c3518cb17d976b8

شناسایی ریخت‌شناختی و مولکولی برخی عامل‏ های شبه قارچی ایجادکنندۀ بیماری پوسیدگی طوقه و ریشۀ درختان بادام در استان‌های اصفهان و چهارمحال بختیاری

نوع مقاله : مقاله پژوهشی

نویسندگان

1 استاد، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه صنعتی اصفهان

2 استادیار، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه یاسوج

3 دانشجوی سابق کارشناسی ارشد، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه صنعتی اصفهان

چکیده

بادام یکی از درختان میوه تیرۀ Rosaceae است که در استان‌های‏‏‏ اصفهان و چهارمحال و بختیاری برای استفاده از دانۀ خوراکی آن نسبت به سایر درختان میوۀ هسته‌دار‏‏‏ بیشتر کشت می‌شود. در این پژوهش، برخی از عامل‏های پوسیدگی ریشه و طوقۀ درختان بادام مورد بررسی قرار گرفتند. بر اساس خصوصیات مورفولوژیکی 53 جدایۀ Phytophthora cactorum (19 جدایه) و Phytophthora cryptogea (21 جدایه) و Phytopythium litorale (13 جدایه) جداسازی و شناسایی شدند. به‌منظور تأیید مولکولی گونه‌های‏‏‏ شناسایی‌شده، تعداد 25 جدایه از بین 53 جدایۀ کل به‌طور تصادفی جهت تعیین توالی، محصول حاصل از تکثیر ناحیه‏های ITS انتخاب شدند. همچنین از جفت آغازگرهای اختصاصی BPhycacL87FRG(F) /BPhycacR87RRG و Cryp1(F)/ Cryp2 برای شناسایی اختصاصی گونه‌ها‏‏‏ استفاده شد. واکنش نهال‌های هشت‌ماهه رقم‏های بادام سنگی، ربیع، شاهرود 12 و پایه GF677 به سه جدایۀ (cactorum P) P1، (P. cryptogea) P32 و (Pp. litorale) P41 در شرایط گلخانه نشان داد که گونه‌های‏‏‏ P. cactorum و P. cryptogea و Pp. litorale به لحاظ بیماری‌زایی‏‏‏ تفاوت معنی‌داری‏‏‏ با یکدیگر دارند. گونه‌های‏‏‏ Pp. litoraleوP. cactorum به ترتیب بیشترین و کمترین بیماری‌زایی را در مقابل همۀ رقم‏های بادام داشتند و رقم‏های سنگی و GF677 به ترتیب بیشترین و کمترین حساسیت را در برابر هر سه گونه از خود نشان دادند. برای پژوهش‌های فیلوژنتیکی، تعداد 9 جدایه (سه جدایه از هر گونۀ P. cactorum، P. cryptogea و Pp. litorale)بر اساس توالی‏یابی‏‏‏ ناحیۀ ITS-rDNA بررسی شدند. این پژوهش اولین گزارش از بیماری‌زایی Pp. litorale روی بادام در ایران است.

کلیدواژه‌ها

عنوان مقاله [English]

Morphological and molecular identification of some causal agents of root and crown rot diseases of almond in Isfahan and Chaharmahal and Bakhtiari provinces

نویسندگان [English]

  • Bahram Sharifnabi 1
  • Fariba Ghaderi 2
  • Nahid Javadi 3
1 Professor, Department of Plant Protection, College of Agriculture, Isfahan University of Technology, Isfahan, Iran
2 Assistant Professor, Department of Plant Protection, College of Agriculture, Yasouj University, Iran
3 Former M. Sc. Student, Department of Plant Protection, College of Agriculture, Isfahan University of Technology, Isfahan, Iran
چکیده [English]

