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.

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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).

Author information

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.

Correspondence to Hui Feng.

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The transcriptome sequencing data have been deposited in the NCBI Gene Expression Omnibus (GEO) Database under Accession Number GSE125485.

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Fig. S1

Distribution of gene coverage in ‘FT’ and msm. (TIFF 2060 kb)

Fig. S2

GO functional classification of DEGs in the msm versus ‘FT’ comparison. (TIFF 2226 kb)

Table S1

Primers sequences used for qRT-PCR analysis. (XLS 17 kb)

Table S2

Read statistics based on the RNA-Seq data of six libraries from ‘FT’ and msm. (XLSX 10 kb)

Table S3

List of all genes detected in the ‘FT’ and msm libraries. (XLS 14592 kb)

Table S4

List of DEGs identified in the msm versus ‘FT’ comparison. (XLS 473 kb)

Table S5

List of SEGs identified in the msm versus ‘FT’ comparison. (XLSX 69 kb)

Table S6

Significantly enriched GO terms identified in the msm versus ‘FT’ comparison. (XLS 38 kb)

Table S7

Summary of SNP types identified in ‘FT’ and msm. (XLS 26 kb)

Table S8

SNPs specifically detected in the ‘FT’ library. (XLSX 1054 kb)

Table S9

SNPs specifically detected in the msm library (XLSX 1063 kb)

Table S10

Insertions detected between ‘FT’ and msm libraries. (XLSX 11490 kb)

Table S11

Supplementary material 13 Deletions detected between ‘FT’ and msm libraries. (XLSX 21675 kb)

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Huang, S., Peng, S., Liu, Z. et al. Investigation of the genes associated with a male sterility mutant (msm) in Chinese cabbage (Brassica campestris ssp. pekinensis) using RNA-Seq. Mol Genet Genomics 295, 233–249 (2020) doi:10.1007/s00438-019-01618-z

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  • Chinese cabbage
  • Male sterility mutant
  • DEGs
  • RNA-Seq