مومنی، آخر درآ در صف رزم که تو را بر آسمان بودست بزم
بر امید راه بالا کن قیام همچو شمعی پیش محراب ای غلام
دم به دم بر آسمان میدار امید در هوای آسمان رقصان چو بید
دم به دم از آسمان می آیدت آب و آتش رزق می افزایدت
کین طلب در تو گروگان خداست زانکه هر طالب به مطلوبی سزاست
جهد کن تا این طلب افزون شود تا دلت زین چاه تن بیرون شود
First report of olive knot caused by Pseudomonas savastanoi pv. savastanoi on olives (Olea europaea)Pseudomonas savastanoi pv. savastanoi was identified as the cause of galling on olives (Olea europaea), cv. Barnea in South Australia in May 2003 from a property at Greenfields ~10 km north of Adelaide. The bacterium was recovered from galls on 2-year-old olive trees (cv. Barnea) showing symptoms consistent with those attributed to olive knot. Following a targeted survey of cv. Barnea, further infections were located on 2 to 5-year-old trees in four other plantings in South Australia. This is the first record of olive knot in Australia
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وجود ويروس تريستزاي مركبات (Citrus tristeza virus, CTV)، براساس علائم روشني رگبرگ در ليموترش و ساقه آبلهاي در نارنگيهاي پيوند شده روي ليموترش ، مشاهدهء پيكرههاي رشتهاي خمشپذير با الكترون ميكروسكپي عصارهء اين گياهان و آزمايش سرولوژيكي اليزا (DAS-ELISA) در استانهاي فارس و بوشهر به اثبات رسيد. ويروس از بافتهاي درختان نارنگي محلي پيوند شده روي ليموترش كه به طور طبيعي آلوده شده بودند، به وسيلهء دوبار رسوب دادن با پلياتيلن گليكول (PEG) و سپس يك بار سانتريفوژ كردن در ستون داراي شيب چگالي مخلوط سوكروز و سولفات سزيوم، خالصسازي شد. آمودهء خالصسازي شده داراي طيف جذبي مشابه طيف جذبي نوكلئوپروتئينها بود و مشاهدهء الكترون ميكروسكپي اين آموده وجود پيكرههاي رشتهاي خمشپذير را نشان داد. همچنين آزمايش اليزا وجود ويروس تريستزا را در اين آموده اثبات كرد. با تزريق زيرجلدي ويروس خالص شده به خرگوش ، آنتيسرمي عليه آن توليد شد. آنتيسرم بدست آمده پس از جذب با عصارهء گياه سالم در آزمايش نشت دوطرفه در آگار حاوي SDS فقط در مقابل نمونههاي آلوده به CTV يك خط رسوب تشكيل داد. اين آنتيسرم پس از جذب با آمودهء گياه سالم و جداسازي گاماگلوبولين و تهيه آنتيبادي متصل به آنزيم در آزمايش DAS-ELISA مورد استفاده قرار گرفت كه در اين آزمايش ، استفاده از رقتهاي 1/900 آنتيبادي و 1/300 آنتيبادي متصل به آنزيم مناسب تشخيص داده شد. الكتروفورز پروتئين آمودههاي خالصسازي شده در ژلپلياكريل آميد داراي دودسيل سولفات سديم (SDS-PAGE) در مورد تمام جدايهها دو نوع پروتئين نشان داد. يكي از پروتئينها با وزن مولكولي 35500-36300 دالتون در عصارهء گياه سالم نيز مشاهده ميشد ولي پروتئيني با وزن مولكولي 22500-23000 دالتون فقط در عصارهء گياهان آلوده وجود داشت . بررسي پوست تنه در قسمت پايه، پوست تنه در قسمت پيوندك ، پوست شاخههاي دوساله، پوست شاخههاي جديد، دمبرگ و آلبيدو ميوهء 11 اصله نارنگي پيوند شده روي ليموترش با روش اليزا نشان داد كه پوست تنه در قسمت پيوندك و پوست شاخههاي دوساله بيشترين مقدار ويروس را دارا هستند. پراكندگي CTV در مناطق مهم مركبات كاري استانهاي فارس و بوشهر، بر روي گونهها و رقمهاي مختلف مركبات با استفاده از روش اليزا مورد بررسي قرار گرفت . از 646 درخت مورد آزمايش 242 درخت آلوده تشخيص داده شد. در ميان مناطق مختلف جهرم، خفر، كازرون، فسارود و جنتشهر داراب از استان فارس و دالكي، رودفارياب ، خائيز و جم از استان بوشهر و در ميان ارقام مختلف پرتقال، ليموشيرين، ليموترش ، نارنگي محلي، آنشو، نارنگي خاري و نارنگي كلمانتين آلوده تشخيص داده شد. آزمايشهاي انجام شده نشان داد پراكنش CTV در مناطق مختلف و روي رقمهاي مختلف مركبات جنوب ايران نسبتا وسيع است . وجود درختان ليموترش آلوده در نقاط دورافتاده نشان ميدهد كه CTV نه تنها احتمالا در سالهاي اخير همراه با رقمهاي جديد مركبات وارد منطقه شده، بلكه ممكن است از زمانهاي خيلي دور نيز در منطقه وجود داشته است . همچنين آلودگي ليموترش امكان وجود ناقل ويروس در منطقهء جنوب كشور را مطرح ميسازد. بنابراين آنچه كه لازم است از اين پس مورد مطالعه قرار گيرد علاوه بر پيگيري نقش ناقلين احتمالي، ميزان خسارت ناشي از CTV در درختان مركبات و نوع سويههايي است كه در منطقهء جنوب وجود دارد. |
خالصسازي، توليد آنتيسرم و توسعه يك سيستم بيوتكنولوژيكي حساس به منظور تشخيص دقيق و سريع ويروس تريست|
يك گياه مركبات با علائم مشكوك به ويروس تريستزا در باغات مهدشت ساري انتخاب و وجود ويروس مزبور در آن به روش Indexing به اثبات رسيد. گياه مزبوربعنوان منبع اوليه ويروس انتخاب و از آن پيوندك آلوده تهيه گرديد. بمنظور تكثير ويروس ، پيوندكهاي مزبور به 150 اصله نهال ليمو آب پيوند زده شد و نهالها در گلخانه نگهداري گرديدند، تكتك نهالها از نظر بيماريهاي ويروسي و شبه ويروسي ديگر از جمله Exocortis، پسورز و نقش حلقوي به روش متداول Indexing مورد آزمايش قرار گرفتند و معلوم شد كه نهالهاي مورد نظر فقط آلوده به ويروس تريستزا هستند و در گياهان محكي كه مورد استفاده قرار گرفته بودند هيچگونه علائم آلودگي ديگر مشاهده نگرديد، در آينده بافت آلوده نهالهاي پيوند زده شده، بمنظور خالصسازي ويروس مورد استفاده قرار خواهند گرفت . |
First report of Clavibacter michiganensis subsp. sepedonicus causing bacterial ring rot of potato inŞ. Altundağ1, A. Karahan1*, A.O. Kılınç1 and M. Özakman2
1 Plant Protection Central Research Institute, Gayret Mahallesi, Fatih Sultan Mehmet Bulvarı No: 66, Yenimahalle 06172, Ankara, Turkey
2 Gennova Pesticide Company, Bekir Saydam Cad., No: 31, Pancar, Torbalı, İzmir, Turkey
* aynur_karahan@zmmae.gov.tr
Accepted for publication 04 Dec 2008
During the 2007 seed potato monitoring programme in Turkey, 336 tuber samples from Kayseri province were tested for the presence of ring rot and brown rot diseases according to the EC Directives 93/85/EEC (Anon., 1993) and 98/57/EC (Anon., 1998). Three samples gave a positive immunofluorescence test (IF) result for the ring rot pathogen, Clavibacter michiganensis subsp. sepedonicus (Spieckermann & Kotthoff) Davis. et al.(Cms). Tuber heel end extracts were plated onto NCP-88 medium. Creamy-white, smooth, mucoid-fluidal colonies were selected and purified. External symptoms were observed on tubers of all three samples (two of cv. Soleia and one of cv. Safrane) as sunken and cracked areas (Fig. 1). When tubers were cut transversely, typical symptoms were seen through the vascular ring of tubers as a brown cheesy decay. Bacterial ooze was expressed when tubers were squeezed (Fig. 2). A positive IF result was obtained directly from the ooze. Isolates hydrolysed aesculin and produced catalase. They did not produce acid from glycerol, lactose, rhamnose and salicin. They gave negative results for oxidase activity, growth at 37°C, urease activity, starch hydrolysis, tolerance of 7 % NaCl, indole production and gelatine liquefaction.
