Rapeseed(Brassica napus L.)is an important source of edible vegetable oil and feed protein;however,seed pigments affect the quality of rapeseed oil and the feed value of the residue from oil pressing.Here,we used a po...Rapeseed(Brassica napus L.)is an important source of edible vegetable oil and feed protein;however,seed pigments affect the quality of rapeseed oil and the feed value of the residue from oil pressing.Here,we used a population of rapeseed recombinant inbred lines(RILs)derived from the black-seeded male parent cultivar Zhongyou 821 and the yellow-seeded female parent line GH06 to map candidate genes controlling seed pigments in embryos and the seed coat.We detected 94 quantitative trait loci(QTLs)for seed pigments(44 for embryos and 50 for seed coat),distributed over 15 of the 19 rapeseed chromosomes.These included 28 QTLs for anthocyanidin content,explaining 2.41–44.66% of phenotypic variation;24 QTLs for flavonoid content,explaining 2.41–20.26% of phenotypic variation;16 QTLs for total phenol content,accounting for 2.74–23.68% of phenotypic variation;and 26 QTLs for melanin content,accounting for 2.37–24.82% of phenotypic variation,indicating that these traits are under multigenic control.Consensus regions on chromosomes A06,A09 and C08 were associated with multiple seed pigment traits,including 15,19 and 10 QTLs,respectively,most of which were major QTLs explaining>10% of the phenotypic variation.Based on the annotation of the B.napus"Darmor-bzh"reference genome,67 candidate genes were predicted from these consensus QTLs regions,and 12 candidate genes were identified as potentially involved in pigment accumulation by RNA-seq and qRT-PCR analysis.These preliminary results provide insight into the genetic architecture of pigment biosynthesis and lay a foundation for exploring the molecular mechanisms underlying seed coat color in B.napus.展开更多
株高和一次有效分枝高度是与甘蓝型油菜结荚层厚度、收获指数紧密关联的重要农艺性状,有关株高的数量性状位点(quantitative trait locus,QTL)和全基因组关联分析(genome-wide association study,GWAS)已有很多报道,但对一次有效分枝高...株高和一次有效分枝高度是与甘蓝型油菜结荚层厚度、收获指数紧密关联的重要农艺性状,有关株高的数量性状位点(quantitative trait locus,QTL)和全基因组关联分析(genome-wide association study,GWAS)已有很多报道,但对一次有效分枝高度的QTL和GWAS定位以及候选基因筛选的研究报道较少。本研究利用已构建的高密度遗传连锁图对2016和2017年2个环境的186个株系组成的重组自交系群体株高和一次有效分枝高度及其最佳线性无偏预测(best linear unbiased prediction,BLUP)值进行QTL定位共检测到8个株高的QTL,分别位于A03、A04和A09染色体,单个QTL解释4.60%~13.29%的表型变异,其中位于A04染色体上的QTL(q-2017PH-A04-2和q-BLUP-PH-A04-2)在2017年和BLUP中均被检测到;检测到9个一次有效分枝高度QTL,分别位于A01、A02、A05、A09、C01和C05染色体上,单个QTL解释5.12%~19.10%的表型变异,其中q-2017BH-A09-1、q-BLUP-BH-A09-2和q-BLUP-BH-A09-3有重叠区段。同时,利用课题组前期完成的588份重测序自然群体进行全基因组关联分析,2年共检测到与株高显著关联的50个SNP位点和与一次有效分枝高度显著关联的12个SNP位点;根据SNP的物理位置,筛选出参与细胞增殖、细胞扩增、细胞周期和细胞壁活动的13个株高候选基因,以及参与赤霉素、亚精胺等合成代谢途径、核糖体组成和在光合、萌发等过程中有一定作用的一次分枝高度的11个候选基因,并利用荧光定量PCR技术验证候选基因在极端材料中的表达情况。本研究结果将为油菜株型改良及后续基因的功能研究提供理论依据。展开更多
