农业生物技术学报
農業生物技術學報
농업생물기술학보
JOURNAL OF AGRICULTURAL BIOTECHNOLOGY
2014年
12期
1514-1524
,共11页
邢丹丹%韩晓敏%冯宗琪%杨杞%齐力旺%刘福军%王瑞刚%李国婧
邢丹丹%韓曉敏%馮宗琪%楊杞%齊力旺%劉福軍%王瑞剛%李國婧
형단단%한효민%풍종기%양기%제력왕%류복군%왕서강%리국청
中间锦鸡儿%CiMYBJ2基因%实时荧光定量PCR%启动子
中間錦鷄兒%CiMYBJ2基因%實時熒光定量PCR%啟動子
중간금계인%CiMYBJ2기인%실시형광정량PCR%계동자
Caragana intermedia%CiMYBJ2%qRT-PCR%Promoter
MYB转录因子是植物体内最大的转录因子家族之一，参与植物生长发育的调控、对逆境的响应等生理过程。本研究通过RACE技术从中间锦鸡儿(Caragana intermedia)中克隆了一个MYB转录因子CiMYBJ2(GenBank登录号：KJ937783)，并对其在植物中的表达及其启动子进行了初步研究。本研究克隆得到该基因cDNA全长1368 bp，3'UTR长196 bp，5'UTR长203 bp。其中开放阅读框长969 bp，编码一个含322个氨基酸的亲水蛋白。gDNA序列长1285 bp，有2个内含子和3个外显子。采用实时荧光定量PCR技术分析了CiMYBJ2的表达模式，结果发现，在高温、脱水、NaCl、干旱等处理条件下，CiMYBJ2基因对不同胁迫均有响应。在高温处理后，CiMYBJ2基因的表达量在1和3 h明显降低，之后维持一个较低值，在不同时间点基因表达量与0 h相比差异均达到极显著水平(P＜0.01)；在脱水处理1 h后CiMYBJ2基因的表达量上升，且与0 h相比差异极显著(P＜0.01)，然后逐渐降低，在3 h CiMYBJ2基因的表达量与0 h相比差异不显著(P＞0.05)，在8和12 h该基因的表达量与0 h相比差异均达到极显著(P＜0.01)，在24 h的表达量是0 h的一半，且差异显著(P＜0.05)；在NaCl处理1 h后CiMYBJ2基因的转录水平开始增加，3 h达到最高，之后总体呈下降趋势，在8h表达量最低，在1、3、8和24h该基因的表达量与0h相比差异极显著(P＜0.01)，在12 h CiMYBJ2基因的表达量与0 h相比差异不显著(P＞0.05)；干旱处理6 d后，CiMYBJ2基因的表达量上升，之后一直下降，且与0 d相比差异达到极显著水平(P＜0.01)，15 d表达量最低。研究结果暗示CiMYBJ2基因可能在植物对胁迫的响应过程中起作用。利用染色体步移技术克隆了中间锦鸡儿CiMYBJ2基因启动子序列，长度为873 bp，分析显示，该启动子具有响应多种非生物胁迫的顺式作用元件。研究结果为进一步研究CiMYBJ2基因功能和表达调控提供了依据。
MYB轉錄因子是植物體內最大的轉錄因子傢族之一，參與植物生長髮育的調控、對逆境的響應等生理過程。本研究通過RACE技術從中間錦鷄兒(Caragana intermedia)中剋隆瞭一箇MYB轉錄因子CiMYBJ2(GenBank登錄號：KJ937783)，併對其在植物中的錶達及其啟動子進行瞭初步研究。本研究剋隆得到該基因cDNA全長1368 bp，3'UTR長196 bp，5'UTR長203 bp。其中開放閱讀框長969 bp，編碼一箇含322箇氨基痠的親水蛋白。gDNA序列長1285 bp，有2箇內含子和3箇外顯子。採用實時熒光定量PCR技術分析瞭CiMYBJ2的錶達模式，結果髮現，在高溫、脫水、NaCl、榦旱等處理條件下，CiMYBJ2基因對不同脅迫均有響應。在高溫處理後，CiMYBJ2基因的錶達量在1和3 h明顯降低，之後維持一箇較低值，在不同時間點基因錶達量與0 h相比差異均達到極顯著水平(P＜0.01)；在脫水處理1 h後CiMYBJ2基因的錶達量上升，且與0 h相比差異極顯著(P＜0.01)，然後逐漸降低，在3 h CiMYBJ2基因的錶達量與0 h相比差異不顯著(P＞0.05)，在8和12 h該基因的錶達量與0 h相比差異均達到極顯著(P＜0.01)，在24 h的錶達量是0 h的一半，且差異顯著(P＜0.05)；在NaCl處理1 h後CiMYBJ2基因的轉錄水平開始增加，3 h達到最高，之後總體呈下降趨勢，在8h錶達量最低，在1、3、8和24h該基因的錶達量與0h相比差異極顯著(P＜0.01)，在12 h CiMYBJ2基因的錶達量與0 h相比差異不顯著(P＞0.05)；榦旱處理6 d後，CiMYBJ2基因的錶達量上升，之後一直下降，且與0 d相比差異達到極顯著水平(P＜0.01)，15 d錶達量最低。研究結果暗示CiMYBJ2基因可能在植物對脅迫的響應過程中起作用。利用染色體步移技術剋隆瞭中間錦鷄兒CiMYBJ2基因啟動子序列，長度為873 bp，分析顯示，該啟動子具有響應多種非生物脅迫的順式作用元件。研究結果為進一步研究CiMYBJ2基因功能和錶達調控提供瞭依據。
MYB전록인자시식물체내최대적전록인자가족지일，삼여식물생장발육적조공、대역경적향응등생리과정。본연구통과RACE기술종중간금계인(Caragana intermedia)중극륭료일개MYB전록인자CiMYBJ2(GenBank등록호：KJ937783)，병대기재식물중적표체급기계동자진행료초보연구。본연구극륭득도해기인cDNA전장1368 bp，3'UTR장196 bp，5'UTR장203 bp。기중개방열독광장969 bp，편마일개함322개안기산적친수단백。gDNA서렬장1285 bp，유2개내함자화3개외현자。채용실시형광정량PCR기술분석료CiMYBJ2적표체모식，결과발현，재고온、탈수、NaCl、간한등처리조건하，CiMYBJ2기인대불동협박균유향응。재고온처리후，CiMYBJ2기인적표체량재1화3 h명현강저，지후유지일개교저치，재불동시간점기인표체량여0 h상비차이균체도겁현저수평(P＜0.01)；재탈수처리1 h후CiMYBJ2기인적표체량상승，차여0 h상비차이겁현저(P＜0.01)，연후축점강저，재3 h CiMYBJ2기인적표체량여0 h상비차이불현저(P＞0.05)，재8화12 h해기인적표체량여0 h상비차이균체도겁현저(P＜0.01)，재24 h적표체량시0 h적일반，차차이현저(P＜0.05)；재NaCl처리1 h후CiMYBJ2기인적전록수평개시증가，3 h체도최고，지후총체정하강추세，재8h표체량최저，재1、3、8화24h해기인적표체량여0h상비차이겁현저(P＜0.01)，재12 h CiMYBJ2기인적표체량여0 h상비차이불현저(P＞0.05)；간한처리6 d후，CiMYBJ2기인적표체량상승，지후일직하강，차여0 d상비차이체도겁현저수평(P＜0.01)，15 d표체량최저。연구결과암시CiMYBJ2기인가능재식물대협박적향응과정중기작용。이용염색체보이기술극륭료중간금계인CiMYBJ2기인계동자서렬，장도위873 bp，분석현시，해계동자구유향응다충비생물협박적순식작용원건。연구결과위진일보연구CiMYBJ2기인공능화표체조공제공료의거。
