抗逆相关基因GmAREB转基因小麦的获得与鉴定

Molecular and Functional Analysis of Transgenic Wheat Transformed with Stress-related Gene GmAREB

从大豆中克隆一个抗逆相关的bZIP类转录因子基因GmAREB,功能分析表明:GmAREB基因的过表达可以显著提高转基因拟南芥和烟草的抗旱、耐盐和耐寒性。为了获得抗逆转基因小麦,本研究利用玉米的Ubiqutin启动子控制GmAREB基因表达,构建了用于小麦转化的载体pSK-GmAREB。采用基因枪共转化法转化小麦品种郑147和济麦22。通过PCR检测共获得T0代的阳性植株70株,转化率为1.37%。其中,郑147阳性植株共31株,转化率为2.14%;济麦22阳性植株39株,转化率为1.08%。目前,已经获得T1代转基因株系18个,其中以郑147为受体的株系4个,以济麦22为受体的株系14个。对部分株系进行Southern blotting分析,进一步证实GmAREB基因已经整合到小麦基因组中。在低温胁迫条件下,3个以济麦22为受体的转基因株系体内脯氨酸的积累与受体小麦相比有显著增加,初步证明在小麦中过表达GmAREB基因,可以促进渗透调节物质脯氨酸的积累,可能有助于转基因小麦抗逆性的提高。本研究为进一步筛选抗逆转基因小麦新材料奠定了基础。

We isolated stress-related bZIP transcription factor gene,GmAREB from soybean previously.The functional analysis indicated that overexpression of GmAREB can significantly improve the stress resistance of transgenic plants.In this study,GmAREB was inserted in downstream of maize Ubiqutin promoter to construct vector,pSK-GmAREB for wheat transformation.The vectors were transformed into wheat varieties Zheng 147 and Jimai 22 by using biolistic method.After transformation,70 T0 transgenic plants were identified using PCR and the transformational efficiency was 1.37%.Among transgenic plants,31 plants came from host Zheng 147 and the transformational efficiency was 2.14%.39 plants came from host Jimai 22 and the transformational efficiency was 1.08%.At present,18 T1 generation of transgenic line had been identificated,and 4 and 14 transgenic plants came from Zheng 147 and Jimai 22,respectively.The southern blot proved that GmAREB was inserted into wheat genome.The stress-tolerance analysis showed that under low temperature stress condition,proline accumulation in three transgenic plants host in Jimai 22 were higher than that of wild type wheat plants,which suggested that overexpression of GmAREB increased accumulation of proline in transgenic wheat.This higher level of proline content might be contributed for stress-tolerance of transgenic wheat plants.We hope that our study can pave the way for selecting stress-tolerance transgenic wheat lines in the future.