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Identification of quantitative trait loci and candidate genes controlling seed pigments of rapeseed

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Journal of Integrative Agriculture 2021年第11期20卷 2862-2879页

作者： ZHU Mei-chen HU Ran ZHAO Hui-yan TANG Yun-shan SHI Xiang-tian JIANG Hai-yan ZHANG Zhi-yuan FU Fu-you XU Xin-fu TANG Zhang-lin LIU Lie-zhao LU Kun LI Jia-na qu cun-min Chongqing Engineering Research Center for Rapeseed/College of Agronomy and Biotechnology Southwest University/Academy of Agricultural SciencesSouthwest UniversityChongqing 400716P.R.China Agriculture and Agri-Food Canada Saskatoon Research CentreSaskatoon S7N02XCanada

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 ***,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 *** detected 94 quantitative trait loci(QTLs)for seed pigments(44 for embryos and 50 for seed coat),distributed over 15 of the 19 rapeseed *** 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 *** 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 *** on the annotation of the ***"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 *** 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 ***.

关键词： Brassica napus L. embryo seed coat pigments QTL

Genome-wide association study and transcriptome analysis reveal integrative RNA-and miRNA-profile analysis reveals a likely role of organic nitrogen compound in thickness of pod canopy regulation of ***

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Genome-wide association study and transcriptome analysis rev...

Transcriptome analysis on dominant GMS homozygous two-type line D3AB in Brassica napus L. by RNA-Seq

中国作物学会油料作物专业委员会第八次会员代表大会暨学术年会

作者： CHEN Zhi-you HUO Qiang YANG Hong qu cun-min LU Kun LI Jia-na Academy of Agricultural Sciences Southwest University

Oilseed rape(Brassica napus L.) is the second largest oilseed crop worldwide and one of the most important oil crops in *** a component of plant architecture,thickness of pod canopy(TPC) plays an important role in yield performance,especially under high-density planting ***,mechanisms underlying TPC regulation are still largely not *** groups of oilseed rape lines with significantly different TPC were used to conduct RNA-and miRNA-profiling at bolting with tiny flower bud stage,identifying differential expression of a large number of genes involved in nitrogen-and carbon-related *** nitrogen-and carbon-response genes,including ASPS,ASP2,ASN3,ATCYSC1,PAI2,APT2,CRTISO,C0 X15,et *** dramatically enriched in the compact *** expressed miRNAs included those involved in regulating nitrogen-and *** patterns of most target genes were fine-tuned byrelated miRNAs,such as miR159,miR6029,and miR827.12 miRNAs were found to be differentially expressed at both sites,including miR319,miR845,*** results provide insight that nitrogen-and carbon-signaling may play a pivotal role in thickness of pod canopy regulation.

关键词： transcriptome Brassica napus pod canopy thickness miRNA

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Transcriptome analysis on dominant GMS homozygous two-type l...

甘蓝型油菜株高、第一分枝高和分枝数的QTL检测及候选基因筛选

中国作物学会油料作物专业委员会第八次会员代表大会暨学术年会

作者： JIA Le-dong XU Dai-xiang MA Guo-qiang DUAN Mou-zheng QIAO Cai-lin ZHOU Xin-tong qu cun-min WANG Rui LI Jia-na Chongqing Rapeseed Engineering Technology Research Center College of Agronomy and BiotechnologySouthwest University Academy of Agricultural Sciences Southwest University

Genie male sterility(GMS) has been widely used in Brassica napus breeding because of its stable and thorough sterility and wide distribution of restorer lines.D3 AB is a near isogenic line(NIL) controlled by a single dominant gene,but the specific molecular mechanisms leading to sterility is still *** this study,the transcriptome data of male sterile D3 A and fertile D3 B at different developmental stages of buds was compared,and some related genes causing abortion were identified.A total of 2 210 differentially expressed genes(DEGs) were detected,of which 374 genes were differentially expressed between young buds(≤1 mm) and middle buds(1-2 mm) in D3 A,and 1 458 in D3 B ***,679 differentially expressed genes were detected in young buds of D3 A and D3 B,and 1 423 in the middle buds ***,based on the results of cytological analysis,90 DEGs were specifically expressed in young buds,23 genes of them were up-regulated and67 were down-regulated,*** the gene expression profile database of Arabidopsis thaliana,10 DEGs were specifically expressed in young flower buds or pollen grains,and the expression patterns of 9 DEGs were consistent with transcriptome sequencing by *** results lay the foundation for further study on the abortive mechanism of D3 AB in Brassica napus.

