Seed weight is a pivotal yield-determining trait in crops,and yet,the genetic and molecular mechanisms underlying its regulation in polyploid species remain underexplored.In a previous study,we identified cqSW.A03-2,a...Seed weight is a pivotal yield-determining trait in crops,and yet,the genetic and molecular mechanisms underlying its regulation in polyploid species remain underexplored.In a previous study,we identified cqSW.A03-2,a QTL that regulates thousand seed weight(TSW)in rapeseed(Brassica napus).Here,we identify BnaA3.AHK2,encoding a histidine kinase,as the causal gene of cqSW.A03-2.BnaA3.AHK2 enhances TSW through maternal control of seed coat cell expansion without significantly compromising other yield-related traits.Protein sequence divergence between parental haplotypes caused functional differentiation,with only the ZY50 allele showing functional kinase activity and rescuing developmental defects in Arabidopsis cytokinin receptor mutants.Strikingly,BnaA3.AHK2 seems to be a cytokinin-independent operator,contrasting with the canonical cytokinin signaling pathway.Transcriptome and protein interaction analyses reveal a signaling module where BnaA3.AHK2 engages BnaAHP-Bna ARR phosphorelay components to regulate downstream targets.Notably,the favorable cqSW.A03-2 haplotype has been historically selected in modern breeding,and its introgression into elite hybrids boosted TSW by 3.6%-9.1%,demonstrating its breeding value.Our findings unveil a non-canonical signaling pathway for seed size regulation,providing a strategic genetic target to break yield trade-offs in polyploid crops.展开更多
为进一步解析甘蓝型油菜温敏细胞核雄性不育系(GMS)TE5A的不育机理,筛选与育性蛋白Bna A.tsMs互作的重要蛋白。提取不育系TE5A的幼蕾(直径≤2mm)总RNA,利用SMART(switching mechanism at 5'end of the RNA transcript)技术构建酵母...为进一步解析甘蓝型油菜温敏细胞核雄性不育系(GMS)TE5A的不育机理,筛选与育性蛋白Bna A.tsMs互作的重要蛋白。提取不育系TE5A的幼蕾(直径≤2mm)总RNA,利用SMART(switching mechanism at 5'end of the RNA transcript)技术构建酵母双杂交cDNA文库和诱饵载体p GBKT7-Bna A.tsMs,转化酵母菌株Y2H gold,检测其毒性和自激活活性,通过Mating方法筛选与蛋白Bna A.tsMs互作的蛋白,并根据基因注释推测其功能。结果成功地构建了甘蓝型油菜温敏细胞核雄性不育系TE5A幼蕾酵母双杂交cDNA文库,文库库容量为2×107cfu,重组率达到100%,插入片段平均长度大于1.8kb。发现诱饵载体p GBKT7-Bna A.tsMs对酵母没有毒性,没有自激活活性。筛选出49个候选互作蛋白,其中有丝分裂纺锤体检测点蛋白(BUBR1)、周期蛋白D3(cyclin-D3)和F-box蛋白(FBW2)三个互作蛋白可能与该不育蛋白相互作用,共同调控该不育系TE5A的育性。展开更多
Oilseed rape (Brassica napus) with yellow flowers is an attractive ornamental landscape plant during the flowering period,and the development of different petal colors has become a breeding objective.Although yellowis...Oilseed rape (Brassica napus) with yellow flowers is an attractive ornamental landscape plant during the flowering period,and the development of different petal colors has become a breeding objective.Although yellowish flower color is a common variant observed in field-grown oilseed rape,the genetics behind this variation remains unclear.We obtained a yellowish-white flower (ywf) mutant from Zhongshuang 9 (ZS9) by ethyl methanesulfonate mutagenesis (EMS) treatment.Compared with ZS9,ywf exhibited a lower carotenoid content with a reduced and defective chromoplast ultrastructure in the petals.Genetic analysis revealed that the yellowish-white trait was controlled by a single recessive gene.Using bulked-segregant analysis sequencing (BSA-seq) and kompetitive allele-specific PCR(KASP),we performed map-based cloning of the ywf locus on chromosome A08 and found that ywf harbored a C-to-T substitution in the coding region,resulting in a premature translation termination.YWF,encoding phytoene desaturase 3 (PDS3),was highly expressed in oilseed rape petals and involved in carotenoid biosynthesis.Pathway enrichment analysis of the transcriptome profiles from ZS9 and ywf indicated the carotenoid biosynthesis pathway to be highly enriched.Further analyses of differentially expressed genes and carotenoid components revealed that the truncated Bna A08.PDS3 resulted in decreased carotenoid biosynthesis in the mutant.These results contribute to an understanding of the carotenoid biosynthesis pathway and manipulation of flower-color variation in B.napus.展开更多
