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禾谷镰刀菌中FgPDE1基因的敲除及其功能的研究 被引量:5
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作者 常玉梅 侯占铭 《中国生物工程杂志》 CAS CSCD 北大核心 2015年第8期59-67,共9页
目的:旨在敲除禾谷镰刀菌Fusarium graminearum Fg PDE1基因,确定其缺失突变体表型,从而分析该基因的生物学功能。方法:应用Split-marker技术构建含有潮霉素基因敲除盒,通过PEG介导原生质体转化,PCR筛查抗潮霉素转化子以获得缺失突变体... 目的:旨在敲除禾谷镰刀菌Fusarium graminearum Fg PDE1基因,确定其缺失突变体表型,从而分析该基因的生物学功能。方法:应用Split-marker技术构建含有潮霉素基因敲除盒,通过PEG介导原生质体转化,PCR筛查抗潮霉素转化子以获得缺失突变体ΔFg PDE1,根据突变体表型变化及致病性的检测对Fg PDE1基因的功能进行分析。结果:采用Split-marker技术,成功构建了Fg PDE1基因敲除盒;PEG介导转化禾谷镰刀菌原生质体后成功获得转化子。经PCR筛查,得到3个PCR确认的敲除突变体;表型观察发现,ΔFg PDE1菌落的外型及菌落生长速度与野生型没有明显差异。孢子侵染西红柿果实实验证明:以西红柿为侵染宿主,相对于野生型,突变体致病性没有明显减弱;但突变体分生孢子产量显著下降。结论:Fg PDE1基因可能与禾谷镰刀菌分生孢子的形成有关。 展开更多
关键词 禾谷镰刀菌 fgpde1 基因敲除 Split-Marker
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FpPDE1 function of Fsarium pseudograminearum on pathogenesis in wheat 被引量:1
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作者 WANG Li-min ZHANG Yi-fan +6 位作者 DU Zhen-lin KANG Rui-jiao CHEN Lin-lin XING Xiao-ping YUAN Hong-xia DING Sheng-li LI Hong-lian 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2017年第11期2504-2512,共9页
Wheat crown rot caused by Fusarium spp. is a common disease worldwide. Both Fusarium pseudograminearum and Fusarium graminearum infect wheat crown and produce mycotoxin leading to grain loss due to white head. F. pseu... Wheat crown rot caused by Fusarium spp. is a common disease worldwide. Both Fusarium pseudograminearum and Fusarium graminearum infect wheat crown and produce mycotoxin leading to grain loss due to white head. F. pseudograminearum (Fp) was reported in wheat from Henan Province of China a couple of years ago. The wheat crown rot (CR) caused by this new pathogen is as an emerging severe disease of wheat, which has recently expanded to several provinces in China and is, therefore, under rapid investigation. Colonization of wheat tissue by Fp is accomplished though the formation of a septated foot-shaped appressoria and generation of a penetration peg to break through the internal cells of leaf sheath. The molecular mechanism by which Fp regulates the pathogenesis on wheat host is unclear. Here, we report FpPDE1, a P-type ATPase-encoding predicted PDE1 orthologue gene of Magnaporthe oryzae, belonging to the DRS2 subfamily of aminophospholipid translocases. The gene deletion of FpPDE1 with the split-marker approach did not obviously affect hyphae growth and conidiation, but led to an attenuated virulence on wheat base stem and root. Our finding indicates that the putative aminophospholipid translocases is not essential for the infectious hyphae development in Fp. 展开更多
关键词 Fusarium pseudograminearum gene knockout fgpde1 PATHOGENICITY
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