甘蓝型油菜TIP4;1基因的克隆与表达分析

Cloning and Expression Analysis of TIP4;1 Gene from Brassica napus

水孔蛋白是位于质膜和液泡膜等生物膜上主要负责水分跨膜运输的通道蛋白。为了分析甘蓝型油菜种子萌发过程水分调控的分子机制,采用RT-PCR法从甘蓝型油菜种子中克隆了液泡膜内在蛋白(Tonop last intrinsic proteins,TIPs)TIP4;1基因片段序列,并对甘蓝型油菜种子TIP4;1基因在种子萌发过程及干旱、低温和盐胁迫下的表达进行了qRT-PCR(Quantitative Real-time PCR)分析。结果表明,TIP4;1基因在干种子中表达水平很低,但是在种子萌发过程中表达量增加。此外,在干旱、低温及盐的胁迫处理下,TIP4;1基因的表达上调,结果暗示着该基因可能参与了甘蓝型油菜种子萌发和逆境响应过程。随着研究的深入,水孔蛋白的水分调控机制将进一步被揭示,这对于解决干旱胁迫和盐胁迫等实际问题具有重大现实意义。

Aquaporins are channel proteins,which located in biological membranes including plasma membrane and tonoplast. It can highly facilitate water transportation across biological membranes. To investigate molecular mechanism of water regulation during seed germination,a tonoplast intrinsic proteins（ TIPs） gene fragment TIP4; 1was isolated from Brassica napus seed by RT-PCR. The expression of TIP4; 1 gene during seed germination and stresses treatment was analyzed by quantitative Real-time PCR（ qRT-PCR）. The transcription levels of TIP4; 1 was analyzed during germination,and these results showed TIP4; 1 expression levels was scarcely detected in dry seed,but was up-regulated during germination as well as early young seedlings. In addition,expression of TIP4; 1 in response to abiotic stress was investigated,and results showed that TIP4; 1 gene expression was upregulated under drought,cold and salt stress,suggesting that TIP4; 1 may be involved in Brassica napus seed germination and stress response process. Aquaporin regulation mechanism would be further revealed in future,which had realistic significance for solving drought and salt stress problems.