تعیین تنوع ژنتیکی و بیماری‌زایی جدایه‌های Fusarium oxysporum از چغندر‌قند در ایران

نوع مقاله: کامل علمی - پژوهشی

نویسندگان

1 دانش‌آموخته کارشناسی ارشد بیماری‌شناسی گیاهی واحد علوم و تحقیقات

2 استادیار دانشگاه آزاد اسلامی واحد علوم و تحقیقات

3 استادیار موسسه تحقیقات چغندرقند

چکیده

تنوع ژنتیکی و بیماری­زایی 13 جدایه قارچ Fusarium oxysporumکه طی سالهای 1387-1378 از مزارع چغندرقند شش استان کشور ایران جمع‌آوری شده بود، مورد بررسی قرار گرفت. تنوع بیماری­زایی آنها با استفاده از روش غوطه ور سازی ریشه گیاهچه‌های چهار‌هفته‌ای در سوسپانسیون اسپور در لوله آزمایش انجام شد. بر این اساس جدایه‌ها به دو گروه بیماری­زای قوی و ضعیف دسته‌بندی شدند. تنوع ژنتیکی جدایه ها با استفاده از ۱۵ آغازگر تصادفی و به روش RAPD ارزیابی شدند. بر این اساس چند شکلی قابل ملاحظه ای بین جدایه‌های مورد بررسی مشاهده شد. گروه بندی خوشه‌ای جدایه ها با استفاده از UPGMA به کمک نرم‌افزار MVSP، جدایه‌ها را به پنج گروه طبقه بندی کرد. رابطه مشخصی بین تنوع ژنتیکی بر اساس RAPD-PCR با بیماری­زایی جدایه‌ها یافت نشد، اما مجزا شدن برخی از جدایه ها از سایرین با مناطق جغرافیایی آنها در ارتباط بود. DNA ژنومی جدایه‌ها با کمک آغازگرهای ITS1 و  ITS4 تکثیر شد. محصول تکثیر شده از DNA جدایه‌های مختلف ، با آنزیم‌های برشگر TaqI ,HaeIII ,EcoRI  هضم گردید. آنالیز ITS-RFLP جدایه ها الگوی باندی مشابهی را نشان داد.

کلیدواژه‌ها


عنوان مقاله [English]

Pathogenic and genetic variation of Iranian isolates of Fusarium oxysporum from sugar beet in Iran

نویسندگان [English]

  • S. Beladi Behbahani 1
  • S. Rezaee 2
  • S.B. Mahmoodi 3
1 M.Sc. of Plant Pathology, Science and Research Branch, Islamic Azad University, Iran
2 Assistant professor of Science and Research Branch, Islamic Azad University, Iran
3 Assistant professor of Sugar Beet Seed Institute (SBSI), Iran
چکیده [English]

Pathogenicity and genetic variability of 13 Fusarium oxysporum isolates collected from sugar beet field of six provinces of Iran during 1999- 2008 were evaluated. Pathogenic variability of the isolates was determined using a root-dip assay in spore suspension in test tubes for four- week plants. The isolates were classified into strong and weak pathogenic groups. A total of 15 RAPD primers were used for the isolates genetic diversity evaluation. On this basis, high polymorphism was observed among the isolates. Cluster analysis (UPGMA) of the isolates using the MVSP software package classified them into five groups. There was a relationship between RAPD results and geographical origins of the isolates. However, no relationship was found between RAPD results and pathogenicity. Genomic DNA of isolates was amplified using ITS1 and ITS4 primers. The amplified DNA products of the isolates was digested by three restriction enzymes EcoR1, Taq1 and HaeIII. ITS- RFLP analysis showed similar banding pattern  the isolates.

کلیدواژه‌ها [English]

  • Fusarium oxysporum
  • sugar beet
  • RAPD- PCR
  • rDNA-RFLP
Abasi Moghadam E, Rastgar M, Jafarpoor B. Etiology of root and crown rot of sugar beet in Khorasan. Proceedings of the 13th Iranian Plant Protection Congress, Karaj, Iran. 1997; P. 125. (In Persian, abstract in English)

Arruda GMT, Miller RNG, Ferreira MASV, Café-fiho AC. Morphological and molecular characterization of the sudden death syndrome pathogen of soybean in Brazil. Plant Pathology. 2005; 54:53-65.

Behdad E. Diseases of Agricultural Plants in Iran. Neshat Esfahan Publishers. 1983. (In Persian)

Bosland PW, Williams PH. An evaluation of Fusarium oxysporum from crucifers based on pathogenicity, isozyme polymorphism, vegetative compatibility, and geographic origin. Can. J. Bot. 1987; 65:2067-2073.

