中国生物防治学报
中國生物防治學報
중국생물방치학보
CHINESE JOURNAL OF BIOLOGICAL CONTROL
2013年
4期
555-560
,共6页
詹晓奂%申屠旭萍%刘卫平%马正%俞晓平
詹曉奐%申屠旭萍%劉衛平%馬正%俞曉平
첨효환%신도욱평%류위평%마정%유효평
脐孢木霉菌%基因tri5%木霉素%基因克隆%实时荧光定量PCR
臍孢木黴菌%基因tri5%木黴素%基因剋隆%實時熒光定量PCR
제포목매균%기인tri5%목매소%기인극륭%실시형광정량PCR
Trichoderma brevicompactum%gene tri5%trichodermin%gene cloning%real-time quantitative PCR
脐孢木霉菌Trichoderma brevicompactum能合成木霉素等单端孢霉烯类化合物，而基因tri5编码的单端孢霉二烯合酶是合成途径中的关键酶，它催化反式法呢基焦磷酸（tFPP）合成单端孢霉二烯。为分析基因tri5与木霉素合成的关系，本研究从脐孢木霉菌0248中克隆了基因tri5片段，并运用实时荧光定量PCR技术分析了基因tri5在不产木霉素培养基和产木霉素培养基中的差异表达。在相同培养时间下，基因tri5在不同产木霉素状态间的表达量差异显著；在培养96 h时基因tri5在2种培养基中均达到最大值，基因tri5在产木霉素状态下的表达量是不产木霉素状态下的107.18倍。通过检测产木霉素培养基中基因tri5表达量与木霉素含量的变化，结果显示木霉素的含量随着基因tri5表达量的增加而不断增加，在培养96 h时达到最大值118.71 mg?L?1，推测基因tri5表达量与木霉素合成相关。该研究为基因tri5功能的进一步研究和对脐孢木霉菌0248进行基因工程菌改造奠定了理论基础。
臍孢木黴菌Trichoderma brevicompactum能閤成木黴素等單耑孢黴烯類化閤物，而基因tri5編碼的單耑孢黴二烯閤酶是閤成途徑中的關鍵酶，它催化反式法呢基焦燐痠（tFPP）閤成單耑孢黴二烯。為分析基因tri5與木黴素閤成的關繫，本研究從臍孢木黴菌0248中剋隆瞭基因tri5片段，併運用實時熒光定量PCR技術分析瞭基因tri5在不產木黴素培養基和產木黴素培養基中的差異錶達。在相同培養時間下，基因tri5在不同產木黴素狀態間的錶達量差異顯著；在培養96 h時基因tri5在2種培養基中均達到最大值，基因tri5在產木黴素狀態下的錶達量是不產木黴素狀態下的107.18倍。通過檢測產木黴素培養基中基因tri5錶達量與木黴素含量的變化，結果顯示木黴素的含量隨著基因tri5錶達量的增加而不斷增加，在培養96 h時達到最大值118.71 mg?L?1，推測基因tri5錶達量與木黴素閤成相關。該研究為基因tri5功能的進一步研究和對臍孢木黴菌0248進行基因工程菌改造奠定瞭理論基礎。
제포목매균Trichoderma brevicompactum능합성목매소등단단포매희류화합물，이기인tri5편마적단단포매이희합매시합성도경중적관건매，타최화반식법니기초린산（tFPP）합성단단포매이희。위분석기인tri5여목매소합성적관계，본연구종제포목매균0248중극륭료기인tri5편단，병운용실시형광정량PCR기술분석료기인tri5재불산목매소배양기화산목매소배양기중적차이표체。재상동배양시간하，기인tri5재불동산목매소상태간적표체량차이현저；재배양96 h시기인tri5재2충배양기중균체도최대치，기인tri5재산목매소상태하적표체량시불산목매소상태하적107.18배。통과검측산목매소배양기중기인tri5표체량여목매소함량적변화，결과현시목매소적함량수착기인tri5표체량적증가이불단증가，재배양96 h시체도최대치118.71 mg?L?1，추측기인tri5표체량여목매소합성상관。해연구위기인tri5공능적진일보연구화대제포목매균0248진행기인공정균개조전정료이론기출。
The filamentous fungus Trichoderma brevicompactum was proved to produce the trichodermin, one of trichothecenes. Trichodiene synthase, which is encoded by the gene tri5, catalyses the conversion of farnesyl pyrophosphate to trichodiene in Fusarium spp. and is the key enzyme in trichothecenes biosynthesis. However, the biosynthetic pathway for trichothecene metabolism is keeping unclear in T. brevicompactum. In order to understand the functions of gene tri5 cloned from T. brevicompactum 0248, which was isolated from garlic Allium sativum in our previous work, the relative expression levels of this gene in two different cultural media were evaluated by the real-time quantitative PCR. The results showed that significantly different expression of gene tri5 occurred in trichodermin producing and non-producing media. The relative expression level of gene tri5 in the trichodermin producing medium was 107.18 times higher than that in non-producing medium after 96 hours incubation, and the production of trichodermin was enhanced with the increasing expression level of gene tri5, which reached to the maximum of 118.71mg?L?1. The results indicated that tri5 gene may have a close relationship with trichodermin metabolism in T. brevicompactum.