Suspensions of pure colonies in water (c. 109 cfu per ml from each of the three samples were used for real-time PCR (Schaad et al. ,1999), with Ct values of 16.53, 16.21 and 32.2. Suspensions (106 cfu/ml) were injected into stems of ten Solanum melongena plants (cv. Black Beauty) at leaf stage 3. Control plants were inoculated with sterile water. Plants were incubated at 21°C and 70-80 % humidity. First symptoms were observed as dark green areas on leaves after ten days followed by wilting and necrosis (Figure 3). Cms was re-isolated and identified.
An intensive survey in 1988-1989 had shown that the potato growing areas of Turkey were free of the ring rot pathogen (Benlioğlu et al., 1991). This is the first report of Cms in Turkey. Strict control measures have been taken in contaminated and potentially contaminated fields since Cms is a regulated quarantine organism in Turkey.
Anon., 1993. Council Directive 93/85/EEC on the control of potato ring rot . Official Journal of the European Communities L 259, 1-25.
Anon., 1998. Council Directive 98/57/EC on the control of Ralstonia solanacearum (Smith) Yabuuchi et al. Official Journal of the European Communities L 235, 1-39.
Benlioğlu K, Öktem Y.E, Özakman M, 1991. Bacterial diseases of potatoes in the major potato-growing areas in Turkey. Bulletin OEPP/EPPO Bulletin 21, 67-72.
Schaad N.W, Berthier-Schaad Y, Sechler A, Knorr D, 1999. Detection of Clavibacter michiganensis subsp. sepedonicus in potato tubers by Bio-PCR and an automated real-time fluorescence detection system. Plant Disease 83, 1095-1100
M. M. Faghihi1, M. Salehi2, A. Bagheri1 and K. Izadpanah3*
1 Hormozgan Agriculture and Natural Resources Research Centre, Iran
2 Fars Agriculture and Natural Resources Research Centre, Iran
3 Department of Plant Protection, College of Agriculture, Shiraz University, Shiraz, Iran
*izadpana@shirazu.ac.ir
Huanglongbing (HLB), also known as greening, is a destructive disease of citrus that may limit production of this crop in many parts of Asia, Africa and South America. HLB is caused by a non-culturable phloem-limited bacterium of the genus ‘Candidatus Liberibacter’. Three Candidatus species of Liberibacter, namely, ‘Ca. L. asiaticus’, ‘Ca. L. africanus’ and ‘Ca. L. americanus’, have been identified (Bové, 2006). The psyllid Diaphorina citri, vector of ‘Ca. L. asiaticus’ was first detected in December 1997 in Iran in an area close to the border with Pakistan (Bové et al., 2000). Since then, high populations of D. citri have been found in citrus plantations of Hormozgan and Kerman provinces in southern Iran (Bové, 2006). Citrus trees in this region exhibit typical HLB symptoms including mottling of leaves and yellowing of shoots. In the present study leaf samples from 20 symptomatic and 20 symptomless Valencia sweet orange trees (Citrus sinensis) and over 50 psyllid samples were collected from various locations in Sistan-Baluchistan and Hormozgan provinces and total DNA was extracted using the CTAB (cetyltrimethylammonium bromide) method. DNA samples were tested for presence of ‘Ca. L. asiaticus’ by nested PCR using primer pairs F1 (5’-TGAATTCTTCGAGGTTGGTGAGC-3’)/R1(5’-GAATTCGACTTAATCCCCACCT-3’) as the first set and F2(5’ GCGTTCATGTAGAAGTTGTG-3’) / R2(5’-CCTACAGGTGGCTGACTCAT-3) as the second set. Both primer pairs were designed based on the published beta-operon of ribosomal protein encoding sequence of ‘Ca. L. asiaticus’ (Villechanoux et al. 1993). Nested PCR resulted in amplification of a 400 bp product from five out of 20 leaf samples and 26 out of 50 psyllid samples. No amplicons were obtained from symptomless sweet orange trees and psyllid samples reared on healthy sweet orange seedlings. The amplified fragment from a psyllid sample was cloned and sequenced (GenBank Accession No. FJ172759). BLAST search showed 100% identity with corresponding sequences of ‘Ca. L. asiaticus’ (M9439, AY34200 and EU078703). This is the first report on the occurrence of HLB disease in Iran. The disease appears to be widely distributed in citrus growing regions of southern Iran and may be a component of citrus decline in this area.
