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Enhanced Accumulation of BnA7HSP70 Molecular Chaperone Binding Protein Improves Tolerance to Drought Stress in Transgenic Brassica napus
Li-Li WAN, Zhuan-Rong WANG, Qiang XIN, Fa-Ming DONG, Deng-Feng HONG, Guang-Sheng YANG
Acta Agronomica Sinica    2018, 44 (04): 483-492.   DOI: 10.3724/SP.J.1006.2018.00483
Abstract   (632 HTML11 PDF(pc) (5501KB)(719)  

The molecular chaperone binding protein gene participates in the constitutive function of plant growth and protects plant cells against stresses. In this study, we found that BnA7HSP70 overexpressed transgenic lines did not wilt and showed only a small decrease in water potential. However, the wild type lines showed a large decrease in leaf water potential. The transgenic plants had higher relative water content, better osmotic adjustment and less lipid membrane peroxidation. In addition, the leaves from the elevated levels of BnA7HSP70 in transgenic lines conferred tolerance to the glycosylation inhibitor tunicamycin during germination. BnA7HSP70 overexpression-mediated attenuation of stress-induced cell death was confirmed by the decreased percentage of dead cell and the reduced induction of the senescence-associated marker gene BnCNX1. These phenotypes were accompanied by a delay in the induction of the cell death marker genes BnNRP, which are involved in transducing a cell death signal generated by ER stress and osmotic stress through the NRP (N-rich protein)-mediated signaling pathway. Enhanced expression of BnA7HSP70 delayed unfold protein response and NRP pathway mediated chlorosis and the appearance of senescence-associated markers BnLSC222 and BnLSC54 in Brassica napus. These results suggest that overexpression of BnA7HSP70 in Brassica napus alleviate ER stress and osmotic stress-integrating cell death response confronted with water stress.


Fig. 8 Gene expression analysis of senescence and cell death-associated genes in wild type and OE plants under tunicamycin treatment
Total RNA were isolated from wild type and transgenic plant leaves at 0, 24, 48, and 72 h of treatment, and the endogenous BnA7HSP70, BnCNX1, BnBiP3, and BnNRP in wild type and OE transgenic plants treatment with tunicamycin and the control DMSO were monitored by qRT-PCR. The bars indicate the confidence interval (P < 0.05, n = 3). BnCNX1, BnBiP3, and BnNRP are ER stress markers.
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图8   衣霉素处理后转基因和非转基因植株内质网胁迫的标签基因的表达分析
取衣霉素处理0、24、48和72 h的非转基因和转基因植株的叶片, 分析内源BnA7HSP70BnCNX1、BnNRPBnBiP3基因的表达量。DMSO处理的植株为对照。BnCNX1、BnBiP3BnNRP基因是内质网胁迫的标志基因。
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