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利用RAPD和ISSR标记对箣竹(Roxb.)Nees进行微繁殖及遗传保真度评估

Micropropagation and assessment of genetic fidelity of Dendrocalamus strictus (Roxb.) nees using RAPD and ISSR markers.

作者信息

Goyal Arvind Kumar, Pradhan Sushen, Basistha Bharat Chandra, Sen Arnab

机构信息

Molecular Cytogenetics Laboratory, Department of Botany, University of North Bengal, Siliguri, 734013, West Bengal, India.

Bamboo Technology, Department of Biotechnology, Bodoland University, Kokrajhar, 783370, Assam, India.

出版信息

3 Biotech. 2015 Aug;5(4):473-482. doi: 10.1007/s13205-014-0244-7. Epub 2014 Aug 9.

Abstract

Dendrocalamus strictus popularly known as 'Male bamboo' is a multipurpose bamboo which is extensively utilized in pharmaceutical, paper, agricultural and other industrial implements. In this study, in vitro regeneration of D. strictus through nodal culture has been attempted. Murashige and Skoog's medium supplemented with 4 mg/l BAP was found to be most effective in shoot regeneration with 3.68 ± 0.37 shoots per explant. The effect of Kn was found to be moderate. These hormones also had considerable effect on the shoot length. The highest shoot length after 6 weeks (3.11 ± 0.41 cm) was noted with 5 mg/l BAP followed by 3.07 ± 0.28 cm with 5 mg/l Kn, while decrease in the shoot length was noted with other treatments. The effect of IBA and NAA individually or in combination at different concentrations on rooting was evaluated. The highest number of root (1.36 ± 0.04) was regenerated on full-strength MS medium supplemented with 3 mg/l NAA, while maximum length of 1.64 ± 0.03 cm of roots was recorded with combination of 1 mg/l IBA and 3 mg/l NAA. Tissue-cultured plants thus obtained were successfully transferred to the soil. The clonal fidelity among the in vitro-regenerated plantlets was assessed by RAPD and ISSR markers. The ten RAPD decamers produced 58 amplicons, while nine ISSR primers generated a total of 66 bands. All the bands generated were monomorphic. These results confirmed the clonal fidelity of the tissue culture-raised D. strictus plantlets and corroborated the fact that nodal culture is perhaps the safest mode for multiplication of true to type plants.

摘要

印度麻竹俗称“雄竹”,是一种多用途竹子,广泛应用于制药、造纸、农业及其他工业器具领域。在本研究中,尝试通过节段培养实现印度麻竹的离体再生。结果发现,添加4 mg/l 苄氨基嘌呤(BAP)的Murashige和Skoog培养基对芽再生最为有效,每个外植体可产生3.68±0.37个芽。发现激动素(Kn)的效果适中。这些激素对芽的长度也有显著影响。6周后芽长最高(3.11±0.41厘米)的是添加5 mg/l BAP的处理,其次是添加5 mg/l Kn的处理,芽长为3.07±0.28厘米,而其他处理的芽长则有所下降。评估了不同浓度的吲哚丁酸(IBA)和萘乙酸(NAA)单独或组合使用对生根的影响。在添加3 mg/l NAA的完全强度MS培养基上再生的根数量最多(1.36±0.04),而1 mg/l IBA和3 mg/l NAA组合处理的根最长,为1.64±0.03厘米。由此获得的组培苗成功移栽到土壤中。通过随机扩增多态性DNA(RAPD)和简单序列重复区间(ISSR)标记评估了离体再生苗之间的克隆稳定性。10个RAPD十聚体产生了58个扩增子,而9个ISSR引物共产生了66条带。所有产生的条带都是单态的。这些结果证实了组培育苗的印度麻竹苗的克隆稳定性,并证实了节段培养可能是繁殖纯种植物最安全的方式这一事实。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/813e/4522732/e8daf0265e53/13205_2014_244_Fig1_HTML.jpg

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