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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   (628 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. 7 BnA7HSP70 overexpression increases resistance against tunicamycin (TUN)-induced cell death
(A) Seeds and seedlings from overexpressed plants (OE) and untransformed wild-type (WT) plants were exposed to 2.5 µg mL-1 or 5.0 µg mL-1 tunicamycin. (B)-(C) Seedlings were monitored for the development of chlorosis and necrotic lesions, and cell viability were measured by the Evans blue dye method. Abs (600 nm) reflects the dead cell content. The values represent the average of three replicates (±SD).
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图7   BnA7HSP70超表达植株表现出对衣霉素诱导细胞死亡的耐受性
(A)转基因和非转基因植株种子和幼苗对2.5 µg mL-1或者5.0 µg mL-1衣霉素的敏感性实验。(B)~(C) Evans Blue染色方法测定转基因和非转基因植株在衣霉素处理后7 d、14 d的细胞死亡程度。Abs (600 nm)测定值反映死细胞含量, 每个实验重复3次。
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