequencing of the various endophytic fun- TS strongly cross-reacted with a protein of gi will provide an opportunity for a compre- the taxol-producing fungus Paraconiothyri- hensive study on the phylogenetic origin of um SSM001 grown in liquid culture, where- fungal and bacterial metabolic pathways.

Exploring endophytes for sustainable tive in stimulating the transcription of atten- and enhanced production of secondary uated biosynthetic gene clusters of endo- metabolites phytic fungi (Vasanthakumari et al., 2015; Magotra et al., 2017), thereby resulting in The discovery of endophytic fungi ca- the enhancement of the production of de- pable of producing the same bioactive sired secondary metabolites. Bioprocess en- compounds as their host medicinal plant gineering strategies such as manipulation has raised the expectation that these com- of media and culture conditions, co-culture pounds could be produced in large scale condition, epigenetic modulation, elicitor through fermentation processes, thus and or chemical induction, mixed fermen- meeting the growing demand of the mar- tation, and fermentation technology, have ket, while relieving the dependence on been proven promising in alleviating to their respective endangered host plants some extent these obstacles (Venugopalan for the metabolites. However, this expecta- and Srivastava, 2015). tion remains hampered primarily by the low Upon availability of the endophytic fun- yields as well as the attenuation of metabo- gal genomes, the putative genes encoding lites production after sub-culturing of fungi the enzymes involved in the biosynthesis (Kusari et al., 2011; Kumara et al., 2014; El-Eli- of bioactive compounds could be identi- mat et al., 2014). The reasons for the attenua- fi ed and their function could be verifi ed tion could be attributed to factors that stem through transcriptomic, proteomic and me- from loss of presumed signals provided by tabolomic, RNA interference, gene knock- the host or co-existing endophytes, result- out, and gene over expression. Genome ing in the silencing of genes in axenic mon- editing technologies implemented for met- ocultures (Sachin et al., 2013). abolic engineering of fi lamentous fungi may Passage of attenuated CPT-producing be applied for triggering the biosynthesis endophytic fungi from the host plants re- of metabolites. Alternatively, the identifi ed stored CPT production in the re-isolated en- biosynthetic pathway of the correspond- dophytic fungi (Vasanthakumari et al., 2015) ing bioactive compounds can be assem- suggesting that a certain critical signaling bled, engineered and then introduced in may be necessary for the fungus to main- other genetically tractable microorganisms tain its endogenous production. Co-culti- to increase their yields (El-Sayed et al., 2017; vation studies of taxol producing fungus Wakai et al., 2017). Paraconiothyrium SSM001 with endophytic fungi isolated from Taxus tree revealed an eightfold increase in fungal Taxol pro- Medicinal plant endophytes in Greece duction from SSM001 (Soliman and Raizada, 2013). Co-cultivation of the endophytic fun- Greece is endowed with a rich biodiver- gus Fusarium tricinctum with the bacterium sity of medicinal plant species with a long Bacillus subtilis, led to an up to 78-fold en- tradition in herbal medicines, and their hancement in the accumulation of the con- complex endomicrobiome may be directly stitutively present fungal metabolites (Ola and indirectly responsible for the produc- et al., 2013). Co-cultivation (mixed fermentation of a wealth of explored and unexplored tion) under optimized conditions of the two bioactive compounds. Thus, it is expected CPT-producing fungal species Colletotri- that many new or known products for medi- chum fruticola and Corynespora cassiicola cine may emerge through the exploration of isolated from the same host tree N. nimmo- the endophytes of these medicinal plants. niana enhanced the yield of produced CPT We are currently isolating fungal and bac- (Bhalkar et al., 2016). terial endophytes from indigenous medici- Epigenetic modifi cations using chemical nal plant species in the genera such as Fritil- inhibitors have also been found to be eff ec- laria, Hypericum, Teucrium, Calendula, Salvia

© Benaki Phytopathological Institute Endophytic fungi & secondary metabolites in medicinal plants 61 as well as Olea europaea and the exotic Ni- Chiang, H.M., Chen, H.C., Wu, C.S., Wu, P.Y. and Wen, gella sativa aiming to identify such bioactive K.C. 2015. Rhodiola plants: chemistry and biological activity. Journal of Food and Drug Anal- compounds. ysis, 23: 359-369. Chithra, S., Jasim, B., Anisha, C., Mathew, J. and Rad- hakrishnan, E.K. 2014. LC-MS/MS based nidenti- Conclusions fi cation of piperine production by endophytic Mycosphaerella sp. PF13 from Piper nigrum. Ap- plied Biochemistry Biotechnology, 173: 30–35. Medicinal plants off er an extensive biore- Chun-Yan, S.U., Qian-Liang, M.I.N.G., Rahman, K., source of new bioactive compounds that Ting, H.A.N., and Lu-Ping, Q.I.N. 2015. Salvia mil- have signifi cant potential as antiparasit- tiorrhiza: Traditional medicinal uses, chemistry, ics, antibiotics, antioxidants, and anticancer and pharmacology. Chinese Journal of Natural agents. During the last 10 years it became Medicines, 13: 163-182. apparent that endophytes are capable to Cui, J., Guo, T., Chao, J., Wang, M. and Wang, J. 2016. Potential of the endophytic fungus Phialo- produce the same bioactive secondary me- cephala fortinii Rac56 found in Rhodiola plants tabolites as their hosts and therefore there to produce salidroside and p-tyrosol. Molecules, is a tremendous interest of the scientifi c 21: 502. community towards isolation, characteriza- Cui, J.L., Guo, T.T., Ren, Z.X., Zhang, N.S. and Wang, tion and exploitation of endophytic fungi M.L. 2015. Diversity and antioxidant activity of culturable endophytic fungi from alpine plants from medicinal plants as was judged by the of Rhodiola crenulata, R. angusta, and R. sacha- amount of publications and number of pat- linensis. PloS one, 10: e0118204. ents (Gokhale et al., 2017). Cui, Y., Yi, D., Bai, X., Sun, B., Zhao, Y. and Zhang, Y. 2012. Ginkgolide B produced endophytic fungus (Fusarium oxysporum) isolated from Ginkgo Literature cited biloba. Fitoterapia, 83: 913-920. Cutignano, A., Villani, G. and Fontana, A. 2012. One Atanasov, A.G., Waltenberger, B., Pferschy-Wenzig, metabolite, two pathways: convergence of E.M., Linder, T., Wawrosch, C., Uhrin, P., Temml, polypropionate biosynthesis in fungi and ma- V., Wang, L., Schwaiger, S, Heiss, E.H., Rollinger, rine molluscs. Organic letters, 14: 992-995. J.M., Schuster, D., Breuss, J.M., Bochkov, V., Mi- Devari, S., Jaglan, S., Kumar, M., Deshidi, R., Guru, S., hovilovic, M.D., Kopp, B., Bauer, R, Dirsch, V.M. Bhushan, S. and Taneja, S.C. 2014. Capsaicin pro- and Rollinger, J. M. 2015. Discovery and resup- duction by Alternaria alternata, an endophytic ply of pharmacologically active plant-derived fungus from Capsicum annum; LC–ESI–MS/MS natural products: A review. Biotechnology ad- analysis. Phytochemistry, 98: 183-189. vances, 33: 1582-1614. Dziggel, C., Schäfer, H., and Wink, M. 2017. Tools of Bhalkar, B.N., Patil, S. M. and Govindwar, S.P. 2016. pathway reconstruction and production of eco- Camptothecine production by mixed fermenta- nomically relevant plant secondary metabolites tion of two endophytic fungi from Nothapodytes in recombinant microorganisms. Biotechnology nimmoniana. Fungal Biology, 120: 873-883. Journal, 2:1666145. Bömke, C. and Tudzynski, B. 2009. Diversity, regu- El-Elimat, T., Raja, H.A., Graf, T.N. Faeth, S.H., Cech, lation, and evolution of the gibberellin biosyn- N.B. and Oberlies, N.H. 2014. Flavonolignans thetic pathway in fungi compared to plants and from Aspergillus iizukae, a fungal endophyte of bacteria. Phytochemistry, 70: 1876-1893. milk thistle (Silybum marianum). Journal of Nat- Butelman, E.R. and Kreek, M.J. 2015. Salvinorin A, a ural Products, 77: 193–199. kappa-opioid receptor agonist hallucinogen: El-Hawary, S.S., Mohammed, R., AbouZid, S.F., Ba- pharmacology and potential template for novel keer, W., Ebel, R., Sayed, A.M., and Rateb, M.E. pharmacotherapeutic agents in neuropsychiat- 2016. Solamargine production by a fungal en- ric disorders. Frontiers in Pharmacology, 6: 285. dophyte of Solanum nigrum. Journal of Applied Carrier, D.J., van Beek, T.A., van der Heijden, R., and Microbiology, 1201: 143–50. Veroorte, R. 1998. Distribution of ginkgolides El-Sayed, A.S., Abdel-Ghany, S.E. and Ali, G.S. 2017. and terpenoids biosynthetic activity in Ginkgo Genome editing approaches: manipulating of biloba. Phytochemistry, 48: 89–92. lovastatin and taxol synthesis of fi lamentous Chen, S.L., Yu, H., Luo, H.M., Wu, Q., Li, C.F. and Stein- fungi by CRISPR/Cas9 system. Applied Microbiol- metz, A. 2016. Conservation and sustainable ogy and Biotechnology, 101: 3953-3976. use of medicinal plants: problems, progress, Feher, J. and Lengyel, G. 2012. Silymarin in the pre- and prospects. Chinese Medicine, 11: 37. vention and treatment of liver diseases and pri-

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Ενδοφυτικοί μύκητες που διαβιούν εντός των φαρμακευτικών φυτών έχουν την ιδιότητα να παράγουν τους ίδιους ή παρόμοιους δευτερογενείς μεταβολίτες με τους ξενιστές τους

Α. Βενιεράκη, Μ. Δήμου και Π. Κατινάκης

Περίληψη Τα φαρμακευτικά φυτά χρησιμοποιούνται εδώ και χιλιάδες χρόνια στην παραδοσιακή φαρ- μακολογία και ιατρική. Στις μέρες μας, τα φυτά αυτά αξιοποιούνται για την απομόνωση ιδιαίτερα απο- τελεσματικών φυτικών φαρμακευτικών ουσιών, με καθόλου ή ελάχιστες παρενέργειες στο χρήστη. Οι φυσικές πηγές φαρμακευτικών φυτών εξαντλούνται σταδιακά με αποτέλεσμα πέραν της οικολογικής διατάραξης από την εξαφάνιση του φυτικού είδους, να κινδυνεύει δραματικά η απόκτηση του βιοδρα- στικού προϊόντος, το οποίο ούτως ή άλλως βρίσκεται σε χαμηλή συγκέντρωση στο φυτό. Επί παρα- δείγματι, η ποσότητα των αλκαλοειδών που προέρχονται από φυτά βίνκας και τα οποία χρησιμοποι- ούνται ως ισχυρά αντικαρκινικά φάρμακα, ανέρχεται στα 3 κιλά ανά έτος δηλαδή απαιτούνται 1.5x106 κιλά ξηρού βάρους φύλλων. Από αυτήν την άποψη, η παρούσα βιβλιογραφική ανασκόπηση αποσκο- πεί στο να τονίσει τη σημασία των ενδοφυτικών μυκήτων που διαβιούν εντός των φαρμακευτικών φυ- τών και οι οποίοι είναι ικανοί να βιοσυνθέτουν τους ίδιους ή παρόμοιους δευτερογενείς μεταβολίτες με τους ξενιστές τους. Επιπλέον, συζητείται η εξελικτική προέλευση των γονιδίων που εμπλέκονται σε αυ- τές τις μεταβολικές οδούς καθώς και οι προσεγγίσεις που αποσκοπούν στην ενίσχυση της παραγωγής αυτών των μεταβολιτών από ενδοφυτικούς μύκητες.

Hellenic Plant Protection Journal 10: 51-66, 2017

SHORT COMMUNICATION

First record of Aphis craccivora Koch (Hemiptera: Aphididae) on aronia crop in Montenegro

N. Latinović1, F. Karamaouna2 and N.G. Kavallieratos3*

Summary The aphid Aphis craccivora was recorded on the crop of aronia, Aronia melanocarpa, in Montenegro, in June 2015 and 2016. This is the fi rst record of A. craccivora in Montenegro on aronia.