植物的丝裂原活化蛋白激酶(Mitogen-Activated Protein Kinases,MAPKs)级联是生长进程中多种信号跨膜传递的共同通路,它可以将外源刺激转入细胞内并引发细胞应答。目前C族MAPKs在甘蓝型油菜中的研究报道还很有限。本研究通过1个甘蓝型...植物的丝裂原活化蛋白激酶(Mitogen-Activated Protein Kinases,MAPKs)级联是生长进程中多种信号跨膜传递的共同通路,它可以将外源刺激转入细胞内并引发细胞应答。目前C族MAPKs在甘蓝型油菜中的研究报道还很有限。本研究通过1个甘蓝型油菜的C族BnMAPK1基因的生物学进程聚类分析发现,BnMAPK1可能参与蛋白磷酸化、生长素介导的信号途径、逆境应答、细胞周期及转录等过程。BnMAPK1具有369个氨基酸残基,其相对分子质量约为42.5 kD,其可溶性蛋白可在原核系统中被诱导表达。BnMAPK1的亚细胞定位结果显示,BnMAPK1主要在细胞核内表达。Bait质粒pGBKT7-BnMAPK1在酵母双杂交系统中无毒性及自激活活性。为深入研究BnMAPK1蛋白参与的生物学进程,从甘蓝型油菜中油821苗期根、茎、叶中分别提取总RNA,分离得到mRNA,利用SMART技术合成并纯化双链cDNA后建立甘蓝型油菜混合cDNA文库。以共转化法筛选与BnMAPK1相互作用的蛋白,对其分析与鉴定显示,BnMAPK1在生长发育、非生物及生物逆境、转录、蛋白合成及代谢、翻译及翻译后修饰等过程中起作用。研究结果为MAPKs级联,尤其是C族MAPKs的研究提供了新的视野,为甘蓝型油菜抗性的机理研究及分子育种奠定了重要的理论依据。展开更多
丝裂原活化蛋白激酶(Mitogen-activated protein kinases,MAPKs)级联途径在植物的生长发育、分裂分化、细胞凋亡以及抗逆等多种生命过程中发挥着极其重要的作用。本研究从甘蓝型油菜中分离克隆了1个C族BnMAPK7基因的启动子,序列长度为16...丝裂原活化蛋白激酶(Mitogen-activated protein kinases,MAPKs)级联途径在植物的生长发育、分裂分化、细胞凋亡以及抗逆等多种生命过程中发挥着极其重要的作用。本研究从甘蓝型油菜中分离克隆了1个C族BnMAPK7基因的启动子,序列长度为1612 bp,命名为ProBnMAPK7。启动子分析工具PlantCARE预测结果表明,ProBnMAPK7含有大量响应光照、激素、防御和损伤等相关顺式作用元件,包括ACE、MRE、ABRE、TGACG-motif和TC-rich repeats等。同时,我们对拟南芥及甘蓝型油菜MAPK7基因的表达模式进行分析发现,MAPK7对生长发育以及生物和非生物胁迫应答过程具有重要调控意义。将ProBnMAPK7逐步分段连接至pCambia1305.1-GUS表达载体筛选启动子的核心区段,GUS组织化学染色分析显示该启动子的核心区段定位于-467~-239 bp(ProBnMAPK7-rPE)。将核心区段3拷贝串联重复后整合至Y1H gold基因组,并进行AbA背景测试,结果显示AbA浓度为500 ng mL^(-1)时,ProBnMAPK7-rPE×3在酵母细胞中的本底表达被完全抑制。利用酵母单杂交技术对BnMAPK7的上游调控因子进行文库筛选,获得3个候选因子BnNAD1B(NADH dehydrogenase 1B)、BnERD6(early response to dehydration 6)和BnPIG3(quinone oxidoreductase PIG3-like)。表明BnNAD1B、BnERD6及BnPIG3蛋白可能通过结合ProBnMAPK7-rPE区段调控BnMAPK7的转录,使得BnMAPK7参与光合作用以及逆境胁迫应答等生物学过程,为进一步阐明甘蓝型油菜BnMAPK7基因的功能奠定了基础,也为MAPKs级联的研究提供了新的思路。展开更多
Rapeseed （Brassica napus L.） oil is the crucial source of edible oil in China. In addition, it can become a major renewable and sustainable feedstock for biodiesel production in the future. It is known that photosyn...Rapeseed （Brassica napus L.） oil is the crucial source of edible oil in China. In addition, it can become a major renewable and sustainable feedstock for biodiesel production in the future. It is known that photosynthesis products are the primary sources for dry matter accumulation in rapeseed. Therefore, increasing the photosynthetic efficiency is desirable for the raise of rapeseed yield. The objective of the present study was to identify the genetic mechanism of photosynthesis based on the description of relationships between different photosynthetic traits and their quantitative trait loci （QTL） by using a recombinant inbred line （RIL） population with 172 lines. Specifically, correlation analysis in this study showed that internal CO2 concentration has negative correlations with other three physiological traits under two different stages. Totally, 