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离体培养植物的微繁殖方案及遗传稳定性

Micropropagation Protocol and Genetic Stability of the Plants Cultivated In Vitro.

作者信息

Parzymies Marzena, Pogorzelec Magdalena, Głębocka Katarzyna, Sliwinska Elwira

机构信息

Institute of Horticultural Production, University of Life Sciences in Lublin, 20-612 Lublin, Poland.

Department of Hydrobiology and Protection of Ecosystems, University of Life Sciences in Lublin, 20-262 Lublin, Poland.

出版信息

Biology (Basel). 2023 Jan 20;12(2):168. doi: 10.3390/biology12020168.

Abstract

L. is a relict species, threatened with extinction in many European countries. To prevent the loss of the species, tissue culture was established to produce plant material for reintroduction in natural habitats. Micropropagation was chosen as a method to obtain new plants. shoots were disinfected with NaOCl, AgNO, or with a two-step disinfection with NaOCl, and then placed on MS medium supplemented with BA at 1 mg·dm and IBA at 0.1 mg·dm. Regenerated shoots were cultivated in presence of BA, KIN, and 2iP to select the treatment with the highest multiplication rate. The obtained plants were acclimatized to ex vitro conditions. Inter-simple sequence repeat (ISSR) and flow cytometric analyses were conducted on in vitro regenerated plants to check their genetic stability. The best disinfection results were obtained when explants were treated with 1.5% NaOCl for 20 min. The highest multiplication rate and good quality plants were noted in the control media, without growth regualtors and in presence of kinetin at 0.5 mg·dm. Flow cytometry and ISSR analyses confirmed genetic stability in plantlets, which indicated the possibility to use the in vitro obtained plants for reintroduction.

摘要

L.是一种残遗物种,在许多欧洲国家面临灭绝威胁。为防止该物种消失,建立了组织培养以生产用于重新引入自然栖息地的植物材料。选择微繁殖作为获取新植物的方法。将芽用次氯酸钠、硝酸银消毒,或用次氯酸钠进行两步消毒,然后置于添加了1毫克·分米⁻³的苄氨基嘌呤和0.1毫克·分米⁻³的吲哚丁酸的MS培养基上。在苄氨基嘌呤、激动素和2-异戊烯基腺嘌呤存在的情况下培养再生芽,以选择增殖率最高的处理。将获得的植物驯化至离体条件。对离体再生植物进行简单序列重复区间(ISSR)和流式细胞术分析,以检查其遗传稳定性。当外植体用1.5%次氯酸钠处理20分钟时,获得了最佳消毒效果。在无生长调节剂且含有0.5毫克·分米⁻³激动素的对照培养基中,观察到最高的增殖率和优质的植物。流式细胞术和ISSR分析证实了小植株的遗传稳定性,这表明有可能将离体获得的植物用于重新引入。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a4bf/9952878/9f92c1fdbf0c/biology-12-00168-g001.jpg

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