بررسی تغییرات ناشی از جهش با اتیل‌متان‌سولفونات (EMS) در لاین‌های موتانت برنج با نشانگر ISSR

نوع مقاله: علمی - پژوهشی

نویسندگان

1 استادیار گروه بیوتکنولوژی، دانشکده مهندسی انرژی و فناوری های نوین، دانشگاه شهید بهشتی تهران

2 دانش‌آموخته کارشناسی ارشد، گروه اصلاح نباتات و بیوتکنولوژی، دانشکده تولید گیاهی دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان

3 دانشیار گروه اصلاح نباتات و بیوتکنولوژی، دانشکده تولید گیاهی دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان

4 دانش‌آموخته کارشناسی‌ارشد، گروه زراعت و اصلاح نباتات، دانشکده علوم زراعی، دانشگاه علوم کشاورزی و منابع طبیعی ساری، ساری

چکیده

    استفاده از جهش جهت ایجاد تنوع ژنتیکی و مواد ژنتیکی جدید از دیرباز متداول بوده و از آن برای بهبود خواص کمّی و کیفی مختلف در گیاهان سود جسته‌اند. در این تحقیق، هجده لاین موتانت برنج که برای پاکوتاهی، تعداد پنجه زیاد، زودرسی و عملکرد بالا از بین یک جمعیت موتانت حاصل از جهش‌زایی رقم الیت ندا با اتیل‌متان‌سولفونات (EMS) انتخاب شده بودند، مورد ارزیابی مولکولی با آغازگرهای ISSR قرار گرفتند. از 10 آغازگر مورد استفاده، 8 آغازگر DNA ژنوتیپ‌های مختلف را تکثیر کردند. تعداد کل نوارهای تولید شده 67 نوار (به ازای هر آغازگر به‌طور متوسط 4/8 نوار) بود که 7/53 درصد آن‌ها چندشکلی بین ژنوتیپ‌ها را نشان دادند. متوسط ضریب تنوع ژنی نئی در جمعیت مورد مطالعه، 1/14 درصد به‌دست آمد. بیشترین شباهت به رقم مادری (8/86 درصد) مربوط به گروه موتانت‌های پرعملکرد و کمترین شباهت (7/76 درصد) مربوط به گروه موتانت‌های پرپنجه بود. تجزیه خوشه‌ای با روش UPGMA، رقم مادری را در یک گروه و 4 دسته لاین‌های موتانت را در گروه جداگانه‌ای قرار داد. تجزیه خوشه‌ای در هر گروه از لاین‌های موتانت نیز به‌طور جداگانه انجام شد که به استثنای دسته موتانت‌های انتخابی برای تعداد پنجه زیاد، در هر سه دسته دیگر رقم مادری به‌تنهایی در یک گروه و همه موتانت‌های انتخابی در گروه جداگانه‌ای قرار گرفتند. براساس نتایج این تحقیق، می‌توان نتیجه‌گیری کرد که تکنیک جهش برای ایجاد تنوع ژنتیکی در برنج مطلوب بوده و از نشانگرهای ISSR می­توان برای شناسایی لاین‌های موتانت در جمعیت‌های حاصل از جهش‌زایی با EMS استفاده کرد.

کلیدواژه‌ها


عنوان مقاله [English]

Assessment of EMS-induced Variations In rice Mutant Lines Using ISSR Marker

نویسندگان [English]

  • Asadollah Ahmadikhah 1
  • Hoda Shojaeian 2
  • M.Hadi Pahlevani 3
  • Leila Nayyeripasand 4
1 Assistant Proffesor, Department of Biotechnology, Faculty of Energy Engineering and Modern Technologies, Shahid Beheshti University, Tehran
2 Former M.Sc. Student, Department of Plant Breeding and Biotechnology, Faculty of Plant Production, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan
3 Associate Professor, Department of Plant Breeding and Biotechnology, Faculty of Plant Production, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan
4 Former Msc. Student, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Sari University of Agricultural Sciences and Natural Resources, Sari
چکیده [English]

Abstract
Using mutation has been relevant to induce genetic variation and new genetic material, and it was used to improve different qualitative and quantitative traits in crop plants. In this research, eighteen rice mutant lines which were selected from a mutant population developed from mutagenesis of rice elite cultivar Neda by using ethyl methane sulfonate (EMS) for reduced plant height, higher tiller number, early maturation and higher yield, were evaluated at molecular level using ISSR markers. Eight out of 10 primers successfully amplified DNA from different genotypes. In total, 67 bands (in average 8.4 bands per primer) were produced, 53.7% of which showed polymorphism between genotypes. Average Nei`s genetic diversity in the studied population was 14.1%. Highest similarity to original cultivar (86.8%) belonged to high-yielding mutant group and lowest similarity (76.7%) belonged to high-tiller mutant group. Cluster analysis using UPGMA method, placed the original cultivar alone in one cluster and 4 mutant groups in another distinct cluster. Cluster analysis was also performed separately in each sub-cluster that except for high-tiller group it could discriminate the original cultivar from any other three mutant groups. On the basis of the results of this research, it can be concluded that mutation technique was a desirable method for inducing genetic variation in rice and also it is possible to use ISSR markers for identification of mutant lines in populations developed from mutagenesis by EMS.
 

