Cloning,Genetic Transformation and Tissue Expression Pattern Analysis of Wheat miR395a Precursor

doi:10.7668/hbnxb.20191103

Abstract

Abstract: miRNA are a group of small non-coding RNAs that play important roles in the process of developmental regulation and biotic and abiotic stress responses in plant. In order to explore the function and mechanism of miR395a in resistance to pathogen in wheat,the mature sequence of miR395a from different species was compared. It was found that miR395a was highly conserved in different species. The mature miR395a in wheat was highly consistent with that in Arabidopsis, tomato and soybean. The pre-miR395a was cloned from wheat leaves and connected to pCambia1301 to construct over-expression vector of pre-miR395a. pre-miR395a was transformed into wheat variety Bainong 207 through Agrobacterium -mediated method. 56 transgenic plants were obtained and 11 positive transformants were determined by using PCR analysis. The tissue expression profiles detected by RT-qPCR indicated that miR395a was differently expressed during the process of grain development among the tested tissue. The expression level of miR395a was highest in leaves at three-leaf and heading stages,followed by that in leaf sheath and the expression level was lowest in the flag leaf. This study obtains wheat transgenic plants overexpressing the precursor of miR395a,and provides gen etic resources for further study of its function in wheat resistance to pathogens.

Key words: Wheat, Pre-miR395a, Construction of over-expression vector, Genetic transformation, Tissue expression profile analysis

CLC Number:

S512.03
Q78

DING Chaojie, GUAN Yuanyuan, LI Chengwei. Cloning,Genetic Transformation and Tissue Expression Pattern Analysis of Wheat miR395a Precursor[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(4): 52-56. doi: 10.7668/hbnxb.20191103.

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http://www.hbnxb.net/EN/Y2020/V35/I4/52

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