发根农杆菌介导龙葵遗传转化体系优化

doi:10.7525/j.issn.1673-5102.2026.01.010

摘要/Abstract

摘要：

该研究旨在通过建立发根农杆菌（Agrobacterium rhizogenes）介导的龙葵（Solanum nigrum）遗传转化体系，提高遗传转化效率及总黄酮含量。研究以龙葵叶片为外植体，探究菌株类型、菌液浓度等因素对龙葵毛状根诱导率的影响。基于该体系，构建重组质粒PBI121-6flag-SnCHI，通过农杆菌介导，诱导转SnCHI基因的毛状根，考察龙葵毛状根转SnCHI基因后的总黄酮含量变化。研究结果表明：菌株Ar.1193在OD₆₀₀为1.0、侵染25 min、共培养24 h条件下，龙葵毛状根诱导率可达88.49%。对转SnCHI基因毛状根进行qRT-PCR检测，发现其基因表达水平是空白组的28.81倍；测定转SnCHI基因毛状根的总黄酮含量，发现转SnCHI基因毛状根的总黄酮含量最高达空白组的4.14倍。成功建立龙葵遗传转化体系，对SnCHI基因的功能进行研究，为同属植物的组织培养和遗传转化研究提供参考。

关键词: 龙葵, 毛状根, 发根农杆菌, 遗传转化体系, 黄酮类化合物

Abstract:

The aim of this study was to establish an Agrobacterium rhizogenes-mediated genetic transformation system for Solanumnigrum（black nightshade） with the goal of enhancing transformation efficiency and increasing its total flavonoid content.Based on this system， the recombinant plasmid PBI121-6flag-SnCHI was constructed， the hairy roots of SnCHI gene transformation were induced by Agrobacterium-mediated pathway， and the content of total flavonoids in the hairy roots of S.nigrum after SnCHI gene transformation was investigated. The results showed that the induction rate of hairy roots of S. nigrum could reach 88.49% under the conditions of OD₆₀₀ value of 1.0， infection for 25 min and co-culture for 24 h. The qRT-PCR detection of transgenic SnCHI hairy roots showed that the gene expression level was 28.81 times that of the blank group. The total flavonoid content of SnCHI transgenic hairy roots was determined， and it was found that the total flavonoid content of SnCHI transgenic hairy roots was up to 4.14 times that of the blank group. The genetic transformation system of S. nigrum was successfully established， and the function of SnCHI gene was studied， which provided a reference for tissue culture and genetic transformation of the same genus.

Key words: Solanum nigrum, hairy root, Agrobacterium rhizogenes, genetic transformation system, flavonoids

中图分类号:

S567

张静静, 骆海霞, 洪艳苹, 赵世成. 发根农杆菌介导龙葵遗传转化体系优化[J]. 植物研究, 2026, 46(1): 111-119.

Jingjing ZHANG, Haixia LUO, Yanping HONG, Shicheng ZHAO. Optimization of Agrobacterium rhizogenes-Mediated Genetic Transformation System of Solanum nigrum[J]. Bulletin of Botanical Research, 2026, 46(1): 111-119.

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引物名称 Primer name	引物序列 Primer sequences（5′→3′）
rolB-F	GAAGGTGCAAGCTACCTCTC
rolB-R	GCTCTTGCAGTGCTAGATTT
rolC-F	CTCCTGACATCAAACTCGTC
rolC-R	TGCTTCGAGTTATGGGTACA

引物名称 Primer name	引物序列 Primer sequences（5′→3′）	产物长度 Product length /bp
qRT-SnActin-F	GCTCCTGAAGAACACCCT	128
qRT-SnActin-R	ACGGCCTGGATAGCAACATA	128
qRT-SnCHI-F	TACTCTGAGAAGGTGGCGGA	140
qRT-SnCHI-R	AAAAGGATGGAGGCACCAGG	140