关键词： Brassica napus dominant GMS two-type line RNA-seq

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作物学报 2015年第7期41卷 1027-1038页

作者：王嘉荆凌云荐红举曲存民谌利李加纳刘列钊西南大学农学与生物科技学院重庆400715

株高、分枝数及第1分枝高是油菜重要的农艺性状。本研究利用甘蓝型油菜GH06和P174杂交,F2通过单粒法连续自交至F11构建重组自交系群体,利用油菜60K芯片对该群体进行基因分型,构建高密度遗传连锁图谱。结果表明,该图谱包含2795个SNP多态... 详细信息

株高、分枝数及第1分枝高是油菜重要的农艺性状。本研究利用甘蓝型油菜GH06和P174杂交,F2通过单粒法连续自交至F11构建重组自交系群体,利用油菜60K芯片对该群体进行基因分型,构建高密度遗传连锁图谱。结果表明,该图谱包含2795个SNP多态性标记位点,总长1832.9 c M,相邻标记间平均距离为0.66 c M。在此图谱基础上采用复合区间作图法（CIM）,检测到3个农艺性状的24个QTL。其中11个株高QTL分别位于A01、A06、A07、A08、A10和C06染色体,单个QTL解释5.00%~15.26%的表型变异;7个第1分枝高QTL分别位于A06、C05和C06染色体,单个QTL解释5.04%~12.99%的表型变异;6个分枝数QTL分别位于A03、A07、C01、C04和C06染色体,单个QTL解释5.95%~8.14%的表型变异。将156个拟南芥株高相关基因、10个拟南芥第1分枝高相关基因和148个拟南芥分枝数相关基因与QTL对应置信区间序列进行同源比较分析（E〈1E–20）,分别找出了20个株高候选基因、3个第1分枝高候选基因以及12个分枝数候选基因。2个环境中在A07染色体上重复检测到的QTL置信区间检测到与株高相关的候选基因ATGID1B/GID1B和WRI1,A08染色体上重复检测到的QTL置信区间检测到SLR/IAA14和AXR2/IAA72个与株高相关的候选基因。在具有部分置信区间重叠的q2013FBH-C05-1和q2014FBH-C05-2区间均检测到第1分枝高候选基因PHT1;8,在A03和C06染色体上的QTL置信区间内,分别检测到4个分枝数候选基因,匹配E值介于0~3E–56之间。

关键词：甘蓝型油菜单核苷酸多态性数量性状位点株高第1分枝高度分枝数

Genome wide identification and comparative expression profile of long-chain acyl-CoA synthetase(LACS)gene family in two different oil content cultivars of Brassica napus

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Genome wide identification and comparative expression profil...

甘蓝型油菜BnMAPK1的原核表达、亚细胞定位及酵母双杂交文库筛选

中国作物学会油料作物专业委员会第八次会员代表大会暨学术年会

作者： XIAO Zhong-chun LI Nan-nan TANG Fang YANG Hong ZHANG Li-yuan ZHANG Chao YANG Bo ZHAO Hui-yan WEI Li-juan DU Hai qu cun-min LU Kun LI Jia-na Chongqing Engineering Research Center for Rapeseed College of Agronomy and BiotechnologySouthwest University Academy of Agricultural Sciences Southwest University Research Center of Bioenergy and Bioremediation College of Resources and EnvironmentSouthwest University

Long-chain acyl-CoA synthetases(LACSs) are one of the key enzymes involved in fatty acid metabolism,including phospholipid biosynthesis,triacylglycerol(TAG) biosynthesis and fatty acid B-oxidation in ***,LACSs play a significant role in lipids formation and are associated with oil content of oil ***,characterization of LACSs family in Brassica napus remains *** the present study,a comprehensive genome wide analysis of this gene family in Brassica napus was performed and 34 *** LACS genes(BnaLACSs) were *** analysis classified BnaLACS proteins into 4 groups(A,B,C and D),which were supported by highly conserved gene structures and consensus ***-Seq(RNA-Sequencing) and qRT-PCR(quantitative real time polymerase chain reaction) combined analysis revealed that 18 BnaLACSs(BnaLACSl-2,1-3,1-4,1-9,1-10,2-1,2-2,4-1,4-2,6-1,6-2,6-4,7-1,7-2,8-1,8-2,9-3 and 9-4) were expressed in developmental *** expression analysis between extremely high-oil-content(P1-HO) and low-oil-content(P2-LO) *** cultivars revealed that BnaLACS6-4,BnaLACS9-3 and BnaLACS9-4 may be involved in fatty acid synthesis in ***-10 and 4-1 may also play vital roles in lipid biosynthesis of *** study provides important information for functional characterization of BnaLACSs in seed oil metabolism of ***.