Additional keywords: aphid, Aphis craccivora, Aronia melanocarpa, southeastern Europe

In recent years aronia, Aronia melanocarpa titudes between 1063 m and 1077 m. They (Michx.) Elliott (Rosales: Rosaceae), has be- were spotted on two plants among a total of come a quite popular fruit crop in Monte- 1600 bushes. One year later, in June 2016, the negro. It is a woody perennial shrub, resis- presence of aphids was recorded on numer- tant to cold and can be successfully grown ous bushes of aronia among a total of 3000 in conditions of severe continental climate plants at the locality Stevanovac of the same (Nikolić and Milivojević, 2010), which domi- Municipality at altitudes between 875 m and nates in the northern part of Montenegro. It 905 m. Samples of aphids were collected in is currently considered as a profi table crop 2016 and were identifi ed as Aphis craccivora due to a relatively high price of the fruit Koch (Hemiptera: Aphididae). To our know- (black chokeberries) and its other uses, in- legde, this is the fi rst record of A. craccivora cluding processed products (i.e., syrup, infesting aronia in Montenegro. Aphids have juice, soft spreads, tea, food colors) (McKay, been previously reported as pests of aro- 2001) and as an ornamental plant (Yovkova nia (infestation of shoot tips) but the conse- et al., 2013). For all these reasons and the fact quent slow down eff ect on the plant growth that it is attacked by a small number of pests is not considered serious because the plants and diseases, aronia has earned a profound are vigorous (McKay, 2001). Recently, Aphis place in the organic production in Montene- spiraecola Patch (Hemiptera: Aphididae) gro, where among the total number of 203 and Aulacorthum circumfl exum (Buckton) registered organic producers, 20 of them (Hemiptera: Aphididae) were identifi ed as grow aronia berries at a surface area of ap- pests of A. melanocarpa from southeastern proximately 10 ha. Europe (Bulgaria) (Yovkova et al., 2013). In June 2015, at the locality of Bojna Nji- Aphis craccivora is a relatively small spe- va, Municipality of Mojkovac, aphids were cies. The apterous viviparous female individ- observed to infest an aronia plantation at al- uals have a shiny black or dark brown body with a prominent cauda and brown to yel- low legs. The immatures are slightly dust- 1 University of Montenegro, Biotechnical Faculty, Mi- ed with wax while adults appear without haila Lalića 15, Podgorica, Montenegro. wax. The antennae have six segments. The 2 Benaki Phytopathological Institute, 8 St. Delta Str., GR-145 61 Kifi ssia, Attica, Greece. distal part of femur, siphunculi and cauda 3 Agricultural University of Athens, Department of are black. The length of apterae individuals Crop Science, 75 Iera Odos Str., GR-118 55, Athens, At- tica, Greece. ranges between 1.4 and 2.2 mm. The alate * Corresponding author: [email protected] viviparous A. craccivora females have abdo-

© Benaki Phytopathological Institute 68 Latinović et al. men with dorsal cross bars. The length of al- Aphidius colemani Viereck, Aphidius matri- atae ranges between 1.4 and 2.1 mm (Black- cariae Haliday, Binodoxys acalephae (Mar- man and Eastop, 2000). shall), Binodoxys angelicae (Haliday), Diaer- Aphis craccivora is associated with about etiella rapae (M’Intosh), Ephedrus pericae 50 crops and weed species belonging to 19 Froggat, Lipolexis gracilis Förster, Lysiphle- plant families (i.e., Amaranthaceae, Araceae, bus confusus Tremblay and Eady, Lysiphle- Asteraceae, Brassicaceae, Caryophyllaceae, bus fabarum (Marshall), Lysiphlebus orientalis Chenopodiaceae, Cucurbitaceae, Fabaceae, Starý and Rakhshani, Lysiphlebus testaceipes Malpighiaceae, Malvaceae, Nyctaginaceae, (Cresson), Praon abjectum (Haliday), Praon Pedaliaceae, Portulacaceae, Ranunculaceae, volucre (Haliday). Important predators in- Rosaceae, Rutaceae, Solanaceae, Sterculi- clude coccinellid beetles [Cheilomenes sex- aceae, Zingiberaceae) from which the aphid maculata (F.), Coccinella septempunctata (L.) mainly attacks Fabaceae (Blackman and Eas- (Coleoptera: Coccinelidae)], syrphid larvae top, 2007; Kavallieratos et al., 2007; Mehrpar- [Ischiodon scutellaris (F.) (Diptera: Syrphidae)] var et al., 2012; Yovkova et al., 2013; CABI data Neuroptera larvae [Micromus timidus Ha- base, 2016). The species is probably palearc- gen (Neuroptera: Hemerobiidae)] and Dip- tic warm temperate in origin but it has now tera larvae [Aphidoletes aphidimyza (Ron- a cosmopolitan distribution; it is abundant dani) (Diptera: Cecidomyidae)]. Spiders may in subtropical and tropical regions, and in also be important in some areas (CABI data the Mediterranean. It is one of the common- base, 2016). Recorded fungal pathogens in- est aphid species with a high pest status in clude Fusarium pallidoroseum (Cooke) Sacc. the tropics (Blackman and Eastop, 2000). (Hypocreales: Nectriaceae) (Hareendranath Aphis craccivora is generally anholocy- et al., 1987) and Neozygites fresenii (Nowak.) clic (wingless and winged females), ovovi- Remaud. and S. Keller (Entomophthorales: viparous. In the tropics the aphid reproduc- Neozygitaceae) (Zhang, 1987; Sewify, 2000). es parthenogenetically throughout the year Most of the major chemical groups of in- while in areas with colder winters, overwin- secticides have been used against this aphid tering may be as egg or hibernation. In Eu- species, including organophosphates, car- rope, males (alate) and sexual forms have bamates and pyrethroids (CABI data base, been recorded in Germany (Falk, 1960). Tem- 2016). However, decisions concerning the peratures that range between 24 and 28.5°C chemical treatment against A. craccivora and 65% relative humidity (= RH) are opti- should take into account the identity and mal conditions for the development of A. abundance of local populations of its natu- craccivora (Réal, 1955; Mayeux, 1984), which ral enemies in the context of an integrated is capable of rapid population development. pest management, so as to avoid outbreaks Formation of winged individuals is triggered of this important pest. by the reduction in the intensity of hydro- carbon translocation (Mayeux, 1984). Young colonies concentrate on growing points of Literature cited plants and are regularly attended by ants (mutualism with ants) (Soans and Soans, Blackman, R.L. and Eastop, V.F. 2000. Aphids on the World’s Crops. An Identifi cation and Informa- 1971; Hamid et al., 1977; Takeda et al., 1982; tion Guide. Second Edition. The Natural Histo- Patro and Behera, 1991). ry Museum, London, 466 pp. The spectrum of natural enemies that Blackman, R.L. and Eastop, V.F. 2007. Taxonom- are associated with A. craccivora is wide. For ic Issues. In: van Emden H.F. and R. Harrington instance, Kavallieratos et al. (2004, 2016) re- (eds.). Aphids as Crop Pests. Wallingford, Oxford- shire, pp 1-30. ported 13 parasitoid species (Hymenoptera: CABI data base. 2016. http://www.cabi.org/isc/ Braconidae: Aphidiinae) that parasitize this datasheet/6192. aphid in agricultural and non-agricultur- Falk, U. 1960. Fber das Auftreten von IntermediSr- al ecosystems in southeastern Europe, i.e., formen zwischen oviparem und gefl ügeltem

viviparem Weibchen bei Aphis craccivora Koch. ent host plants. North-Western Journal of Zoolo- Zoologischer Anzeiger, 165: 388-392. gy, 8(1): 172-180. Hamid, S., Sha, M.A. and Anwar, M.A. 1977. Some Nikolić, M.D. and Milivojević, J.M. 2010. Jagodaste ecological and behavioural studies on Aphis voćke - Tehnologoja gajenja. Naučno voćarsko craccivora Koch (Hemi.: Aphididae). Technical društvo Srbije, Čačak. Bulletin, Commonwealth Institute of Biological Patro, B. and Behera, M.K. 1991. Mutualism between Control, 18: 99-111. the bean aphids (Aphis craccivora Koch) and ants. Hareendranath, V., Nair, K.P.V. and Paulos, S. 1987. Orissa Journal of Agricultural Research, 4: 238. Fusarium pallidoroseum (Cooke) Sacc. as a fun- Réal, P. 1955. Le cycle annuel du puceron de l’arachi- gal pathogen of Aphis craccivora Koch. Ento- de (Aphis leguminosae Theob.) en Afrique noire mon, 12: 392-394. française et son déterminisme. Revue de Patho- Kavallieratos, N.G., Tomanović, Ž, Starý, P., Athanas- logie Végétale et d’Entomologie agricole de Fran- siou, C.G., Sarlis, G.P., Petrović, O., Niketić, M. ce, 34(1-2): 1-122. and Anagnou-Veroniki, M. 2004. A survey of Sewify, G.H. 2000. Neozygites fresenii causing epi- aphid parasitoids (Hymenoptera: Braconidae: zootic in aphids (Aphis craccivora Koch.) popula- Aphidiinae) of southeastern Europe and their tion on faba bean in Egypt. Bulletin of Faculty of aphid - plant associations. Applied Entomology Agriculture, University of Cairo, 51: 85-94. and Zoology, 39: 527-563. Soans, A.B. and Soans, J.S. 1971. Proximity of the col- Kavallieratos, N.G., Tomanović, Ž., Sarlis, G.P, Vayias, onies of the tending ant species as a factor de- B.J., Žikić, V. and Emmanouel, N.E. 2007. Aphids termining the occurrence of aphids. Journal of (Hemiptera: Aphidoidea) on cultivated and self- the Bombay Natural History Society, 68: 850-851. sown plants in Greece. Biologia, 62: 335-344. Takeda, S., Kinomura, K. and Sakurai, H. 1982. Eff ects Kavallieratos, N.G., Tomanović, Ž., Petrović, A., of ant-attendance on the honeydew excretion Kocić, K., Janković, M. and Starý, P. 2016. Para- and larviposition of the cowpea aphid, Aphis sitoids (Hymenoptera: Braconidae: Aphidiinae) craccivora Koch. Applied Entomology and Zoolo- of aphids feeding on ornamental trees in south- gy, 17: 133-135. eastern Europe: key for identifi cation and tri- trophic associations. Annals of the Entomologi- Yovkova, M., Petrović-Obradović, O., Tasheva- cal Society of America, 109: 473-487. Terzieva, E. and Pencheva, A. 2013. Aphids (Hemiptera, Aphididae) on ornamental plants in Mayeux, A. 1984. The groundnut aphid. Biology and greenhouses in Bulgaria. ZooKeys, 319: 347–361. control. Oleagineux, 39: 425-434. Zhang, X.L. 1987. Processes of infection and patho- McKay, S.A. 2001. Demand increasing for aronia genesis of Entomophthora fresenii on aphids. and elderberry in North America. New York Fruit Chinese Journal of Biological Control, 3: 121-123. Quarterly, 9: 2-3. Mehrpavar, M., Madjdzadeh, S.M., Mahdavi Arab, N., Esmaeilbeygi, M. and Ebrahimbour, E. 2012. Morphometric discrimination of black legume aphid, Aphis craccivora Koch (Hemiptera: Aphididae), populations associated with diff er- Received: 18 January 2017; Accepted: 8 February 2017

ΣΥΝΤΟΜΗ ΑΝΑΚΟΙΝΩΣΗ

Πρώτη καταγραφή της αφίδας Aphis craccivora Koch (Hemiptera: Aphididae) σε καλλιέργεια αρωνίας στο Μαυροβούνιο

N. Latinović, Φ. Καραμαούνα και Ν.Γ. Καβαλλιεράτος

Περίληψη Η αφίδα Aphis craccivora καταγράφηκε να προσβάλλει την καλλιέργεια της αρωνίας, Aronia melanocarpa, στο Μαυροβούνιο, τον Ιούνιο των ετών 2015 και 2016. Πρόκειται για την πρώτη καταγρα- φή του A. craccivora επί του A. melanocarpa στο Μαυροβούνιο.

Hellenic Plant Protection Journal 10: 67-69, 2017

Reaction of the native Greek tomato varieties ‘Chondrokatsari Messinias’ and ‘Katsari Santorinis’ to Fusarium oxysporum f. sp. lycopersici and Rhizoctonia solani infection

A.I. Darras1*, A. Kotsiras1, C. Delis1, K. Nifakos1, E. Pavlakos2 and V. Demopoulos1

Summary Plants have to cope with a number of biotic stresses among which, infectious diseases. The present study was conducted to investigate the reaction of two native Greek tomato vars, ‘Chon- drokatsari Messinias’ and ‘Katsari Santorinis’, to infection by Fusarium oxysporum f. sp. lycopersici and Rhizoctonia solani. Disease symptoms, disease incidence and severity were recorded and the eff ects of infection on the number of fl owers, the biomass production (fresh and dry weight), CO2 assimilation, stomatal conductance and transpiration were also evaluated. Both tomato varieties were susceptible to F. oxysporum f. sp. lycopersici and R. solani infection. However, ‘Chondrokatsari Messinias’ was found to be less susceptible to F. oxysporum f. sp. lycopersici compared to ‘Katsari Santorinis’. Both pathogens negatively aff ected biomass production of var. ‘Chondrokatsari Messinias’ but not that of ‘Katsari San- torinis’. The number of fl owers produced by ‘Chondrokatsari Messinias’ was negatively aff ected by R. solani but not by F. oxysporum f. sp. lycopersici. Infection of both varieties by R. solani also caused reduction in the CO2 assimilation, stomatal conductance and transpiration.

