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. 2022 Apr 15;22(1):199.
doi: 10.1186/s12870-022-03581-1.

Transcriptome profiling of flower buds of male-sterile lines provides new insights into male sterility mechanism in alfalfa

Bo Xu  1 , Rina Wu  1 , Fengling Shi  2 , Cuiping Gao  1 , Jia Wang  1
Affiliations

Transcriptome profiling of flower buds of male-sterile lines provides new insights into male sterility mechanism in alfalfa

Bo Xu et al. BMC Plant Biol. .

Abstract

Background: The use of heterosis to produce hybrid seeds is a challenge to breeding for improved crop yield. In previous studies, we isolated a male sterile alfalfa hybrid and successfully obtained a genetically stable alfalfa male sterile line through backcrossing, henceforth named MS-4. In this study, we used RNA-seq technology to analyze the transcriptome profiles of the male sterile line (MS-4) and the male fertile line (MF) of alfalfa to elucidate the mechanism of male sterility.

Results: We screened a total of 11,812 differentially expressed genes (DEGs) from both MS-4 and MF lines at three different stages of anther development. Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed that these DEGs are mainly involved in processes such as energy metabolism, lipid and amino acid metabolism, carbohydrate metabolism, in addition to cell synthesis and aging. The results from protein-protein interaction (PPI) network analysis showed that the ribosomal protein (MS.Gene25178) was the core gene in the network. We also found that transcriptional regulation was an influential factor in the development of anthers.

Conclusions: Our findings provide new insights into understanding of the fertility changes in the male sterile (MS-4) of alfalfa.

Keywords: Alfalfa; DEGs; Male sterility; Ribosomal protein; Transcriptome.

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Conflict of interest statement

The authors declare that they have no competing interests. Table S1. An overview of the transcriptome data. Fig. S1. GO functional enrichment analysis of the 2,834 DEGs. Fig. S2. Pathway classification statistics histogram of all DEGs. Table S2. PPI network information. Table S3. Statistics of transcription factor families. Fig. S3. Transcription factors distribution among 2,834 DEGs. Table S4. 17 genes related to tapetum and pollen development. Fig. S4. Relative expression of MS.gene78221 via qRT-PCR. Fig. S5. Relative expression of MS.gene25178 via qRT-PCR.

Figures

Fig. 1
Fig. 1
Statistics of DEGs. A The number of up- and down-regulated DEGs in three developmental stages. B Veen diagram analyses of differentially and stage-specific expression genes at three developmental stages
Fig. 2
Fig. 2
Clustering analysis of the 2,834 DEGs between the MF line and the MS-4 line. A Heat map diagram of the 2,834 DEGs. B Expression patterns of the 2,834 DEGs in the six clusters. The number of genes in each cluster was shown in parentheses
Fig. 3
Fig. 3
GO functional enrichment analysis of DEGs at the three stages of bud development
Fig. 4
Fig. 4
KEGG pathway enrichment analysis of the 2,834 DEGs between MF and MS-4
Fig. 5
Fig. 5
Co-expression network analysis of the partially predicted PPIs. Different colors and shapes represent different groups; the size of the nodes represents the importance in the network, the larger the node, the greater the importance of the gene in the network; the red-framed circle represents the hub gene
Fig. 6
Fig. 6
Distribution of differentially expressed transcription factors
Fig. 7
Fig. 7
Verification of the expression of the selected DEGs was carried out via qRT-PCR. Error bars indicate the standard deviation of three biological replicates
See this image and copyright information in PMC

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