کلیدواژه‌ها [English]

  • EMS
  • Molecular marker
  • Mutant
  • Rice
Ahloowalia, B. S., Maluszynski, M. and Nichterlein, K. 2004. Global impact of mutation-derived varieties. Euphytica, 135: 187-204.

Ahmadikhah, A. 2009. A rapid mini-prep DNA extraction method in rice. African Journal of Biotechnology, 8(2): 323-27.

Ahmadikhah, A. 2012. Advanced genetics. Publication of Gorgan University of Agricultural Sciences and Natural Resources. Gorgan, Iran. 354 p.

Anbarasan, K., Sivalingam, D., Rajendran, R., Anbazhagan, M. and.Chidambaram, A. A. 2013. Studies on the mutagenic effect of EMS on seed germination and seedling characters of Sesame (Sesamum indicum L.) Var.T MV3. International Journal of Research in Biological Sciences, 3(1): 68-70.

Anderson, J. A., Churchill, J. E., Autrique, S. D., Tanksley, S. and Sorrells, M. E. 1993. Optimizing parental selection for genetic linkage maps. Genome, 36: 181-188.

Benjavad Talebi, A., and Shahrokhifar, B. 2012. Ethyl methane sulphonate (EMS) induced mutagenesis in Malaysian rice (cv. MR219) for lethal dose determination. American Journal of Plant Science, 3: 1661-1665.

Bughio, H. R., Asad, M. A., Odhano, I. A., Bughio, M. S., Khan, M. A. and Mastoi, N. N. 2007. Sustainable rice production through the use of mutation breeding. Pakistan Journal of Botany, 39(7): 2457-2461.

Fotookian, M. H. and Esfahani M. 2001. Induction of short culm mutant in Domsiah rice (Oryza sativa L.) variety. Iranian Journal of Crop Science, 3(3): 31-41.

Hoang, T. M. L., De Filippis, L. F. and Le, X. T. 2009. Salt tolerance and screening for genetic changes in rice mutants after gamma irradiation using RAPD and microsatellite (RAMP) markers. The Open Horticulture Journal 2: 62-69.

Human, S. and Sihono, H. 2010. Sorghum breeding for improved drought tolerance using induced mutation with gamma irradiation. Journal of Agronomy of Indonesia, 2: 95-99.

Ilirjana, S., Ariana, Y. and Andon, D. 2007. Induced mutations for improving production on bread and durum wheat. AIP Conference Proceedings, 899: 747.

Jabeen, N. and Mirza, B. 2002. Ethyl methane sulfonate (EMS) enhances genetic variability in Capsicum annum. Asian Journal of Plant Science, 1(4): 425-428.

Jana, M. K., and Roy, K. 1973. Induced quantitative mutations in rice. Radiation Botany, 13: 245-257.

Khademian, R., Babaeian Jelodar, N. and Kianoosh, Gh. 2004. Study of gamma radiation mutagenesis effects on some Iranian rice cultivars. Khazar Researches on Agricultural Sciences and Natural Resources, 2(4): 16-26.

Khan, A.J., Hassan, S., Tariq, M. and Khan, T. 2001. Haploidy breeding and mutagenesis for drought tolerance in wheat. Euphytica, 120: 409-414.

Kinnear, P. R. and Colin, D. G. 2000. SPSS for Windows made simple: Release 10. Hove, UK: Psychology Press. 456 p.

Lee, G. H. and Lee, S. Y. 2002. Selection of stable mutants from cultured rice anthers treated with ethyl methane sulfonic acid. Plant Cell Tissue and Organ Culture, 71: 165-17.

Majd, F., Rahimi, M. and Rezazadeh, M. R. 2002. Development of resistant lines to logging and high-yielding in rice gamma-irradiation- induced mutagenesis. Journal of Nuclear Technology, 26: 37-43.

Micke, A. 1999. Mutation and in vitro mutation breeding. Bahar Samiullah Khan, Kalani Publishers, Ludhiana, India, pp: 1-19.

