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在三叶黄芪中过表达查尔酮异构酶 A 基因以刺激芹菜素的产生。

Overexpression of chalcone isomerase A gene in Astragalus trigonus for stimulating apigenin.

机构信息

Department of Genetics, Faculty of Agriculture, Cairo University, Giza, 12613, Egypt.

National Biotechnology Network of Expertise, Cairo, Egypt.

出版信息

Sci Rep. 2021 Dec 17;11(1):24176. doi: 10.1038/s41598-021-03704-y.

Abstract

Apigenin is one of the most studied flavonoids and is widely distributed in the plant kingdom. Apigenin exerts important antioxidant, antibacterial, antifungal, antitumor activities, and anti-inflammatory effects in neurological or cardiovascular disease. Chalcone isomerase A (chiA) is an important enzyme of the flavonoid biosynthesis pathway. In order to enhance the apigenin production, the petunia chi A gene was transformed for Astragalus trigonus. Bialaphos survived plants were screened by PCR, dot blot hybridization and RT-PCR analysis. Also, jasmonic acid, salicylic acid, chitosan and yeast extract were tested to evaluate their capacity to work as elicitors for apigenin. Results showed that yeast extract was the best elicitor for induction of apigenin with an increase of 3.458 and 3.9 fold of the control for calli and cell suspension culture, respectively. Transformed cell suspension showed high apigenin content with a 20.17 fold increase compared to the control and 6.88 fold more than the yeast extract treatment. While, transformed T calli derived expressing chiA gene produced apigenin 4.2 fold more than the yeast extract treatment. It can be concluded that the highest accumulation of apigenin was obtained with chiA transgenic cell suspension system and it can be utilized to enhancement apigenin production in Astragalus trigonus.

摘要

芹菜素是研究最多的类黄酮之一,广泛分布于植物界。芹菜素在神经或心血管疾病中具有重要的抗氧化、抗菌、抗真菌、抗肿瘤活性和抗炎作用。查尔酮异构酶 A(chiA)是类黄酮生物合成途径中的重要酶。为了提高芹菜素的产量,将矮牵牛 chiA 基因转化为三叶草。通过 PCR、点杂交和 RT-PCR 分析筛选出 Bialaphos 存活的植株。还测试了茉莉酸、水杨酸、壳聚糖和酵母提取物,以评估它们作为芹菜素诱导剂的能力。结果表明,酵母提取物是诱导芹菜素产生的最佳诱导剂,对愈伤组织和细胞悬浮培养的诱导分别增加了 3.458 和 3.9 倍。与对照相比,转化细胞悬浮液中的芹菜素含量增加了 20.17 倍,与酵母提取物处理相比,增加了 6.88 倍。而表达 chiA 基因的转化 T 愈伤组织产生的芹菜素比酵母提取物处理增加了 4.2 倍。可以得出结论,chiA 转基因细胞悬浮体系获得了最高的芹菜素积累量,可用于提高三叶草中芹菜素的产量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be8e/8683443/d2d39356f1a3/41598_2021_3704_Fig1_HTML.jpg

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