转TPSP融合基因小麦植株的获得及抗旱性初步鉴定

Obtaining of Transgenic Wheat Plants with TPSP Fusion Gene and Preliminary Assay of Drought Tolerance

为了培育抗旱小麦新材料,在构建含大肠杆菌海藻糖-6-磷酸合成酶基因(otsA)和海藻糖-6磷酸磷酸酯酶基因(otsB)融合基因TPSP的表达载体pBS-ATPSP的基础上,利用花粉管通道法对小麦品种豫麦34和豫麦18进行了遗传转化。两个品种共处理2 495朵小花,获得T0代种子1 611粒。两个品种分别获得T1代小麦植株857和659株,经PCR鉴定转基因植株分别为11株和7株。通过半定量RT-PCR技术对4个T2代转基因株系中融合基因TPSP的表达分析显示,3个株系中TPSP基因受水分胁迫而诱导表达,其中2个株系在温室进行的干旱胁迫鉴定中表现出较强的抗旱能力。

In order to obtain drought resistant wheat materials based on the expression vector pBS-ATPSP harboring a fusion gene TPSP （fused by the otsA and otsB gene which encodes trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase respectively）, two wheat cultivars, Yumai 34 and Yumai 18, were transformed Via pollen tube pathway. In total, 2 495 florets were treated with plasmid DNA and 1 611 To seeds were harvested, 857 and 659 T1 plantlets were obtained from Yumai 34 and Yumai 18, respectively, and the PCR assay showed that the TPSP gene had been integrated into 11 and 7 wheat plantlets of the two wheat varieties, respectively. The semi-quantitative RT-PCR analysis showed that the expression of the fusion gene TPSP was highly induced by water-stress in 3 of 4 detected T2 transgenic wheat lines, and 2 lines of them were strongly tolerant to drought stress under the green house test condition. The transgenic wheat materials possibly can be further used in molecular breeding of wheat for drought tolerance.