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利用RAPD标记对菠萝蜜树(Artocarpus heterophyllus L.)微繁殖炼苗植株进行克隆保真度研究。

Clonal fidelity investigation of micropropagated hardened plants of jackfruit tree (Artocarpus heterophyllus L.) with RAPD markers.

作者信息

Kader Abdul, Sinha Sankar Narayan, Ghosh Parthadeb

机构信息

Department of Botany, University of Kalyani, Kalyani, Nadia, West Bengal, India.

出版信息

J Genet Eng Biotechnol. 2022 Oct 20;20(1):145. doi: 10.1186/s43141-022-00426-0.

Abstract

BACKGROUND

Artocarpus heterophyllus is an important tropical agroforestry species that bears multiple applications. However, the population of this species is reduced due to various anthropogenic activities. For this reason, in vitro approach is needed to propagate or conserve this species as in vivo propagation methods face various obstacles. In this respect, the present investigation was undertaken to produce genetically stable jackfruit trees through in vitro technology. In vivo grew shoot tips were harvested on Murashige and Skoog (MS) medium containing several plant growth regulators to achieve this.

RESULTS

The 6-benzylaminopurine (BAP) at the concentration of 1.5 mg L, indole-3-butyric acid (IBA) at 0.5 mg L, and α-naphthaleneacetic acid (NAA) at 0.1 mg L in combination on MS media yielded superior shoot response (94.44%), longest shoot length (4.02 cm), and the maximum number of shoots per explant (4.78). They were further multiplied by repeated subculturing on the same media composition and the third subculture resulted in a maximum number of shoots (5.92) with the largest shoot length (5.85 cm). Among the different media screened for rooting, the ¼ MS media yielded 94.44% rooting response, the longest root length (3.78 cm), and the maximum number of roots per shoot (8.44) with 0.1 mg L NAA, 0.5 mg L IBA and 0.1 mg L BAP in combination. Primary hardening showed 88.89% of plant survival under greenhouse conditions after 4 weeks of incubation having a sterilized mixture of garden soil and vermiculite mixture (1:1, w/w). It increased to 90.60% after the secondary hardening process in a vermiculite-soil mixture (2:1; w/w). No polymorphism was detected on random amplification of polymorphic DNA (RAPD) profiling between the mother plant and hardened plants, indicating high genetic stability among the clones.

CONCLUSIONS

This is the first report of the genetic fidelity study of in vitro grown regenerants of A. heterophyllus. This study established a micropropagation protocol for genetically uniform in vitro regeneration of this species to supply plant resources to various industries or conservation of elite germplasm.

摘要

背景

波罗蜜是一种重要的热带农林物种,具有多种用途。然而,由于各种人为活动,该物种的种群数量减少。因此,由于体内繁殖方法面临各种障碍,需要采用体外方法来繁殖或保护该物种。在这方面,本研究旨在通过体外技术培育遗传稳定的波罗蜜树。为实现这一目标,在含有多种植物生长调节剂的Murashige和Skoog(MS)培养基上采集体内生长的茎尖。

结果

在MS培养基上,浓度为1.5 mg/L的6-苄基腺嘌呤(BAP)、0.5 mg/L的吲哚-3-丁酸(IBA)和0.1 mg/L的α-萘乙酸(NAA)组合产生了优异的芽响应(94.44%)、最长的芽长度(4.02 cm)和每个外植体的最大芽数(4.78)。通过在相同培养基组成上反复继代培养进一步增殖,第三次继代培养产生了最大数量的芽(5.92)和最大的芽长度(5.85 cm)。在筛选的不同生根培养基中,¼ MS培养基在含有0.1 mg/L NAA、0.5 mg/L IBA和0.1 mg/L BAP组合时,生根响应率为94.44%,最长根长度为3.78 cm,每个芽的最大根数为8.44。初次炼苗显示,在温室条件下,经过4周培养后,使用灭菌的园土和蛭石混合物(1:1,重量/重量),植株存活率为88.89%。在蛭石-土壤混合物(2:1;重量/重量)中进行二次炼苗后,存活率提高到90.60%。在母株和炼苗植株之间的随机扩增多态性DNA(RAPD)分析中未检测到多态性,表明克隆之间具有高度的遗传稳定性。

结论

这是关于体外培养的波罗蜜再生植株遗传保真度研究的首次报道。本研究建立了该物种遗传一致的体外再生微繁殖方案,可为各行业提供植物资源或保护优良种质。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d77/9583975/25d139a2f8e7/43141_2022_426_Fig1_HTML.jpg

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