Rapid Generation of Barley Homozygous Transgenic Lines Based on Microspore Culture: HvPR1 Overexpression as an Example
文献类型： 外文期刊
作者： Chen, Zhiwei 1 ; Jiang, Qi 1 ; Guo, Guimei 1 ; Shen, Qiufang 2 ; Yang, Jun 3 ; Wang, Ertao 3 ; Zhang, Guoping 2 ; Lu, Ruiju 1 ; Liu, Chenghong 1 ;
作者机构： 1.Shanghai Acad Agr Sci, Biotechnol Res Inst, Shanghai Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China
2.Zhejiang Univ, Inst Crop Sci, Coll Agr & Biotechnol, Hangzhou 310058, Peoples R China
3.Chinese Acad Sci, Natl Key Lab Plant Mol Genet, CAS Ctr Excellence Mol Plant Sci, Inst Plant Physiol & Ecol,SIBS, Shanghai 200032, Peoples R China
关键词： Hordeum vulgare L; microspore culture; transformation; doubled haploids; pathogenesis-related-1 gene
期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.6； 五年影响因子：6.2 ）
ISSN：
年卷期： 2023 年 24 卷 5 期
页码：
收录情况： SCI
摘要： Obtaining homozygous lines from transgenic plants is an important step for phenotypic evaluations, but the selection of homozygous plants is time-consuming and laborious. The process would be significantly shortened if anther or microspore culture could be completed in one generation. In this study, we obtained 24 homozygous doubled haploid (DH) transgenic plants entirely by microspore culture from one T-0 transgenic plant overexpressing the gene HvPR1 (pathogenesis-related-1). Nine of the doubled haploids grew to maturity and produced seeds. qRCR (quantitative real-time PCR) validation showed that the HvPR1 gene was expressed differentially even among different DH1 plants (T-2) from the same DH0 line (T-1). Phenotyping analysis suggested that the overexpression of HvPR1 inhibited nitrogen use efficiency (NUE) only under low nitrogen treatment. The established method of producing homozygous transgenic lines will enable the rapid evaluation of transgenic lines for gene function studies and trait evaluation. As an example, the HvPR1 overexpression of DH lines also could be used for further analysis of NUE-related research in barley.
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