Cloning and Stress Resistance Analysis of ADF Gene in Oryza officinalis

doi:10.7668/hbnxb.2017.06.006

Abstract

Abstract: In order to investigate the effect of ADF gene on abiotic stress in Oryza officinalis. Based on the analysis on the drought-stressed transcriptome of Oryza officinalis, OoADF1 was cloned by RT-PCR technique and sequence analysis showed that OoADF1 contains 453 bp-length ORF and was intronless,and OoADF1 was predicted to contain 150 amino acids and have conserved ADF structure.Quantitative RT-PCR analysis showed that OoADF1 was expressed in all seven tissues,including leaf blade,leaf sheath and root at four-leaves stage,leaf blade,leaf sheath,stem and young panicle at heading stage in Oryza officinalis.The highest expression level of OoADF1 was detected in the root at four-leaves stage,while leaf sheath at heading stage.The fold of expression of OoADF1 amounted to 17.9 and 40.0 in the drought-or high NaCl-stressed young seedling,respectively.The coding sequence of OoADF1 was fused to the prokaroytic expression vector pET-28a(+), and the combinant vectors were induced by IPTG and detected by SDS-PAGE.The molecular weight of protein OoADF1 was detected to be 16 kDa,which was consistent with theoretical one.The results of stress tolerance assay showed that the recombinant strain producing OoADF1 fusion protein improved the resistance to the high concentration,indicating that OoADF1 may exert a role in respond to salt stress and protect cells from high-salt harm.These results provides that OoADF1 has a certain degree of response to high salt stress,so that cells from high salt poisoning.

Key words: Oryza officinalis, Rice, Resistance, ADF

CLC Number:

Q78
S511.03

LI Peigang, LI Hongmei, ZHANG Shuai, LI Jinsong, CHEN Zhixiong, LIU Xiangdong. Cloning and Stress Resistance Analysis of ADF Gene in Oryza officinalis[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(6): 37-44. doi: 10.7668/hbnxb.2017.06.006.

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