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利用体细胞融合技术开发利用澳大利亚指橘（Citrus australasica）培育耐黄龙病资源。

Utilization of somatic fusion techniques for the development of HLB tolerant breeding resources employing the Australian finger lime (Citrus australasica).

机构信息

Citrus Research and Education Center, University of Florida, Lake Alfred, FL, United States of America.

Faculty of Agriculture, Pomology Department, Mansoura University, Mansoura, Egypt.

出版信息

PLoS One. 2021 Aug 10;16(8):e0255842. doi: 10.1371/journal.pone.0255842. eCollection 2021.

DOI:10.1371/journal.pone.0255842
PMID:34375348
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8354479/
Abstract

The Australian finger lime is a unique citrus species that has gained importance due to its unique fruit characteristics and perceived tolerance to Huanglongbing (HLB), an often-fatal disease of citrus trees. In this study, we developed allotetraploid finger lime hybrids and cybrids by utilizing somatic cell fusion techniques to fuse diploid 'OLL8' sweet orange or 'Page' tangelo callus-derived protoplasts with finger lime (FL) mesophyll-derived protoplasts. Six somatic fusions were regenerated from the 'OLL8' + FL fusion, while three putative cybrids were regenerated from the 'Page' + FL fusion. Ploidy levels and nuclear-expressed sequence tag derived simple sequence repeat (EST-SSR) markers confirmed the somatic hybrid production, and mitochondrial DNA primer sets confirmed the cybrid nature. Several trees produced by the somatic fusion remained HLB negative even after 6 years of growth in an HLB-endemic environment. Pathogenesis related (PR) and other genes that are often upregulated in HLB-tolerant trees were also upregulated in our somatic fusions. These newly developed somatic fusions and cybrids could potentially be used as breeding parents to develop the next generation of improved HLB-tolerant rootstocks and scions.

摘要

澳大利亚指橙是一种独特的柑橘属物种，由于其独特的果实特性和对黄龙病（HLB）的耐受性而受到重视，HLB 是柑橘树的一种常致命疾病。在这项研究中，我们利用体细胞融合技术，将二倍体 'OLL8' 甜橙或 'Page' 橘柚愈伤组织衍生原生质体与指橙（FL）叶肉衍生原生质体融合，产生了同源四倍体指橙杂种和胞质杂种。从 'OLL8' + FL 融合中再生了 6 个体细胞杂种，而从 'Page' + FL 融合中再生了 3 个可能的胞质杂种。染色体倍性和核表达序列标签衍生的简单重复序列（EST-SSR）标记证实了体细胞杂种的产生，而线粒体 DNA 引物证实了胞质杂种的性质。在 HLB 流行环境中生长 6 年后，一些由体细胞融合产生的植株仍然没有 HLB。在 HLB 耐受植株中常上调的病程相关（PR）和其他基因在我们的体细胞融合中也上调了。这些新开发的体细胞杂种和胞质杂种可能被用作育种亲本，以开发下一代改良的 HLB 耐受砧木和接穗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b025/8354479/da0cf66e8948/pone.0255842.g008.jpg
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