Additional Keywords: dry weight, Fusarium wilt, native tomato varieties, photosynthesis, stem canker, transpiration

Introduction ly studied (Terzopoulos and Bebeli, 2008; Mazzucato et al., 2010; Cebolla-Cornejo et Native plant varieties have been extensive- al., 2013; Corrado et al., 2014). For example, ly examined throughout the modern hu- seven out of 33 native Greek tomato variet- man history (Teshome et al., 1997; Zeven, ies comprise 27 diff erent morphotypes (Ter- 1998). Such plant material is usually select- zopoulos and Bebeli, 2008). However, most ed and maintained by traditional farmers as of them have not yet been genetically clas- part of their social, economic, cultural and sifi ed or morphologically described. ecological history. Louette et al. (1997) de- Plants have to cope with a number of bi- scribed a native variety as a farmer’s variety otic and abiotic stresses during their growth which has not been improved by any formal and development (Kai et al., 2007). Fusari- breeding programme. Native varieties con- um wilt diseases, caused by the pathogen- tain much more genetic diversity than mod- ic soil-inhabiting fungus Fusarium oxyspo- ern cultivars or hybrids (Zeven, 1998; Terzo- rum Schlectend.:Fr., can cause severe losses poulos and Bebeli, 2008; Terzopoulos and in a wide range of cultivated and non-culti- Bebeli, 2010). Therefore, they are among the vated plants (Larkin et al., 1998). On tomato, most important sources of genetic variation two forms of the pathogen, F. oxysporum f. for breeders. So far, a large number of native sp. lycopersici W.C. Snyder & H.N. Hans. and F. varieties grown in the Mediterranean region oxysporum f. sp. radicis-lycopersici W.R. Jarvis have been morphologically and genetical- & Shoemaker, cause two symptomatologi- cally distinct diseases, i.e. vascular wilt and crown and root rot, respectively. F. oxyspo- 1 Department of Agricultural Technology, Technologi- cal Educational Institute (TEI) of Peloponnese, GR-241 rum f. sp. lycopersici invades the vascular sys- 00 Antikalamos, Kalamata, Greece. tem of the plant through natural openings 2 COMPO HELLAS S.A. Egialias 54, GR-151 25 Marousi Attica, Greece. or damaged tissue of the roots (Bishop and * Corresponding author: [email protected] Cooper, 1983; Agrios, 1997; Di Pietro et al.,

2003). Initial symptoms of Fusarium wilt are from local growers were sown in 60 × 20 cm described as vein clearing of the younger plastic trays (INA plastics, Athens, Greece) leaves and leaf epinasty, followed by stunt- fi lled with sterile white peat moss (TS2 Klas- ing, yellowing of the lower leaves, progres- mann-Deilmann, Geeste, Germany; pH 6.0). sive wilting of leaves and stem, defoliation Tomato seedlings were grown inside a non- and fi nally plant death. In cross-sections of heated greenhouse located at the premis- the stem, a brown ring is evident in the area es of the Technological Educational Institute of the vascular bundles (Bishop and Cooper, of Peloponnese (lat. 37° 20΄ 20΄΄N, long. 22° 1983; Di Pietro et al., 2003). 60΄ 51΄΄E) for 35 d and until they reached the The soil-borne pathogen Rhizoctonia 4-true-leaf stage (approx. 30 cm in height). solani Kühn [teleomorph: Thanatephorus cu- The young plants were then transplanted cumeris (A.B. Frank) Donk] causes serious individually into 5 lt plastic pots fi lled with a damage to many economically important mixture of white peat moss (TS2 Klasmann- horticultural crops (Baker, 1970; Anderson, Deilmann, Geeste, Germany; pH 6.0) and 1982; Sneh et al., 1996). In the past few years, perlite (Perlofl or, Isocon SA, Athens, Greece) the importance of the disease caused by this at 1:1 (v/v). The pots were then placed on alu- pathogen has increased dramatically in Eu- minium benches (0.2 m width x 15 m length rope (Grosch et al., 2005). R. solani strains oc- x 0.5 m height) in a non-heated greenhouse cur ubiquitously and are either saprophytic in a completely randomised design. Stan- or pathogenic to more than 500 plant spe- dard cultivation practices, such as plant tie- cies. Damping-off diseases caused by R. solani up, irrigation and fertilization, were applied in greenhouse-grown vegetables are com- to all plants. The nutrient solution used for monly encountered (Lumsden and Locke, the fertilization of the plants consisted of (in 1989). Symptoms develop as dark brown to mmol/lt) 5.10 Ca2+, 2.40 Mg2+,, 7.00 K+,, 1.50 + 2 - - black cankers on the base of the plant, which NH4 , 3.60 SO4 , 14.30 NO3 , 1.50 H2PO4 and increase in size over time resulting in plant (in μmol/lt) 20 Fe 10 Mn, 5 Zn, 0.80 Cu, 35 collapse (Baker, 1970; Agrios, 1997). B and 0.5 Mo. Electrical conductivity (EC) No information is available in the litera- and pH of the nutrient solution ranged be- ture with respect to the reaction of the na- tween 2.4-2.5 mS/cm and 5.8-6.0, respective Greek tomato varieties ‘Chondrokatsari tively. Three hundred ml of the nutrient so- Messinias’ and ‘Katsari Santorinis’ to the in- lution was provided to the plants every two fection by soil-borne fungal pathogens or on days during the experimental period. the eff ects of infection on plant growth and Two individual experiments, Experi- development. The objectives of the pres- ment 1 and Experiment 2, were conduct- ent study were to investigate F. oxysporum f. ed starting out at the end of February 2015 sp. lycopersici and R. solani infection process and fi nishing 95 d later. In Experiment 1, to- on the native tomato vars ‘Chondrokatsari mato plants of vars ‘Chondrokatsari Messini- Messinias’ and ‘Katsari Santorinis’, record the as’ and ‘Katsari Santorinis’ were challenged symptomatology of the diseases and corre- with F. oxysporum f. sp. lycopersici, where- late disease intensity (incidence and severi- as in Experiment 2, the same varieties were ty) with plant growth decline after infection. challenged with R. solani. In each experiment, six plants per variety and time of assessment [i.e. 40 or 60 days post inoculation Materials and Methods (dpi)] were used as replicates.

Plant material, cultivation practices and Pathogen isolates, inoculum prepara- experimental design tion and plant inoculation Untreated tomato (Lycopersicon escu- For the inoculation of experimental lentum L.) seeds of vars ‘Chondrokatsari plants, strain BPIC2550 of F. oxysporum f. sp. Messinias’ and ‘Katsari Santorinis’ obtained lycopersici isolated from tomato plants (Lyc-

© Benaki Phytopathological Institute 72 Darras et al. opersicon esculentum L.) and strain BPIC2531 a distance of approximately 1 cm from the of R. solani isolated from potato plants (Sola- stem base. Control plants were treated with num tuberosum L.) were used. Both strains non-inoculated PDA plugs. were provided by the Benaki Phytopatho- logical Institute (Kifi ssia, Athens, Greece). Disease assessments Tomato plants at the stage of 4 true In both experiments, disease symptoms leaves (approx. 30 cm in height) were inoc- were recorded 40 and 60 dpi. In Experiment ulated with F. oxysporum f. sp. lycopersici by 1, disease severity index (DSI) and disease applying a conidial suspension at the basal incidence (DI) on tomato plants inoculated stem-end of each plant (Dhingra and Sinclair with F. oxysporum f. sp. lycopersici were as- 1995; Akköprü and Demir, 2005). The fungal sessed on the root system and stem base. inoculum was prepared as follows: initially DSI was determined using the arbitrary the fungus was cultured on potato dextrose scale of: 0: no symptoms, 1: 1% of roots with agar (PDA, Oxoid Ltd., Basingstoke, Hamp- symptoms, 2: >1-5% of roots with symp- shire, UK) medium in Petri plates at 26°C in toms, 3: 6-10% of roots with symptoms, and the dark for 12 d. The conidial suspension, 4: >10% of roots with symptoms. DI was cal- which consisted of both micro- and mac- culated according to the following formula: roconidia, was prepared by pouring 20 ml Disease incidence (DI) = (number of symp- of sterile distilled water containing 0.01% tomatic plants/total number of inoculated Tween 80 (Sigma, St. Louis, USA) in each plants) x 100 (1) plate. The conidia were dislodged by gen- In Experiment 2, DI, number of cankers tly rubbing the fungal colony surface with a (CN) and average canker diameter (ACD) sterile razor blade. The suspension was fi l- were recorded. DI was calculated according tered through two layers of fi ne, nylon, ster- to formula (1) above. Canker diameter was ile cheesecloth to remove mycelia. The fi nal measured in cm using a digital micrometer conidial concentration was adjusted to 4.5 (Stock No. 600-880, Mitutoyo, Japan). x 106 conidia/ml using a haemocytometer. Twenty ml of the conidial suspension were Biomass production, number of fl owers applied to each plant approx. 3 cm below and physiological parameters of toma- the surface of the growing substrate and at to plants a contact with the stem base using a 5 ml Plant biomass production was record- plastic syringe (i.e. 4 applications around the ed 40 and 60 dpi. Prior to assessment, the plant stem) without wounding the roots (Ak- growing substrate was completely removed köprü and Demir, 2005). Control plants were by gentle washing the root system of the treated with 20 ml of sterile distilled water. plants under running tap water. Biomass Tomato plants at the stage of 4 true was determined by measuring the fresh leaves (approx. 30 cm in height) were inoc- weight (FW; gr) of the aerial plant parts (i.e. ulated with R. solani using mycelium plugs stems, leaves and infl orescences) and the (Dhingra and Sinclair, 1995). Initially, R. solani root system using a digital balance (Kern & cultures were prepared by placing mycelium Sohn GmbH, Balingen, Germany). Then, the plugs cut from the edges of 12-d-old cultures same plant parts were dried separately in at the centre of PDA (Oxoid Ltd., Basingstoke, an oven (Daihan Labtech Co. Ltd, Gagok-ri, Hampshire, UK) plates. The inoculated plates Korea) at 75°C for 72 h and the dry weights were incubated at 25°C for 12 d in the dark. (DW; gr) were also measured. The number of Mycelium plugs, 5 mm in diameter, were fl owers was recorded once every week (total then cut from the edges of the growing colo- of eight counts over the 60 dpi period). nies using a cork borer. Inoculation of toma- The physiological parameters of CO2 as- 2 to plants was carried out by placing three, 5 similation (As; μmol CO2/m /s), stomatal con- 2 mm in diameter, mycelium plugs 3 cm below ductance (gs; mmol/m /s) and transpiration the surface of the growing substrate and at (E; mmol/m2/s) were recorded 16 and 27 dpi

© Benaki Phytopathological Institute Fusarium and Rhizoctonia infecting native Greek tomato varieties 73 at anthesis and fruit set, respectively. As, gs Alabouvette (1999), F. oxysporum f. sp. lyco- and E were measured using a LCpro+ port- persici was able to perform a vascular infec- able photosynthesis system (ADC BioScien- tion of tomato root tissue producing lesions tifi c Ltd. Great Amwell, Herts, UK). Record- on the roots. However, these lesions had lim- ings were made between 10:00 and 12:00 ited expansion probably due to intense de- a.m on fully expanded young leaves of sim- fense reactions occurring in the superficial ilar size. Photosynthetic photon fl ux density cell layers (Olivain and Alabouvette, 1999). (PPFD) in the leaf chamber was set at 1100 Brown discoloration of the vascular system μmol/m2/sec with a halogen lamp at 25°C, of the plants was also observed in the pres- while CO2 reference ranged between 380 ent study indicating colonization of xylem and 437 ppm. vessels by the pathogen (Figure 1B). This is considered a typical symptom of infection Statistical analysis of tomato plants by F. oxysporum f. sp. lyco- Both experiments were factorial with va- persici following root tissue penetration and riety and time of assessment (i.e. dpi) as the colonization of the vascular system by the main factors. Experimental data were sub- pathogen (Bishop and Cooper, 1983; Agrios, jected to one-way ANOVA and means were 1997; Olivain and Alabouvette, 1999). separated using the Duncan’s multiple range R. solani infected the plants of both vari- test at P = 0.05. Prior to analysis, DI percent- eties at the stem base (Figure 2). Symptoms age data were transformed to logarithmic were recorded as volcano-like cankers of values (i.e. Log10) to highlight signifi cant dif- various sizes, with a brown centre and dark ferences between means, although, the un- brown to black margin (Figure 2A). Cankers transformed data are presented in the tables. increased in size with time resulting in plant Scale data of DSI were analysed using the collapse 40 dpi (Figure 2B). Usually, R. so- Kruskal-Wallis non-parametric test. Statistical lani infection progresses quickly, especial- analysis was performed with SPSS for Win- ly when conditions are favourable (i.e. low dows, Version 12.0 (Chicago, SPSS Inc., USA). temperatures and increased soil humidity) (Baker, 1970; Agrios, 1997).