Bové JM, 2006. Huanglongbing: A destructive, newly–emerging, century-old disease of citrus. Journal of Plant Pathology 88, 7-37.
Bové JM, Danet G L, Bananej K, Hassanzadeh N, Taghizadeh M, Salehi M, Garnier M, 2000. Witches' broom disease of lime (WBDL) in Iran. In Yokomi, RH, da Graça, JV, Lee, RF, eds. Proceedings of the 14th Conference of the International Organization of Citrus Virologists, 300-309. Riverside, CA, USA: IOCV.
Villechanoux S, Garnier M, Laigret F, Renaudin J, Bové JM, 1993. The genome of the non-cultured, bacterial-like organism associated with citrus greening disease contains the nusG-rplKAJL gene cluster and the gene for a bacteriophage type DNA polymerase. Current Microbiology 26, 161-166.
E. Postnikova1, I. Agarkova2, S. Altundag3, F. Eskandari1, A. Sechler1, A. Karahan3 A.K. Vidaver2 W. Schneider1, M. Ozakman3 and N.W. Schaad1*
1 ARS-USDA, Foreign Disease-Weed Science Research Unit, Ft. Detrick, Maryland, USA
2 Department of Plant Pathology, University of Nebraska, Lincoln, Nebraska, USA
3 Plant Protection Research Institute, P.O. Box 49, Ankara, Turkey
Rathayibacter iranicus (Ri), originally reported in Iran in 1961 (Sharif, 1961), has not been reported outside Iran and only one strain is known to exist. Like R. tritici (Rt), Ri causes a gumming disease of wheat in association with the nematode Anguina tritici (Paruthi et al., 1989). During 2003, a survey of wheat seed for Rathayibacter species (RS) in Turkey using samples from 799 farmers in six provinces in Central Anatolia was conducted. The samples showed neither the brown to black galls typical of Anguina nor the yellowish galls typical of Ri and Rt. To determine the presence of RS, 120 g samples were washed in 100 ml of sterile 0.85% NaCl with 0.02% Tween 20 and plated onto 523 agar (Schaad et al., 2001) modified by adding 30 mg nalidixic acid, 15 mg polymixin B sulphate and 100 mg cycloheximide per litre. After incubation at 27º C for seven days, several typical colonies of RS were cloned by streaking onto YDC agar (Schaad et al., 2001). Twenty five strains were presumptively identified as RS based on a Gram positive reaction, oxidase negative reaction and yellow growth.
Three strains, TRS2 from Konya, TRS10 from Hisarkikaya, and TRS25 from Ankara, were used for further identification and speciation. The other 22 strains were archived for later studies. The type strains of Rt, (International Collection of Phytopathogenic Bacteria [ICPB] 70004T; FH-5; CT 102T) and Ri (ICPB 70005T; FH-6 T; CI 148T), were included as controls. All three strains were biochemically typical of Ri and Rt by producing acid from D-mannose, inulin, galactose, and mannitol.However, the strains failed to utilize acetate, or to hydrolyze casein, failed to grow in greater than 1% sodium chloride amended NBY, or on CNS agar, which is characteristic of Ri but not Rt (Davis & Vidaver, 2001). The new strains contained the fatty acids iso-14:0 and 16:0; anteiso 15:0 and 15:1, and 17:0, and 16:0, also typical of Ri. Identification of the three strains as Ri was further supported by 16s rDNA sequence similarities to Ri and Rt of 100%and 99.3%, respectively (GenBank Accession Nos. FJ595101 - Ri, ICPB 70005 and FJ595102 – Rt, ICPB 70004). Identification asRi was confirmed by AFLP analysis (EcoRI+0 and MseI+C) showing a pattern of 90% similarity to Ri but only 46% to Rt. This is the first report of Ri outside of Iran. Cultures have been deposited in the ICPB at Ft. Detrick, MD, USA, as ICPB 70146 (FH-154; TRS2), 70154 (FH-162; TRS10), and 70169 (FH-177; TRS25).
