大豆GmbZIP16的抗旱功能验证及分析

doi:10.3864/j.issn.0578-1752.2018.15.001

Abstract

Abstract: 【Objective】Soybean GmbZIP16 protein was screened by analyzing soybean drought transcriptome. Functions of soybean GmbZIP16 were verified by analyzing the phenotypic characterization of transgenic GmbZIP16 Arabidopsis and soybean hairy root complexes. On the basis of experimental result analyses above-mentioned, we could determine that GmbZIP16 was involved in the drought resistance process.【Method】soybean GmbZIP16 was found out by analyzing soybean drought transcriptome and cloned by PCR, which soybean cDNA as a template, and then ligated the fragment to pCAMBIA1302 and pCAMBIA3301 expression vectors by in-fusion ligase. The recombinant pCAMBIA1302-GmbZIP16 and pCAMBIA3301-GmbZIP16 vectors were transferred into Agrobacterium GV3101 and K599 competent cells by liquid nitrogen freeze-melt method, respectively. The transgenic Arabidopsis plants and transgenic soybean hairy root complexes were generated by Agrobacterium-mediated transformation method. The transgenic GmbZIP16 Arabidopsis plants were identified by semi quantitative PCR and quantitative real time PCR. Which demonstrated GmbZIP16 could over express in transgenic Arabidopsis and transgenic soybean hairy root complex plants. 2 weeks old transgenic Arabidopsis and WT plants grew under normal conditions were transferred into MS0 solid medium supplement with 6% and 8% PEG for 7d. Biomass differences between transgenic Arabidopsis and WT plants were investigated and analyzed. The different expression of stress- related genes between Arabidopsis and WT plants were analyzed by qRT-PCR. The transgenic GmbZIP16 soybean hairy root complexes and control group plants were treated with 25% PEG for 7 days, and then the leaf samples of transgenic GmbZIP16 soybean hairy root complex and control group plants were taken. The proline content, MDA content and chlorophyll content of leaf samples were measured by Multiskan Spectrum Microplate Spectrophotometer.【Result】The GmbZIP16 gene was isolated by PCR technology. The two transgenic GmbZIP16 Arabidopsis lines were obtained by Agrobacterium-mediated transformation method. Compared with the WT plants, the transgenic GmbZIP16 Arabidopsis lines had higher biomass (the fresh weight and the root length) and survival rate under drought stress by phenotypic characterization experiment. The expression levels of some relative genes such as RD29B, DREB2A and P5CS were improved in transgenic GmbZIP16 Arabidopsis, compared with the WT plants. The leaves of transgenic GmbZIP16 soybean hairy root complex plants had the higher proline and chlorophyll content and a lower MDA content than that of the control group plants under the deal with 25% PEG. 【Conclusion】The drought resistance of transgenic Arabidopsis thaliana was improved by expressing soybean GmbZIP16 in Arabidopsis thaliana. Over expression of GmbZIP16 could enhance the drought resistance of transgenic soybean hairy root complex. GmbZIP16 can improve the drought resistance of plants mainly by affecting the expression of genes related to stress tolerance.

Key words: soybean, GmbZIP16, drought resistance, qRT-PCR

ZHAO WanYing, YU TaiFei, YANG JunFeng, LIU Pei, CHEN Jun, CHEN Ming, ZHOU YongBin, MA YouZhi, XU ZhaoShi, MIN DongHong. Verification and Analyses of Soybean GmbZIP16 Gene Resistance to Drought[J].Scientia Agricultura Sinica, 2018, 51(15): 2835-2845.

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Verification and Analyses of Soybean GmbZIP16 Gene Resistance to Drought

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