基金supported by the National Natural Science Foundation of China(U23A20190 and 31971977)the Project 2662022ZKYJ004 supported by the Fundamental Research Funds for the Central Universities and the Fund for CAR-12。
文摘Seed weight is a pivotal yield-determining trait in crops,and yet,the genetic and molecular mechanisms underlying its regulation in polyploid species remain underexplored.In a previous study,we identified cqSW.A03-2,a QTL that regulates thousand seed weight(TSW)in rapeseed(Brassica napus).Here,we identify BnaA3.AHK2,encoding a histidine kinase,as the causal gene of cqSW.A03-2.BnaA3.AHK2 enhances TSW through maternal control of seed coat cell expansion without significantly compromising other yield-related traits.Protein sequence divergence between parental haplotypes caused functional differentiation,with only the ZY50 allele showing functional kinase activity and rescuing developmental defects in Arabidopsis cytokinin receptor mutants.Strikingly,BnaA3.AHK2 seems to be a cytokinin-independent operator,contrasting with the canonical cytokinin signaling pathway.Transcriptome and protein interaction analyses reveal a signaling module where BnaA3.AHK2 engages BnaAHP-Bna ARR phosphorelay components to regulate downstream targets.Notably,the favorable cqSW.A03-2 haplotype has been historically selected in modern breeding,and its introgression into elite hybrids boosted TSW by 3.6%-9.1%,demonstrating its breeding value.Our findings unveil a non-canonical signaling pathway for seed size regulation,providing a strategic genetic target to break yield trade-offs in polyploid crops.
文摘为进一步解析甘蓝型油菜温敏细胞核雄性不育系(GMS)TE5A的不育机理,筛选与育性蛋白Bna A.tsMs互作的重要蛋白。提取不育系TE5A的幼蕾(直径≤2mm)总RNA,利用SMART(switching mechanism at 5'end of the RNA transcript)技术构建酵母双杂交cDNA文库和诱饵载体p GBKT7-Bna A.tsMs,转化酵母菌株Y2H gold,检测其毒性和自激活活性,通过Mating方法筛选与蛋白Bna A.tsMs互作的蛋白,并根据基因注释推测其功能。结果成功地构建了甘蓝型油菜温敏细胞核雄性不育系TE5A幼蕾酵母双杂交cDNA文库,文库库容量为2×107cfu,重组率达到100%,插入片段平均长度大于1.8kb。发现诱饵载体p GBKT7-Bna A.tsMs对酵母没有毒性,没有自激活活性。筛选出49个候选互作蛋白,其中有丝分裂纺锤体检测点蛋白(BUBR1)、周期蛋白D3(cyclin-D3)和F-box蛋白(FBW2)三个互作蛋白可能与该不育蛋白相互作用,共同调控该不育系TE5A的育性。
基金supported by the National Key Research and Development Program Of China (2016YFD0101007 and 2018YFE0108000)National Natural Science Foundation of China (31770250)+3 种基金the Natural Science Foundation of Hubei Province (2019CFB628)China Agriculture Research System (CARS-12)Agricultural Science and Technology Innovation Program (ASTIP) of Chinese Academy of Agricultural SciencesThe Agricultural Scientific and Technological Research Projects of Guizhou Province (No. Qiankehezhicheng [2019] 2397)。
文摘Oilseed rape (Brassica napus) with yellow flowers is an attractive ornamental landscape plant during the flowering period,and the development of different petal colors has become a breeding objective.Although yellowish flower color is a common variant observed in field-grown oilseed rape,the genetics behind this variation remains unclear.We obtained a yellowish-white flower (ywf) mutant from Zhongshuang 9 (ZS9) by ethyl methanesulfonate mutagenesis (EMS) treatment.Compared with ZS9,ywf exhibited a lower carotenoid content with a reduced and defective chromoplast ultrastructure in the petals.Genetic analysis revealed that the yellowish-white trait was controlled by a single recessive gene.Using bulked-segregant analysis sequencing (BSA-seq) and kompetitive allele-specific PCR(KASP),we performed map-based cloning of the ywf locus on chromosome A08 and found that ywf harbored a C-to-T substitution in the coding region,resulting in a premature translation termination.YWF,encoding phytoene desaturase 3 (PDS3),was highly expressed in oilseed rape petals and involved in carotenoid biosynthesis.Pathway enrichment analysis of the transcriptome profiles from ZS9 and ywf indicated the carotenoid biosynthesis pathway to be highly enriched.Further analyses of differentially expressed genes and carotenoid components revealed that the truncated Bna A08.PDS3 resulted in decreased carotenoid biosynthesis in the mutant.These results contribute to an understanding of the carotenoid biosynthesis pathway and manipulation of flower-color variation in B.napus.
