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BrWAX2 plays an essential role in cuticular wax biosynthesis in Chinese cabbage (Brassica rapa L. ssp. pekinensis)

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Map-based cloning was used to identify the BrWAX2 gene, which was involved in the cuticular wax biosynthesis. The malfunction of BrWAX2 together with other reduced expression of genes in alkane-forming pathway caused the glossy phenotype.

Abstract

Cuticular wax covering the outer plant surface plays various roles in protecting against biotic and abiotic stresses. Wax-less mutant shows glossy in stem and leaf surface and plays important roles in enriching Chinese cabbage germplasm resources for breeding brilliant green varieties. However, genes responsible for the glossy trait in Chinese cabbage are rarely reported. In this study, we identified a glossy Chinese cabbage line Y1211-1. Genetic analysis indicated that the glossy trait in Y1211-1 was controlled by a single recessive locus, BrWAX2 (Brassica rapa WAX 2). Using bulked segregant sequencing (BSA-Seq) and kompetitive allele-specific PCR (KASP) assays, BrWAX2 was fine-mapped to an interval of 100.78 kb. Functional annotation analysis, expression analysis, and sequence variation analysis revealed that Bra032670, homologous to CER1 in Arabidopsis, was the most likely candidate gene for BrWAX2. The gene Bra032670 was absent in glossy mutant. Cuticular wax composition analysis and RNA-Seq analysis suggested that the absence of BrWAX2 together with the decreased expression of other genes in alkane-forming pathway reduced the wax amount and caused the glossy phenotype. Furthermore, we developed and validated the functional marker BrWAX2-sp for BrWAX2. Overall, these results provide insight into the molecular mechanism underlying cuticular wax biosynthesis and reveal valuable information for marker-assisted selection (MAS) breeding in Chinese cabbage.

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Acknowledgements

This work was financially supported by Zhongyuan Scholar Program (202101510003), the Modern Agro-Industry Technology Research System (CARS-23-G-16), the National Science Foundation of China (31872945), Sci-Tech Innovation Team of Henan Academy of Agricultural Sciences (2021TD06), and Programs for Science and Technology Development of Henan Province (212102110126).

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  1. Shuangjuan Yang and Honglei Liu contributed equally to this work.

Authors and Affiliations

  1. Institute of Horticulture, Henan Academy of Agricultural Sciences, Graduate T & R Base of Zhengzhou University, Zhengzhou, 450002, China

    Shuangjuan Yang, Honglei Liu, Xiaochun Wei, Yanyan Zhao, Zhiyong Wang, Henan Su, Xiaobin Zhao, Xiao-Wei Zhang & Yuxiang Yuan

  2. School of Agricultural Sciences, Zhengzhou University, Zhengzhou, 450001, China

    Shuangjuan Yang, Honglei Liu, Baoming Tian, Xiao-Wei Zhang & Yuxiang Yuan

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  1. Shuangjuan Yang
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  2. Honglei Liu
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Contributions

YY, X-WZ, and BT conceptualized and designed the experiments. SY, HL, and XW performed the experiments and analyzed the data. YZ, ZW, HS, and XZ participated in phenotypic assays and genetic analysis. SY and HL drafted the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Baoming Tian, Xiao-Wei Zhang or Yuxiang Yuan.

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The authors declare that they have no conflict of interest.

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The authors declare that this study complies with the current laws of the countries in which the experiments were performed.

Accession codes

The raw data from BSA-Seq analysis have been deposited into the Sequence Read Archive (https://www.ncbi.nlm.nih.gov/sra/) with accession number PRJNA751715. The raw data from transcriptome analysis were deposited in the Sequence Read Archive under accession number PRJNA751924. The CDS sequence of Brwax2 from waxy R16-11 was deposited in GenBank under accession number MW707248.

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Communicated by Carlos F. Quiros.

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Yang, S., Liu, H., Wei, X. et al. BrWAX2 plays an essential role in cuticular wax biosynthesis in Chinese cabbage (Brassica rapa L. ssp. pekinensis). Theor Appl Genet 135, 693–707 (2022). https://doi.org/10.1007/s00122-021-03993-x

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