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伊朗大蒜(Allium sativum L.)基因型的微繁殖与体细胞克隆变异

Micropropagation and somaclonal variation in Iranian genotypes of garlic (Allium sativum L.).

作者信息

Afzaz Mohammad-Hossein, Mozafari Javad, Sanjari Sepideh

机构信息

Genetic Research Department, Seed and Plant Improvement Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran.

Agricultural Research, Education and Extension Organization, Tehran, Iran.

出版信息

PLoS One. 2025 Sep 10;20(9):e0331782. doi: 10.1371/journal.pone.0331782. eCollection 2025.

Abstract

Garlic is an important bulb vegetable which is used for both culinary and medical purposes worldwide. In vitro propagation is considered a promising technic for production and conservation of disease-free garlic seed. The efficiency of in vitro culture was studied for micropropagation of native Iranian garlic genotypes. A factorial experiment based on a completely randomized design with three replications was conducted to optimize the in vitro culture media components for establishment, regeneration and conservation of four Iranian garlic genotypes. The highest number of bulblets were obtained and established on MS medium supplemented with 1.5 mgL-1 BA (benzyl adenine) + 0.5 mgL-1 IBA (indole-3-butyric acid) and 0.5 mgL-1 2-iP (2-isopentenyl adenine) + 0.25 mgL-1 NAA (naphthalene acetic acid), respectively. In the regeneration phase, however, the highest number of bulblets regenerated from in vitro grown plants on MS medium supplemented with 0.5 mgL-1 2-iP + 0.1 mgL-1 NAA. A drastic increase in bulblet formation was also observed on this culture medium during conservation phase. At least 20 bulblets were formed per each explant in all genotypes in the first subculture. Interestingly, the bulblet formation in the second subculture was 4-5 times more than the first subculture, indicating a very efficient regeneration rate in micropropagation of Iranian garlic genotypes. Moreover, RAPD (Randomly Amplified Polymorphic DNA) markers were used to evaluate the genetic stability in regenerated garlic plantlets. All fragments amplified by five RAPDs were the same in regenerated plantlets and their mother plants showing no somaclonal variation in micropropagated Iranian garlics. Our results indicated that in vitro protocols used in this study can provide an efficient system for regeneration and conservation of garlic germplasm in an in vitro gene-bank.

摘要

大蒜是一种重要的鳞茎类蔬菜,在全球范围内用于烹饪和医疗目的。离体繁殖被认为是生产和保存无病大蒜种子的一种有前景的技术。研究了伊朗本土大蒜基因型微繁殖的离体培养效率。进行了一项基于完全随机设计、有三次重复的析因实验,以优化四种伊朗大蒜基因型的离体培养基成分,用于其建立、再生和保存。分别在添加了1.5 mgL-1 BA(苄基腺嘌呤)+ 0.5 mgL-1 IBA(吲哚-3-丁酸)和0.5 mgL-1 2-iP(2-异戊烯基腺嘌呤)+ 0.25 mgL-1 NAA(萘乙酸)的MS培养基上获得并建立了最多数量的小鳞茎。然而,在再生阶段,在添加了0.5 mgL-1 2-iP + 0.1 mgL-1 NAA的MS培养基上,从离体生长的植株再生出的小鳞茎数量最多。在保存阶段,在这种培养基上也观察到小鳞茎形成急剧增加。在第一次继代培养中,所有基因型的每个外植体至少形成20个小鳞茎。有趣的是,第二次继代培养中的小鳞茎形成比第一次继代培养多4至5倍,表明伊朗大蒜基因型微繁殖中的再生率非常高。此外,使用RAPD(随机扩增多态性DNA)标记来评估再生大蒜植株的遗传稳定性。由五个RAPD扩增的所有片段在再生植株及其母本中相同,表明微繁殖的伊朗大蒜中没有体细胞克隆变异。我们的结果表明,本研究中使用的离体方案可以为在离体基因库中再生和保存大蒜种质提供一个有效的系统。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4792/12422459/482620c97e9a/pone.0331782.g001.jpg

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