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珍稀物种东北茶藨子的离体保存及遗传多样性分析

In Vitro conservation and genetic diversity analysis of rare species Ribes janczewskii.

作者信息

Nurtaza Aidana, Dyussembekova Damira, Islamova Symbat, Samatova Indira, Zhanybekova Zhanargul, Umirzakova Alima, Magzumova Gulmira, Muranets Anna, Kakimzhanova Almagul

机构信息

National Center for Biotechnology, 13/5, Korgalzhyn Road, 010000, Astana, Kazakhstan.

Department of Science, Information and Monitoring, Sairam-Ugam State National Nature Park, 24 1, Ilyaev Street, 160011, Shymkent, Kazakhstan.

出版信息

Sci Rep. 2024 Dec 28;14(1):31117. doi: 10.1038/s41598-024-82320-y.

Abstract

Ribes janczewskii is a rare and valuable plant known for its resistance to spring frosts, pests, and diseases. It is used in hybridization to develop resistant currant varieties but is on the verge of extinction, listed in Kazakhstan Red Book. This study developed a micropropagation and slow-growth storage protocol for conservation. Genotypes were identified through DNA barcode analysis (rbcL, ITS, and matK) and sequences uploaded to the National Center for Biotechnology Information database. Genetic diversity was assessed using iPBS primers, generating 98 fragments with 88-94% polymorphic bands. Biochemical analysis of fruits showed vitamin C content from 4.64 to 5.61 mg/100 g, vitamin E from 2.26 to 3.16 mg/100 g, vitamin B5 from 3.18 to 4.93 mg/100 g, and quercetin up to 12.5 mg/100 g. Micropropagation stages were optimized with 12% hydrogen peroxide for surface sterilization, achieving up to 73.3% explant viability. Effective hormonal combinations for in vitro culture included WPM with BAP 0.2 mg L and GA 0.5 mg L, and for propagation, BAP 0.25 mg L, GA 0.5 mg L, and IBA 0.5 mg L. Mannitol (20 g L) was used for slow-growth storage, keeping explants viable for 4 months without re-cultivation.

摘要

扬氏茶藨子是一种珍稀且有价值的植物,以其抗春霜、抗虫害和抗病能力而闻名。它被用于杂交培育抗性醋栗品种,但已濒临灭绝,被列入哈萨克斯坦红皮书。本研究开发了一种用于保护的微繁殖和缓慢生长保存方案。通过DNA条形码分析(rbcL、ITS和matK)鉴定基因型,并将序列上传至国家生物技术信息中心数据库。使用iPBS引物评估遗传多样性,产生了98个片段,多态性条带比例为88 - 94%。果实的生化分析表明,维生素C含量为4.64至5.61毫克/100克,维生素E为2.26至3.16毫克/100克,维生素B5为3.18至4.93毫克/100克,槲皮素含量高达12.5毫克/100克。微繁殖阶段用12%过氧化氢进行表面灭菌优化,外植体存活率高达73.3%。体外培养的有效激素组合包括添加0.2毫克/升BAP和0.5毫克/升GA的WPM培养基,用于增殖的激素组合为0.25毫克/升BAP、0.5毫克/升GA和0.5毫克/升IBA。使用甘露醇(20克/升)进行缓慢生长保存,外植体无需继代培养可存活4个月。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c65/11680832/5e0b8a20bf0d/41598_2024_82320_Fig1_HTML.jpg

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