11 and 12 QTLs of the four physiological traits measured at the stages 1 and 2 were detected by using a high-density single nu- cleotidepolymorphism （SNP） markers linkage map with composite interval mapping （CIM）, respectively. Three co-localized QTLs on A03 were detected at stage 1 with 5, 5, and 10% of the phenotypic variation, respectively. Other two co-localized QTLs were located on A05 at stage 2, which explained up to 12 and 5% of the phenotypic variation, respectively. The results are beneficial for our understanding of genetic control of photosynthetic physiological characterizations and improvement of rapeseed yield in the future.展开更多
基金supported by the National Natural Science Foundation of China(31830067)the Major S&T Projects on the Cultivation of New Varieties of Genetically Modified Organisms,China(2018ZX08020001)+3 种基金the Fundamental Research Funds for the Central Universities,China(XDJK2020B030)the China Agriculture Research System of MOF and MARA(CARS-12)the Natural Science Foundation of Chongqing,China(cstc2017jcyjAX0321)the Higher Education Discipline Innovation Project,China(B12006)。
文摘Rapeseed(Brassica napus L.)is an important source of edible vegetable oil and feed protein;however,seed pigments affect the quality of rapeseed oil and the feed value of the residue from oil pressing.Here,we used a population of rapeseed recombinant inbred lines(RILs)derived from the black-seeded male parent cultivar Zhongyou 821 and the yellow-seeded female parent line GH06 to map candidate genes controlling seed pigments in embryos and the seed coat.We detected 94 quantitative trait loci(QTLs)for seed pigments(44 for embryos and 50 for seed coat),distributed over 15 of the 19 rapeseed chromosomes.These included 28 QTLs for anthocyanidin content,explaining 2.41–44.66% of phenotypic variation;24 QTLs for flavonoid content,explaining 2.41–20.26% of phenotypic variation;16 QTLs for total phenol content,accounting for 2.74–23.68% of phenotypic variation;and 26 QTLs for melanin content,accounting for 2.37–24.82% of phenotypic variation,indicating that these traits are under multigenic control.Consensus regions on chromosomes A06,A09 and C08 were associated with multiple seed pigment traits,including 15,19 and 10 QTLs,respectively,most of which were major QTLs explaining>10% of the phenotypic variation.Based on the annotation of the B.napus"Darmor-bzh"reference genome,67 candidate genes were predicted from these consensus QTLs regions,and 12 candidate genes were identified as potentially involved in pigment accumulation by RNA-seq and qRT-PCR analysis.These preliminary results provide insight into the genetic architecture of pigment biosynthesis and lay a foundation for exploring the molecular mechanisms underlying seed coat color in B.napus.
