TAI Lian-mei,ZUO Yu-hu,ZHANG Ya-ling,et al.Optimization of RAPD-PCR Reaction System on Alternaria solani Causing Potato Early Blight[J].HEILONGJIANG AGRICULTURAL SCIENCES,2016,(05):4-7.[doi:10.11942/j.issn1002-2767.2016.05.0004]
马铃薯早疫病菌RAPD-PCR反应体系优化
- Title:
- Optimization of RAPD-PCR Reaction System on Alternaria solani Causing Potato Early Blight
- 文章编号:
- 1002-2767（2016）05-0004-04
- 分类号:
- S432.44
- 文献标志码:
- A
- 摘要:
- 为了应用RAPD分子标记技术分析马铃薯早疫病菌遗传多样性，采用单因素水平对影响RAPD反应的Mg2+，dNTPs、TaqDNA聚合酶、引物浓度及引物退火温度等条件进行了优化。结果表明：建立了适宜于早疫病菌的RAPD最佳反应体系，25 μL的反应体系中，Mg2+ 2.5 mmo1·L-1，dNTPs 0.20 mmol·L-1，Taq DNA聚合酶1 U，引物0.40 μmol·L-1。
- Abstract:
- In order to analyze the population genetic diversity of Alternaria solani causing potato early blight by RAPD,single factor design was used to optimize RAPD -PCR reaction on Mg2+，dNTPs,Taq DNA polymerise,primer and annealing temperature.The optimized RAPD system consisted of 2.5 mmo1?L-1 Mg2+，0.20 mmol?L-1 dNTPs,1 U Taq DNA polymerise，0.40 μmol?L-1 primer.
参考文献/References:
[1] 台莲梅，郑寰宇，左豫虎，等 . 黑龙江省马铃薯早疫病菌生物学特性研究 [J]. 植物保护， 2012,38(1):85-89. [2]郑寰宇，马力，左豫虎，等.马铃薯早疫病菌分生孢子萌发条件的研究[J].植物保护，2010,36(6):91-95. [3]左豫虎，史洁， 韩青梅，等.茄链格孢菌侵染马铃薯叶片过程的细胞学观察[J].中国蔬菜，2011(14):35-40. [4]Shahbazi H,Aminian H,Sahebani N,et al.Biochemical evaluation of resistance responses of potato to different isolates of Alteraria solani[J].Phytopathology,2010,100(5):454-459. [5]台莲梅，梁伟伶，左豫虎，等.马铃薯不同品种感染早疫病菌后防御酶活性变化[J].植物生理学通讯，2010,46(11):1147-1150. [6]台莲梅，张宗敏，张亚玲，等.混配杀菌剂对马铃薯早疫病的防效及对产量的影响[J].江苏农业科学，2014,42(9):110-112. [7]Weir T L，Huff D R，Christ B J,et al．RAPD-PCR analysis of genetic variation among isolates of Alternaria solani and Alternaria alternata from potato and tomato[J]．Mycologia，1998,90(5):813-821. [8]Kumar V,Haldar S,Pandey K K,et al.Cultural morphological,pathogenic and molecular variability amongst tomato isolates of Alternaria solani in India [J].World Journal of Microbiology and Biotechnology,2008,24:1003-1009. [9]桂秀梅，张金林，董金皋，等.玉米大斑病菌RAPD分析最佳反应体系的建立[J].河北农业大学学报，2005，28（1）：56-58.〖ZK)〗 [10]陈瑞辉，周晓云，王克荣.大丽轮枝菌RAPD反应组分的优化与应用[J].农业生物技术学报，2002,10（3）：291-294. [11]郭小敏，段会军，温小杰，等.西瓜枯萎病菌最优RAPD体系的建立[J].河北农业大学学报，2004,27（3）:64-67.
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备注/Memo
收稿日期： 2016-04-04 基金项目：黑龙江省教育厅科学技术研究资助项目（12541587）