Results and Discussion Disease assessments DSI on tomato plants of vars ‘Chon- Disease symptoms drokatsari Messinias’ and ‘Katsari Santorinis’ F. oxysporum f. sp. lycopersici infected the inoculated with F. oxysporum f. sp. lycopersi- root and the vascular system of tomato vars ci was low and ranged between 0.5 and 2.5 ‘Chondrokatsari Messinias’ and ‘Katsari San- (on a 0-4 scale). In general, DSI and DI did torinis’ (Figure 1). No visual symptoms were not signifi cantly (P < 0.05) increase with time observed on the experimental plants 40 dpi. (from 40 to 60 dpi) with the exception of DSI However, tomato plants of ‘Chondrokatsa- on var. Katsari Santorinis (Table 1). More spe- ri Messinias’ and, to a lesser extent, ‘Katsa- cifi cally, DSI on ‘Katsari Santorinis’ increased ri Santorinis’ showed a limited degree of leaf by 5-fold from 40 to 60 dpi (Table 1). epinasty and leaf yellowing 60 dpi. Di Pietro In general, CN, ACD and DI on ‘Chon- et al. (2003) described the symptoms caused drokatsari Messinias’ and ‘Katsari Santorin- on tomato plants infected by F. oxysporum is’ tomato plants inoculated with R. solani as leaf epinasty, followed by stunting, yel- signifi cantly (P < 0.05) increased with time lowing of the lower leaves, progressive wilt- (from 40 to 60 dpi) with the exception of CN ing, defoliation and fi nally plant death. In on ‘Katsari Santorinis’ (Table 2). More spe- the present study, symptoms were also ob- cifi cally, CN, ACD and DI on ‘Chondrokatsa- served on the surface of the roots as dark ri Messinias’ increased with time by 7-, 23- brown to black, necrotic, circular or irregular and 67%, respectively (Table 2). ACD on var. lesions (Figure 1A). According to Olivain and ‘Katsari Santorinis’ increased by 4-fold from

40 to 60 dpi. Nevertheless, 40 dpi, all ex- Eff ects of F. oxysporum f. sp. lycopersici perimental plants of var. ‘Katsari Santorinis’ infection of tomato plants on biomass showed disease symptoms (DI=100%) (Ta- production, number of fl owers As, gs and ble 2). E Based on the above-mentioned results, The results of the present study showed var. ‘Katsari Santorinis’ was found to be that FW and DW of the aerial parts of var. more susceptible to F. oxysporum f. sp. lyco- ‘Chondrokatsari Messinias’ inoculated with F. persici infection compared to ‘Chondrokat- oxysporum f. sp. lycopersici were signifi cant- sari Messinias’, as the former showed sig- ly (P < 0.05) lower compared to those of the nifi cantly (P < 0.05) higher disease levels 60 control plants 60 dpi (Figure 3). However, FW dpi compared to the latter (Table 1). The results of the present study also showed that, 60 dpi, vars ‘Chondrokatsari Messinias’ and ‘Katsari Santorinis’ showed similar susceptibility to infection by R. solani (Table 2).

Figure 1. Light to dark brown lesions (arrows) on roots of Figure 2. Dark brown cankers (arrows) on the stem base of ‘Katsari Santorinis’ tomato plants (A) and discoloration (ar- ‘Chondrokatsari Messinias’ tomato plants as a result of their rows) of the vascular system of ‘Chondrokatsari Messinias’ to- infection by Rhizoctonia solani (A) 60 dpi. Collar rot symptoms mato plants (B) inoculated with Fusarium oxysporum f. sp. ly- on ‘Katsari Santorinis’ tomato plants inoculated with R. solani copersici 60 dpi. (B).

Table 1. Disease severity index (DSI; scale 0–4) and disease incidence (DI; % plants with symptoms) on tomato plants of vars ‘Chondrokatsari Messinias’ and ‘Katsari Santorinis’ inoculated with Fusarium oxysporum f. sp. lycopersici. DSI and DI are means of six replicates and were recorded 40 and 60 days post- inoculation (dpi). Means followed by diff erent letters are statistically diff erent according to the Duncan’s Multiple Range test (P = 0.05)).

DSI DI Variety Treatment (scale 0–4) (%) dpi 40 60 40 60 ‘Chondrokatsari Messinias’ Control 0 0 0 0 F. oxysporum f. sp. lycopersici 0.67 a 1.00 a 33 a 50 a

‘Katsari Santorinis’ Control 0 0 0 0 F. oxysporum f. sp. lycopersici 0.50 a 2.50 b 67 ab 100 b

Table 2. Number of cankers (CN), average canker diameter (ACD; cm) and disease incidence (DI; % plants with symptoms), on tomato plants of vars ‘Chondrokatsari Messinias’ and ‘Kat- sari Santorinis’ inoculated with Rhizoctonia solani. CN, ACD and DI are means of six replicates and were recorded 40 and 60 days post inoculation (dpi). Means followed by diff erent letters are statistically diff erent according to the Duncan’s Multiple Range test (P = 0.05).

ACD DI Variety Treatment CN (cm) (%) dpi 40 60 40 60 40 60 ‘Chondrokatsari Messinias’ Control 0 0 0 0 0 0 R. solani 0.67 a 4.67 b 0.13 a 3.00 bc 33 a 100 b

‘Katsari Santorinis’ Control 0 0 0 0 0 0 R. solani 3.00 b 4.50 b 1.00 ab 4.00 c 100 b 100 b and DW of the root system of ‘Chondrokat- (Lorenzini et al., 1997). In the present study, sari Messinias’ were not signifi cantly aff ected As, gs and E were measured earlier (i.e. 16 and by the infection of the pathogen. Infection of 27 dpi) than DS and DI (i.e. 40 and 60 dpi). var. ‘Katsari Santorinis’ by F. oxysporum f. sp. Even at 40 dpi, DSI and DI on both varieties lycopersici did not signifi cantly (P < 0.05) af- were very low (Table 1) which may explain fect FW and DW of the aerial parts and the the insignifi cant eff ect of F. oxysporum f. sp. root system of the experimental plants (Fig- lycopersici infection on As, gs and E. ure 3). The number of fl owers of ‘Katsari San- Eff ects of R. solani infection of tomato torinis’ plants inoculated with F. oxyspo- plants on biomass production, number rum f. sp. lycopersici was signifi cantly (P < of fl owers As, gs and E 0.05) lower compared to that of the con- FW and DW of the aerial parts of var. trol plants (Table 3). However, F. oxysporum ‘Chondrokatsari Messinias’ inoculated with f. sp. lycopersici did not signifi cantly aff ect R. solani were signifi cantly (P < 0.05) low- the number of fl owers of var. ‘Chondrokat- er compared to those of the control plants sari Messinias’ (Table 3). 60 dpi (Figure 4). However, R. solani did not

As, gs and E of both tomato varieties were signifi cantly aff ect FW and DW of the aerial not signifi cantly (P < 0.05) aff ected by F. ox- parts of var. ‘Katsari Santorinis’ even 60 dpi ysporum f. sp. lycopersici infection (Table 3). (Figure 4). No signifi cant (P < 0.05) diff erenc- Pshibytko et al. (2006) showed that Fusari- es in FW and DW of roots were observed 60 um wilt led to suppression of the photosyn- dpi between the inoculated plants of both thetic activity of 4- to 6-month-old tomato varieties and the controls (Figure 4). plants of var. Kunera. Although, only in the The number of fl owers of var. ‘Chon- case of a slowly developed pathogen could drokatsari Messinias’ inoculated with R. so- damage the photosystem. Signifi cant diff er- lani was signifi cantly (P < 0.05) lower com- ences in As, gs and E between tomato plants pared to the controls (Table 4). However, inoculated with F. oxysporum f. sp. lycopersi- tomato plants of var. ‘Katsari Santorinis’ in- ci and the non-inoculated controls were re- oculated with R. solani produced signifi cant- ported by Lorenzini et al. (1997). As, gs and E, ly (P < 0.05) more fl owers than the control were negatively aff ected by F. oxysporum plants (Table 4). f. sp. lycopersici infection and correlations As, gs and E of both tomato varieties inoc- were made between the time post-inocula- ulated with R. solani were signifi cantly (P < tion (i.e. dpi) and the values of As, gs and E. 0.05) lower than those of the control plants

As dpi increased, As, gs and E were reduced (Table 4).

Figure 3. Fresh weight (FW) and dry weight (DW) of the aerial parts (A, C) and the root system (B, D) of ‘Chondrokatsari Messinias’ (A, B) and ‘Katsari Santorinis’ (C, D) tomato plants inoculated with Fusarium oxysporum f. sp. lycopersici. Data are means of six replicates. FW and DW were measured 40 and 60 dpi. Columns followed by diff erent letters are statistically dif- ferent according to the Duncan’s Multiple Range test (P = 0.05).

Table 3. Eff ect of Fusarium oxysporum f. sp. lycopersici on the number of fl owers, CO2 assimilation (As), stomatal conductance (gs) and transpiration (E) of tomato plants of vars ‘Chon- drokatsari Messinias’ and ‘Katsari Santorinis’. Data are means of six replicates and were recorded 16 and 27 dpi. Means followed by diff erent letters are statistically diff erent according to the Duncan’s Multiple Range test (P = 0.05).

Variables

Variety Treatment Number of As gs E 2 2 2 fl owers (μmol CO2/m /s) (mmol /m /s) (mmol /m /s)

‘Chondrokatsari Control 0.65 a 9.60 a 0.23 ab 2.58 ab Messinias’ F. oxysporum f. sp. lycopersici 1.68 a 8.84 a 0.21 a 2.36 a

‘Katsari Santorinis’ Control 8.85 c 10.43 a 0.32 b 2.98 b F. oxysporum f. sp. lycopersici 5.00 b 9.89 a 0.30 b 3.32 b

Figure 4. Fresh weight (FW) and dry weight (DW) of the aerial parts (A, C) and the root system (B, D) of ‘Chondrokatsari Messinias’ (A, B) and ‘Katsari Santorinis’ (C, D) tomato plants inoculated with Rhizoctonia solani. Data are means of six replicates. FW and DW were measured 40 and 60 dpi. Columns followed by diff erent letters are statistically diff erent according to the Duncan’s Multiple Range test (P = 0.05).

Table 4. Eff ect of Rhizoctonia solani on the number of fl owers, CO2 assimilation (As), stomatal conductance (gs) and transpiration (E) of tomato plants of vars ‘Chondrokatsari Messinias’ and ‘Katsari Santorinis’. Data are means of six replicates and were recorded 16 and 27 dpi. Means followed by diff erent letters are statistically diff erent according to the Duncan’s Mul- tiple Range test (P = 0.05).

Variables Variety Treatment Number of As gs E 2 2 2 fl owers (μmol CO2/m /s) (mmol/m /s) (mmol/m /s)

‘Chondrokatsari Messinias’ Control 2.40 b 9.08 b 0.19 bc 2.77 b R. solani 1.55 a 7.80 a 0.15 a 2.30 a

‘Katsari Santorinis’ Control 7.95 c 11.01 c 0.23 c 3.01 c R. solani 8.80 d 8.35 ab 0.18 ab 2.50 ab

Conclusions their occurence. In ‘Rhizoctonia solani, Biology and Pathology’ J.R. Parmeter ed. University of California Press, USA. pp.125-148. The present study was the fi rst attempt to Bishop, C.D. and Cooper, R.M. 1983. An ultrastruc- investigate the reaction of two native Greek tural study of vascular colonization in three vas- tomato varieties, ‘Chondrokatsari Messini- cular wilt diseases I. Colonization of susceptible as’ and ‘Katsari Santorinis’, to infection by cultivars. Physiological Plant Pathology, 23(3): F. oxysporum f. sp. lycopersici and R. solani. 323-343. Results showed that both tomato variet- Cebolla-Cornejo, J., Rosello, S. and Nuez, F. 2013. Phe- notypic and genetic diversity of Spanish tomato ies were susceptible to F. oxysporum f. sp. landraces. Scientia Horticulturae, 162: 150-164. lycopersici and R. solani infection. Howev- Corrado, G., Caramante, M., Piff anelli, P. and Rao, R. er, ‘Chondrokatsari Messinias’ was found to 2014. Genetic diversity in Italian tomato landrac- be less susceptible to F. oxysporum f. sp. ly- es: implications for the development of a core copersici compared to ‘Katsari Santorinis’. collection. Scientia Horticulturae, 168: 138-144. Both of the pathogens negatively aff ected Dhingra, O.D. and Sinclair, J.B. 1995. Basic Plant Pa- thology Methods. 2nd edition. CRC Press, Lewis biomass production of var. ‘Chondrokatsari publishers. Messinias’ but not that of ‘Katsari Santorinis’. Di Pietro, A., Madrit, M.P., Caracuel, Z., Delgado-Ja- The number of fl owers produced by ‘Chon- rana, J. and Roncero, M.I.G. 2003. Fusarium ox- drokatsari Messinias’ was negatively aff ect- ysporum: exploring the molecular arsenal of a ed by R. solani but not by F. oxysporum f. sp. vascular wilt fungus. Molecular Plant Pathology, lycopersici. Infection of both tomato variet- 4(5): 315-325. ies by R. solani also caused reduction in the Grosch, R., Faltin, F., Lottmann, J., Kofoet, A., and Berg, G. 2005. Eff ectiveness of three antago- CO2 assimilation, stomatal conductance and nistic bacterial isolates to suppress Rhizocto- transpiration. nia solani Kühn on lettuce and potato. Canadian Additional work is required on the inter- Journal of Microbiology, 51: 345–353. action between the two native Greek toma- Kai, M., Eff mert, U., Berg, G. and Piechulla, B. 2007. to vars ‘Chondrokatsari Messinias’ and ‘Kat- Volatiles of bacterial antagonists inhibit myce- lial growth of the plant pathogen Rhizoctonia sari Santorinis’ and the soil-borne fungi F. solani. Archives of Microbiology, 187: 351-360. oxysporum f. sp. lycopersici and R. solani as Larkin, R.P. and Fravel, D.R. 1998. Effi cacy of vari- well as on the management of the diseases ous fungal and bacterial biocontrol organisms caused by these pathogens. for control of fusarium wilt of tomato. Plant Dis- ease, 82: 1022-1028. Lorenzini, G., Guidi, L., Nali, C., Ciompi, S. and Sol- datini, G. 1997. Photosynthetic response of to- The Benaki Phytopathological Institute is mato plants to vascular wilt diseases. Plant Sci- gratefully acknowledged for providing the ence, 124: 143-152. strains of Fusarium oxysporum f. sp. lycopersici Louette, D., Charrier, A. and Berthaud, J. 1997. In situ (BPIC2550) and Rhizoctonia solani (BPIC2531) conservation of maize in Mexico: genetic diver- used in the present study. sity and maize seed management in a traditional community. Economic Botany, 51: 20–38. Lumsden, R.D. and Locke, J.C. 1989. Biological control of damping-off caused by Pythium ultimum Literature Cited and Rhizoctonia solani with Gliocladium virens in soiless mix. Phytopathology, 79: 361-366. Agrios, G.N. 1997. Plant Pathology. Academic Press. London, UK. Mazzucato, A., Ficcadenti, N., Caioni, M., Mosconi, P., Piccinini, E., Rami, R., Sanampudi, R., Sestili, Akköprü, A. and Demir, S. 2005. Biological control of S. and Ferrari, V. 2010. Genetic diversity and dis- Fusarium wilt in tomato caused by Fusarium ox- tinctiveness in tomato (Solanum lycopersicum ysporum f. sp. lycopersici by AMF Glomus intra- L.) landraces: The Italian case study of ‘A pera radices and some rhizobacteria. Journal of Phy- Abruzzese’. Scientia Horticulturae, 125: 55-62. topathology, 153(9): 544-550. Olivain, C. and Alabouvette, C. 1999. Process of to- Anderson, N.A. 1982. The genetics and pathology mato root colonization by a pathogenic strain of Rhizoctonia solani. Annual Review of Phytopa- of Fusarium oxysporum f. sp. lycopersici in com- thology, 20: 329–374. parison with a non-pathogenic strain. New Phy- Baker, K.F. 1970. Types of Rhizoctonia diseases and tologist, 141(3): 497-510.

