应用与环境生物学报
應用與環境生物學報
응용여배경생물학보
CHINESE JOURNAL OF APPLIED & ENVIRONMENTAL BIOLOGY
2009年
5期
591-595
,共5页
刘继恺%高永峰%牛向丽%刘永胜
劉繼愷%高永峰%牛嚮麗%劉永勝
류계개%고영봉%우향려%류영성
HP1%HP2%RNA干涉%果实特异启动子%农杆菌介导转化%叶绿素%番茄
HP1%HP2%RNA榦涉%果實特異啟動子%農桿菌介導轉化%葉綠素%番茄
HP1%HP2%RNA간섭%과실특이계동자%농간균개도전화%협록소%번가
HP1%HP2%RNA interference%fruit-specific promoter%Agrobacterium-mediated transformation%chlorophyll%tomato
番茄HP1和HP2是色素积累的负调控因子,在光形态建成和色素积累调控中起着重要作用.将番茄HP1、HP2基因片段导入到植物表达载体pBI121,用番茄果实特异表达的TMF7基因的启动子替换原有的CaMV 35S启动子,构建果实特异表达HP1、HP2双基因RNA共干涉植物表达载体pBI121-TMF7-HP1HP2.通过根癌农杆菌介导转入番茄子叶,经组织培养成功获得转基因植株.半定量RT-PCR分析显示,转基因植株果实内HP1、HP2的表达量均明显低于野生型植株果实.转基因植株果实叶绿素含量比野生型明显升高,而叶片中的叶绿素含量无明显差异.该研究结果为采用基因工程的方法改善番茄果实营养品质作出了新的尝试和提出了新的思路.图6表2参15
番茄HP1和HP2是色素積纍的負調控因子,在光形態建成和色素積纍調控中起著重要作用.將番茄HP1、HP2基因片段導入到植物錶達載體pBI121,用番茄果實特異錶達的TMF7基因的啟動子替換原有的CaMV 35S啟動子,構建果實特異錶達HP1、HP2雙基因RNA共榦涉植物錶達載體pBI121-TMF7-HP1HP2.通過根癌農桿菌介導轉入番茄子葉,經組織培養成功穫得轉基因植株.半定量RT-PCR分析顯示,轉基因植株果實內HP1、HP2的錶達量均明顯低于野生型植株果實.轉基因植株果實葉綠素含量比野生型明顯升高,而葉片中的葉綠素含量無明顯差異.該研究結果為採用基因工程的方法改善番茄果實營養品質作齣瞭新的嘗試和提齣瞭新的思路.圖6錶2參15
번가HP1화HP2시색소적루적부조공인자,재광형태건성화색소적루조공중기착중요작용.장번가HP1、HP2기인편단도입도식물표체재체pBI121,용번가과실특이표체적TMF7기인적계동자체환원유적CaMV 35S계동자,구건과실특이표체HP1、HP2쌍기인RNA공간섭식물표체재체pBI121-TMF7-HP1HP2.통과근암농간균개도전입번가자협,경조직배양성공획득전기인식주.반정량RT-PCR분석현시,전기인식주과실내HP1、HP2적표체량균명현저우야생형식주과실.전기인식주과실협록소함량비야생형명현승고,이협편중적협록소함량무명현차이.해연구결과위채용기인공정적방법개선번가과실영양품질작출료신적상시화제출료신적사로.도6표2삼15
Tomato HP1 and HP2 genes are negative regulators of pigment accumulation, and they play an important role in photomorphogenesis and pigmentation regulation. In this study, a RNA interference (RNAi) vector composing of inversely-repeated sequence fragments derived from both tomato HP1 and HP2 was constructed. The CaMV 358 promoter in pBI121 was replaced by fruit-specific promoter TFM7. The construct was introduced into tomato Lysopersicon esculentum Mill. cv. Ailsa Craig by means of Agrobacterium-mediated transformation. The analysis of semi-quantitative RT-PCR revealed a distinct reduction of endogenous HP1 and HP2 expression levels in the RNAi transgenic lines compared to that of wild plants. The chlorophyll content in green fruit of the RNAi repression lines was significantly elevated, whereas the chlorophyll in their leaves remained unaltered compared with that of wild plants. The results show a new way to modify fruit nutritional quality in tomato through gene engineering. Fig 6, Tab 2, Ref 15