茶多酚对农杆菌介导的植物遗传转化体系的影响

李晶; 林彩容; 黄艳; 邓旭铭; 王艺清; 孙威江

doi:10.13305/j.cnki.jts.2022.04.003

茶叶科学 >

2022 , Vol. 42 >Issue 4: 477 - 490

DOI: https://doi.org/10.13305/j.cnki.jts.2022.04.003

研究报告

茶多酚对农杆菌介导的植物遗传转化体系的影响

李晶 ,
林彩容 ,
黄艳 ,
邓旭铭 ,
王艺清 ,
孙威江

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1.福建农林大学园艺学院,福建福州 350002;
2.福建农林大学安溪茶学院,福建泉州 362400;
3.福建省茶产业工程技术研究中心,福建福州 350002;
4.海峡两岸特色作物安全生产省部共建协同创新中心,福建福州 350002

李晶,女,硕士研究生,主要从事茶树栽培育种与生物技术研究。

收稿日期: 2021-12-30

修回日期: 2022-03-29

网络出版日期: 2022-08-23

基金资助

国家自然科学基金（31770732）、福建省高校产学合作项目（2019N5007）、福建主要茶类原产地溯源与标准体系研究（K1520005A04）、福建张天福茶叶发展基金会科技创新基金（FJZTF01）

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

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

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1. College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, China;
2. Anxi College of Tea Science, Fujian Agriculture and Forestry University, Quanzhou 362400, China;
3. Fujian Tea Industry Engineering Technology Research Center, Fuzhou 350002, China;
4. Ministerial and Provincial Joint Innovation Centre for Safety Production of Cross-Strait Crops, Fujian Agriculture and Forestry University, Fuzhou 350002, China

Received date: 2021-12-30

Revised date: 2022-03-29

Online published: 2022-08-23

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摘要

为研究在农杆菌介导的茶树遗传体系中,茶多酚对农杆菌侵染效率的影响,以LBA4404、EHA105、ATCC15834和K599 4个农杆菌菌株为研究对象,分析其在不同浓度茶多酚处理下的耐酚能力、被膜完整率、吸附性、vir和chv基因表达,以及遗传转化差异。结果显示,4个菌株的耐酚能力依次为LBA4404>K599>EHA105>ATCC15834,其中EHA105和ATCC15834菌株对茶多酚较为敏感;ATCC15834菌株被膜完整率与茶多酚的浓度、静置时间呈负相关,EHA105在600 mg·L^-1茶多酚处理48 h后,其被膜完整率最低;ATCC15834和EHA105对烟草叶肉细胞的吸附能力随着茶多酚浓度的升高而降低,在经茶多酚处理24 h后,ATCC15834中chvB和EHA105中chvB2的表达受到显著抑制,virA表达量与低浓度酚类的敏感性呈正相关性;烟草经农杆菌菌液（含茶多酚）侵染后,瞬时转化效率和稳定转化效率均受到不同程度的抑制,发状根诱导率大大降低。1 000 mg·L^-1茶多酚处理后,ATCC15834的发根诱导率最低,仅为13.85%,坏死率高达33.85%。综上所述,茶多酚会降低农杆菌活力和被膜完整率,chv基因通过降低表达量影响其吸附性,最终导致烟草转化体系中瞬时转化效率和稳定转化效率均受到不同程度的抑制。

关键词： 茶多酚; 农杆菌介导; 吸附性; chv和vir基因; 遗传转化

本文引用格式

李晶 , 林彩容 , 黄艳 , 邓旭铭 , 王艺清 , 孙威江 . 茶多酚对农杆菌介导的植物遗传转化体系的影响[J]. 茶叶科学, 2022 , 42(4) : 477 -490 . DOI: 10.13305/j.cnki.jts.2022.04.003

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： tea polyphenol; adsorbability; chv and vir gene; genetic transformation

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