禾谷镰刀菌Homeobox转录因子FgHtf1敲除突变体基因差异表达分析  

Differential analysis of gene expression in the Homeobox transcription factor FgHtf1 deletion mutant of Fusarium graminearum

作　　者：张红红[1] 郑文辉[1] 骆琼[1] 乔向威[1] 陈阿海[1] 余文英[1] 王宗华[1] 

机构地区：[1]福建农林大学生物农药与化学生物学教育部重点实验室,福州350002

出　　处：《中国科技论文》2015年第12期1392-1397,共6页China Sciencepaper

基　　金：高等学校博士学科点专项科研基金资助项目(20113515130001)

摘　　要：禾谷镰刀菌是引起小麦赤霉病的主要病原真菌。分生孢子在侵染过程中起关键作用。先前研究表明同源异型盒(Homeobox)转录因子FgHtf1特异调控分生孢子的产生。为明确其调控产孢的分子机制,本研究采用高通量RNA测序技术(RNA-Seq),对ΔFghtf1突变体菌株和野生型PH-1菌株进行测序,结果显示其中591个基因转录上调,549个基因下调。对这些基因进行功能预测并聚类分析发现:上下调基因中分别有约30%和22%参与新陈代谢和能量转运。运用荧光定量PCR,检测10个基因的表达,验证了RNA-Seq的结果是可靠的。运用同源重组方法敲除明显下调的基因FGSG_10565和FGSG_07629,表型分析结果显示,相比野生型,ΔFGSG_10565和ΔFGSG_07629的生长速率和产孢量均不变,说明两个基因的表达受FgHtf1调控,但功能上不直接参与分生孢子的形成。Fusarium graminearum is the major causal agent of Fusarium head blight （FHB）on wheat.The conidia play a key role in the disease cycle.Our previous work has shown that the homeobox transcription factor FgHtf1 specifically controls the produc-tion of spores in F.graminearum.In order to further investigate the molecular mechanisms of FgHtf1 in regulating conidiation, the high-throughput RNA-sequencing technology has been developed to analyze the differential expressions between wild-type strain PH-1 andΔFghtf1 mutant strain at whole genome level.The results showed that 591 genes were up-regulated and 549 genes were down-regulated inΔFghtf1 compared with the wild-type.According to the functional annotation,about 30% up-reg-ulated genes are involved in metabolism and energy.6.7% and 3% are implicated in cell cycle and cellular differentiation path-way,respectively.Among the down-regulated genes,22%,3.8% and 2.2% are involved in metabolism,cell cycle and cellular differentiation,respectively.Further quantitative PCR of 10 genes was conducted to validate that the gene expression pattern was consistent with the change patterns of RNA-Seq transcriptional profiling.In order to understand the biological function of these genes,we knock out two genes FGSG 10565 and FGSG 07629 which were significantly down-regulated in theΔFghtf1 .The re-sults showed that there is no significant difference in conidiation and growth between the mutants and the wild type,suggesting that although the expressions of these two genes are down-regulated by FgHtf1 but are not involved in the conidiogenesis directly.

关 键 词：禾谷镰刀菌 产孢 同源异型盒基因 RNA-Seq 基因差异表达 

分 类 号：S432.44[农业科学—植物病理学]