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BcHTT4 Inhibits Branching of Non-Heading Chinese Cabbage at the Vegetative Stage.

ABSTRACT: Branching is speculated to contribute to the plant architecture and crop yield. As a quantitative trait, branching is regulated by multiple genes in non-heading Chinese cabbage (NHCC). Several related candidate genes have been discovered in previous studies on the branching of NHCC, but their specific functions and regulatory mechanisms still need to be verified and explored. In this study, we found that the expression of BcHTT4, the ortholog to HEAT-INDUCED TAS1 TARGET4 (HTT4) in Arabidopsis, was significantly different between 'Suzhouqing' (common type) and 'Maertou' (multiple shoot branching type) in NHCC, which was consistent with the previous transcriptome sequencing results. The silencing of BcHTT4 expression in non-heading Chinese cabbage promotes axillary bud growth at the vegetative stage. When BcHTT4 is overexpressed in Arabidopsis, branching will decrease. In further study, we found that BcHTT4 interacts with immunophilin BcFKBP13 in vivo and in vitro through yeast two-hybrid analysis and bimolecular fluorescence complementation (BiFC) assays. Moreover, quantitative real-time PCR analysis showed that when the expression of BcHTT4 was silenced in 'Suzhouqing', the expression of BcFKBP13 also decreased significantly. Our findings reveal that BcHTT4 is involved in the branching mechanism and interacts with immunophilin BcFKBP13 in NHCC.

SUBMITTER: Guo M

PROVIDER: S-EPMC7999546 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundNon-heading Chinese cabbage (Brassica rapa ssp. chinensis) is an important leaf vegetable grown worldwide. However, there has currently been not enough transcriptome and small RNA combined sequencing analysis of cold tolerance, which hinders further functional genomics research.ResultsIn this study, 63.43 Gb of clean data was obtained from the transcriptome analysis. The clean data of each sample reached 6.99 Gb, and the basic percentage of Q30 was 93.68% and above. The clean reads of each sample were sequence aligned with the designated reference genome (Brassica rapa, IVFCAASv1), and the efficiency of the alignment varied from 81.54 to 87.24%. According to the comparison results, 1,860 new genes were discovered in Pak-choi, of which 1,613 were functionally annotated. Among them, 13 common differentially expressed genes were detected in all materials, including seven upregulated and six downregulated. At the same time, we used quantitative real-time PCR to confirm the changes of these gene expression levels. In addition, we sequenced miRNA of the same material. Our findings revealed a total of 34,182,333 small RNA reads, 88,604,604 kinds of small RNAs, among which the most common size was 24 nt. In all materials, the number of common differential miRNAs is eight. According to the corresponding relationship between miRNA and its target genes, we carried out Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis on the set of target genes on each group of differentially expressed miRNAs. Through the analysis, it is found that the distributions of candidate target genes in different materials are different. We not only used transcriptome sequencing and small RNA sequencing but also used experiments to prove the expression levels of differentially expressed genes that were obtained by sequencing. Sequencing combined with experiments proved the mechanism of some differential gene expression levels after low-temperature treatment.ConclusionIn all, this study provides a resource for genetic and genomic research under abiotic stress in Pak-choi.

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BcHTT4 Inhibits Branching of Non-Heading Chinese Cabbage at the Vegetative Stage.

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