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Analysis of the Candidate Genes and Underlying Molecular Mechanism of P198, an RNAi-Related Dwarf and Sterile Line

文献类型： 外文期刊

作者： Zhao, Shengbo 1 ; Luo, Junling 1 ; Tang, Min 1 ; Zhang, Chi 1 ; Song, Miaoying 1 ; Wu, Gang 1 ; Yan, Xiaohong 1 ;

作者机构： 1.Chinese Acad Agr Sci, Key Lab Biol & Genet Improvement Oil Crops, Minist Agr & Rural Affairs, Oil Crops Res Inst, Wuhan 430062, Peoples R China

2.Huazhong Agr Univ, Coll Life Sci & Technol, Wuhan 430070, Peoples R China

3.Chinese Acad Agr Sci, Oil Crops Res Inst, Key Lab Agr Genet Modified Organisms Traceabil, Minist Agr & Rural Affairs, Wuhan 430062, Peoples R China

4.Minist Agr & Rural Affairs, Supervis & Test Ctr Wuhan Plant Ecol Environm Safe, Wuhan 430062, Peoples R China

关键词： RNAi; dwarfism; sterility; T-DNA; molecular characteristic; ONT sequencing; transcriptome

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.6； 五年影响因子：6.2 ）

ISSN： 1661-6596

年卷期： 2024 年 25 卷 1 期

页码：

收录情况： SCI

摘要： The genome-wide long hairpin RNA interference (lhRNAi) library is an important resource for plant gene function research. Molecularly characterizing lhRNAi mutant lines is crucial for identifying candidate genes associated with corresponding phenotypes. In this study, a dwarf and sterile line named P198 was screened from the Brassica napus (B. napus) RNAi library. Three different methods confirmed that eight copies of T-DNA are present in the P198 genome. However, only four insertion positions were identified in three chromosomes using fusion primer and nested integrated polymerase chain reaction. Therefore, the T-DNA insertion sites and copy number were further investigated using Oxford Nanopore Technologies (ONT) sequencing, and it was found that at least seven copies of T-DNA were inserted into three insertion sites. Based on the obtained T-DNA insertion sites and hairpin RNA (hpRNA) cassette sequences, three candidate genes related to the P198 phenotype were identified. Furthermore, the potential differentially expressed genes and pathways involved in the dwarfism and sterility phenotype of P198 were investigated by RNA-seq. These results demonstrate the advantage of applying ONT sequencing to investigate the molecular characteristics of transgenic lines and expand our understanding of the complex molecular mechanism of dwarfism and male sterility in B. napus.

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