中草药

鸡血藤无色花色素还原酶基因SsLAR2克隆及功能分析

PDF下载

中草药 | 药材与资源 2026,57(14): 5648-5657

收起

中草药 |药材与资源 2026 , 57 (14) : 5648 -5657

鸡血藤无色花色素还原酶基因SsLAR2克隆及功能分析

全屏

熊峥¹, 林泉¹, 李莹¹, 柯芳¹, 乔柱¹, 余丽莹¹, 秦双双¹

作者信息

通讯作者:

余丽莹

作者简介:

熊峥: 熊峥（1988—），硕士，研究实习员，研究方向为中药资源。E-mail:369606419@qq.com

Cloning and functional analysis of leucoanthocyanidin reductase gene SsLAR2 from Spatholobi Caulis

XIONG Zheng, LIN Quan, LI Ying, KE Fang, QIAO Zhu, YU Liying, QIN Shuangshuang

Affiliations

doi: 10.7501/j.issn.0253-2670.2026.14.024

文章导航

摘要

收起

目的克隆鸡血藤（密花豆Spatholobus suberectus的干燥藤茎）无色花色素还原酶基因SsLAR2，开展生物信息学及基因功能研究，以期解析其在儿茶素生物合成中的作用。方法以鸡血藤cDNA作为模板，通过RT-PCR技术克隆SsLAR2，并对其开展系统的生物信息学分析。构建SsLAR2过表达载体，遗传转化本氏烟草，测定转基因植株中总黄酮含量、儿茶素含量以及儿茶素合成途径中关键酶基因的表达水平。结果 SsLAR2基因全长1 092 bp，编码363个氨基酸，相对分子质量为40 513.75，等电点为5.75。SsLAR2蛋白为亲水性蛋白，存在跨膜区域，与木豆、毒羊豆及相思子等多种植物中的LAR蛋白同源性较高。SsLAR2启动子序列中包含光响应元件，以及茉莉酸甲酯、赤霉素、水杨酸等多种植物激素的响应元件。基因功能分析表明，过表达SsLAR2能显著提高转基因植株中总黄酮和儿茶素的含量以及儿茶素合成途径中关键酶基因的表达水平。结论鸡血藤SsLAR2基因能够促进植物的儿茶素合成，为深入解析鸡血藤儿茶素生物合成的调控机制，开展相关的分子育种工作提供重要的基因资源和理论依据。

关键词

鸡血藤 / SsLAR2 / 儿茶素 / 生物信息学 / 基因功能

Abstract

收起

Objective To clone the leucoanthocyanidin reductase gene SsLAR2 from Spatholobi Caulis (dried rattan stem of Spatholobus suberectus), and conduct bioinformatics and function verification, in order to elucidate the role of SsLAR2 gene in the biosynthesis of catechin. Methods SsLAR2 was amplified from the cDNA of Spatholobi Caulisvia RT-PCR and conducted systematic bioinformatics analysis. The SsLAR2 overexpression vector was constructed and genetically transformed into Nicotiana benthamiana. The total flavonoid content, catechin content, and the expression levels of key enzyme genes in the catechin synthesis pathway were determined in the transgenic plants. Results The full length of SsLAR2 gene was 1 092 bp in length, encoding 363 amino acids. The molecular weight of SsLAR2 protein was 40 513.75, and the isoelectric point was 5.75. The SsLAR2 protein was a hydrophilic protein with a transmembrane region, showing high homology with LAR proteins from various plants such as Cajanus cajan, Gastrolobium bilobum and Abrus precatorius. The promoter sequence of SsLAR2 contained photoresponsive elements, as well as responsive elements for multiple plant hormones such as methyl jasmonate, gibberellin, and salicylic acid. Gene functional analysis revealed that overexpression of SsLAR2 significantly increased the total flavonoid and catechin contents, as well as the expression levels of key enzyme genes in the catechin biosynthetic pathway in the transgenic plants. Conclusion Heterologous expression of SsLAR2 can promote catechin biosynthesis in plants. This gene provides important genetic materials and a theoretical basis for in-depth dissection of the regulatory mechanisms underlying catechin biosynthesis and targeted molecular breeding of Spatholobi Caulis.

