福建农业学报
福建農業學報
복건농업학보
FUJIAN JOURNAL OF AGRICULTURAL SCIENCES
2014年
3期
197-203
,共7页
毛小辉%魏毅东%张建福%谢华安
毛小輝%魏毅東%張建福%謝華安
모소휘%위의동%장건복%사화안
云引%近等基因系%谷胱甘肽S-转移酶%序列分析
雲引%近等基因繫%穀胱甘肽S-轉移酶%序列分析
운인%근등기인계%곡광감태S-전이매%서렬분석
‘Yunyin’%near-isogenic lines%glutathione S-transferase%sequence analysis
在对粳稻‘云引’受稻瘟病菌诱导的近等基因系蛋白质组学分析的基础上,通过实时定量PCR等方法,以普感稻瘟病品种丽江新团黑谷(LTH)为遗传背景,研究含有云引抗稻瘟病菌株“四川-43”基因的近等基因系的抗病和感病材料,预测基因谷胱甘肽转移酶OsGST(Genbank序列号:Os05g0116100)对稻瘟病菌诱导的响应及响应程度的差别。利用RACE技术克隆目的基因cDNA全长序列,并在近等基因系抗病材料的抗病株系和感病株系中进行序列比对,寻找差异位点。结果表明,该基因与短柄二叶草中编码谷胱甘肽转移酶的基因亲缘关系最接近,结合其编码的蛋白质序列分析进一步论证该基因可能作为一个相对保守的抗病防卫相关基因、编码谷胱甘肽转移酶,将基因命名为OsGST。同时构建了该基因的过表达载体、干扰载体,为后续揭示其在感抗材料中响应稻瘟病菌程度差异的原因、研究其功能及抗病机制奠定基础。
在對粳稻‘雲引’受稻瘟病菌誘導的近等基因繫蛋白質組學分析的基礎上,通過實時定量PCR等方法,以普感稻瘟病品種麗江新糰黑穀(LTH)為遺傳揹景,研究含有雲引抗稻瘟病菌株“四川-43”基因的近等基因繫的抗病和感病材料,預測基因穀胱甘肽轉移酶OsGST(Genbank序列號:Os05g0116100)對稻瘟病菌誘導的響應及響應程度的差彆。利用RACE技術剋隆目的基因cDNA全長序列,併在近等基因繫抗病材料的抗病株繫和感病株繫中進行序列比對,尋找差異位點。結果錶明,該基因與短柄二葉草中編碼穀胱甘肽轉移酶的基因親緣關繫最接近,結閤其編碼的蛋白質序列分析進一步論證該基因可能作為一箇相對保守的抗病防衛相關基因、編碼穀胱甘肽轉移酶,將基因命名為OsGST。同時構建瞭該基因的過錶達載體、榦擾載體,為後續揭示其在感抗材料中響應稻瘟病菌程度差異的原因、研究其功能及抗病機製奠定基礎。
재대갱도‘운인’수도온병균유도적근등기인계단백질조학분석적기출상,통과실시정량PCR등방법,이보감도온병품충려강신단흑곡(LTH)위유전배경,연구함유운인항도온병균주“사천-43”기인적근등기인계적항병화감병재료,예측기인곡광감태전이매OsGST(Genbank서렬호:Os05g0116100)대도온병균유도적향응급향응정도적차별。이용RACE기술극륭목적기인cDNA전장서렬,병재근등기인계항병재료적항병주계화감병주계중진행서렬비대,심조차이위점。결과표명,해기인여단병이협초중편마곡광감태전이매적기인친연관계최접근,결합기편마적단백질서렬분석진일보론증해기인가능작위일개상대보수적항병방위상관기인、편마곡광감태전이매,장기인명명위OsGST。동시구건료해기인적과표체재체、간우재체,위후속게시기재감항재료중향응도온병균정도차이적원인、연구기공능급항병궤제전정기출。
Based on the proteomic analysis of blast-resistant near-isogenic lines derived from j aponica rice‘Yunyin’ ,infected with M . oryz ae ,real-time quantitative PCR and other methods were used to explore the blast resistant and susceptible materials of near-isogenic lines using LTH as genetic background and containing blast‘Sichuan-43’ resistance strain to forecast for rice blast fungus-induced response and the different degrees between the responses of Glutathione S-transferase gene OsGST (Genbank sequence number :Os05g0116100) .Using RACE technology to get the full cDNA Sequence ,and comparing the sequence between the blast resistant lines and the blast susceptible lines .The phylogenetic tree showed the gene had the closest genetic relationship with glutathione transferase encoding gene of brachypodium distachyon .Combined with the analysis of the protein sequence ,OsGST was suggested to be a conserved gene that encoding glutathione transferase . The overexpression and RNAi expression vector were constructed for subsequent study of its functional identification and resistant mechanisms .