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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   (626 HTML11 PDF(pc) (5501KB)(718)  

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. 6 BnA7HSP70 overexpression delays drought-induced leaf senescence in OE lines confronted with stress
Drought was induced in wild type (WT) and OE transgenic plants (OE2, OE3, OE7, and OE8) at four-week old stage by withholding irrigation for 10 days. Control: normally irrigated plants. Stress: drought-stressed plants. Values are given as mean SD from three replicates. The experiment of senescence-associated genes BnLSC45 and BnLSC222 was induced by drought treatment. Total RNA was isolated from irrigated and drought-stressed mature leaves of wild plants and OE lines, and gene induction was monitored by quantitative RT-PCR using gene-specific primers.
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图6   BnA7HSP70超表达能够延迟逆境下转基因植株的叶片衰老相关基因表达
野生型(WT)和超表达(OE)转基因家系(OE2、OE3、OE7和OE8)在生长4周后断水10 d, 取样检测油菜叶片衰老相关基因BnLSC45BnLSC222的表达。Control: 正常灌溉条件下的植株。Stress: 受到干旱胁迫后的植株。每次实验3次重复。干旱条件下诱导衰老相关基因BnLSC45BnLSC222基因的表达。从正常灌溉和断水胁迫处理下的非转基因和转基因植株成熟叶片中提取RNA, 用qRT-PCR实验检测基因的表达量。
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