Miri, S. M., Mousavi, A., Naghavi, M. R., Mirzaii, M., Talaei, A. R. and Naserian Khiabani, B. 2009. Analysis of induced mutants of salinity resistant banana (Musa acuminata cv. Dwarf Cavendish) using morphological and molecular markers. Iranian Journal of Biotechnology, 7(2): 86-92.

Mohamad, O., Loo, M. W., Mohd Nazir, B., Rusli, I., Herman, S. and Bakhendri, S. 2009. Drought tolerance in sorghum and soybean. Forum for Nuclear Cooperation in Asia (FNCA).

Naseri Tafti, M., Yousefi, F., Rezazadeh, M., Sabzi, H. and Oujani, R. 2003. Assessment of mutant lines developed from gamma irraradiation in soybean. Journal of Nuclear Sciences and Technologies, 27: 17-22.

Nei, M. 1978. Analysis of gene diversity in subdivided populations. Proceeding of National Academy of Science USA 70(12): 3321-3323.

Nogueira, P. K., Amorim, E. P., Ferreira, C. U. F., Amorim, V. B. O., Santos, L., Silva Ledo, C. A. and Silva, S. O. 2011. Agronomic and molecular characterization of gamma ray induced banana (Musa sp.) mutants using a multivariate statistical algorithm. Euphytica, 178:151-158.

Patnaik, A., Chaudhary, D. and Rao, G. J. N. 2006. Genetic improvement of long grain aromatic rices through mutation approach. Plant Mutation, 1(1): 7-10.

Penmetsa, R. V. and Cook, D. R. 2000. Production and characterization of diverse developmental mutants of Medicago truncatula. Plant physiology, 123: 1387-1398.

Rakshit, S., Kanzaki, H., Matsumura, H., Rakshit, A., Fujibe, T., Okuyama, Y., Yoshida, K., Oli, M., Shenton, M., Utsushi, H., Mitsuoka, C., Abe, A., Kiuchi, Y. and Terauchi, R. 2010. Use of TILLING for reverse and forward genetics of rice, P 187-197, In: Meksem, K., and Kahl, G. (eds), The Handbook of Plant Mutation Screening, Wiley-VCH, Germany.

Reddi, S. T. V. V. and Reddi, V. R. 1984. Frequency and spectrum of chlorophyll mutants induced in rice by chemical mutagens. Theoreticah and Applied Genetics, 67: 231-233.

Reddy, P. M., Sarla, N. and Siddiq, E. A. 2002. Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding. Euphytica, 128: 9-17.

Shehata, S. M., Allah, A. A. and Zayed, B. A. 2009. Development of salt tolerant rice lines through mutation breeding. Journal of Agriculture Research, 35(4): 954-963.

Shehzad, T., Allah, A. B. D., Ammar, M. H. and Abdelkhalik, A. F. 2011. Agronomic and molecular evaluation of mutant rice (Oryza sativa L.) lines in Egypt Megahed Helmy and Amr Farouk. Pakistan Journal of Botany, (2): 1183-1194.

Shojaeian, H. 2012. Study on phenotypic and molecular diversity induced by ethyl methane sulphonate (EMS) mutagen in ric. Msc. Dissertation, Gorgan University of Agricultural Sciences and Natural Resources, Goragn, Iran. 122 p.

Shu, Q. Y. and Lagoda, P. J. L. 2007. Mutation techniques for gene discovery and crop improvement. Mol. Plant Breed, 2: 193-195.

Singh, N. K. and Balyan, H. S. 2009. Induced mutations in bread wheat (Triticum aestivum L.) CV. ‘Kharchia 65’ for reduced plant height and improve grain quality traits. Advances in Biological Research, 3 (5-6): 215-221.

Van Harten, A. M. 1998. Mutation breeding: theory and practical applications. Cambridge University Press, London, UK.

Vasline, Y. A. 2013. An investigation on induced mutations in rice (Oryza sativa L.). Plant Archives 13(1)-555-557.

Wongsawad, P., Wongsawad, C., Mahadtanapuk, S., Kantawong, S., Chariyavidhawat, P. and Paratasilpin, T. 2005. Induced mutation in Adenium obesum Balf. using ethyl methane sulphonate. Proceedings of Congress on Science and Technology of Thailand. Suranaree University of Technology, 31: 18-20.

Yeh, F. C., Yang, R. C., Boyle, T. B. J., Ye, Z. H. and Mao, J. X. 1997. POPGENE, the User-friendly shareware for population genetic analysis.molecular biology and biotechnology centre, University of Alberta, Canada.Ahloowalia, B. S. and Maluszynski, M. 2001. Induced mutations–A new paradigm in plant breeding. Euphytica, 118:167-173.