Key words

Spatholobus suberectus Dunn / SsLAR2 / catechin / bioinformatics / gene function

引用本文

熊峥, 林泉, 李莹, 柯芳, 乔柱, 余丽莹, 秦双双. 鸡血藤无色花色素还原酶基因SsLAR2克隆及功能分析. 中草药, 2026 , 57 (14) : 5648 -5657 . DOI: 10.7501/j.issn.0253-2670.2026.14.024

XIONG Zheng, LIN Quan, LI Ying, KE Fang, QIAO Zhu, YU Liying, QIN Shuangshuang. Cloning and functional analysis of leucoanthocyanidin reductase gene SsLAR2 from Spatholobi Caulis[J]. Chinese Traditional and Herbal Drugs, 2026 , 57 (14) : 5648 -5657 . DOI: 10.7501/j.issn.0253-2670.2026.14.024

基金

收起

国家重点研发计划 (2024YFC3506703); 广西自然科学基金项目 (2025GXNSFAA069354,2020GXNSFBA159006); 广西中医药适宜技术开发与推广项目 (GZSY2025005)

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

中国药典[S]. 一部. 2025: 209-210.
秦双双, 朱艳霞, 韦坤华, 等. 鸡血藤的本草沿革与黄酮类成分及其药理学研究进展[J]. 中国中药杂志, 2018, 43(11): 2216-2223.
Zhang F, Ganesan K, Liu Q Q, et al. A review of the pharmacological potential of Spatholobus suberectus Dunn on cancer [J]. Cells, 2022, 11(18): 2885.
Zhou Z Y, Huan L Y, Zhao W R, et al. Spatholobi Caulis extracts promote angiogenesis in HUVECs in vitro and in zebrafish embryos in vivo via up-regulation of VEGFRs [J]. J Ethnopharmacol, 2017, 200: 74-83.
廖佳伟, 金晨, 陈志, 等. 鸡血藤化学成分、药理作用研究进展及其质量标志物(Q-Marker)预测[J]. 中草药, 2023, 54(20): 6866-6877.
Araguirang G E, Richter A S. Activation of anthocyanin biosynthesis in high light–what is the initial signal? [J]. New Phytol, 2022, 236(6): 2037-2043.
Liu W X, Feng Y, Yu S H, et al. The flavonoid biosynthesis network in plants [J]. Int J Mol Sci, 2021, 22(23): 12824.
Jiang L N, Gao Y F, Han L Q, et al. Designing plant flavonoids: Harnessing transcriptional regulation and enzyme variation to enhance yield and diversity [J]. Front Plant Sci, 2023, 14: 1220062.
Khusnutdinov E, Sukhareva A, Panfilova M, et al. Anthocyanin biosynthesis genes as model genes for genome editing in plants [J]. Int J Mol Sci, 2021, 22(16): 8752.
Qin S S, Wei K H, Cui Z H, et al. Comparative genomics of Spatholobus suberectus and insight into flavonoid biosynthesis [J]. Front Plant Sci, 2020, 11: 528108.
Yan H L, Pei X N, Zhang H, et al. MYB-mediated regulation of anthocyanin biosynthesis [J]. Int J Mol Sci, 2021, 22(6): 3103.
Jiang L Y, Yue M L, Liu Y Q, et al. A novel R2R3-MYB transcription factor FaMYB5 positively regulates anthocyanin and proanthocyanidin biosynthesis in cultivated strawberries (Fragaria×Ananassa) [J]. Plant Biotechnol J, 2023, 21(6): 1140-1158.
Stafford H A. Pathway to proanthocyanidins (condensed tannins), flanvan-3-ols, and unsubstituted flavones [M]. Florida: CRC Press, 1990: 63-100.
Wang P Q, Zhang L J, Jiang X L, et al. Evolutionary and functional characterization of leucoanthocyanidin reductases from Camellia sinensis [J]. Planta, 2018, 247(1): 139-154.
张芷苓, 张媛媛, 林晓蓉, 等. 茶儿茶素合成关键酶基因CsANS和CsLAR的功能鉴定[J]. 园艺学报, 2024, 51(4): 804-814.
Li J L, Zhang Z X, Wang X, et al. Functional identification of LAR genes in apple (Malus halliana) demonstrates that it enhances saline-alkali stress tolerance [J]. Plant Growth Regul, 2024, 104(3): 1575-1588.
Sun S H, Qi X J, Zhang Z Z, et al. A structural variation in the promoter of the leucoanthocyanidin reductase gene AaLAR1 enhances freezing tolerance by modulating proanthocyanidin accumulation in kiwifruit (Actinidia arguta) [J]. Plant Cell Environ, 2024, 47(10): 4048-4066.