Almond (Prunus amygdalus L.) is a deciduous tree in the Rosaceae family and is grown mostly for its edible seed. It is one of the most popular nuts on a worldwide basis and widely grown in Iran in Isfahan and Chaharmahal and Bakhtiari provinces. Root and crown rot causal agents which are caused by several pathogens were studied. According to morphological characteristics, 53 isolates were identified as Phytopythium litorale (13 isolates), Phytophthora cactorum (19 isolates), and P. crytogea (21 isolates). Out of 53 isolates, 25 isolates were selected for molecular studies. Specific primer pairs of BPhycacL87FRG (F)/BPhycacR87RRG and Cryp1(F)/Cryp2 were used to confirm morphological identification of species. The reaction of crown and root of 8-month-old almondcultivars Sangi, Rabee, Shahrood12 and GF677 to virulent isolates of P. cactorum, P. cryptogea,and Pp. litorale was evaluated under greenhouse conditions. Comparison of pathogenicity means revealed that there are significant differences among P. cactorum, P. cryptogea,and Pp. litorale isolates.The results alsoshowedthat Pp. litorale was more aggressive than P. cactorum. On all tested almond cultivars, disease severity percentage showed that Sangi is the most susceptible cultivar and GF677 showed the least susceptibility. In this study, phylogenetic relationships of some Phytophthora and Phytopythium species based on sequencing of the rDNA-ITS region is provided. This is the first report of isolation and pathogenicity of Phytopythium litorale from almond orchards in Iran.

کلیدواژه‌ها [English]

