ارزیابی تنوع ژنتیکی برخی جمعیت‌های هندی نماتد سیستی لوبیای سودانی Heterodera cajani با استفاده از RAPD

نویسندگان

چکیده

یکی از مهم‌ترین عوامل خسارت‌زای لوبیای سودانی، نماتد سیستی Heterodera cajani Koshy است که در هندوستان خسارت اقتصادی قابل توجهی به این محصول و برخی حبوبات دیگر وارد می‌کند. اطلاع از تنوع ژنتیکی جمعیت‌های این عامل خسارت‌زا در مناطق جغرافیایی مختلف، به انتخاب استراتژی‌های مؤثر در کنترل آن کمک بسیاری خواهد کرد. در تحقیق حاضر، با استفاده از روش RAPD، تنوع ژنتیکی ده جمعیت از نماتد H. cajani به دست آمده از مناطق مختلف تحت کشت لوبیای سودانی ارزیابی گردید. در این مطالعه از پرایمرهای ده تایی به عنوان سری‌های آغازکننده برای بررسی چندشکلی ملکول DNA استفاده گردید. تعداد قطعات تکثیر یافته به ازای هر پرایمر از 7 (در مورد OPA 16 و OPD 4) تا 24 (در مورد OPB 2) متغیر بود در حالی که اندازه‌ها بین 300 جفت باز تا 1/3 کیلو باز تغییر نشان دادند. از 32 پرایمر مورد آزمایش، 23تای آن‌ها (OPA 1, 2, 3, 4, 5, 6, 10 و OPB 2, 3, 4, 12 و OPC 1, 4, 5, 7, 8, 9, 10, 11, 16 و OPD 4, 10) 100 درصد چندشکلی از خود نشان دادند. در مجموع 451 قطعه تکثیر گردید که از این تعداد 434 چندشکلی و 17 تک‌شکلی بودند. ضریب جاکارد و تجزیه خوشه‌ای داده‌های به دست آمده نشان داد که جمعیت به دست آمده از منطقه ایندور از جمعیت سایر مناطق متمایز است. جمعیت‌های به دست آمده از منطقه سمستیپور و ایستگاه منطقه‌ای پوسا بیشترین خویشاوندی را دارا بودند.

کلیدواژه‌ها


عنوان مقاله [English]

Evaluation of genetic variation among Indian populations of the pigeonpea cyst nematode, Heterodera cajani, by RAPD

نویسندگان [English]

  • Mohammad Abdollahi
  • Ajoy Kumar Ganguly
چکیده [English]

One of the most important pathogen attacking pigeonpea root, the nematode Heterodera cajani Koshy, is causing significant economic losses in India. Knowledge of genetic variability present among different geographical populations is important for the selection of suitable control strategies. Molecular diversity among ten populations of H. cajani in India from pigeonpea growing areas is demonstrated by random amplified polymorphic DNA (RAPD). Operon series of decamer primers were used for exploring the polymorphism. The number of amplified fragments per primer varied from 7 (with OPA 16 and OPD 4) to 24 (with OPB 2) whereas the size varied from 300 bp to 3.1 kb. Out of the 32 primers tested 23 revealed 100% polymorphism (OPA 1, 2, 3, 4, 5, 6, 10, OPB 2, 3, 4, 12, OPC 1, 4, 5, 7, 8, 9, 10, 11, 16, OPD 4 and 10). Overall, 451 amplification products were obtained out of which, 434 were polymorphic and 17 were monomorphic. The Jaccard’s coefficient and cluster analysis showed that the nematode population from Indore was distinct from the other populations of H. cajani in India. Samastipur and Pusa R.S. populations showed high degree of similarity in this experiment.

کلیدواژه‌ها [English]

  • RAPD
  • Pigeonpea
  • Heterodera cajani
  • India
  • Genetic variation
  • Cyst nematode
Abdollahi, M. 2006. Intraspecific variations and race status of Heterodera cajani, infecting pigeonpea, Ph.D. Thesis, Indian Agricultural Research Institute, New Delhi, India.

Abdollahi, M. 2008a. Comparison of some Indian populations of cereal cyst nematode, Heterodera avenae (Wollenweber, 1924) using RAPD. Proceedings of the Pakistan Academy of Sciences, 45: 1-10.

Abdollahi, M. 2008b. Morphology and Morphometrics of Heterodera filipjevi (Madzhidov, 1981) Steller, 1984 From Kohgiluyeh and Boyer-Ahmad Province, Iran. Pakistan Journal of Biological Sciences, 11: 1864-1867.

Abdollahi, M. 2009a. Analysis of cyst and cone top morphometrics of Indian populations of maize cyst nematode. Journal of Plant Protection Research, 49: 41-47.

Abdollahi, M. 2009b. Hierarchical cluster analysis of Indian populations of Heterodera zeae based on second stage juveniles and egg morphometrics. International Journal of Agriculture and Biology, 11: 756–760.

Abdollahi, M., Ganguly, A.K. & Kaushal, K.K. 2006. Morphometric variations in Indian populations of Heterodera cajani I- Second stage juveniles and males. Indian Journal of Nematology, 36: 247-255.

Abdollahi, M., Ganguly, A.K. & Kaushal, K.K. 2007. Morphometric variations in Indian populations of Heterodera cajani II- Eggs, cysts and cone top structure. Indian Journal of Nematology, 37: 8-14.

Andersen, S. & Andersen, K. 1982. Suggestions for determination and terminology of pathotypes and genes for resistance in cyst‑forming nematodes, especially Heterodera avenae. EPPO Bulletin, 12: 379‑386.

Ayala, F.J. 1976. Molecular genetic Evolution. In: Ayala F.J. (Ed.), Molecular Evolution. Sunderland, MA. USA; Sinauer Association pp. 1-20.