Davis M.J., Vidaver A.K., 2001. Gram Positive Bacteria, Coryneform Plant Pathogens. In: Schaad N.W., Jones J.B., Chun W., eds, Laboratory Guide for Identification of Plant Pathogenic Bacteria, 3rd Edn, St. Paul, MN, USA: APS Press, 218-234.
Paruthi I. J., Bajaj H.K., Bhatti D.S., 1989. Further observations on Anguina tritici (Nematoda) and Corynebacterium michiganense pv. tritici causing earcockle and yellow ear rot in wheat. Nematologica 35, 491-493.
Schaad N.W., Jones J.B., Chun W., eds, 2001. Laboratory Guide for Identification of Plant Pathogenic Bacteria, 3rd Edn, St. Paul, MN, USA: APS Press, 373 pp.
Scharif G., 1961. Corynebacterium iranicum sp. nov. on wheat (Triticum vulgare L.) in Iran, and a comparative study of it with C. tritici and C. rathayi. Entomologie et Phytopathologie Appliqués 19, 1-24.
N. Alabdalla1, F. Valentini1, C. Moretti2*, S. Essa3, R. Buonaurio2 and M. Abu-Ghorra4
1 Istituto Agronomico Mediterraneo, Via Ceglie 9, I-70010 Valenzano (Bari), Italy
2 Dipartimento di Scienze Agrarie e Ambientali, sez. Arboricoltura e Protezione delle Piante, Via Borgo XX Giugno, 74, I-06121 Perugia, Italy
3 General Commission for Scientific and Agriculture Research, Lattakia Center, Lattakia, Syria
4 Division of Plant Pathology and Plant Protection, Faculty of Agriculture, University of Damascus, Syria
*chiaraluce.moretti@unipg.it
Accepted for publication 25 Mar 2009
During field surveys carried out in 2007 in the main Syrian olive (Olea europaea) growing areas, bacterial knot symptoms were observed on olive twigs and branches, with the highest incidence (70%) in the coastal region (Lattakia and Tartous). Bacterial colonies isolated from knots resembled those of Pseudomonas savastanoi pv. savastanoi. Ten selected representative bacterial strains and the reference strains LMG 2209T, CFBP 6012 and 6013 of P. savastanoi pv. savastanoi were subjected to identification tests. All strains were Gram negative, fluorescent on King’s medium B and had oxidative but not fermentative metabolism. They were negative for levan, oxidase, potato rot and arginine dihydrolase and positive for tobacco hypersensitivity. One-year-old olive plants (cvs. Nebali and Jlot) were inoculated by introducing bacterial suspensions (108 cfu/ml) into wounds made in the bark with a sterile scalpel. All strains induced knots at the site of inoculation from 20 days onwards. Bacteria with characteristics identical to the original strains were re-isolated from inoculated plants. PCR analysis using primers specific for P. savastanoi pv. savastanoi, which amplify fragments of iaaL (Penyalver et al., 2000) and ptz (Powell & Morris, 1986) genes, generated amplicons of the expected size from all strains. Using BOX-, ERIC- and REP-PCR, we demonstrated that the isolates have a similarity of 87-98.9 % between themselves and with the reference strains. Based on morphological, biochemical, physiological and pathogenicity tests as well as molecular analyses, we can conclude that the Syrian strains belongs to P. savastanoi pv. savastanoi.
Although strains from Syria have previously been characterised with respect to their ability to produce auxin (Gardan et al., 1992), this is the first authoritative report of olive knot disease symptoms in Syria caused by P. savastanoi pv. savastanoi.