Pshibytko, N.L., Zenevich, L.A. and Kabashnikova, L. to (Solanum lycopersicum L.) landraces. Scientia F. 2006. Changes in the photosynthetic appa- Horticulturae, 116: 354-361. ratus during Fusarium wilt of tomato. Russian Terzopoulos, P.J. and Bebeli, P.J. 2010. Phenotipic di- Journal of Plant Physiology, 53(1): 25-31. versity in Greek tomato (Solanum lycopersicum Sneh, B., Jabaji-Hare, S., Neate, S.M. and Dijst, G. L.) landraces. Scientia Horticulturae, 126: 138- 1996. Rhizoctonia species: taxonomy, molecular 144. biology, ecology; pathology and disease con- Zeven, A.C. 1998. Landraces: a review of defi nitions trol. Kluwer, Dordrecht. and clarifi cations. Euphytica, 104:127-139. Teshome, A., Baum, B.R., Fahrig, L., Torrance, J.K., Arnason, T.J. and Lambert, J.D. 1997. Sorghum (Sorghum bicolor (L.) Moench) landrace variation and classifi cation in North Shewa and South Welo, Ethiopia. Euphytica, 97: 255–263. Terzopoulos, P.J. and Bebeli, P.J. 2008. DNA and mor- phological diversity of selected Greek toma- Received: 8 August 2016; Accepted: 26 April 2017

Προσβολή φυτών τομάτας των Ελληνικών παραδοσιακών ποικιλιών ‘Χοντροκατσαρή Μεσσηνίας’ και ‘Κατσαρή Σαντορίνης’ από τους μύκητες Fusarium oxysporum f. sp. lycopersici και Rhizoctonia solani

Α.Ι. Δάρρας, Α. Κώτσιρας, Κ. Δελής, Κ. Νηφάκος, Ε. Παυλάκος και Β. Δημόπουλος

Περίληψη Τα φυτά συχνά πρέπει να αντιμετωπίσουν καταπονήσεις που οφείλονται σε βιοτικούς παράγοντες μεταξύ των οποίων είναι και οι ασθένειες. Η παρούσα μελέτη πραγματοποιήθηκε με σκοπό να εξετάσει την αντίδραση δύο Ελληνικών παραδοσιακών ποικιλιών τομάτας, της ‘Χοντροκατσαρής Μεσσηνίας’ και της ‘Κατσαρής Σαντορίνης’, στην προσβολή από τους μύκητες Fusarium oxysporum f. sp. lycopersici και Rhizoctonia solani. Καταγράφηκαν τα συμπτώματα και η ένταση κάθε ασθένειας και εκτιμήθηκε η επίδραση των προσβολών στην παραγωγή βιομάζας, στον αριθμό των ανθέων, στη φωτοσυνθετική δραστηριότητα, στη στοματική αγωγιμότητα και στη διαπνοή των φυτών. Και οι δύο ποικιλίες ήταν ευπαθείς στη μόλυνση από τους μύκητες F. oxysporum f. sp. lycopersici και R. solani. Εντούτοις η ποικ. ‘Χοντροκατσαρή Μεσσηνίας’ ήταν λιγότερο ευπαθής στην προσβολή από το μύκητα F. oxysporum f. sp. lycopersici σε σχέση με την ‘Κατσαρή Σαντορίνης’. Και τα δύο παθογόνα επηρέασαν αρνητικά την παραγωγή βιομάζας των φυτών της ποικ. ‘Χοντροκατσαρή Μεσσηνίας’ αλλά όχι της ποικ. ‘Κατσαρή Σαντορίνης’. Ο αριθμός των ανθέων της ποικ. ‘Χοντροκατσαρή Μεσσηνίας’ επηρεάστηκε αρνητικά από την προσβολή από το μύκητα R. solani αλλά όχι από τον F. oxysporum f. sp. lycopersici. Η προσβολή από το μύκητα R. solani είχε ως αποτέλεσμα τη μείωση της φωτοσυνθετικής δραστηριότητας, της στοματικής αγωγιμότητας και της διαπνοής των φυτών και των δύο ποικιλιών.

Hellenic Plant Protection Journal 10: 70-79, 2017

SHORT COMMUNICATION

The pink hibiscus mealybug Maconellicoccus hirsutus (Green) (Hemiptera: Pseudococcidae) in Greece

P.G. Milonas* and G.K. Partsinevelos

Summary The invasive pink hibiscus mealybug, Maconellicoccus hirsutus (Green) (Hemiptera: Pseudo- coccidae), is reported for fi rst time in Greece. Individuals of the mealybug were found infesting Hibiscus rosa-sinensis (Linnaeus) (Malvaceae) in private and public gardens in the urban environment in Rhodes, Dodecanese islands, East Greece. This is the fi rst report of genus Maconellicoccus in Greece.

Additional Keywords: hibiscus, invasive species, mealybug

Introduction the world, i.e. North America, the Caribbean and Africa. Overall M. hirsutus distribution The genus Maconellicoccus Ezzat (Hemipte- includes 75 countries in all over the world ra: Pseudococcidae) includes eight species (EPPO, 2005). In Europe, it was reported that are distributed in Australian, Oriental for fi rst time in Cyprus in 2011 (EPPO, 2011). and Ethiopian regions but only Maconellicoc- Upon its introduction into several countries, cus hirsutus has been reported in Palaearctic it has caused substantial economic damag- region (García Morales et al., 2016). Maconel- es through the cost of control operations licoccus hirsutus Green, known as pink hi- and impact on trade. In the US, it has been biscus mealybug, is a highly polyphagous estimated that without control, it may cause species native to southern Asia, that feeds a damage of 163 million dollars only in Flori- on 212 genera in 75 host plant families, in- da (Chong et al., 2015). cluding important crops such as bean (Pha- Adult females are 2.5-4 mm long, wing- seolus vulgaris), chrysanthemum (Chrysan- less, soft-bodied, elongate oval and fl at- themum spp.), hibiscus (Hibiscus spp.), rose tened. Females can lay more than 500 eggs. (Rosa spp.), pumpkin (Cucurbita pepo), avo- Eggs are orange initially but turn into pink cado (Persea americana), citrus (Citrus spp.), before hatching. Crawlers are 0.3 mm long, coconut (Cocos nucifera), coff ee (Coff ea spp.), pink, oval in shape with well-defi ned legs cotton (Gossypium spp.), corn (Zea mays), and antennae, and lack the waxy body vegetables, grape (Vitis vinifera) and pea- coating; young adult females turn greyish- nuts (Arachis hypogaea) (Chong et al., 2015; pink, dusted with mealy white wax that cov- García Morales et al., 2016). For a complete ers their bodies; adult males are gnat-like 1 list of M. hirsutus host plants see Chong et mm long, pink to orange, with a single pair al. (2015). of wings and two pairs of fi laments. They Pink hibiscus mealybug is considered a are weak fl yers, lack mouthparts and live highly invasive species. Although it is be- only one day or two. Entire colony is cov- lieved to originate from India, it has been ered by white, waxy ovisac material (Chong accidentally introduced into other parts of et al., 2015; García Morales et al., 2016). One generation is completed in approximately fi ve weeks in warm conditions. In Jordan, Laboratory of Biological Control, Benaki Phytopatho- nymphs have three peaks and adult females logical Institute, 8 St. Delta Str., GR-145 61, Kifi sia, Atti- ca, Greece. two peaks, in early February and mid-Ju- * Corresponding author: [email protected] ly, respectively (Al-Fwaeer et al., 2014). Here

© Benaki Phytopathological Institute Maconellicoccus hirsutus in Greece 81 we report the fi rst presence of M. hirsutus in M. hirsutus have been reported outside Rho- Greece. des. The specifi c pathway of introduction of Feeding by larvae and adults causes char- the species into Rhodes is unknown. How- acteristic symptoms on the damaged plants. ever, long distance dispersal of the pest is During feeding both larvae and adults inject likely to occur through movement of infest- toxic saliva that causes curling and contor- ed plant material and to a lesser extend with tion of leaves. Infested plants become stunt- infested fruits and cut fl owers (EPPO 2005). ed, swollen with leaf curl, shortened inter- The introduction of the pink hibiscus nodes or malformed stems. Damage varies mealybug in Rhodes without its natural en- according to the susceptibility of each host emies could pose a serious threat for sev- species; in highly susceptible plants feeding eral crops of high economic importance, can ultimately cause the death of the plant such as vegetables, vineyards and orna- (EPPO, 2005; Vitullo et al., 2009; Hoy et al., mental plants in urban areas and in nurser- 2014; García Morales et al., 2016). ies. Control eff orts of the mealybug should principally focus on the identifi cation and mapping of the actual infested area on the Materials and Methods island of Rhodes in order to design an effi - cient management plan that would restrict Infested hibiscus plants were found on further spread of the pest in the area. A sus- the island of Rhodes (36°26’1.49”N and tainable solution should defi nitely include 28°13’28.54”E) in September 2014 and sam- the careful introduction of its natural ene- ples were sent to BPI. New samples were mies following all the appropriate proce- sent to BPI collected from the Municipali- dures for such an approach. ty of Rhodes coming from Hibiscus sp. (Mal- Because of the minimum tolerance lev- vaceae), Ceratonia silique (Fabaceae), Erythri- el for M. hirsutus damage, intense man- na sp. (Fabaceae) and Bauhinia sp. (Fabaceae) agement approaches are often required. in autumn of 2016. All of them are known Management tactics include monitoring, host plants of M. hirsutus. Microscopic slides cultural, biological and chemical treatments. were prepared following the procedure de- Observations for typical damage symptoms scribed by Kosztarab and Kozár (1988) and like bunchy top, honeydew and sooty-mold identifi ed according to description and illus- presence can help in the identifi cation of in- tration by Williams and Watson (1988). Spec- festation spots. The pheromone of M. hirsu- imens are deposited at the Biological Con- tus has been identifi ed and can be used in trol Laboratory of BPI. pheromone traps for monitoring and detec- Material examined: Rhodes (Dodeca- tion especially in areas with low infestation nese islands); 02.ix.2014, 2 ♀♀, Hibiscus sp. density. Monitoring with pheromone traps (Malvaceae); Rhodes, 02.ix.2014 1 ♀♀ Hi- is also useful for timing insecticide applica- biscus sp. (Malvaceae); Rhodes; 13.xii.2016, tions. 2 ♀♀, Hibiscus sp (Malvaceae); Rhodes; After its introduction into a new area 13.xii.2016, 1 ♀♀, Hibiscus sp. (Malvaceae). M. hirsutus is usually diffi cult to eradicate due to its high reproductive ability and polyphagy. In areas where it has been estab- Results and Discussion lished, long standing sustainable management has been provided by biological con- This is the fi rst record of the pink hibiscus trol. Especially, classical biological control mealybug, M. hirsutus in Greece. Reports attempts have been quite successful (Kai- from local authorities from the island of ro et al., 2000). Several natural enemies have Rhodes had pointed out that infestation has been associated with M. hirsutus, includ- expanded within the island during the years ing specialist and generalist parasitoids and 2014-2016. No other outbreaks or records of predators. In classical biological control the

