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.
Highlights
With the increasing population in the world, the problem of food production is becoming more and more prominent and needs to be solved urgently
The quantitative real-time PCR analysis (Figure 1) showed that BcHTT4 was expressed in all the samples tested from
Through further quantitative real-time PCR analysis, we found that pTY-BcHTT4 plasmids, but there was no axillary bud in ‘Suzhouqing’ which grew natthe expression of BcHTT4 decreased significantly in the plants infected with pTY-BcHTT4 urally and was infected by pTY-S plasmids (Figure 3B)
Summary
With the increasing population in the world, the problem of food production is becoming more and more prominent and needs to be solved urgently. Multiceps Hort has unique growth characteristics and a large number of branches are formed at the vegetative stage. Hort provides an ideal material for the study of branching mechanisms of NHCC at the vegetative stage. In previous transcriptome sequencing analysis of mixed tissues, including roots, stems, leaves, flowers, and pods of the five varieties of NHCC, we found that the expression of BcHTT4, the ortholog to HEAT-INDUCED TAS1 TARGET4 (HTT4) in Arabidopsis, in var. The expression of BcHTT4 was down-regulated in var. Multiceps Hort, but up-regulated in the other four varieties This suggests that BcHTT4 may participate in the branching regulation of NHCC to some extent, but this needs further verification. We found that BcHTT4 plays a negative role in branching through utilizing VIGS technology in ‘Suzhouqing’ and overexpression in Arabidopsis. When the expression of HTT4 was silenced in ‘Suzhouqing’, the expression of FKBP13 was down-regulated
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