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园艺学报 ›› 2014, Vol. 41 ›› Issue (7): 1467-1475.

• 研究报告 • 上一篇    下一篇

抗菌肽基因Gnk2-1经花粉管通道法导入西瓜的初步研究

许珺然,张 显*,张 勇,马建祥   

  1. 西北农林科技大学园艺学院,陕西杨凌 712100
  • 出版日期:2014-07-25 发布日期:2014-07-25
  • 基金资助:

    国家自然科学基金项目(31372086)

Introduction of Antimicrobial Peptide Gene Gnk2-1 into Watermelon via
Pollen-tube Pathway

XU Jun-ran,ZHANG Xian*,ZHANG Yong,and MA Jian-xiang   

  1. College of Horticulture,Northwest A & F University,Yangling,Shaanxi 712100,China
  • Online:2014-07-25 Published:2014-07-25

摘要: 通过花粉管通道介导银杏抗菌肽基因Gnk2-1 于西瓜自交系‘04-1-2’中,以浓度为100、200、
300 和400 ng · μL-1 的DNA 为供体,分别于授粉后24、27、30 和33 h 导入,对子代进行PCR 检测。结
果表明:T0 代坐果率和单瓜结籽数随授粉后处理时间的增加而升高,但在各个处理时间下均显著低于对
照;外源DNA 的导入对授粉后27 h 处理的T1 代种子发芽率和授粉后24 h 处理的T1 代种子出苗率均造成
显著降低;不同DNA 浓度转化的T1 代种子的发芽率和出苗率与对照相比差异不显著;对1 280 株T1 幼
苗进行PCR 检测,得到32 株阳性转化植株,总体转化率为2.5%,最佳转化时间为授粉后24 ~ 27 h,最佳
DNA 转化浓度为100 ~ 200 ng · μL-1;PCR 阳性植株抗病鉴定表明转基因植株对西瓜枯萎病的抗性有所增强。

关键词: 西瓜, 花粉管通道法, 分子检测, 抗菌肽

Abstract: In order to study the feasibility of transferring exogenous DNA into watermelon by
pollen-tube pathway,the antimicrobial peptide gene Gnk2-1 was introduced into watermelon accession
‘04-1-2’by pollen-tube pathway with different concentration(100,200,300 and 400 ng · μL-1)at different
time(24 h,27 h,30 h,33 h)after self-pollination. Seeds from treated fruits were harvested and tested by
PCR. The results were as follows:The fruit setting rate and average number of seeds of T0 generation
increased with the increment of treatment time after self-pollination,though the two figures both showed a
significant decrease under all treatments compared with control. The germination rate of T1 seeds treated
27 h after self-pollination and the seedling emergence rate of T1 seeds treated 24 h after self-pollination
decreased significantly. Compared with control,there was no distinct difference illustrated in the rate of
germination and seedling emergence of T1 seeds between different concentration treatments. The PCR
results of 1 280 tested T1 seedlings showed 32 positive plants and the total transformation rate was 2.5%.
The optimum transformation time was 24–27 h after self-pollination and the optimum transformation concentration were 100–200 ng · μL-1. The positive plants were inoculated with Fusarium and showed a
stronger resistance compared with non transgenic plants.

Key words: watermelon, pollen-tube pathway, molecular detection, antimicrobial peptide

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