predatory species Cryptolaemus montrouz- Chong, J.H. Roda, LA and Mannion, M.C. 2008. Life ieri Mulsant (Coleoptera: Coccinellidae) and History of the Mealybug, Maconellicoccus hirsu- the parasitoids Anagyrus kamali Moursi and tus (Hemiptera: Pseudococcidae), at Constant Temperatures. Environmental Entomology, 37(2): Gyranusoidea indica Shafee, Alam and Agar- 323-332. wal (Hymenoptea: Encyrtidae) are regard- EPPO, 2005. Data sheets on quarantine pests. Maco- ed as most commonly used biological con- nellicoccus hirsutus. OEPP/EPPO, Bulletin, 35: 413- trol agents (Chong et al., 2015). The releases 415. of the above parasitoids and predators have EPPO, 2011. First report of Maconellicoccus hirsutus in resulted in very successful control of M. hir- Cyprus. EPPO Reporting Service, No 4 Paris, 2011- 04-01. sutus in Central America (Chong et al., 2015). García Morales, M., Denno, B.D., Miller, D.R., Miller, The use of contact insecticides may re- G.L., Ben-Dov, Y. and Hardy, N.B. 2016. ScaleNet: sult in scarce control of the scale population A literature-based model of scale insect biolo- due to the cryptic behaviour and the waxy gy and systematics. Database. doi: 10.1093/da- covering of the mealybug bodies. Therefore, tabase/bav118. http://scalenet.info. any applications should follow after care- Hoy, A.M., Avas, H. and Nguyen, Ru, 2014. Pink hibiscus mealybug Maconellicoccus hirsutus. UF/IFAS ful monitoring for the presence of crawlers University of Florida. Database http://entnemd- which are the most susceptible stage. Sys- ept.ufl .edu/creatures/ (Assessed 28/12/2016). temic insecticides might have a higher ef- Kairo, T.K.M., Pollard, V.G., Peterkin, D.D. and Lo- fi cacy on reducing M. hirsutus populations. pez, V.F. 2000. Biological control of the hibis- Nevertheless, application of insecticides cus mealybug, Maconellicoccus hirsutus Green (Hemiptera: Pseudococcidae) in the Caribbe- should be avoided when biological control an. Integrated Pest Management Reviews, 5: 241– eff orts are taking place. At present, no in- 254. secticide against M. hirsutus is registered in Kosztarab, M. and Kozár, F. 1988. Scale insects of Greece. central Europe. Boletin del Museo de Entomolo- gia de la Universidad del Valle Akademiai Kiado. Budapest. 456 p. We would like to thank the two anonymous re- Vitullo, J., Zhang, A., Mannion, C. and Bergh, J.C. 2009. Expression of feeding symptoms from viewers for their valuable comments. pink hibiscus mealybug (Hemiptera: Pseudo- coccidae) by commercially important cultivars of hibiscus. Florida Entomologist, 92(2): 248-254. Literature Cited Williams, D.J. and Watson, G.W. 1988. The Scale In- sects of the Tropical South Pacifi c Region. Pt. 2: Al-Fwaeer, M., Abu-Obaid, I., Al-Zyoud, F., Abo- The Mealybugs (Pseudococcidae). CAB Interna- Alosh, A. and Halaybeh, M. 2014. Population Dy- tional Wallingford, U.K. 260 pp. namics of the Hibiscus Mealybug Maconellicoc- cus hirsutus Green (Hom., Pseudococcidae) and Its Parasitoid on Guava Trees in Madaba-Jordan. International Journal of Agriculture and Forestry, 4(3): 171-177. Chong, J.H., Aristizabal, F.L. and Arthurs, P.S. 2015. Biology and Management of Maconellicoccus hirsutus (Hemiptera: Pseudococcidae) on Orna- mental Plants. Journal of Integrated Pest Man- agement, 6 (1): 1-14. Received: 10 January 2017; Accepted: 26 April 2017

Πρώτη καταγραφή του Maconellicoccus hirsutus (Green) (Hemiptera: Pseudococcidae) στην Ελλάδα

Π.Γ. Μυλωνάς και Γ.Κ. Παρτσινέβελος

Περίληψη Το είδος Maconellicoccus hirsutus (Green) (Hemiptera: Pseudococcidae), καταγράφεται για πρώτη φορά στην Ελλάδα. Ενήλικα άτομα του ψευδόκοκκου συλλέχθηκαν σε φυτά ιβίσκου Hibiscus rosa-sinensis (Linnaeus) (Malvaceae) σε ιδιωτικούς και δημόσιους κήπους στο αστικό περιβάλλον, στη Ρόδο. Αυτή είναι η πρώτη καταγραφή του γένους Maconellicoccus στην Ελλάδα.

Hellenic Plant Protection Journal 10: 80-83, 2017

SHORT COMMUNICATION

A catalogue of the Coleoptera of the G.P. Moazzo Collection in the Goulandris Natural History Museum. Part III (Scarabaeidae)

J. Tylianakis1, M. Dimaki2* and V. Perdiou2

Summary This is a detailed list of 223 species (727 specimens) of the family Scarabaeidae, the subfamilies Dynastinae (21 species), Melolonthinae (11 species), Rutelinae (29 species), Aphodiinae (104 species), Cetoniinae (59 species), represented in G.P. Moazzo’s collection at the Goulandris Natural History Museum. All label data for each specimen are given. The aim of this paper is to present this collection as reference data for comparison with recent entomological material.

Additional keywords: Scarabaeidae, Dynastinae, Melolonthinae, Rutelinae, Aphodiinae, Cetoniinae

Introduction erence information regarding insect specimens of Scarabaeidae. The Entomological Collection of the Gou- The material is of historical importance, landris Natural History Museum (GNHM) was with some specimens over 100 years old established in 1973. One particularly signifi - with a wide geographical distribution across cant collection is that of Georgi Polychronis 5 continents (Europe, Africa, Asia, America, Moazzo, which contains insects (Coleoptera Australia). It is published with the informa- in their majority) collected since 1910 (Gou- tion given exactly as it appears on the in- landris, 1977). Moazzo‘s entomological col- dividual labels. In some cases data may be lection of GNHM includes 5500 specimens. missing such as collection date and exact lo- Among them there are 1312 beetle species cality. An eff ort has been made to provide belonging to 58 families (Dimaki and Tyli- the current nomenclature, where possible. anakis 2006; Tylianakis and Dimaki 2006, Di- The specimens are in a very good condition maki et al., 2016). This is part III of the afore- and their identifi cation is still possible. mentioned collection, part I containing the families Carabaeidae, Cicindelidae, and Dy- tiscidae (Dimaki and Tylianakis, 2006) and Materials and methods part II containing part of the Scarabaeidae family (Tylianakis and Dimaki, 2006). The specimens were collected during fi eld- The aim of this paper is to present the work and have all been mounted on pins species of the Scarabaeidae subfamilies Dy- and arranged in unit trays within cabinet nastinae, Melolonthinae, Rutelinae, Aphodi- drawers at the GNHM. A determination label inae and Cetoniinae kept in the entomologi- accompanies each specimen as well as a da- cal collection of GNHM (Moazzo’s collection). tabase number. This number corresponds to The catalogue would be of particular inter- the information of each specimen as well as est for use by entomologists seeking for ref- the current name. The material has been examined and described mainly by G.P. Moaz- zo. 1 Zoology Department, University of Canterbury, Pri- This collection includes material from vate Bag 4800, Christchurch, New Zealand. collectors such as A. Carneri, G. Louvet, B. 2 Goulandris Natural History Museum, 13 Levidou, GR- 145 62 Kifi ssia, Attica, Greece. Alpes, Talbiele, Petroff , Schuster, Winkler, * Corresponding author: [email protected] McMaygiore, Effl atoun and G.P. Moazzo

© Benaki Phytopathological Institute Scarabaeidae Collection in Goulandris Natural History Museum 85 himself. Tucuman We followed the systematics of Ratc- Oryctes rhinoceros 2 spm, INDONESIA: Am- liff e and Jameson (2004) and Triplehorn and boine, Iles Moluques, leg. A. Carneri Johnson (2005). Oryctes spp. 5 spm, ARGENTINA: Tucuman Oryctes sp. 2 spm, ITALY, 1950, leg. Gentile Catalogue of Coleoptera, Scarabaei- Oryctes nasicornis 1 spm dae In the list below, we record the species and the Melolonthinae (11 species, 30 spm) relevant metadata: the number of specimens Europtron gracile 4 spm, ALGERIA: Ghardaia, (spm), the place of the collection, and the year leg. A. Chobaut and collector’s name, when available. Haplidia nitidula 7 spm Hoplia coerulea 1 spm, FRANCE: Prats de Dynastinae (21 species, 105 spm) Mollo, leg. Waegner Pentodon dispar (syn. P. algerinum sbsp. Hoplia dubia 5 spm, ITALY: Macarese, Rome, dispar) 23 spm, EGYPT: Alexandria, Victo- 1919, leg. P. Crozet ria?, Mariout, Ramleh, Rond-Point, Roun- Hoplia africanum 4 spm, MOROCCO: Casa- leh, Abou el Chekark, 1917, 1921, 1924, 1929, blanca, leg. Antoine 1935, 1938 leg. Effl atoun Hoplia farinosa 2 spm, AUSTRIA: Graz; Pentodon punctatus 1 spm, ITALY: Gerace Ca- FRANCE: Besançon labria, leg. Schuster Hoplia philanthus 1 spm Pentodon idiota 1 spm, AUSTRIA: Neusiedi Hoplia graminicola 1 spm lake, leg. Schuster Hoplia praticola 1 spm, ALGERIA: Ain Sefra, Pentodon bispinosus 1spm, MOROCCO: Casa- 1923, leg. A. Chobaut blanca, 1919, leg. Antoine Hoplia pubicollis 1 spm, CORSICA: Ajaccio Pentodon sp.1 spm, TURKESTAN: Samarkand Hoplia spp. 3 spm, ALGERIA: Bou-berak; MO- Pentodon sp.1 spm, TURKESTAN: Samarkand ROCCO: Casablanca, leg. A. Chobaut, An- Dyscinetus rugifron 2 spm, ARGENTINA: Bue- toine nos Aires Pachononyx fuscodeneus 2 spm, ARGENTINA: Rutelinae (29 species, 71 spm) Rio Salodo Anisoplia fl avipennis1 spm Phyllognathus silenus 12 spm, FRANCE: Tou- Anisoplia segetum var. straminea (syn. louse; TURKEY: Adana, Cilicie; SPAIN: Tener- Chaetopteroplia segetum straminae) 1 spm, ife; EGYPT: Mariout, Ramleh, Palais; ALGERIA: TURKEY: Konya, 1899, leg. Korh Bou-Berak, 1921, 1924, 1935, 1953, leg.V.M. Anisoplia segetum (syn. Chaetopteroplia sege- Duchon, Effl atoun, A. Chobaut, Thery, A. tum) 1 spm, IRAN: Kopet-Dagh, Descht, 1902, Carneri leg. F. Hauser Heteronychus parumpunctatus 16 spm, Anisoplia pallidipennis 2 spm, ALGERIA: Ain EGYPT: Alexandria, 1921, 1922, 1924, 1929 Sefra, 1923, leg. A. Chobaut Heteronychus dilatatus 2 spm, ARGENTINA: Anisoplia cyathigera 1 spm, BOSNIA: Sanski- Tucuman most Heteronychus spp. 3 spm, EGYPT: Mariout, Anisoplia biguttata 1 spm, INDONESIA: Java 1933 Breanger Tjigembong, leg. Corporaai Oryctomorphus maculicollis 1 spm, CHILE Anisoplia lata 2 spm, AUSTRIA: Graz, CROA- Oryctomorphus sp. 2 spm, SOUDAN: FRANCE TIA: Pola Istria Temnorrhynchus baal 21 spm, EGYPT: Alex- Anisoplia leucaspis 1 spm, IRAN: Kerman- andria, Victoria?, Mazarita, Mandara Cairo, schah, 1909, leg. F. Hauser Rond Point, Nousha, 1919, 1921, 1923, 1924, Anisoplia agricola 1 spm, GERMANY: Off en- 1932, 1942, leg. A. Carneri bach Phileurus valgus 4 spm, VENEZUELA Anisoplia sp.1 spm, CROATIA: Velezgb, leg. Scaptophilus dasipleurus 2 spm, ARGENTINA: Krauss

