Effects of Tea Polyphenols on Agrobacterium-mediated Plant Genetic Transformation System

LI Jing; LIN Cairong; HUANG Yan; DENG Xuming; WANG Yiqing; SUN Weijang

doi:10.13305/j.cnki.jts.2022.04.003

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Journal of Tea Science ›› 2022, Vol. 42 ›› Issue (4) : 477-490. DOI: 10.13305/j.cnki.jts.2022.04.003

Research Paper

Effects of Tea Polyphenols on Agrobacterium-mediated Plant Genetic Transformation System

LI Jing^1,3,4, LIN Cairong^1,3,4, HUANG Yan^2,3,4, DENG Xuming^1,3,4, WANG Yiqing^1,3,4, SUN Weijang^1,3,4,*

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Abstract

In order to study the effect of tea polyphenols on the infection efficiency of Agrobacterium in tea plant genetic system mediated by Agrobacterium tumefaciens, four Agrobacterium strains LBA4404, EHA105, ATCC15834 and K599 were used as research objects. The phenolic resistance, membrane integrity, adsorption, vir gene and chv gene expression and genetic transformation of Agrobacterium tumefaciens under different concentrations of tea polyphenols were analyzed. The results show that: (1) the phenolic resistance of the four strains followed the order of LBA4404>K599>EHA105>ATCC15834, and EHA105 and ATCC15834 were more sensitive to tea polyphenols. (2) The ratio of ATCC15834 with intact membrane was negatively correlated with the concentration of tea polyphenols and the standing time, and the lowest intact membrane rate of EHA105 appeared 48 h after treatment with 600 mg·L^-1 tea polyphenols. (3) The adsorption capacities of ATCC15834 and EHA105 on tobacco mesophyll cells decreased with the increase of tea polyphenol concentrations. After treatment with tea polyphenols for 24 h, the expressions of chvB and chvB2 in ATCC15834 and EHA105 were significantly inhibited. The expression of virA was positively correlated with the sensitivity to low concentration phenols. (4) After infecting tobacco by the Agrobacterium tumefaciens solution (containing tea polyphenols), the transient and stable transformation efficiencies were inhibited to varying degrees, and the induction rate of hairy roots was greatly reduced. When the concentration of tea polyphenols was 1 000 mg·L^-1, ATCC15834 achieved the lowest root induction rate of 13.85% and the highest necrosis rate of 33.85%. In conclusion, tea polyphenols could reduce the activity and membrane integrity rate of Agrobacterium tumefaciens, and reduce the expression of chv gene to affect its adsorption, which finally suppress the instantaneous conversion efficiency and the stable conversion efficiency in tobacco conversion system to varying degrees.

Key words

chv and vir gene / adsorbability / genetic transformation / tea polyphenol

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LI Jing, LIN Cairong, HUANG Yan, DENG Xuming, WANG Yiqing, SUN Weijang. Effects of Tea Polyphenols on Agrobacterium-mediated Plant Genetic Transformation System[J]. Journal of Tea Science. 2022, 42(4): 477-490 https://doi.org/10.13305/j.cnki.jts.2022.04.003

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