Establishment and Optimization of SRAP-PCR Reaction System for Elymus breviaristatus

doi:10.11924/j.issn.1000-6850.2014-1190

Chinese Agricultural Science Bulletin ›› 2014, Vol. 30 ›› Issue (30): 259-264.doi: 10.11924/j.issn.1000-6850.2014-1190

Establishment and Optimization of SRAP-PCR Reaction System for Elymus breviaristatus

Received:2014-04-23 Revised:2014-04-23 Accepted:2014-08-06 Online:2014-10-31 Published:2014-10-31

Abstract

Abstract: The SRAP- PCR for Elymus breviaristatus was optimized using the single factor test and the orthogonal design test in the study. The optimized SRAP- PCR system for E. breviaristatus was: 2.00 mmol/L MgCl₂, 0.40μmol/L primers, 1.0 U Taq DNA polymerase, 250μmol/L dNTPs, 30 ng DNA template, 2μL 10×PCR buffer in the total volume of 20μL. The results showed that the order for the most influenced factor to the least influenced factor was: Mg²⁺ , primer, Taq polymerase, dNTPs and DNA template. The optimized PCR system was confirmed using six E. breviaristatus accessions. The electrophoresis results showed that the amplified bands were highly polymorphic and clear, further indicating that the optimized system was reliable. The optimized SRAP- PCR system could be used for genetics and breeding studies in E. breviaristatus or other Elymus species.

References

[1] 周永红,郑有良,杨俊良,等.10种披碱草属植物的 RAPD分析及其系统学意义[J].植物分类学报,1999,37(5):425-432.
[2] 郭本兆.中国植物志[M].北京:科学出版社,1987,9(3):7-104.
[3] 曾植虎.短芒披碱草与老芒麦、垂穗披碱草的对比试验[J].青海草业,2012,21(1):20-23.
[4] 张桥英,何兴金.四川省珍稀濒危植物及其保护[J].武汉植物学研究,2002,20(5):387-394.
[5] Ma X., Zhang X Q, Zhou Y H, et al. Assessing genetic diversity of Elymus sibiricus (Poaceae: Triticeae) populations from QinghaiTibet Plateau by ISSR markers[J]. Biochemical Systematics and Ecology, 2008, 365: 514-522.
[6] Gaudett M., Salomon B., and Sun G. Molecular variation and population structure in Elymus trachycaulus and comparison with its morphologically similar E. alaskanus[J]. Plant Systematics and Evolution, 2005, 250: 81-91.
[7] Stevens Laci, Salomon Bj?rn, Sun Genlou. Microsatellite variability and heterozygote excess in Elymus trachycaulus populations from British Columbia in Canada[J]. Biochemical Systematics and Ecology, 2007, 35: 725-736.
[8] Mizianty M., Bieniek W., Czech A., et al. Variability and structure of natural populations of Elymus caninus (L.) L. and their possible relationship with Hordelymus europaeus (L.) Jess. ex Harz as revealed by AFLP analysis[J]. Plant Systematics and Evolution, 2005, 256: 193-200.
[9] Li G, Quiros C. F. Sequence- related amplified polymorphism (SRAP), a new marker system based on a simple PCR reaction: Its application to mapping and gene tagging in Brassica[J]. Theoretical and Applied Genetics, 2001, 103(3): 455-461.
[10] Budak H., Shearman R.C., Parmaksiz I., et al. Molecular characterization of Buffalograss germplasm using sequence related amplified polymorphism markers[J]. Theor. Appl. Genet, 2004, 108: 328-334.
[11] 陈智华,苗佳敏,钟金城,等.野生垂穗披碱草种质遗传多样性的SRAP研究[J].草业学报,2009,18(5):192-200.
[12] 苗佳敏,张新全,陈智华,等.青藏高原和新疆地区垂穗披碱草种质的 SRAP及 RAPD分析[J].草地学报,2011,19(2):127-134.
[13] 鄢家俊,白史且,张新全,等.青藏高原老芒麦种质基于 SRAP标记的遗传多样性研究[J].草业学报,2010,19(1):173-183.
[14] Dong P., Wei Y. M., Chen G. Y., Li W., Wang J. R., Eviatar Nevo, Zheng Y. L. Sequence-related amplified polymorphism (SRAP) of wild emmer wheat (Triticum dicoccoides) in Israel and its ecological association[J]. Biochemical Systematics and Ecology, 2010, 38: 1-11.
[15] 许绍斌,陶玉,杨昭庆,等.简单快速的 DNA银染和胶保存方法[J].遗传,2002,24(3):335-336.
[16] 薛丹丹,郑轶琦,王志勇,等.结缕草属植物 SRAP-PCR体系的建立和优化[J].草业学报,2008,17(6):93-101.
[17] 郑轶琦,王志勇,郭海林,等.正交设计优化假俭草 SRAP-PCR反应体系及引物筛选[J].草业学报,2008,17(4):110-117.
[18] 王志勇,袁学军,刘建秀,等.狗牙根 SRAP-PCR反应体系优化及引物筛选[J].草业学报,2008,17(3):79-85.
[19] 何正文,刘运生,陈立华,等.正交设计直观分析法优化 PCR 条件[J].湖南医科大学学报,1998,23(4):403-404

Establishment and Optimization of SRAP-PCR Reaction System for Elymus breviaristatus

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