文摘株高和一次有效分枝高度是与甘蓝型油菜结荚层厚度、收获指数紧密关联的重要农艺性状,有关株高的数量性状位点(quantitative trait locus,QTL)和全基因组关联分析(genome-wide association study,GWAS)已有很多报道,但对一次有效分枝高度的QTL和GWAS定位以及候选基因筛选的研究报道较少。本研究利用已构建的高密度遗传连锁图对2016和2017年2个环境的186个株系组成的重组自交系群体株高和一次有效分枝高度及其最佳线性无偏预测(best linear unbiased prediction,BLUP)值进行QTL定位共检测到8个株高的QTL,分别位于A03、A04和A09染色体,单个QTL解释4.60%~13.29%的表型变异,其中位于A04染色体上的QTL(q-2017PH-A04-2和q-BLUP-PH-A04-2)在2017年和BLUP中均被检测到;检测到9个一次有效分枝高度QTL,分别位于A01、A02、A05、A09、C01和C05染色体上,单个QTL解释5.12%~19.10%的表型变异,其中q-2017BH-A09-1、q-BLUP-BH-A09-2和q-BLUP-BH-A09-3有重叠区段。同时,利用课题组前期完成的588份重测序自然群体进行全基因组关联分析,2年共检测到与株高显著关联的50个SNP位点和与一次有效分枝高度显著关联的12个SNP位点;根据SNP的物理位置,筛选出参与细胞增殖、细胞扩增、细胞周期和细胞壁活动的13个株高候选基因,以及参与赤霉素、亚精胺等合成代谢途径、核糖体组成和在光合、萌发等过程中有一定作用的一次分枝高度的11个候选基因,并利用荧光定量PCR技术验证候选基因在极端材料中的表达情况。本研究结果将为油菜株型改良及后续基因的功能研究提供理论依据。
文摘植物的丝裂原活化蛋白激酶(Mitogen-Activated Protein Kinases,MAPKs)级联是生长进程中多种信号跨膜传递的共同通路,它可以将外源刺激转入细胞内并引发细胞应答。目前C族MAPKs在甘蓝型油菜中的研究报道还很有限。本研究通过1个甘蓝型油菜的C族BnMAPK1基因的生物学进程聚类分析发现,BnMAPK1可能参与蛋白磷酸化、生长素介导的信号途径、逆境应答、细胞周期及转录等过程。BnMAPK1具有369个氨基酸残基,其相对分子质量约为42.5 kD,其可溶性蛋白可在原核系统中被诱导表达。BnMAPK1的亚细胞定位结果显示,BnMAPK1主要在细胞核内表达。Bait质粒pGBKT7-BnMAPK1在酵母双杂交系统中无毒性及自激活活性。为深入研究BnMAPK1蛋白参与的生物学进程,从甘蓝型油菜中油821苗期根、茎、叶中分别提取总RNA,分离得到mRNA,利用SMART技术合成并纯化双链cDNA后建立甘蓝型油菜混合cDNA文库。以共转化法筛选与BnMAPK1相互作用的蛋白,对其分析与鉴定显示,BnMAPK1在生长发育、非生物及生物逆境、转录、蛋白合成及代谢、翻译及翻译后修饰等过程中起作用。研究结果为MAPKs级联,尤其是C族MAPKs的研究提供了新的视野,为甘蓝型油菜抗性的机理研究及分子育种奠定了重要的理论依据。
基金supported by the National Natural Science Foundation of China (31171584, 31371655)National Basic Research Program of China (973 Program, 2015CB150201)+1 种基金the Fundamental Research Funds for the Central Universities, China (XDJK2013B015)the Earmarked Fund for Modern Agro-industry Technology Research System, China (CARS-13)
文摘Rapeseed （Brassica napus L.） oil is the crucial source of edible oil in China. In addition, it can become a major renewable and sustainable feedstock for biodiesel production in the future. It is known that photosynthesis products are the primary sources for dry matter accumulation in rapeseed. Therefore, increasing the photosynthetic efficiency is desirable for the raise of rapeseed yield. The objective of the present study was to identify the genetic mechanism of photosynthesis based on the description of relationships between different photosynthetic traits and their quantitative trait loci （QTL） by using a recombinant inbred line （RIL） population with 172 lines. Specifically, correlation analysis in this study showed that internal CO2 concentration has negative correlations with other three physiological traits under two different stages. Totally, 11 and 12 QTLs of the four physiological traits measured at the stages 1 and 2 were detected by using a high-density single nu- cleotidepolymorphism （SNP） markers linkage map with composite interval mapping （CIM）, respectively. Three co-localized QTLs on A03 were detected at stage 1 with 5, 5, and 10% of the phenotypic variation, respectively. Other two co-localized QTLs were located on A05 at stage 2, which explained up to 12 and 5% of the phenotypic variation, respectively. The results are beneficial for our understanding of genetic control of photosynthetic physiological characterizations and improvement of rapeseed yield in the future.
