分子植物育种
分子植物育種
분자식물육충
MOLECULAR PLANT BREEDING
2013年
1期
62-71
,共10页
吴广锡%魏崃%唐晓飞%王伟威%王鹏飞%王兴宇%刘丽君
吳廣錫%魏崍%唐曉飛%王偉威%王鵬飛%王興宇%劉麗君
오엄석%위래%당효비%왕위위%왕붕비%왕흥우%류려군
大豆%热激转录因子%GmHsFA1%农杆菌介导%转基因%耐热性%抗旱性
大豆%熱激轉錄因子%GmHsFA1%農桿菌介導%轉基因%耐熱性%抗旱性
대두%열격전록인자%GmHsFA1%농간균개도%전기인%내열성%항한성
Soybean%Heat shock transcription factor%GmHsFA1%Agrobacterium tumefaciens%Transgene%Heat tolerance%Drought resistance
本研究将热激转录因子GmHsFA1基因构建在植物表达载体pCAMBIA3300中,以大豆子叶节作为受体,通过农杆菌介导法将热激转录因子GmHsFA1基因导入到高产品种黑农53中,获得一批转基因株系.采用Real-time PCR的方法对各代株系GmHsFA1基因的转录水平进行相对定量分析,确定转基因大豆中热激转录因子GmHsFA1的表达量明显提高.利用转热激转录因子GmHsFA1大豆研究了高温和干旱胁迫下,大豆热激转录因子GmHsFA1应对高温和干旱信号的基因表达,生理生化特性,光合特性及产量性状的变化反应,采用胁迫系数与灰色关联分析相结合方法,对转GmHsFA1基因大豆的耐热性和抗旱性进行综合鉴定和评价.结果表明:转热激转录因子GmHsFA1大豆品系在高温和干旱诱导条件下,大豆叶片中GmHsFA1、GmHSP70、GmHSP22和GmHSP17.9基因的表达量明显高于非转基因受体黑农53,叶肉细胞中的脯氨酸含量和可溶性糖含量明显高于受体,丙二醛含量增幅较小,叶片的光合特性受胁迫变化较小,光合特性能力和产量性状表现高于受体,表明过量表达的热激转录因子GmHsFA1大豆植株的耐热能力和抗干旱能力得到明显的提高.
本研究將熱激轉錄因子GmHsFA1基因構建在植物錶達載體pCAMBIA3300中,以大豆子葉節作為受體,通過農桿菌介導法將熱激轉錄因子GmHsFA1基因導入到高產品種黑農53中,穫得一批轉基因株繫.採用Real-time PCR的方法對各代株繫GmHsFA1基因的轉錄水平進行相對定量分析,確定轉基因大豆中熱激轉錄因子GmHsFA1的錶達量明顯提高.利用轉熱激轉錄因子GmHsFA1大豆研究瞭高溫和榦旱脅迫下,大豆熱激轉錄因子GmHsFA1應對高溫和榦旱信號的基因錶達,生理生化特性,光閤特性及產量性狀的變化反應,採用脅迫繫數與灰色關聯分析相結閤方法,對轉GmHsFA1基因大豆的耐熱性和抗旱性進行綜閤鑒定和評價.結果錶明:轉熱激轉錄因子GmHsFA1大豆品繫在高溫和榦旱誘導條件下,大豆葉片中GmHsFA1、GmHSP70、GmHSP22和GmHSP17.9基因的錶達量明顯高于非轉基因受體黑農53,葉肉細胞中的脯氨痠含量和可溶性糖含量明顯高于受體,丙二醛含量增幅較小,葉片的光閤特性受脅迫變化較小,光閤特性能力和產量性狀錶現高于受體,錶明過量錶達的熱激轉錄因子GmHsFA1大豆植株的耐熱能力和抗榦旱能力得到明顯的提高.
본연구장열격전록인자GmHsFA1기인구건재식물표체재체pCAMBIA3300중,이대두자협절작위수체,통과농간균개도법장열격전록인자GmHsFA1기인도입도고산품충흑농53중,획득일비전기인주계.채용Real-time PCR적방법대각대주계GmHsFA1기인적전록수평진행상대정량분석,학정전기인대두중열격전록인자GmHsFA1적표체량명현제고.이용전열격전록인자GmHsFA1대두연구료고온화간한협박하,대두열격전록인자GmHsFA1응대고온화간한신호적기인표체,생리생화특성,광합특성급산량성상적변화반응,채용협박계수여회색관련분석상결합방법,대전GmHsFA1기인대두적내열성화항한성진행종합감정화평개.결과표명:전열격전록인자GmHsFA1대두품계재고온화간한유도조건하,대두협편중GmHsFA1、GmHSP70、GmHSP22화GmHSP17.9기인적표체량명현고우비전기인수체흑농53,협육세포중적포안산함량화가용성당함량명현고우수체,병이철함량증폭교소,협편적광합특성수협박변화교소,광합특성능력화산량성상표현고우수체,표명과량표체적열격전록인자GmHsFA1대두식주적내열능력화항간한능력득도명현적제고.
In this study, we inserted GmHsFA1 gene to dicotyledon expression vector pCAMBIA3300 that contains the selection marker gene bar, and pCAMBIA3300-GmHsFA1 has been transferred into new soybean lines Heinong5337 by Agrobacterium tumefaciens. The expression of GmHSFA1 have been analyzed in each generations of transgenic soybean by Real-time PCR, and overexpressing GmHsFA1 gene was demonstrated in transgenic plant. Photosynthetic characteristics (Pn, Cond, Ci, Tr), Physiological, biochemical (Pro, MDA, Wss) and yield traits of the GmHsFA1 and GmHSP70, GmHSP22, GmHSP17.9 were tested in transgenic soybean under the high temperature stress and drought stress. Stress coefficient analysis and grey related correlation analysis were introduced into this experiment, which provided comprehensive evaluation on heat tolerance and drought resistance of soybean materials. The results showed that the expression levels of GmHsFA1, GmHSP17.9, GmHSP22 and GmHSP70 in leaf blade of transgenic plants were notably higher than that in Heinong53 that is non-transgenic plant growing under normal condition or heat and drought introduction. Under the condition of stress, the content of soluble sugar (wss) and free proline (pro) of the transgenic lines were significant higher than those growing in the normal conditions, while the increment of malondialdehyde (MDA) appeared. The transgenic lines also had little change in photosynthetic characteristics and yield traits, Whereas the heat tolerance and drought resistance were remarkably enhanced in transgenic plants.m.
