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中国农学通报 ›› 2016, Vol. 32 ›› Issue (3): 100-107.doi: 10.11924/j.issn.1000-6850.casb15020112

所属专题: 生物技术

• 生物技术科学 • 上一篇    下一篇

尖孢镰刀菌古巴专化型SIX7基因分析

杨腊英,郭立佳,刘磊,梁昌聪,王国芬,汪军,陈平亚,黄俊生   

  1. 中国热带农业科学院环境与植物保护研究所/农业部热带农林有害生物入侵检测与控制重点实验室,中国热带农业科学院环境与植物保护研究所/农业部热带农林有害生物入侵检测与控制重点实验室,中国热带农业科学院环境与植物保护研究所/农业部热带农林有害生物入侵检测与控制重点实验室,中国热带农业科学院环境与植物保护研究所/农业部热带农林有害生物入侵检测与控制重点实验室,中国热带农业科学院环境与植物保护研究所/农业部热带农林有害生物入侵检测与控制重点实验室,中国热带农业科学院环境与植物保护研究所/农业部热带农林有害生物入侵检测与控制重点实验室,中国热带农业科学院环境与植物保护研究所/农业部热带农林有害生物入侵检测与控制重点实验室,中国热带农业科学院环境与植物保护研究所/农业部热带农林有害生物入侵检测与控制重点实验室
  • 收稿日期:2015-02-28 修回日期:2015-09-29 接受日期:2015-11-24 出版日期:2016-01-29 发布日期:2016-01-29
  • 通讯作者: 黄俊生
  • 基金资助:
    国家自然科学基金项目“尖孢镰刀菌古巴专化型4 号生理小种特有SIX基因的功能分析”(31301629)。

Analysis of SIX7 Gene of Fusarium oxysporum f. sp. cubense

  • Received:2015-02-28 Revised:2015-09-29 Accepted:2015-11-24 Online:2016-01-29 Published:2016-01-29

摘要: SIX蛋白编码基因最早由尖孢镰刀菌番茄专化型入侵的番茄木质部蛋白质组中发现,且成功用于区分尖孢镰刀菌番茄专化型的3个生理小种。一些SIX编码基因亦在引起香蕉枯萎病的Fusarium.oxysporum f. sp. cubense(简称Foc)中有报道,为区分Foc不同生理小种或亚型,比较分析了Foc中的SIX7基因序列及其特异性。以现有的SIX7基因序列设计合成引物,通过PCR扩增并测序,比较分析国内不同地理区域及来源于澳大利亚与南非的Foc1、Foc2、Foc3、Foc4共56株菌株中的SIX7基因,并以8个以上其他专化型或其他种或属病原菌共39株菌株分析了SIX7基因序列的特异性。研究发现所设计合成的SIX7基因引物序列仅能从供试的亚热带4号生理小种(ST4)中扩增出663 bp的目的条带。亚热带4号生理小种(ST4)中仅有的SIX7基因序列结合Foc4特异性引物Foc-1/Foc-2可快速区分Foc4亚型,提供了快速区分香蕉枯萎病病样内的Foc4亚型的分子鉴定手段,可指导蕉农及时采取有效防控香蕉枯萎病的措施。

关键词: 羊肚菌, 羊肚菌, 温度, pH, 碳源, 氮源, 菌落干重, 生长速度

Abstract: SIX genes were first detected in the xylem sap proteome of F.oxysporum f. sp. lycopersici (Fol)-infected tomato plants, and had been successfully used to differentiate the three races of F.oxysporum f. sp. lycopersici. Some SIX-genes have been reported in Fusarium oxysporum f. sp. cubense (Foc) which is the ca usal agent of fusarium wilt of banana. To differentiate the races or subtypes of Foc, the SIX7 gene was analyzed.Based on the known SIX7 gene sequences, primers were designed and synthesized. And PCR and sequencing approach were used to detect the SIX7 in a collection of 56 Foc isolates from different domestic banana growing areas and from Australia or South Africa. And also the specific of SIX7 gene was screened with totally 39 isolates including eight other F.oxysporum formae speciales and non- pathogen F.oxysporum and F. monilifarme and F.solani and some other genus pathogens of tropical crops. The study found that the designed and synthesized SIX7 gene primers were only amplified to a 663 bp target band from the tested subtropical species of Foc race 4 (ST4). Together with the known Foc4 specific primers Foc-1/Foc-2, Foc-SIX7 gene could be allowed to differentiate subtropical race 4 from tropical race 4 isolates. It would provide a quick molecular identification method to distinguish Foc4 subtypes in disease affected banana plants, and would guide banana farmers to take timely and effective measures to prevent and control banana Fusarium wilt disease.

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