The authors would like to thank Mr. L. Bonciarelli for his excellent technical assistance.
Gardan L, David C, Morel M, Glickmann E, Abu-Ghorra M, Pettit A, Dessaux Y, 1992. Evidence for a correlation between auxin production and host plant species among strains of Pseudomonas syringae subsp. savastanoi. Applied and Environmental Microbiology 58, 1780-1783.
Penyalver R, García A, Ferrer A, Bertolini E, López MM, 2000. Detection of Pseudomonas savastanoi pv. savastanoi in olive plants by enrichment and PCR. Applied and Environmental Microbiology 66, 2673-2677.
Powell GK, Morris RO, 1986. Nucleotide sequence and expression of a Pseudomonas savastanoi cytokinin biosynthetic gene: homology with Agrobacterium tumefaciens tmr and tzs loci. Nucleic Acids Research 14, 2555-2565.
A. Z. Mirabadi1*, K. Rahnama2 and A. Esmaailifar3
1 Oilseeds Research & Development Company, Pasdaran Avenue Sari province, Iran
2 Department of Plant Protections, GorganUniversity Agricultural Sciences&Natural Resources, Iran
3 Department of Plant Protections, Faculty of Agriculture and Natural Resources Islamic AzadUniversity, Arak
*alizaman@arc-ordc.ir
Accepted for publication 28 Apr 2009
Rapeseed (Brassica napus) is one of the most important oilseed crops in Iran with more than 200,000 ha planted in 2008. Phoma blackleg (Leptosphaeria biglobosa), pathogenicity group 1 (PG-1) or non-aggressive type, has been reported on rapeseed from Golestan province (Fernando et al., 2007). Recently, in some regions of Mazandaran and Golestan provinces of northern Iran,typical stem canker symptoms were observed with incidence ranging from 20 to 60%. During October and November 2008, ascospores were isolated from pseudothecia on infected rapeseed debris and cultured on V8-agar medium (Mengistu et al. 1993). Based on the description of Punithalingam & Holliday (1972), sporulating colonies were identified as Leptosphaeria maculans (Desmaz.) Ces. & de Not. (Anamorph Phoma lingam (Tode: Fr.) Desmaz.).
Eleven isolates of L. maculans were used for determining the pathogenicity group according to phenotypic interaction (PI) on rapeseed cultivars including Westar, Quinta and Glacier. Nine seven-day old seedlings of each cultivar were inoculated for each isolate and the test was repeated three times. Wounded cotyledons were each inoculated with 10µl of conidial suspensions at 2 x 107 spores/ml. All plants were maintained in a growth chamber at 21° C (light) to 16° C (dark), with a 16-hour photoperiod and relative humidity of 95%. After 10 days, disease severity wasrated on a 0-9 scale (Williams, 1985) (Fig. 1).
Two isolates (Es-5 and Es-7) were classified as belonging to pathogenicity group PG2 and nine isolates as PG1. PG2 isolates showed PI reactions 0 to 2, 7 to 9 and 7 to 9 on Glacier, Quinta and Westar, respectively. In addition, cultivars Hyola401 and Okapi were highly sensitive to PG-2 isolates in the cotyledon assay. This is the first report ofthe occurrence of Leptosphaeria maculans PG-2 in Iran.
Special thanks to the Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) for providing the seeds needed for this testing.
Fernando WGD, Ghanbarnia K, Salati M, 2007. First report on the presence of phoma blackleg pathogenicity group 1 (Leptosphaeria biglobosa) on Brassica napus (canola/ rapeseed) in Iran. Plant Disease 91, 465.
Mengistu A, Rimmer RS, Williams PH, 1993. Protocols for in vitro sporulation, ascospore release, sexual mating, and fertility in crosses of Leptosphaeria maculans. Plant Disease 77, 538-540.
Leptosphaeria maculans. CMI descriptions of pathogenic Fungi and Bacteria. No. 331. Kew, UK: Commonwealth Mycological Institute.
Williams PH, 1985. Crucifer Genetics Cooperatives (CrGC) Resource Book. Madison, WI, USA: University of Wisconsin.
