背景:禾谷镰刀菌(Fusarium graminearum)是小麦及其它粮食作物小麦赤霉病(Fusarium head blight)的主要病原菌。目前,分子分析在研究病原菌的致病机理中已有广泛应用。MGV1菌株是禾谷镰刀菌MAPK信号通路基因MGV1缺失后的突变体,MGV1敲...背景:禾谷镰刀菌(Fusarium graminearum)是小麦及其它粮食作物小麦赤霉病(Fusarium head blight)的主要病原菌。目前,分子分析在研究病原菌的致病机理中已有广泛应用。MGV1菌株是禾谷镰刀菌MAPK信号通路基因MGV1缺失后的突变体,MGV1敲除突变体的差异基因表达(DGE)分析显示,MGV1基因缺失后,MDT1基因表达量显著下调。GO注释预测是参与ATP结合蛋白二聚作用的活性因子。推测该基因涉及MGV1通路。目的:探讨MDT1基因(MGV1dependent transcripts)的生物学功能。方法:应用Split-Marker技术构建基因敲除盒,并通过常规方法检测缺失突变体的表型和致病性。结果:与禾谷镰刀菌PH-1相比,MDT1基因缺失突变体的分生孢子量和子囊壳量显著下降,表明该基因对于无性繁殖和有性生殖能力至关重要。同时发现,在固体培养基上,缺失突变体的营养生长能力大大下降。该缺失突变体对细胞壁降解酶也很敏感,液体培养基中32℃下,菌丝顶端膨大,并有菌丝断裂,表明该基因与禾谷镰刀菌的细胞壁整合有关。互补实验证实了敲除突变体表型改变确实是由于MDT1基因缺失所致。结论:MDT1基因与禾谷镰刀菌的分生孢子和营养生长能力有关。展开更多
Fusarium wilt of banana, caused by Fusarium oxysporum f. sp. cubense tropic race 4(Foc TR4), is a typical vascular and soil-borne disease which has significantly threatened the sustainable development of banana indust...Fusarium wilt of banana, caused by Fusarium oxysporum f. sp. cubense tropic race 4(Foc TR4), is a typical vascular and soil-borne disease which has significantly threatened the sustainable development of banana industry. In order to reveal the infection process and pathogenesis of Foc TR4, the young mycelia(66.7 mg/ml) of wild-type strain of Foc TR4(WT-Foc TR4) cultured for 18-20 h were lysed with enzyme mixture for protoplast formation, which consisted of 25 mg/ml driselase, 0.4 mg/ml chitinase, 15 mg/ml lysing enzyme and 1.2 mol/L potassium chloride. The resulted protoplasts of 2×10~7 cells/ml were used to test the efficiency of transformation mediated by polyethylene glycol, and up to 9 transformants per microgram of DNA were obtained. AmCyan, RFP and YFP genes were stably transferred into the WT-Foc TR4, separately, using the protoplast transformation system. The gene FoOCH1 encoding α-1, 6-mannosyltransferase in the WT-Foc TR4 was knocked out using the split-marker recombination technology. The genetic transformation and gene knockout system in this pathogen lays a foundation for the study of functional genomics and plant-pathogen interactions.展开更多
在Government and Binding Theory框架下讨论汉语中表示强调的“是”字句的词性及其所处位置.在树形图的描述上,运用LuigiRizzi关于splitComp假说中的理论,印证了“是”是焦点标记词的观点;同时,根据splitComp假说发展“是”是焦点标记...在Government and Binding Theory框架下讨论汉语中表示强调的“是”字句的词性及其所处位置.在树形图的描述上,运用LuigiRizzi关于splitComp假说中的理论,印证了“是”是焦点标记词的观点;同时,根据splitComp假说发展“是”是焦点标记词的观点,认为“是”是FocusP的中心语,位于“是”之前的成分是从TP生成然后显性移位到TopP位置上,而被强调成分隐性移位到[Spec,FocusP]的位置获得解释.展开更多
基金National Natural Science Foundation of China(30840014)SRF for ROCS,SEM(Scientific Research Foundation for the Returned Overseas Chinese Scholars,State Education Ministry)Natural Science Foundation of Shandong Province,China(ZR2010CM011)~~
文摘背景:禾谷镰刀菌(Fusarium graminearum)是小麦及其它粮食作物小麦赤霉病(Fusarium head blight)的主要病原菌。目前,分子分析在研究病原菌的致病机理中已有广泛应用。MGV1菌株是禾谷镰刀菌MAPK信号通路基因MGV1缺失后的突变体,MGV1敲除突变体的差异基因表达(DGE)分析显示,MGV1基因缺失后,MDT1基因表达量显著下调。GO注释预测是参与ATP结合蛋白二聚作用的活性因子。推测该基因涉及MGV1通路。目的:探讨MDT1基因(MGV1dependent transcripts)的生物学功能。方法:应用Split-Marker技术构建基因敲除盒,并通过常规方法检测缺失突变体的表型和致病性。结果:与禾谷镰刀菌PH-1相比,MDT1基因缺失突变体的分生孢子量和子囊壳量显著下降,表明该基因对于无性繁殖和有性生殖能力至关重要。同时发现,在固体培养基上,缺失突变体的营养生长能力大大下降。该缺失突变体对细胞壁降解酶也很敏感,液体培养基中32℃下,菌丝顶端膨大,并有菌丝断裂,表明该基因与禾谷镰刀菌的细胞壁整合有关。互补实验证实了敲除突变体表型改变确实是由于MDT1基因缺失所致。结论:MDT1基因与禾谷镰刀菌的分生孢子和营养生长能力有关。
基金Supported by Yunnan Science and Technology Innovation Talent Program(2015HA034)National Natural Science Foundation of China(NSFC31560505)
文摘Fusarium wilt of banana, caused by Fusarium oxysporum f. sp. cubense tropic race 4(Foc TR4), is a typical vascular and soil-borne disease which has significantly threatened the sustainable development of banana industry. In order to reveal the infection process and pathogenesis of Foc TR4, the young mycelia(66.7 mg/ml) of wild-type strain of Foc TR4(WT-Foc TR4) cultured for 18-20 h were lysed with enzyme mixture for protoplast formation, which consisted of 25 mg/ml driselase, 0.4 mg/ml chitinase, 15 mg/ml lysing enzyme and 1.2 mol/L potassium chloride. The resulted protoplasts of 2×10~7 cells/ml were used to test the efficiency of transformation mediated by polyethylene glycol, and up to 9 transformants per microgram of DNA were obtained. AmCyan, RFP and YFP genes were stably transferred into the WT-Foc TR4, separately, using the protoplast transformation system. The gene FoOCH1 encoding α-1, 6-mannosyltransferase in the WT-Foc TR4 was knocked out using the split-marker recombination technology. The genetic transformation and gene knockout system in this pathogen lays a foundation for the study of functional genomics and plant-pathogen interactions.
文摘在Government and Binding Theory框架下讨论汉语中表示强调的“是”字句的词性及其所处位置.在树形图的描述上,运用LuigiRizzi关于splitComp假说中的理论,印证了“是”是焦点标记词的观点;同时,根据splitComp假说发展“是”是焦点标记词的观点,认为“是”是FocusP的中心语,位于“是”之前的成分是从TP生成然后显性移位到TopP位置上,而被强调成分隐性移位到[Spec,FocusP]的位置获得解释.