Anisoplia monticola 2 spm, CROATIA: Pola Is- out, Dekhela, Palais, 1919, 1921, 1923, leg. A. tria Carneri Anisoplia syriaca 1 spm, TURKEY: Ephesos Aphodius spp. 5 spm EGYPT: Palais, Alexan- Anisoplia fl avocincta1 spm, ITALY: Emilia, dria, 1923, 1925, leg. A. Carneri 1897, leg. Fiori Aphodius (Bodolius) hydrochoeris 17 spm, Anisoplia graminivora (syn. Anisoplia tempes- EGYPT: Palais, Victoria?, Bulgley, 1919, 1923, tiva) 2 spm, FRANCE: Ste Baume, 1920, leg. leg. A. Carneri, Fleming, M. Gantes A. Chobaut Aphodius (Volinus) hieroglyphicus 10 spm, Anisoplia sabulicola (syn. Brancoplia pumila) EGYPT: Mariout, Dekhela, Rond Point, 1921, 2 spm, EGYPT: Saba Pacha, 1918 1923, 1925 Anisoplia deserticola 4 spm, ALGERIA: Bou Aphodius granarius (syn. Calamosternus gra- Saada, Ain Sefra, 1923, leg. A. Chobaut, Mar- narius) 29 spm, EGYPT: Mariout, Dekhela, tin Rond Point, Cairo, Victoria?, Smouha, Sidi Anisoplia fl oricola4 spm, ALGERIA: Bou-Be- Bishr, Alexandria, ALGERIA: Bou-Berak; RUS- rak; MOROCCO: Casablanca; SPAIN: Valen- SIA: Transcaspia Kisil Arwat, 1898,1920, 1921, cia, leg. Antoine 1923, 1939, 1944, leg. A. Chobaut, F. Hauser Anisoplia austriaca 1 spm, Nd. Tesders? Aphodius varians 9 spm, EGYPT: Mariout, Anisoplia sp. 2 spm, TURKEY: Marmara, leg. Rond Point, Smouha 1920,1923, 1938,1939 Thery Aphodius (Erytus) sp. 29 spm, EGYPT: Mariout, Anisoplia bromicola var. nigra 1 spm, ITALY: Dekhela, Alexandria, Saba Pacha Montaza; Cerchio ALGERIA: Batra; RUSSIA: Transcaspia, 1900, Anisoplia bromicola 1 spm, CROATIA: Velezgb, 1919, 1920, 1921, 1922, 1925, leg. F. Hauser leg. Krauss Aphodius (Erytus) brunneus 3 spm, 1919, leg. Anisoplia austriaca var. major 1 spm A. Chobaut Anisoplia tempestiva 1 spm, FRANCE: Ste Aphodius (Erytus) lividus (syn. Labarrus livi- Baume, 1920, leg. A. Chobaut dus) 5 spm, leg. Montaza, 1921, 1922 Anisoplia sabulicola 24 spm, EGYPT: Abou Aphodius (Erytus) leucopterus (syn. Mecyn- kir, Rond Point, Cherbine, Mariout, Smouha, odes leucopterus) 4 spm, EGYPT: Mariout, Nouzha, Alexandria, Sidi Bishu 1919, 1920, 1925, 1944, leg. Garbaniat 1938, 1940, 1943, 1944 Aphodius (Eremazus) sp. 3 spm Pelidnota paraguayensis 1 spm, ARGENTINA: Aphodius (Eremazus) punctatus 2 spm Tucuman Aphodius (Eremazus) sp.1 spm, leg. A. Cho- Pelidnota sp. 1spm, SUDAN baut Blitopertha horticola (syn. Phyllopertha horti- Aphodius (Eremazus) unistriatus 2 spm, cola) 6 spm, FRANCE: Rouen; BOSNIA, 1921 EGYPT: Mariout, leg. A. Chobaut Blitopertha horticola (var. ustulatipennis) 1 Aphodius thermicola 8 spm, ITALY; AUSTRIA; spm, AUSTRIA: Wien MOROCCO: Rabat, 1917, 1922, leg. A. Cho- Blitopertha arenaria (syn. Blitopertha lineola- baut, P. Crozet ta) 1 spm, GREECE: Korinth Aphodius luridus var. nigripes 6 spm, ITALY, Blitopertha hirtella 1 spm, GREECE: Attica 1917, leg. P. Crozet Blitopertha lineolata 1 spm, IRAN: Dagh, Aphodius erraticus 6 spm, FRANCE: Cote D’Or; 1902, leg. F. Hauser ALGERIA: Bou-Berak Aphodius erraticus var. fumigatus 1 spm, ITA- Aphodiinae (103 species, 294 spm) LY: Cerchio Aphodius contaetus 3 spm, EGYPT: Barages Aphodius subterraneus 6 spm, FRANCE: de Caire Rennes, Hungbor? 1920, leg. P. Crozet, V. Fo- Aphodius punctifer 1 spm ufal Aphodius sordidus (syn. Bodilopsis rufus) 1 Aphodius bonnaizei (Type) 13 spm, FRANCE: spm, SWITZERLAND: Geneva Carrieres des Angles; ITALY: Valgares; TUNIS, Aphodius punctipennis 4 spm, EGYPT: Mari- 1922, leg. A. Chobaut

Aphodius merdarius 1 spm, PORTUGAL: Lei- Aphodius rufi pes 1 spm, AUSTRIA: Trifail Styr- ria Lusitania ia Aphodius scrofa (syn. Trichonotulus scrofa) 1 Aphodius lividus 5 spm, ALGERIA: Beni Ounif spm, GERMANY?: Marburg, leg. Krauss de Figule, 1923, leg. A. Chobaut Aphodius pusillus 1 spm, SLOVENIA: Carniole, Aphodius spp. 2 spm, GERMANY: Marburg; leg. Krauss TURKEY: Adana Aphodius maculatus 1 spm, GREECE: Nemea Aphodius depressus var. atramentarius (syn. Aphodius obscurus 1 spm, SWITZERLAND: Aphodius atramentarius) 1 spm, AUSTRIA: Faulhorn Koralpe, leg. Penecke Aphodius obscurus var. olichrous 1 spm, AUS- Aphodius licardi 1 spm, ALGERIA TRIA: Obir Carinthia Aphodius sp. 1 spm, TUNISIA Aphodius guttatus (syn. Agolinus guttatus) 2 Aphodius vitellinus (syn. Subrinus vitellinus) 2 spm, ALGERIA: Bou-Berak, leg. A. Chobaut spm, MOROCCO: Casablanca, Rabat, leg. An- Aphodius lineolatus (syn. Chilothorax lineola- toine tus) 2 spm, SPAIN: Cadiz, ALGERIA: Bou-be- Aphodius unicolor (=eastaneus) 4 spm, ALGE- rak, leg. A. Chobaut RIA: Bou-Berak, leg. A. Chobaut Aphodius spp. 9 spm, MOROCCO: Rabat, Taja; Aphodius (Bodilus) longispina 1 spm, MO- ALGERIA: Ougda Thery ROCCO: Casablanca, 1920, leg. Antoine Aphodius stricticus (syn. Volinus stricticus) 1 Aphodius serutator (syn. Teuchestes serutator) spm 3 spm, ITALY: Maggiore, leg. P. Crozet Aphodius depressus 1 spm, AUSTRIA: Stiria Aphodius fossor 1 spm, AUSTRIA: Graz, leg. bor., leg. Krauss Krauss Aphodius distinctus (syn. Chilothorax distinc- Aphodius brevis 1 spm, AUSTRIA: Neusiedler tus) 3 spm, FRANCE: Les Angles; AUSTRIA: Aphodius conjugatus 1 spm, ROMANIA: Umgeb. Graz, 1922, leg. A. Chobaut Klausenberg Aphodius tessulatus 1 spm, GERMANY, 1922, Aphodius (Otophorus) haemorrhoidolis 1 leg. A. Chobaut spm Aphodius obliteratus 1 spm, GERMANY: Re- Aphodius (Otophorus) sanguinolentus (syn. ichenau island, leg. Ponecke Aphodius (Otophorus) haemorrhoidolis) 4 Aphodius prodromus 3 spm, ITALY: Valcar- spm, Marrier de la Mer; TURKEY: Kizil-Dagh, es; PORTUGAL: Leiria Lusitania, 1922, leg. A. Taurus, leg. A. Chobaut Chobaut Aphodius fi metarius1 spm, ALGERIA: Bou- Aphodius punctatosulcatus 1 spm, SPAIN: Berak Ronda Aphodius foetens 3 spm, BOSNIA; CYPRUS Aphodius punctatosulcatus var. marginalis 1 Aphodius scybalarius var. confl agratus1 spm, spm, SPAIN: Valencia TUNIS: Bel, 1900 Aphodius limbatus 1 spm, AUSTRIA: Neusie- Aphodius (Bodilus) ictericus (=nitidulus) 6 spm, dlersee FRANCE: Le Cailar, CROATIA: Sebenico, 1923 Aphodius maenlatus 1 spm, Valegb Hz., leg. Aphodius hydrochoeris 2 spm, ALGERIA: Ghar- Krauss daia; MOROCCO: Tanger Aphodius zenkeri 2 spm, LIBERIA; GERMANY, Aphodius sordidus 1 spm, FRANCE leg. Grunwld Aphodius rufus 1 spm, HUNGARY Aphodius miaetus 1 spm, ROMANIA: Paring, Aphodius rufus var. arcuatus 1 spm, ITALY: leg. Krauss Mte Baldo Aphodius miaetus var. unicolor 1 spm, ROMA- Aphodius lugeus 1 spm, GREECE: Nemea NIA: Paring, leg. Krauss Aphodius immundus 1 spm, GERMANY: Mar- Aphodius praecose 1 spm, Penecke st. leg. H. burg, leg. Krauss Reichart Aphodius ater 1 spm Aphodius satellitius 1 spm, CROATIA: Pola Is- Aphodius piceus 1spm, FINLAND tria Aphodius gibbus 1 spm

Aphodius nemoralis 1 spm, AUSTRIA: Reka- Cetoniinae (59 species, 227 spm) winkel Stalagmosoma abbela 1 spm, Wadioff , 1927, Aphodius putridus 1spm leg. Petroff Aphodius alpinus 1 spm, SWITZERLAND Faul- Tropinota squalida 37 spm, EGYPT: Saba Pa- horn cha, Alexandria, Mex, Mariout, Siouf, Avi- Aphodius plagiatus var. immaculatus 1 spm, gnon, St. Genes, Sidi Bishr, Mandara, 1918, ROMANIA: Mezo-Zah 1923, 1924, 1934, 1935, 1936, 1943, 1944, Aphodius sturmi 1 spm, AUSTRIA: Umgeb. 1949, leg. A. Chobaut Zara, leg. Muller Tropinota hirta (syn. Epicometis hirta) 2 spm, Aphodius niger1 spm, AUSTRIA: Neusiedler, St Gemes de lo Molus; GREECE: Rhodus, leg. Krauss 1924, leg. Plason Aphodius lapponum 1 spm Lasiotrichius succinctus 4 spm, EGYPT: Mon- Aphodius corvinus 1 spm, AUSTRIA: Umgeb. taza Dekhela, Mandara, Agami, 1919, 1927, Graz 1938, 1940 Aphodius varians (=ambiguus) 2 spm BOS- Oxythyrea abigail 22 spm, EGYPT: Avignon, NIA, GREECE: Korfu TURKEY: Istanbul, SLOVENIA: Carniolia, GER- Aphodius tristis 1 spm GERMANY MANY: Wippach, FRANCE: St. Genie’s de Co., Aphodius biguttatus 1 spm Rouen, 1924 Aphodius cribricollis 1 spm, TUNIS: Le kef Oxythyrea funesta 9 spm, MOROCCO: Casa- Aphodius (Amidorus) cribarius 1 spm, CROA- blanca, 1921, leg. Antoine TIA: Kistange, leg. D. Muller Oxythyrea funesta var. Wagner 3 spm, UNIT- Aphodius contaminatus 1 spm,, TURKEY ED ARAB EMIRATES: Wadi, SWITZERLAND: Aphodius tingens 3 spm, ALGERIA: Bou-Be- Stadt?, TURKEY: Istanbul, 1927 rak, Philippevile Oxythyrea cinctella 3 spm, SOUDAN, MO- Aphodius consputus 1 spm, TURKEY ROCCO: Casablanca, 1918, leg. Antoine Aphodius affi nis 1 spm, TURKEY: Adana Oxythyrea spp. 3 spm, FRANCE: Tonkin, AUS- Aphodius kraatzi (syn. Liothorax kraatzi) 1 TRIA: Wien , leg. H Fruhsturfer spm, KAZAKHSTAN: Thian – Shan, Musart Trichius bifasciatus 5 spm Aphodius rhododactylus 1 spm, CROATIA: Trichius bifasciatus var. dubius 1 spm, TURKE- Velezgb, leg. Krauss STAN: Djarkent Semirjetschensk, leg. Win- Aphodius sp. 2 spm kler Aphodius lividus 1 spm Trichius bifasciatus var. vulgaris 1 spm, AUS- Enpleureus subterraneus 2 spm, Marrier de la TRIA: Dobratsch Mer, leg. A. Chobaut Trichius bifasciatus var. succinctus 1 spm, Euheptaulacus sus 1 spm, GERMANY IRAN: Astrabad=Gorgan, 1899, leg. Hauser Heptaulacus alpinus (syn. Oromus alpinus) 1 Trichius abdominalis 1 spm, SLOVENIA: spm, CROATIA: Brokovo Wochein Carniolia Heptaulacus porcellus 1 spm AUSTRIA: Wien Trichius sexualis 1 spm, FRANCE: Rouen Psammobius porcicollis 1 spm, HUNGARY Trichius gallicus 1 spm, USA: Pennsylvania Pleurophorus baebus1 spm, ALGERIA: Bou- Trichius affi nis 2 spm, ALGERIA: Bou-Berak Berak leg. D. Chobaut Pleurophorus caesus 1 spm, Zoppa Tr. Trichius zonatus var. fortunatarum 2 s p m , Aegialia sabuleti 1 spm FRANCE: St. Saurent Htes Pyrenees Aegialia arenaria 1 spm, GERMANY: Borkum, Gnorimus nobilis 2 spm, AUSTRIA, leg. Schus- leg. Schneider ter Rhyssemus germanus 1 spm, EGYPT: Kabush- Gnorimus variabilis 1 spm, CHILE ia leg. Alfi eri Euphoria lurida 2 spm, MEXICO Rhyssemodes reilleri 1 spm, ALGERIA: Ghar- Euphoria bascalis 1 spm, MADAGASCAR: Ta- daia, leg. A. Chobaut nanarive, ERITREA, 1918, leg. A. Mochi Gnorimus variabilis 16 spm, AUSTRIA: Prze-

