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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. 2 BnA7HSP70 overexpressed (OE) plants confer tolerance under a restricted water regime and 20% PEG treatment
(A) For the fast soil drying treatment, wild type (WT) and BnA7HSP70 overexpressed (OE) lines were allowed to reach four to five weeks leaves stage of development when drought was rapidly induced by withholding irrigation for 10 days. The stress condition was prolonged until the leaves of wild type plants completely wilted. (B) For the fast soil drying treatment, wild type (WT) and BnA7HSP70 overexpressed (OE) lines were allowed to reach four to five weeks leaves stage of development when drought was rapidly induced by withholding irrigation for 15 days. The stress condition was prolonged until the leaves of wild type plants completely wilted. (C) After rewatering for 3 days, most wild type plants were unable to recover, while OE plants survived continued to grow. (D) 40-day-old WT and OE transgenic plants grown in nutrient solution. (E) For drought stress, 40-day-old WT and OE transgenic plants were transferred into nutrient solution containing 20% (w/v) PEG-6000 for 48 h. The concentration of PEG was maintained daily by changing the nutrient solution. (F) Leaf relative water content from WT and OE plants after 10 days, 15 days drought stress and 3 days rewater treatment. (G) Leaf relative water content from WT and OE plants after 20% PEG-6000 treatment for 48 h.
Extracts from the Article
图2    BnA7HSP70基因超表达植株(OE)和非转基因对照(WT)在干旱和20% PEG渗透胁迫处理后表现出耐旱性
(A)对生长4~5周油菜苗进行断水10 d处理, 可见转基因植株表现出更好的耐受性; (B)对生长4~5周油菜苗进行断水15 d处理, 可见转基因植株表现出更好的耐受性; (C)干旱10 d后复水3 d, 大多数非转基因植株不能恢复正常生长, 而转基因植株能够恢复正常生长。(D)在营养液中生长40 d的非转基因和转基因植株; (E)将生长在营养液中40 d的非转基因和转基因植株转移到20% (w/v) PEG-6000的培养液中48 h。通过处理过程中每天更换新鲜的PEG溶液来保证处理时的浓度保持不变; (F)转基因和非转基因植株在干旱10 d、15 d和复水3 d后叶片的相对含水量测定; (G)转基因和非转基因植株从营养液中正常生长转移到20% PEG-6000渗透胁迫处理48 h后叶片的相对含水量。
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