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Fine-mapping and transcriptome analysis of BoGL-3, a wax-less gene in cabbage (Brassica oleracea L. var. capitata)

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Abstract

The great majority of terrestrial plants produce epicuticular wax that is used to protect plants from a variety of biotic and abiotic stresses. Cabbage epicuticular wax is a white crystalline compound of various lipids. Wax-less cabbage has the characteristics of lustrous green leaves and beautiful exterior, which facilitates the brilliant green cabbage breeding. CGL-3 is a spontaneous wax-less mutant identified from cabbage. Genetic analysis indicated that the waxy deficiency of the mutant was controlled by a single dominant gene. To clarify the mechanism of the waxy deficiency, fine-mapping and transcriptome analysis of the wax-less gene, BoGL-3, were carried out in this study. The result of fine mapping showed that the wax-less gene, BoGL-3, was delimited in a 33.5-kb interval which is between the flanking marker C08-98 and the end of chromosome 8. Two cDNA libraries, constructed with wax-less cabbage CGL-3 and the wild-type cabbage WT, were sequenced for screening of the target gene BoGL-3. A total of 8340 genes were identified with significant differential expression between CGL-3 and WT. Among these genes, 3187 were up-regulated and 5153 were down-regulated in CGL-3. With homologous analysis, four differential expressed genes related to wax metabolism were obtained. Among these four genes, only Bol018504 is located within the region of fine-mapping. Bol08504 is homologous to CER1, which encodes fatty acid hydroxylase and plays an important role in wax synthesis in Arabidopsis. However, there was no difference of Bol08504 sequence between CGL-3 and WT. We suggested that Bol018504 was regulated by BoGL-3. The suppression of Bol018504 leads to the reduction of wax. These findings will be helpful to reveal the mechanism of the wax metabolism in cabbage and develop lustrous green cabbage germplasm material.

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Acknowledgements

This work was financially supported by grants from the National Key Research and Development Program (2017YFD0101804), National Natural Science Foundation of China (31672155), the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences (CAAS-ASTIP-2013-IVFCAAS), 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), the Modern Agro-Industry Technology Research System (CARS-25-B-01), and the Project of Science and Technology Commission of Beijing Municipality (Z141105002314020-1).

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  1. Xin Dong and Jialei Ji contributed equally to this work.

Authors and Affiliations

  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

    Xin Dong, Jialei Ji, Limei Yang, Zhiyuan Fang, Mu Zhuang, Yangyong Zhang, Honghao Lv, Yong Wang, Peitian Sun, Jun Tang, Dongming Liu, Yumei Liu & Zhansheng Li

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  1. Xin Dong
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Correspondence to Limei Yang.

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Fig. S1.

Statistics of gene ontology (TIFF 196 kb)

Fig. S2.

Sequence alignment of gDNA of Bol018504 between WT and CGL-3 (TIFF 461 kb)

Fig. S3.

Sequence alignment of cDNA of Bol018504 between WT and CGL-3 (TIFF 775 kb)

Fig. S4.

Sequence alignment of upstream of Bol018504 between WT and CGL-3 (TIFF 271 kb)

Fig. S5.

Sequence alignment of downstream of Bol018504 between WT and CGL-3 (TIFF 1423 kb)

Fig. S6.

Sequence alignment of gDNA of Bol018505 between WT and CGL-3 (TIFF 126 kb)

Fig. S7.

Sequence alignment of gDNA of Bol018506 between WT and CGL-3 (TIFF 173 kb)

Fig. S8.

Sequence alignment of gDNA of Bol018507 between WT and CGL-3 (TIFF 505 kb)

Fig. S9.

Sequence alignment of gDNA of Bol018508 between WT and CGL-3 (TIFF 158 kb)

Table S1.

SSR polymorphism primers obtained from scanning by parents and F1 (DOCX 22 kb)

Table S2.

Sequences of the primers for detecting candidate gene (DOCX 14 kb)

Table S3.

Gene annotation of candidate region (DOCX 17 kb)

Table S4.

Data filtering statistics analysis (DOCX 14 kb)

Table S5.

Statistics of different genes expression among groups (XLSX 5203 kb)

Table S6.

The significant KEGG pathways (XLSX 9 kb)

Table S7.

DEGs concerning wax biosynthesis (XLSX 20 kb)

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Dong, X., Ji, J., Yang, L. et al. Fine-mapping and transcriptome analysis of BoGL-3, a wax-less gene in cabbage (Brassica oleracea L. var. capitata). Mol Genet Genomics 294, 1231–1239 (2019). https://doi.org/10.1007/s00438-019-01577-5

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