Investigation of the genes associated with a male sterility mutant (msm) in Chinese cabbage (Brassica campestris ssp. pekinensis) using RNA-Seq
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In Chinese cabbage, hybrid seed production is performed using male sterility lines, an important approach to heterosis utilization. In this study, a stably inherited male sterile mutant msm was obtained from the ‘FT’-doubled haploid line of Chinese cabbage using isolated microspore culture combined with 60Co γ-ray mutagenesis. The genetic backgrounds of ‘FT’ and msm were highly consistent; however, compared with wild-type ‘FT’, msm exhibited completely degenerated stamens and no pollen phenotype. Other characters showed no significant differences. Cytological observations revealed that stamen abortion in msm begins during the tetrad period and that tapetum cells were abnormally expanded and highly vacuolated, leading to microspore abortion. Genetic analysis indicated that the msm mutant phenotype is controlled by a single recessive nuclear gene. Comparative transcriptome analysis of ‘FT’ and msm flower buds using RNA-Seq technology revealed 1653 differentially expressed genes, among which, a large number associated with male sterility were detected, including 64 pollen development- and pollen tube growth-related genes, 94 pollen wall development-related genes, 11 phytohormone-related genes, and 16 transcription factor-related genes. An overwhelming majority of these genes were down-regulated in msm compared with ‘FT’. Furthermore, KEGG pathway analysis indicated that a variety of carbohydrate metabolic and lipid metabolic pathways were significantly enriched, which may be related to pollen abortion. The expression patterns of 24 male sterility-related genes were analyzed using qRT-PCR. In addition, 24,476 single-nucleotide polymorphisms and 413,073 insertion–deletion events were specifically detected in msm. These results will facilitate elucidation of the regulatory mechanisms underlying male sterility in Chinese cabbage.
KeywordsChinese cabbage Male sterility mutant DEGs RNA-Seq
This work was supported by grants from the National Natural Science Foundation of China (31801854) and the Doctoral Start-up Foundation of Liaoning Province (20170520096).
HF, SNH, and ZYL conceived and designed the research; SLP, CYL, and TC performed the experiments; SNH, RPY, DYL, XL, and LH analyzed the data; SNH wrote the paper. All authors approved the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
The transcriptome sequencing data have been deposited in the NCBI Gene Expression Omnibus (GEO) Database under Accession Number GSE125485.
This article does not contain any studies with human participants or animals performed by any of the authors.
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