Bekel, S., Gauthier, J.R. & Rivoal, R. 1997. Genetic diversity among a complex of cereal cyst nematodes inferred from RFLP analysis of the ribosomal internal transcribed spacer (ITS) region. Genome, 40: 479-496.

Bossis, M. & Rivoal, R. 1996. Protein variability in cereal cyst nematodes from different geographic regions assessed by two-dimensional gel electrophoresis. Fundamental and Applied Nematology, 19:25-34.

Dice, L.R. 1945. Measures of the amount of ecological association between species. Ecology, 26: 297-302.

Ferris, V.R., Faghihi, J., Ireholm, A. & Ferris, J.M. 1989. Two-dimensional protein patterns of cereal cyst nematode. Phytopathology, 79: 927-933.

Ferris, V.R., Miller, L.I., Faghihi, J. & Ferris, J.M.1994. Comparisons of isolates of Heterodera avenae using 2-D PAGE protein patterns and ribosomal DNA. Journal of Nematology, 26: 144-51.

Ganguly, A.K. & UmaRao. 2003. Application of molecular biology in nematology. In: Trivedi, P.C. (Ed.) Advances in Nematology. Scientific Publication, New Delhi, India. pp. 1-14.

Goodwin, P.H. & Annis, S.L. 1991. Rapid identification of genetic variation and pathotype of Leptosphaaeria maculans by random Amplified Polymorphic DNA assay. Applied Environmental & Microbiology, 57: 2452-2486.

Koshy, P.K. 1967. A new species of Heterodera from India. Indian Phytopathology, 20: 272-274.

Lopez-Brana, I., Romero, M.D. & Delibes, A. 1996. Analysis of Heterodera avenae populations by the random amplified polymorphic DNA technique. Genome, 39:118-122.

Lynch, M. & Milligan, B.G. 1994. Analysis of population genetic structure with RAPD markers. Molecular Ecology, 3: 91-99.

Nadler, S.A., Lindquist, R.L. & Near, T.J. 1995. Genetic structure of mid-western Ascaris suum populations: a comparison of isoenzymes and RAPD markers. Journal of Parasitology, 81: 385–394.

Nei, M. & Li, W.H. 1979. Mathematical model for studying genetic variation in terms of restriction endonucleases. Proceeding of National Academic of Sciences of USA, 76: 5269-5270.

Nobbs, J.M., Ibrahim, S.K. & Rowe, J. 1992. A morphological and biochemical comparison of the four cyst nematode species, Heterodera elachista, H. oryzicola, H. oryzae and H. sacchari (Nematoda: Heteroderidae) known to attack rice (Oryza sativa). Fundamental and Applied Nematology, 15: 39-41.

Pinochet, P., Cennis, J.L., Fernandez, C., Doucet, M. & Marull, J. 1994. Reproductive fitness and random amplified polymorphic DNA variation among isolates of Pratylenchus vulnus. Journal of Nematology, 26: 271-277.

Podzol, R.R. & Noel, G.R. 1984. Comparative electrophoretic analyses of soluble proteins from Heterodera glycines races 1-4 and three other Heterodera species. Journal of Nematology, 16: 332-340.

Rohlf, F.J. 1998. NTSYS-pc: Numerical taxonomy and multivariate analysis system. version 2.0. Applied Biostatistics, New York.

Sharma, S.B., Reddy, B.M.R. & Krishnappa, K. 1992. Incidence and importance of plant parasitic nematodes on pigeonpea and groundnut in Karnataka state in Southern India. Afro-Asian Journal of Nematology, 2: 21-26.

Siddiqui, Z.A. & Mahmood, I. 1993. Occurrence of races of Heterodera cajani in Uttar Pradesh, India. Nematologica mediterranea, 21: 185-186.

Subbotin, S.A., Waeyenberge, L., Irina, A.M. & Moens, M. 1999. Identification of Heterodera avenae group species by morphometrics and rDNA-RFLPs. Nematology, 1:195-207.

Subbotin, S.A., Waeyenberge, L. & Moens, M. 2000. Identification of the cyst forming nematodes of the genus Heterodera (Nematoda: Heteroderidae) based on ribosomal DNA-RFLP. Nematology, 2: 153-164.

Subbotin, S.A., Vierstraete, A. DeLey, P., Rowe, J. Waeyenberge, L., Moens, M. & Vanfleteren, J.R. 2001. Phylogenetic relationships within the cyst-forming nematodes (Nematoda, Heteroderidae) based on analysis of sequences from ITS regions of ribosomal DNA. Molecular Phylogenetics and Evolution, 21: 1-16.

Swain, S.C., Ganguly, A.K., Sahoo, P. & Mohapatra, T. 1999. RAPD analysis distinguishes four races of root-knot nematode, Meloidogyne incognita. Indian Journal of Nematology, 29: 1-7.

Tanha Maafi, Z., Subbotin, S.A. & Moens, M. 2003. Molecular identification of cyst-forming nematodes (Heteroderidae) from Iran and a phylogeny based on ITS-rDNA sequences. Nematology, 5: 99-111.

Walia, R.K. & Bajaj, H.K. 1986. Existence of host races in pigeonpea cyst nematode, Heterodera cajani. Nematologica, 32: 117-119.

Walia, R.K. & Bajaj, H.K. 1988. Further studies on the existence of races in pigeonpea cyst nematode, Heterodera cajani. Indian Journal of Nematology, 18: 269-272.

Welsh, J. & McClelland, M. 1990. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Research, 18: 7213-7218.

Williams, J.G.K., Kubelik, A.R., Livak, K.J., Rafalski, J.A. & Tingey, S.V. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research, 18: 6531-6535.