Cramer RA. Characterization of Fusarium oxysporum isolates from common bean and sugar beet using pathogenicity assays and random-amplified polymorphic DNA markers. Journal of Phytopathology. 2003; 151: 352-360.

Elias KS, Zamir D, Lichtman-Pleban T, Katan T. Population structure of Fusarium oxysporum f. sp. lycopersici. Mol. Plant-Microb Intract. 1993; 6:565-572.

Ferderick MA, Roger Brent R, Kingston D, Seidamn J, Smith A, Strunl K. Preparation of plant DNA using CTAB. Short Protocol Molecular Biology. 2002; 2/4: 2/10.

Fisher GA, Gerik JS. Genetic diversity of Fusarium oxysporum isolates pathogenic to sugar beets. (Abstr) Phytopathology. 1994; 84:1098.

Hanson LE. Beet root-rot inducing isolates of Fusarium oxysporum from Colorado and Montana. Plant Disease. 2006; 90: 247.

Harveson RM, Rush CM. Genetic variation among Fusarium oxysporum isolates from sugar beet as determined by vegetative compatibility. Plant Disease. 1997; 81:85-88p.

Kistler HC. Genetic diversity in the plant pathogenic fungus Fusarium oxysporum. Phytopathology. 1997; 87:474-479.

Lee YM, Choi YK, Min BY. PCR-RFLP and sequence analysis of the rDNA ITS region in the Fusarium spp. The Journal of Microbiology. 2000; 38: 66-73.

Mahmoodi SB, Mesbah MR, Rahimian H, Alizadeh A, Noroozi P. Genetic diversity of sugar beet of Rhizoctonia solani revealed by RAPD-PCR and ITS-PCR and ITS-rDNA analysis. Iranian Journal of Plant Pathology. 2005; 41: 523-542. (In Persian, abstract in English)

Martin FN, English JT. Population genetics of soil born Fungal plant pathogens. Phytopathology. 1997; 87: 446-447.

Martyn RD, Rush CM, Biles CL, Bake EH. Etiology of a root rot disease of sugar beet in Texas. Plant Disease. 1989; 73: 879-883.

MacDonald B. The population genetics of Fungi: tools and techniques. Phytopathology. 1997; 87: 448-453.

Namiki F, Shiomi T, Nishi K, Kayamura T, Tsuge T. Pathogenic and genetic variation in the Japanese strains of Fusarium oxysporum f. sp. melonis. Phytopathology. 1998; 88:804-810.

Noorifar N, Etebarian HR, Mahmoodi SB, Hashemi M. Separation and identification of dsRNA related to some isolates of three species of genus Fusarium and study of their effects on isolates pathogenicity. Proceedings of the 6th National Biotechnology Congress of Iran, 13-15 Aug, Tehran, Iran; 2009. (In Persian)

Panella LW, Ruppel EG, Hecker RJ. Registration of four multigerm sugar beet germplasm resistant to Rhizoctonia root rot. Crop Science. 1995; 35: 291-292.

Pavaanen-Huhtala S, Hyvonen J, Bulat SA, Yli-Maltila T. RAPD-PCR, isozyme, rDNA RFLP and rDNA sequence analyses in identification of finfish Fusarium oxysporum isolates. Mycological Research. 1999;103:625-634.

Puhalla JE. Classification of strains of Fusarium oxysporum on the basis of vegetative compatibility. Can. J. Bot. 1985; 63:179-183.

Raufi M, Farokhinejad R, Mahmoodi SB. Identification and pathogenicity of Fusarium species associated with sugar beet root and crown rot in Iran. Journal of Sugar Beet. 2003; 19(2): 109-122. (In Persian, abstract in English)

Ruppel EG. Pathogenicity of Fusarium spp. from diseased sugar beets and variation among sugar beet isolates of Fusarium oxysporum. Plant Disease. 1991; 75:486-489.

Salazar O, Julian MC, Rubio V. Primers based on specific rDNA-ITS sequence for PCR detection of Rhizoctonia solani, R. solani :AG-2 subgroups and ecological types and binucleate Rhizoctonia. Mycological Research. 2000; 104: 281-285.

Toda T, Hyakumachi M, Suga H, Kageyama K, Tanaka A, Tani T. Differentiation of Rhizoctonia AG-D isolated from turf grass in to subgroups I and II based on rDNA and RAPD analyses. European Journal of Plant Pathology. 1999; 105: 835-846.

Woo SL, Zonia A, Del Sorbo G, Lorito M, Nanni B, Scala F, Noviello P. Characterization of Fusarium oxysporum f. sp. phaseoli by pathogenic races, VCGs, RFLP and RAPD. Phytopathology. 1996; 88:966-973.