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    李佳欣, 刘绍梅, 任燕娜, 周祥山, 蔡孟浩, 张元兴. 海洋灰绿曲霉AgveA基因响应盐度胁迫的功能分析[J]. 华东理工大学学报(自然科学版), 2018, (5): 650-660. DOI: 10.14135/j.cnki.1006-3080.20170809003
    引用本文: 李佳欣, 刘绍梅, 任燕娜, 周祥山, 蔡孟浩, 张元兴. 海洋灰绿曲霉AgveA基因响应盐度胁迫的功能分析[J]. 华东理工大学学报(自然科学版), 2018, (5): 650-660. DOI: 10.14135/j.cnki.1006-3080.20170809003
    LI Jia-xin, LIU Shao-mei, REN Yan-na, ZHOU Xiang-shan, CAI Meng-hao, ZHANG Yuan-xing. Function Analysis of AgveA Responsed to Salt-Stress in Marine-Derived Aspergillus glaucus[J]. Journal of East China University of Science and Technology, 2018, (5): 650-660. DOI: 10.14135/j.cnki.1006-3080.20170809003
    Citation: LI Jia-xin, LIU Shao-mei, REN Yan-na, ZHOU Xiang-shan, CAI Meng-hao, ZHANG Yuan-xing. Function Analysis of AgveA Responsed to Salt-Stress in Marine-Derived Aspergillus glaucus[J]. Journal of East China University of Science and Technology, 2018, (5): 650-660. DOI: 10.14135/j.cnki.1006-3080.20170809003

    海洋灰绿曲霉AgveA基因响应盐度胁迫的功能分析

    Function Analysis of AgveA Responsed to Salt-Stress in Marine-Derived Aspergillus glaucus

    • 摘要: 通过染色体步移方法克隆了基因AgveA、AgfphA、AglreAAglreB的上下游侧翼序列,并通过逆转录确定了各个基因的内含子序列。通过实时荧光定量PCR发现,基因AgveA转录受盐度胁迫的作用下调较明显,而AgfphA、AglreAAglreB上调较明显。通过构建打靶质粒以敲除AgveA,最终成功构建了基因缺失株ΔAgveA,并从菌丝形态、菌落形态、菌丝生长速率和细胞发育表型4个方面进行了基因功能分析。结果表明,菌株ΔAgveA的菌丝形态并未产生明显变化,而菌落的边缘存在残缺,菌丝生长速率在生长后期加快;细胞发育模式产生明显改变,AgveA基因的缺失导致海洋灰绿曲霉在高盐度胁迫条件下子囊果的合成完全受阻,而在低盐度胁迫条件下产生分生孢子的同时也形成了极少量的子囊果。

       

      Abstract: Marine environment with high pressure, high salt, hypoxia and low light intensity is quite different from that of the land, which allows that marine microorganisms often have special physiology, growth and production behaviors. Aspergillus glaucus HB1-19 is a typical marine-derived fungus that can tolerate high salt stress. The developmental pattern of filamentous fungi usually switches between an asexual and sexual mode, correlating with secondary metabolism and switching in response to numerous environmental signals including light, ambient pH, nutrient availability, temperature, and osmolality. Our previous results showed that salt stress inhibited its sexual development but promoted asexual one at the same time. Filamentous fungi easily response to light signal and regulate cell development. A global regulation factor VeA and its protein complex related proteins of FphA, LreA and LreB usually function in this physiological process. This work aims to investigate whether or not the VeA protein responses to salt stress signal and regulates the cell development transitions. Genome walking is firstly used to clone the flanking gene sequences of AgveA, AgfphA, AglreA and AglreB, and then introns of these genes are confirmed by reverse transcription. The results of qRT-PCR show that the transcription of AgveA is obviously up-regulated by salt stress, while AgfphA, AglreA and AglreB are down-regulated. Afterwards, AgveA is knocked out and a gene deficient strain of ΔAgveA is constructed to further study the functions of AgveA in A. glaucus. The morphology of mycelial morpholo and colony, mycelial growth rate and cell development phonotypes are evaluated. The mycelial morphology is little changed while some edge defects are found in colonies. Mycelial growth rate accelerates in the late stages of development. Deletion of AgveA results in the blockage in production of cleistothecia under high salt stress, while it produces many conidiosprores and simultaneously a spot of cleistothecia under low salt stress.

       

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