MYB transcription factor is one of the largest family of transcription factors in plants which play impotant roles in regulation of plant growth and development, and response to many environmental stimuli. In this study, a gene named CiMYBJ2 (GenBank accession No.KJ937783) was cloned by RACE technique from Caragana intermedia and its expression level along with the promoter was analysed. The full-length cDNA of CiMYBJ2 was 1 368 bp with the 3'UTR of 196 bp and the 5'UTR of 203 bp. The ORF of CiMYBJ2 was 969 bp, encoding a protein of 322 amino acids which was hydrophilia. The full-length gDNA was 1 285 bp, including two introns and three exons. The expression pattern of CiMYBJ2 analysed by qRT-PCR showed that it was altered by heat, dehydration, NaCl and drought treatments. After heat treatment, the expression of CiMYBJ2 decreased at 1 and 3 h, and then maintained a lower level expression. the change of gene expression at different time points was extremely significant compared with the 0 h (P<0.01). After dehydration treatment for 1 h, the expression of CiMYBJ2 increased shortly, and then gradually decreased at 3 h, and the expression difference was not significant compared with the 0 h (P>0.05). After 8 h , the expression of the gene showed extremely significant comapred with that of the 0 h (P<0.01), and the expression level of the gene at 24 h reached only half of that at 0 h (P<0.05). Under NaCl treatment, the transcript level of CiMYBJ2 began to increase after 1 h, and reached the maximum at 3 h. After that the overall trend kept downward, the expression dropped to the lowest level at 8 h, and expression of the gene was extremely significant at different time points except for 12 h (P<0.01). After drought treatment of 6 d, the expression of CiMYBJ2 increased slightly, and dropped significanly after compared with 0 d (P<0.01), reached the minimum expression at 15 d. The promoter sequence of CiMYBJ2 with 873 bp in length was obtained by genome walking technique and many abiotic stress-related cis-elements were found. These results indicated that CiMYBJ2 might be involved in response of C. intermedia to abiotic stresses. The research results provide a basis for the further study of gene function and expression regulation.