© Benaki Phytopathological Institute Scarabaeidae Collection in Goulandris Natural History Museum 89 mysl Galizieu, FRANCE: Rouen, ITALY, leg. Kalgan Vogel Potosia fl oricola var. daurica 1 spm Valgus hemipterus 4 spm Potosia fl oricola var. metallica 1 spm, CROA- Osmoderma eremita 1 spm, CANADA: Joli- TIA: Pola Istria, 1920 ette, 1906 Potosia fl oricola var. obscura 2 spm, ITALY: Osmoderma scabra 2 spm, ARGENTINA: Rome, GREECE: Itea, 1918 Tucuman Potosia fl oricola var. fl o r e n t i n e 3 spm, IRAN: Diplognatha gagates 1 spm, SRI LANKA: Kopet-Dagh south, 1889, leg. H Fruhsturfer Potosia fl oricola var. hieroglyphica 1 spm Glycyphana versicolor var. luduosa 2 spm, Potosia angustata 1 spm, SYRIA: Damascus, East Africa, leg. A. Heyne leg. Schuster Smaragdesthes viridis (syn. Smaragdesthes Potosia affl icta 2 spm, FRANCE: St Lucie, leg. africana viridis) 1 spm, VIETNAM: Saigon Schuster Protaetia sp. 2 spm, east Africa Potosia oblonga 1 spm, TURKEY: Istanbul, Gnathocera cruda 2 spm, MADAGASCAR leg. Chobaut, F. Charles Euryomia argentea 2 spm, GREECE: Vrilissia, Potosia vidua 4 spm, SYRIA: Aleppo, FRANCE: ITALY: Ragusa, 1958, leg. Schuster Bouchat (Drome), leg. Winkler, Chobaut Potosia speciosissima (=aeruginosa) 2 spm, Potosia opaca 2 spm, Bumfacio, ITALY: Rome, SLOVENIA: Carniolia GERMANY: Wippach leg. Chobaut Potosia affi nis 1 spm, TURKESTAN: Djarkent Potosia morio 4 spm, MOROCCO, 1921, leg. Semirjetschensk, leg. Winkler Antoine, Thery Potosia aurata var. viridiventris 1 spm, SYRIA Potosia morio var. punctate 2 spm, NIGIRIA: Potosia chrysosoma 1 spm, IRAN: Kopet- east, leg. E.V. Bodemeyer Dag Potosia funeraria 3 spm, FRANCE: Bouchat Potosia aeratula 1 spm, FRANCE: Bonifacio (Drome), leg. Antoine Potosia carthami (syn. Cetonia carthami) 2 Potosia cardui 2 spm, (syn. Potosia spm, MOROCCO: Casablanca, leg. Antoine opaca), EGYPT: Avignon, leg. Chobaut Aethiessa fl oralis var. barbara 2 spm, TUR- Potosia marmorata (liocala) 1 spm, ITALY: Sic- KEY: Istanbul, leg. F. Charles, Yervanh ily, leg. Schuster Potosia hungariga 4 spm, Caucasus, TURKEY: Aethiessa inhumata 2 spm, ARGENTINA: Istanbul, leg. Yervanh Tucuman Potosia hungariga var. armeniaca 3 spm, AR- Stephanorrhina guttata 2 spm, GREECE: Par- MENIA, MOROCCO, leg. Chobaut, Thery nitha, 1921, leg. Sophrone, Schuster Aethiessa fl oralis 4 spm, EGYPT: Alexandria, Diplognatha gagates 2 spm, AUSTRALIA: Rond-Point, 1919 Freshwater Queensland Aethiessa inhumata 1 spm, EGYPT: Nouzha, Celidota stephensis 2 spm, KENYA: Nairobi Alexandria, TURKEY: Taurus, 1920, 1922, Pachnoda fasciata 7 spm, ZAMBIA: Katona, 1927 1909 Potosia cuprea 1 spm, KYRGHYZSTAN: Ket- Pachnoda petersi 2 spm, ERITREA men Tjube, TURKEY: Sussamyr Potosia cuprea ssp. ignicollis 9 spm, TURKEY: Taurus Literature cited Potosia fl oricola var. phoebe (syn. Poto- Dimaki, M., Anagnou-Veroniki, Μ. and Tylianakis, sia phoebe) 1 spm, TURKEY: Istanbul, leg. F. J. 2016. A catalogue of Coleoptera specimens Charles with potential forensic interest in the Goulan- Potosia fl oricola (syn. Cetonia fl oricola) 5 spm, dris Natural History Museum collection. Ento- EGYPT: Avignon mologia Hellenica, 25: 31-38. Potosia fl oricola var. marginicollis 1 spm, IT- Dimaki, M. and Tylianakis, J. 2006. A catalogue of the G.P. Moazzo collection in the Goulandris Natu- ALY ral History Museum. Part I. Annales Musei Gou- Potosia fl oricola var. incerta 3 spm, CHINA: landris, 11: 281-287.

Goulandris, N. 1977. Georgi P. Moazzo (1893-1975), Seventh Edition.Publisher P.Marshall Brooks/ Greek conchologist. Annales Musei Goulandris, Cole CENGAGE Learning, p 367-416. 3: 105-122. Tylianakis, J. and Dimaki, M. 2006. A catalogue of the Ratcliff e, B.C. and Jameson, M.L. 2004. The Re- G.P. Moazzo collection in the Goulandris Natu- vised Classifi cation for Sarabaeoidea: What ral History Museum. Part II. Annales Musei Gou- the Hell is going on? Papers in Entomology. Pa- landris, 11: 289-296. per 25, 10 p. http://digitalcommons.unl.edu/ entomologypapers/25/ Triplehorn C.A. and Johnson N.F. 2005. Borror and DeLong’s Introduction to the Study of Insects, Received: 22 May 2017; Accepted: 26 June 2017

ΣΥΝΤΟΜΗ ΑΝΑΚΟΙΝΩΣΗ

Καταγραφή ειδών Κολεοπτέρων εντόμων της Συλλογής G.P. Moazzo του Μουσείου Γουλανδρή Φυσικής Ιστορίας. Μέρος III (Scarabaeidae)

J. Tylianakis, Μ. Δημάκη και Β. Περδίου

Περίληψη Σε αυτήν την εργασία προυσιάζεται ο κατάλογος 223 ειδών (727 δείγματα) της οικογένει- ας Scarabaeidae των υποοικογενειών Dynastinae (21 είδη), Melolonthinae (11 είδη), Rutelinae (29 είδη), Aphodiinae (104 είδη) και Cetoniinae (59 είδη), της Συλλογής του G.P. Moazzo του Μουσείου Γουλαν- δρή Φυσικής Ιστορίας. Παρέχονται όλες οι πληροφορίες των δειγμάτων που υπάρχουν. Σκοπός αυτής της εργασίας είναι η παρουσίαση της εντομολογικής συλλογής ως βάση αναφοράς για συγκριτικές με- λέτες με σύγχρονο εντομολογικό υλικό.

Hellenic Plant Protection Journal 10: 84-90, 2017

P. Jaikaew, F. Malhat, J. Boulange and H. Watanabe Aspect of the degradation and adsorption kinetics of atrazine and metolachlor in andisol soil 1-14

I.A. Mohamed, G.S. Mohamed, E.Y. Abdul-Hafeez and O.H.M. Ibrahim Effi ciency of sweet fl ag and curly parsley volatile oils compared with synthetic insecticides against Ceroplastes rusci on Ruellia plants 15-24

F.-S. Tabatabaei and A. Saeedizadeh Rhizobacteria cooperative eff ect against Meloidogyne javanica in rhizosphere of legume seedlings 25-34

A. Adam, M.I.E. Arabi, I. Idris and E. Al-Shehadah Eff ect of several rhizobacteria strains on barley resistance against Pyrenophora graminea under fi eld conditions 35-45

O. Melita, V. Gkounti, D. Kontodimas, D. Papachristos and F. Karamaouna Can high pest pressure of the red palm weevil Rhynchophorus ferrugineus beat the defense of Phoenix theophrasti? 46-50

A. Venieraki, M. Dimou and P. Katinakis Endophytic fungi residing in medicinal plants have the ability to produce the same or similar pharmacologically active secondary metabolites as their hosts 51-66

Ν. Latinović, F. Karamaouna and N.G. Kavallieratos First record of Aphis craccivora Koch (Hemiptera: Aphididae) on aronia crop in Montenegro 67-69

A.I. Darras, A. Kotsiras, C. Delis, K. Nifakos, E. Pavlakos and V. Demopoulos Reaction of the native Greek tomato varieties ‘Chondrokatsari Messinias’ and ‘Katsari Santorinis’ to Fusarium oxysporum f. sp. lycopersici and Rhizoctonia solani infection 70-79

J. Tylianakis, M. Dimaki and V. Perdiou A catalogue of the Coleoptera of the G.P. Moazzo Collection in the Goulandris Natural History Museum. Part III (Scarabaeidae) 84-90

P. Jaikaew, F. Malhat, J. Boulange και H. Watanabe Κινητική αποδόμησης και ρόφησης των ζιζανιοκτόνων ατραζίνη και metolachlor σε έδαφος ηφαιστειακής τέφρας 1-14

I.A. Mohamed, G.S. Mohamed, E.Y. Abdul-Hafeez και O.H.M. Ibrahim Αποτελεσματικότητα αιθέριων ελαίων των φυτών Acorus calamus και Petroselinum crispum, σε σύγκριση με συνθετικά εντομοκτόνα, κατά του κηροπλάστη, Ceroplastes rusci, σε φυτά Ruellia simplex 15-24

F.-S. Tabatabaei και A. Saeedizadeh Συνδυαστική δράση ριζοβακτηρίων κατά του κομβονηματώδη Meloidogyne javanica στη ριζόσφαιρα ψυχανθών 25-34

A. Adam, M.I.E. Arabi, I. Idris και E. Al-Shehadah Επίδραση στελεχών ριζοβακτηρίων στην ανθεκτικότητα του κριθαριού έναντι του Pyrenophora graminea σε συνθήκες αγρού 35-45

Ο. Μελιτά, Β. Γκουντή, Δ. Κοντοδήμας, Δ. Παπαχρήστος και Φ. Καραμαούνα Μπορεί η υψηλή πίεση πληθυσμού του ρυγχοφόρου των φοινικοειδών, Rhynchophorus ferrugineus, να υπερνικήσει την άμυνα του φοίνικα του Θεόφραστου Phoenix theophrasti; 46-50

Α. Βενιεράκη, Μ. Δήμου και Π. Κατινάκης Ενδοφυτικοί μύκητες που διαβιούν εντός των φαρμακευτικών φυτών έχουν την ιδιότητα να παράγουν τους ίδιους ή παρόμοιους δευτερογενείς μεταβολίτες με τους ξενιστές τους 51-66

N. Latinović, Φ. Καραμαούνα και Ν.Γ. Καβαλλιεράτος Πρώτη καταγραφή της αφίδας Aphis craccivora Koch (Hemiptera: Aphididae) σε καλλιέργεια αρωνίας στο Μαυροβούνιο 67-69

Α.Ι. Δάρρας, Α. Κώτσιρας, Κ. Δελής, Κ. Νηφάκος, Ε. Παυλάκος και Β. Δημόπουλος Προσβολή φυτών τομάτας των Ελληνικών παραδοσιακών ποικιλιών ‘Χοντροκατσαρή Μεσσηνίας’ και ‘Κατσαρή Σαντορίνης’ από τους μύκητες Fusarium oxysporum f. sp. lycopersici και Rhizoctonia solani 70-79

J. Tylianakis, Μ. Δημάκη και Β. Περδίου Καταγραφή ειδών Κολεοπτέρων εντόμων της Συλλογής G.P. Moazzo του Μουσείου Γουλανδρή Φυσικής Ιστορίας. Μέρος ΙΙΙ (Scarabaeidae) 84-90

Περιεχόμενα

Π.Γ. Μυλωνάς και Γ.Κ. Παρτσινέβελος Πρώτη καταγραφή του Maconellicoccus hirsutus (Green) (Hemiptera: Pseudococcidae) στην Ελλάδα 80-83

Περιεχόμενα Τόμου 10 (2017)

Hellenic Plant Protection Journal www.hppj.gr © Benaki Phytopathological Institute Volume 10, Issue 2, July 2017 ISSN 1791-3691 H Hellenic Plant Protection Journal Contents e l l e

A. Venieraki, M. Dimou and P. Katinakis n Endophytic fungi residing in medicinal plants have the ability i

to produce the same or similar pharmacologically active c

secondary metabolites as their hosts 51-66 P

Ν. Latinović, F. Karamaouna and N.G. Kavallieratos l a First record of Aphis craccivora Koch (Hemiptera: Aphididae) on aronia crop in Montenegro 67-69 n t

A.I. Darras, A. Kotsiras, C. Delis, K. Nifakos, E. Pavlakos P

and V. Demopoulos r

Reaction of the native Greek tomato varieties ‘Chondrokatsari o

Messinias’ and ‘Katsari Santorinis’ to Fusarium oxysporum f. sp. t

lycopersici and Rhizoctonia solani infection 70-79 e c

P.G. Milonas and G.K. Partsinevelos t

The pink hibiscus mealybug Maconellicoccus hirsutus (Green) i (Hemiptera: Pseudococcidae) in Greece 80-83 o n

J. Tylianakis, M. Dimaki and V. Perdiou J A catalogue of the Coleoptera of the G.P. Moazzo Collection in the o Goulandris Natural History Museum. Part III (Scarabaeidae) 84-90 u

Contents of Volume 10 (2017) r n a l