华北农学报 ›› 2011, Vol. 26 ›› Issue (2): 124-127. doi: 10.7668/hbnxb.2011.02.027

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南瓜矮生性研究的AFLP体系的建立

武峻新1,2, 畅志坚1,3   

  1. 1. 山西大学生命科学学院, 山西太原, 030006;
    2. 山西省农业科学院蔬菜研究所, 山西太原, 030031;
    3. 山西省农业科学院作物科学研究所, 山西太原, 030031
  • 收稿日期:2011-01-10 出版日期:2011-04-28
  • 作者简介:武峻新(1966-),男,山西沁县人,在读博士,主要从事南瓜属的分子遗传育种和示范研究.
  • 基金资助:
    山西省回国留学人员科研项目(2004087;2010112)

A Construction of AFLP Analysis System for Dwarf Cucurbita moschata Research

WU Jun-xin1,2, CHANG Zhi-jian1,3   

  1. 1. College of Life Science, Shanxi University, Taiyuan 030006, China;
    2. Institute of Vegetables, Shanxi Academy of Agriculture Sciences, Taiyuan 030031, China;
    3. Institute of Crop Sciences, Shanxi Academy of Agriculture Sciences, Taiyuan 030032, China
  • Received:2011-01-10 Published:2011-04-28

摘要: 以南瓜矮生和蔓生的自交系为材料,通过对影响南瓜AFLP反应体系中的关键因素(如DNA的提取方法、酶切和连接反应时间、预扩增产物稀释倍数和银染检测方法等)进行研究,结果表明,采用加入PVP和β-巯基乙醇后的改良CTAB提取法获得的南瓜嫩叶基因组DNA质量较好;5.5 h可以满足酶切和连接反应;酶切连接产物稀释5倍,预扩增产物稀释30倍,可以满足试验要求.应用该体系,对96对引物进行筛选,在南瓜矮生和蔓生自交系间获得的引物扩增出了重复性好、清晰的南瓜矮生和蔓生的自交系多态性条带.为进一步进行南瓜矮生基因定位奠定了良好的基础.

关键词: 南瓜, 矮生, AFLP体系

Abstract: Based on dwarf and vine inbred lines,key factors of AFLP analysis system(such as genomic DNA extraction,time requirement of digestion and ligation reactions,dilution of pre-amplification and silver staining method et a1)were studied.The result showed:the best quality genomic DNA were adding PVP and B-mercaptoethanol modified CTAB method;5.5 h can be meet digestion and ligation reactions;Diluted 5 times for products of digestion and ligation reactions and diluted 30 times for products of pre-amplification can be meet experimental conditions.96 primers screened,reproduce,clear polymorphic bands were obtained between dwarf and vine inbred lines.It laid good foundation to tropic pumpkin dwarf vine mapping.

Key words: Tropic pumpkin, Dwarf, AFLP analysis system

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引用本文

武峻新, 畅志坚. 南瓜矮生性研究的AFLP体系的建立[J]. 华北农学报, 2011, 26(2): 124-127. doi: 10.7668/hbnxb.2011.02.027.

WU Jun-xin, CHANG Zhi-jian. A Construction of AFLP Analysis System for Dwarf Cucurbita moschata Research[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(2): 124-127. doi: 10.7668/hbnxb.2011.02.027.

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