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一种由……介导的用于……的有效体细胞再生和遗传转化方法

An Effective Somatic-Cell Regeneration and Genetic Transformation Method Mediated by for L.

作者信息

Xu Mengyun, Zhao Xinyu, Fang Jiahui, Yang Qinwen, Li Ping, Yan Jian

机构信息

Key Laboratory of Agro-Environment in the Tropics, College of Natural Resources and Environment, South China Agricultural University, Guangzhou 510642, China.

出版信息

Plants (Basel). 2024 Aug 27;13(17):2390. doi: 10.3390/plants13172390.

Abstract

Purslane ( L.) is highly valued for its nutritional, medicinal, and ecological significance. Genetic transformation in plants provides a powerful tool for gene manipulation, allowing for the investigation of important phenotypes and agronomic traits at the genetic level. To develop an effective genetic transformation method for purslane, various organ tissues were used as explants for callus induction and shoot regeneration. Leaf tissue exhibited the highest dedifferentiation and regeneration ability, making it the optimal explant for tissue culture. By culturing on Murashige and Skoog (MS) medium supplemented with varying concentrations of 6-benzyleaminopurine (6-BA) and 1-naphthaleneacetic acid (NAA), somatic cells from leaf explants could be developed into calli, shoots, and roots. The shoot induction results of 27 different purslane accessions elucidated the impact of genotype on somatic-cell regeneration capacity and further confirmed the effectiveness of the culture medium in promoting shoot regeneration. On this basis, a total of 17 transgenic plants were obtained utilizing the genetic transformation method mediated by . The assessment of GUS staining, hygromycin selection, and polymerase chain reaction (PCR) amplification of the transgenic plants as well as their progeny lines indicated that the method established could effectively introduce foreign DNA into the purslane nucleus genome, and that integration was found to be stably inherited by offspring plants. Overall, the present study demonstrates the feasibility and reliability of the -mediated genetic transformation method for introducing and integrating foreign DNA into the purslane genome, paving the way for further research and applications in purslane genetic modification.

摘要

马齿苋(Portulaca oleracea L.)因其营养、药用和生态意义而备受重视。植物基因转化为基因操作提供了强大工具,能够在基因水平上研究重要表型和农艺性状。为开发一种有效的马齿苋基因转化方法,使用了各种器官组织作为外植体进行愈伤组织诱导和芽再生。叶片组织表现出最高的去分化和再生能力,使其成为组织培养的最佳外植体。通过在添加不同浓度6-苄基腺嘌呤(6-BA)和1-萘乙酸(NAA)的Murashige和Skoog(MS)培养基上培养,叶片外植体的体细胞可发育成愈伤组织、芽和根。27个不同马齿苋种质的芽诱导结果阐明了基因型对体细胞再生能力的影响,并进一步证实了该培养基在促进芽再生方面的有效性。在此基础上,利用[具体介导方法]介导的基因转化方法共获得了17株转基因植株。对转基因植株及其后代株系进行的GUS染色、潮霉素筛选和聚合酶链反应(PCR)扩增评估表明,所建立的方法能够有效地将外源DNA导入马齿苋核基因组,并且发现整合可稳定地遗传给后代植株。总体而言,本研究证明了[具体介导方法]介导的基因转化方法将外源DNA导入并整合到马齿苋基因组中的可行性和可靠性,为马齿苋基因改良的进一步研究和应用铺平了道路。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/244a/11396874/1e4d5f64703d/plants-13-02390-g001.jpg

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