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. 2022 Mar 10:13:850330.
doi: 10.3389/fpls.2022.850330. eCollection 2022.

Fine Mapping and Candidate Gene Analysis of BnC08.cds, a Recessive Gene Responsible for Sepal-Specific Chlorophyll-Deficiency in Brassica napus L

Wei Zhang  1 , Xiaoying Zhou  1 , Feng Chen  1 , Hongli Zhu  1 , Rui Shi  1   2 , Chengming Sun  1 , Song Chen  1 , Maolong Hu  1 , Jiefu Zhang  1   2 , Xiaodong Wang  1
Affiliations

Fine Mapping and Candidate Gene Analysis of BnC08.cds, a Recessive Gene Responsible for Sepal-Specific Chlorophyll-Deficiency in Brassica napus L

Wei Zhang et al. Front Plant Sci. .

Abstract

Chloroplast development is crucial for photosynthesis and plant growth and many factors are involved in its regulation. The regulatory mechanism differs in different green tissues, and previous studies have focused on chloroplasts in leaves. In this study, a mutant with sepal-specific chlorophyll-deficiency was observed in Brassica napus and named as df74. Genetic analysis indicated that the phenotype was controlled by a single recessive nuclear gene. The gene was located on chromosome C08 by bulked-segregant analysis with whole-genome sequencing, which was designated as BnC08.cds. To fine-map the BnC08.cds, a F2 population was created from the cross of df74 and Zhongshuang11 (ZS11). Finally, the BnC08.cds was fine-mapped in the region between the single-nucleotide polymorphism (SNP) markers M5 and M6, corresponding to a 228.72 kb interval of the B. napus "ZS11" genome. Eighteen genes were predicted in the target region, and it was speculated that BnaC08G0442100ZS was the most likely candidate gene based on the results of transcriptome analyses and sequence variation analyses. These results provide a foundation to explore the regulation of chloroplast development in sepals.

Keywords: Brassica napus L; RNA-sequencing; candidate gene; chlorophyll-deficient mutant; fine-mapping; yellow-sepal.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Phenotypic characterization of Ningyou 18 (NY18) and df74. (A) NY18 and df74 at seedling stage; (B) inflorescence of NY18 and df74; (C–F) sepal, petal, stamen, and carpel of NY18 and df74.
Figure 2
Figure 2
Chlorophyll (Chl) contents in sepals of NY18 and df74.
Figure 3
Figure 3
Transmission electron micrographs of chloroplasts from NY18 (A,C) and df74 (B,D). Examples of chloroplast (Cp), plastoglobule (Pg), starch granules (SG), and thylakoid (Thy). Bars: 20 μm (A,B), 2 μm (C,D).
Figure 4
Figure 4
SNP-index (G-pool, Y-pool) and Δ(SNP-index; G-Y) graphs from the BSA-seq analysis. The x axes represent the Brassica napus chromosomes, and the y axes represent the SNP-index or Δ(SNP-index).
Figure 5
Figure 5
Fine-mapping of BnC08.cds. Using 900 recessive individuals of F2 population derived from the cross between df74 and ZS11, the BnC08.cds was mapped to a 228.72 kb interval on Chromosome C08, restricted by PARMS SNP markers M5 and M6.
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