Over-expression of a grafting-responsive gene from hickory increases abiotic stress tolerance in Arabidopsis
Over-expression of a grafting-responsive gene from hickory increases abiotic stress tolerance in Arabidopsis
复制标题
山核桃嫁接响应基因的过度表达提高了拟南芥的非生物胁迫耐受性
DOI：
10.1007/s00299-018-2250-4
复制
发表时间：
2018-03-01
影响因子：
6.2
通讯作者：
Zheng, Bingsong
中科院分区：
文献类型：
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作者：
Kumar, R. M. Saravana;Ji, Guocun;Zheng, Bingsong
从山核桃中克隆了一个嫁接反应基因CcPIP 1;2，并通过在非洲爪蟾和拟南芥中的异源表达，对该基因的水分转运活性和非生物胁迫耐受性进行了进一步的功能鉴定。质膜内在蛋白（Plasma membrane intrinsic proteins，PIP）是膜内在蛋白（membrane intrinsic protein，MIP）家族的一种多功能通道蛋白。本研究克隆了山核桃嫁接反应基因（CcPIP 1;2），并对其功能进行了研究。应用非选择性水分抑制剂（HgCl 2和根皮素）导致嫁接后30天（DAG）的山核桃嫁接植株死亡。此外，在抑制剂处理条件下，所选CcPIP 1;2基因的转录物积累在接枝样品中从0至14 DAG逐渐降低。GFP-CcPIP 1;2融合蛋白的瞬时表达分析表明，CcPIP 1;2定位于细胞质膜。CcPIP 1;2蛋白在非洲爪蟾卵母细胞系统中的异源表达有助于水进入细胞。CcPIP 1过表达; 2在拟南芥中的种子发芽率提高种子生长在H2 O2-，阿坝-，和甘露醇的培养基中，但在含盐介质中生长时没有效果。CcPIP 1;2转基因植株在干旱条件下生长较好。各种ABA相关的胁迫标记基因以及细胞壁扩展蛋白标记基因的表达在CcPIP 1;2过表达拟南芥系中显著高于野生型（WT）。
A grafting response gene CcPIP1;2 was cloned from hickory plant, further functional characterization of the gene for water transport activity and abiotic stress tolerances were carried out through heterologous expression in Xenopus and Arabidopsis. Plasma membrane intrinsic proteins (PIPs) are multifunctional channel proteins belonging to the membrane intrinsic protein (MIP) family. In this study, a grafting-responsive gene from hickory (CcPIP1;2) was cloned and functionally characterized. Application of non-selective water inhibitors (HgCl2and phloretin) led to the death of grafted hickory plants at 30 days after grafting (DAG). Furthermore, the transcript accumulation of the selected CcPIP1;2 gene was gradually decreased from 0 to 14 DAG in the grafted samples under inhibitor treatment conditions. Transient expression analysis of the GFP-CcPIP1;2 fusion protein showed that CcPIP1;2 was located at plasma membrane. Heterologous expression of CcPIP1;2 protein in the Xenopus oocyte system helped the access of water into the cells. Over-expression of CcPIP1;2 in Arabidopsis improved the percentage of seed germination when the seeds were grown in H2O2-, ABA-, and mannitol-containing media, but had no effect when grown in the salt containing media. CcPIP1;2 transgenic plants grew better under drought conditions. The expression of various ABA-related stress marker genes as well as cell wall expansin marker genes was significantly higher in CcPIP1;2 over-expression Arabidopsis lines than in the wild type (WT).