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Recessive male sterility in cabbage (Brassica oleracea var. capitata) caused by loss of function of BoCYP704B1 due to the insertion of a LTR-retrotransposon

Theoretical and Applied Genetics volume 130, pages 1441–1451 (2017)Cite this article

Abstract

Key message

The LTR-retrotransposon insertion in BoCYP704B1 is proved to be the primary cause of the male sterility in cabbage. Effective allele-specific markers were developed for marker-assisted selection of male sterile gene.

Abstract

83121A is a spontaneous male sterile mutant identified from cabbage. Genetic analysis indicated that male sterility is controlled by a single recessive gene. Pollen wall formation in the 83121A mutant was severely defective, with a lack of sporopollenin or exine. To understand the mechanisms of male sterility in 83121A, transcription analysis using RNA-Seq was carried out in the buds of the male sterile line 83121A and the male fertile line 83121B, which are near-isogenic lines differing only in the fertility trait. Via expression analysis of differentially expressed genes involved in pollen exine development before the bicellular pollen stage, BoCYP704B1 was identified as a candidate gene, which was approximately downregulated 30-fold in 83121A. BoCYP704B1 is a member of the evolutionarily conserved CYP704B family, which is essential for sporopollenin formation. The BoCYP704B1 transcript is specifically detected in the developing anthers of wild-type cabbage. Further sequence analysis revealed that a 5424-bp long terminal repeat-retrotransposon (LTR-RT) was inserted into the first exon of BoCYP704B1 in 83121A, which is not found in wild-type plants. The insertion of LTR-RT not only reduced the expression of BoCYP704B1 but also altered structure of protein encoded by BoCYP704B1. Moreover, linkage analysis showed that the homozygotic mutational BoCYP704B1 always cosegregated with male sterility. These data suggest that the LTR-RT insertion in BoCYP704B1 hinders sporopollenin formation in 83121A leading to male sterility. The allele-specific markers developed in this study were effective for marker-assisted selection of the male sterile gene.

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Acknowledgements

This work was funded by the National Key Research and Development Program (SQ2017ZY030004), the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences (CAAS-ASTIP-2013-IVFCAAS), the Modern Agro-Industry Technology Research System (CARS-25-B-01), the National High Technology Research and Development Program of China (863 Program, 2012AA100101), the Key Projects in the National Science and Technology Pillar Program during the Twelfth Five-Year Plan Period (2012BAD02B01), and the Project of Science and Technology Commission of Beijing Municipality (Z141105002314020-1). The authors thank LetPub (http://www.letpub.com) for its linguistic assistance during the preparation of this manuscript.

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  1. Key Laboratory of Biology and Genetic Improvement of Horticultural Crops, Ministry of Agriculture/Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China

    Jia-lei Ji, Li-mei Yang, Zhi-yuan Fang, Mu Zhuang, Yang-yong Zhang, Hong-hao Lv, Yu-mei Liu & Zhan-sheng Li

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  1. Jia-lei Ji
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Communicated by Maria Laura Federico.

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122_2017_2899_MOESM1_ESM.tif

Supplementary Fig. S1 SEM micrographs of pollen from 83121A plants (a, c) and 83121B plants (b, d). Scale bars = 10 μm (a, b) and 2 μm (TIF 643 KB)

Supplementary Fig. S2 Correlation analysis of gene expression levels between replicate samples (TIF 1840 KB)

122_2017_2899_MOESM3_ESM.tif

Supplementary Fig. S3 Protein phylogeny of the CYP704 family and related P450 enzymes. Phylogenetic analysis was performed using MrBayes v3.2.1 based on the Dayhoff model for 100,000 generations. At, Arabidopsis; Bn, Brassica napus; Mt, Medicago truncatula; Os, rice; Pp, Physcomitrella patens; Pta, Pinus taeda; Ptr, Populus trichocarpa; Sm, Selaginella moellendorffii; Vv, Vitis vinifera; Zm, Zea mays (TIF 532 KB)

122_2017_2899_MOESM4_ESM.tif

Supplementary Fig. S4 Predicted proteins encoded by BoCYP704B1 in wild-type and 83121A. BoCYP704B1 sequence in 83121A is completely changed with the exception of the fragment in black box (TIF 447 KB)

Supplementary Table 1 (DOCX 12 KB)

Supplementary Table 2 (DOCX 14 KB)

Supplementary Table 3 (DOCX 16 KB)

Supplementary Table 4 (DOCX 12 KB)

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Ji, Jl., Yang, Lm., Fang, Zy. et al. Recessive male sterility in cabbage (Brassica oleracea var. capitata) caused by loss of function of BoCYP704B1 due to the insertion of a LTR-retrotransposon. Theor Appl Genet 130, 1441–1451 (2017). https://doi.org/10.1007/s00122-017-2899-z

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