  • Pathogenicity
  • phylogeny
  • Phytophthora
  • Phytopythium
  • Taxonomy

مراجع

  1. Afzali, H. & Khabbazjolfai, H. (2004). Phytophthora iranica a new causal agent of almond trees death in Tehran province In: Proceedings of the 16th Iranian Plant Protection Congress, 28 August-1 September, Tabriz, Iran, pp. 397.
  2. Altschul, S. F., Madden, T. L., Schaffer, A. A., Zhang, J. & Lipman, D. J. (1997). Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Nucleic Acids Research, 25, 3389-3402.
  3. Banihashemi, Z. (1995). Role of Phytophthora cactorum on decline of fruit and nut trees in Fars province. In: Proceedings of the 12th Iranian plant protection congress, 21–26 August, Karaj, Iran, pp. 224.
  4. Fatemi, G. (1977). Almond decline in Fars province and it’s relation with the Phytophthora Fungus species. In: Proceedings of 6th Iranian plant protection congress, 12-14 September, Karaj, Iran, pp. 34.
  5. Banihashemi, Z. & Sartipi, A. (2004). Identification of Phytophthora species associated with stone fruits crown rot in Fars province and reaction of certain rootstock to P. cactorum. Journal of Sciences and Technology of Agriculture and Natural Resources, 8, 241-249
  6. Banihashemi, Z. (2004). A method to monitor the activity of Phytophthora spp. in the root zone of Pistacia spp. Phytopathologia Mediterranea, 43, 411-414.
  7. Bouket, A. C., Babai-Ahari, A., Arzanlou, M. & Tojo, M. (2016). Morphological and molecular characterization of Phytopythium litorale and Pp. oedochilum from Iran. Nova Hedwigia, 102, 257-270.
  8. De Cock, A. W. A. M. & Levesque, C. A. (2004). New species of Pythium and Phytophthora. Studies in Mycology, 50, 481-487.
  9. De Cock, A. W. A. M., Lodhi, A. M., Rintoul, T. L., Bala, K., Robideau, G. P., Abad, Z. G., Coffey, M. D., Shahzad, S. & Levesque, C. A. (2015). Phytopythium: Molecular phylogeny and systematics. Persoonia, 34, 25-39.
  10. El-Hamalawi, Z. A. & Menge, J. A. (1994). Effect of wound, age and fungicide treatment of wound on susceptibility of stems to infection by Phytophthora citricola. Plant Disease, 78, 700-704.
  11. Ershad, D. (1992). Phytophthora species in Iran (Isolation, Purification, Identification). Agriculture Research Organization. 215pp.
  12. Ersek, T., Schoelz, J. E. & English, J. T. (1994). PCR Amplification of species-specific DNA sequences can distinguish among Phytophthora species. Applied Environmental Microbiology, 60, 2616-2621.
  13. Erwin, D. C. & Ribeiro, O. K. (1996). Phytophthora: Diseases worldwide. APS Press.St. Paul, Minn. 562p.
  14. Food and Agriculture Organization. (2012). Biodiversity: Agricultural biodiversity in FAO. Retrieved January 12, 2009. From http://www.fao.org/biodiversity.
  15. Ghaderi, F. & Banihashemi, Z. (2006). Distribution and relative importance of Phytophthora species isolated from declining walnut tree in Fars, Kerman, and Kohgilueh and BoyerAhmad provinces and reaction of walnut genotype to them. Iranian Journal of Plant Pathology, 43, 163-183. (in Farsi)
  16. Grasselly, C. & Duval, H. (1997). Ľ aimandier: Centre technique interprofessionnel des fruits et legumes (Ctifl), 167 pp.
  17. Hall, T. A. (1999). BioEdit: A User-Friendly Biological Sequence Alignment Editor and Analysis Program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95-98.
  18. Hansen, E. M., Hamm, P. B., Julis, A. J. & Roth, L. F. (1979). Isolation, incidence and management of Phytophthora in forest tree nurseries in the Pacific Northwest. Plant Disease Report, 63, 607-611.
  19. Haygood, R. A., Graves, C. H. & Riding, W. H. (1986). Phytophthora root rot and stem canker of peach trees in Missisippi. Plant Disease, 70, 866-868.
  20. Kannwischer, M. E. & Mitchell, D. J. (1981). Relationships of numbers of spores of Phytophthora parasitica var nicotianae to infection and mortality of tobacco. Phytopathology, 71, 69-73.
  21. Katoh, K. & Standley, D. M. (2013). MAFFT multiple sequence alignment software. Version 7: improvements in performance and usability. Molecular Biology and Evolution, 30, 772-780.
  22. Kimura, M. (1980). A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution, 16, 111-120.
  23. Li, M., Asano, T., Suga, H. & Kageyama, K. (2011). A multiplex PCR for the detection of Phytophthora nicotianae and Phytophthora cactorum. Plant Disease, 95, 1270-1278.
  24. Minerdi, D., Yuan, L., Garibaldi, A., Lodovica, M., Moretti, M. & Gaggero, L. (2007). Conventional PCR and Real time quantitative PCR detection of Phytophthora cryptogea on Gerbera jamesonii. Plant Pathology, 122, 227-237.
  25. Murray, M. G. & Thompson, W. F. (1980). Rapid isolation of high molecular weight plant DNA. Nucleic Acids Research, 8, 4321-4326.
  26. Nei, M. (1973). Analysis of gene diversity in subdivided populations. Science, 70, 3321-3323.
  27. Robideau, G., Pde Cock, A. W. A. M., Coffey, M. D., Voglmayr, H., Brouwer, K., Bala, D. W., Chitty, N., Desaulniers, Q. A., Eggertson, C. M., Gachon, C. H., Rintoul, T. L., Sarhan, E., Verstappen, E.C., Zhang, Y., Bonants, P. J., Ristaino, G B. & Levesque, C. A. (2011). DNA barcoding of oomycetes with cytochrome c oxidase subunit I and internal transcribed spacer. Molecular Ecology Resources, 11, 1002-1011.
  28. Saitou, N. & Nei, M. (1987). The Neighbor-Joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4, 406-425.
  29. Sahragard, N. & Banihashmi, Z. (2006). Evaluation of resistance of some almond genotypes and cultivars to Phytophthora cactorum. Iranian Jouranal of Plant Pathology, 42, 309-322.
  30. Schena, L., Duncan, J. M. & Cooke, D. E. L. (2008). Development and application of a PCR-based molecular tool box for the identification of Phytophthora species damaging. Plant Pathology, 57, 64-75.
  31. Tamura, K., Dudley, J., Nei, M. & Kumar, S. (2007). Molecular Evolutionary Genetics Analysis (MEGA) Software Version 4.0. Molecular Biology and Evolution, 24, 1596-1599.
  32. Weste, G. (1983). Population dynamics and survival of Phytophthora. Pp.: 237-259. In: Phytophthora: Its Biology, Taxonomy, Ecology, and Pathology. Erwin, D.C., Bartnicki-Garcia, S., and Tsao, P.H. (eds.). APS Press. 329 P.
  33. Zakii, Z. & Ershad, D. (1994). Isolation of Phytophthora cactorum from cherry trees in Karaj. Iranian Journal of Plant Pathology, 30, 78. (in Farsi)

فایل ها

سابقه مقاله

  • تاریخ دریافت: 27 دی 1396
  • تاریخ بازنگری: 03 آبان 1397
  • تاریخ پذیرش: 25 آذر 1397

هم رسانی

ارجاع به این مقاله

آمار