Zhong R H, Liu B, Wei J B, et al. Molecular and functional characterization of the key proanthocyanidin pathway enzymes anthocyanidin reductases and leucoantho- cyanidin reductases in Litchi chinensis [J]. J Agric Food Chem, 2024, 72(46): 25972-25986.
张娟娟, 刘雪梅, 李美欢, 等. 基于WGCNA挖掘鸡血藤儿茶素和表儿茶素合成途径关键基因[J]. 中草药, 2025, 56(24): 9124-9134.
Qin S S, Wu L Q, Wei K H, et al. A draft genome for Spatholobus suberectus [J]. Sci Data, 2019, 6: 113.
Qin S S, Wei G L, Lin Q, et al. Analysis of the Spatholobus suberectus full-length transcriptome identified an R2R3-MYB transcription factor-encoding gene SsMYB158 that regulates flavonoid biosynthesis [J]. Plant Physiol Biochem, 2024, 214: 108929.
Qin S S, Liang Y, Wei F, et al. The light-regulated SsMYB106 transcription factor promotes flavonoids in Spatholobus suberectus [J]. Int J Mol Sci, 2025, 26(11): 5292.
Qin S S, Liang Y, Wei G L, et al. Shade responses and resistant mechanisms in Spatholobus suberectus [J]. Heliyon, 2024, 10(6): e28077.
Tanner G J, Francki K T, Abrahams S, et al. Proanthocyanidin biosynthesis in plants purification of legume leucoanthocyanidin reductase and molecular cloning of its cDNA [J]. J Biol Chem, 2003, 278(34): 31647-31656.
Shirley B W, Kubasek W L, Storz G, et al. Analysis of Arabidopsis mutants deficient in flavonoid biosynthesis [J]. Plant J, 1995, 8(5): 659-671.
Maugé C, Granier T, d’Estaintot B L, et al. Crystal structure and catalytic mechanism of leucoanthocyanidin reductase from Vitis vinifera [J]. J Mol Biol, 2010, 397(4): 1079-1091.
Zhang W J, Zhou Y W, Zhang Y, et al. Protein phosphorylation: A molecular switch in plant signaling [J]. Cell Rep, 2023, 42(7): 112729.
Sharma A, Shahzad B, Rehman A, et al. Response of phenylpropanoid pathway and the role of polyphenols in plants under abiotic stress [J]. Molecules, 2019, 24(13): 2452.
Duan S C, Kim J H, Kim C K, et al. Role of methyl jasmonate on flavonoid pathway of UV-B-irradiated apple fruit [J]. J Agric Food Chem, 2024, 72(49): 27139-27149.
Karapınar L, Fereydouni R, Al-Khafaji S. Influence of salicylic acid on phytochemical constituents of halwani and kamali grapevine leaves [J]. J Pharmacogn Phytochem, 2025, 14(2): 605-609.
Cheng J, Yu K J, Zhang M Y, et al. The effect of light intensity on the expression of leucoanthocyanidin reductase in grapevine calluses and analysis of its promoter activity [J]. Genes, 2020, 11(10): 1156.
贾展慧, 贾晓东, 许梦洋, 等. 薄壳山核桃原花青素合成关键酶基因的克隆与表达分析[J]. 南京林业大学学报: 自然科学版, 2022, 46(5): 49-57.
Liao L, Vimolmangkang S, Wei G C, et al. Molecular characterization of genes encoding leucoanthocyanidin reductase involved in proanthocyanidin biosynthesis in apple [J]. Front Plant Sci, 2015, 6: 243.
周平, 范雨昕, 姚红, 等. 中国水仙LAR基因启动子的克隆及功能初步分析[J]. 西北植物学报, 2019, 39(8): 1353-1360.
李军, 王崇敏, 周涛, 等. 血人参原花青素合成关键酶基因IsANR、IsLAR的鉴定及分析[J]. 分子植物育种, 2024, 22(24): 8048-8056.

2026年第57卷第14期

PDF下载

引用本文

BibTeX

文章信息

doi: 10.7501/j.issn.0253-2670.2026.14.024

接收时间：2026-02-20
首发时间：2026-09-10

补充材料

相关文章

文章信息

作者

出版历史

收稿日期：2026-02-20

基金

作者信息

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/zcy/CN/10.7501/j.issn.0253-2670.2026.14.024

分享至

全文二维码

扫描看全文

引用本文

BibTeX

本文的引用情况

2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

关闭全屏

BibTeX
EndNote
RefWorks
TxT