关键词： LACS B.napus long-chain acyl-CoA synthetases lipid biosynthesis

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作物学报 2020年第9期46卷 1312-1321页

作者：王珍姚梦楠张晓莉曲存民卢坤李加纳梁颖西南大学农学与生物科技学院/油菜工程研究中心/西南大学现代农业科学研究院重庆400715

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... 详细信息

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。

关键词：甘蓝型油菜 BnMAPK1 原核表达亚细胞定位酵母双杂交

甘蓝型油菜不同抗倒性材料中木质素代谢途径关键基因表达特点

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作物学报 2013年第8期39卷 1339-1344页

作者：黄杰恒李威曲存民刘列钊徐新福王瑞李加纳西南大学农学与生物科技学院/南方山地农业教育部工程研究中心重庆400716

植物体内木质素的含量不仅与植物的抗病性有关,也与植物的抗倒伏性状密切联系。本试验以15个抗倒性等级不同的甘蓝型油菜品种为材料,分别在初花期和青荚期测定茎秆中部木质素含量,同时用qRT-PCR的方法分析了木质素代谢途径中6个关键基因... 详细信息

植物体内木质素的含量不仅与植物的抗病性有关,也与植物的抗倒伏性状密切联系。本试验以15个抗倒性等级不同的甘蓝型油菜品种为材料,分别在初花期和青荚期测定茎秆中部木质素含量,同时用qRT-PCR的方法分析了木质素代谢途径中6个关键基因PAL、4CL、C4H、CCR1、CCR2和CAD的表达量差异。结果表明,油菜茎秆木质素从初花期到青荚期平均增加了28%,抗倒性强的材料木质素增加最明显,达到33.5%;不同时期,不同抗倒伏性的油菜茎秆中部木质素含量差异均极显著;各基因表达量在两个时期间差异均显著或极显著,但在不同抗倒性材料间,只有基因PAL、4CL和CCR1表达量间差异显著;基因PAL的表达量与木质素含量相关性最强,4CL基因的表达量与其他关键酶基因表达量相关性最强。综上所述,木质素的合成积累由代谢途径中各个环节关键基因所共同决定,但基因PAL和4CL对木质素的代谢影响更明显。

关键词：木质素荧光定量PCR 代谢途径抗倒性

甘蓝型油菜BnTT3基因的表达与eQTL定位分析

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作物学报 2015年第11期41卷 1758-1766页

作者：卢坤曲存民李莎赵会彦王瑞徐新福梁颖李加纳西南大学农学与生物科技学院重庆400715 重庆市油菜工程技术研究中心重庆400715

类黄酮途径中, TT3编码的4-二氢黄铜醇还原酶是参与原花色素和花青素合成的关键酶。为了明确该基因可能的上游调控网络,利用黄籽母本 GH06和黑籽父本 ZY821构建的遗传图谱,以 BnTT3基因在高世代重组自交系群体中随机选取的94个株系花后4... 详细信息

类黄酮途径中, TT3编码的4-二氢黄铜醇还原酶是参与原花色素和花青素合成的关键酶。为了明确该基因可能的上游调控网络,利用黄籽母本 GH06和黑籽父本 ZY821构建的遗传图谱,以 BnTT3基因在高世代重组自交系群体中随机选取的94个株系花后40 d种子的表达量作为性状,采用复合区间作图法进行eQTL分析。结果共检测到5个表达量相关的eQTL,分别位于A03、A08、A09和C01染色体,单个eQTL解释表型变异的5.22%~24.05%。A09染色体上存在2个主效 eQTL,单个 eQTL 分别解释24.05%和16.55%的表型变异,分别位于标记 KS10260~KBrB019I24.15和B055B21-5~KS30880之间,微效eQTL分布于A03、A08和C01染色体上。A09染色体上的2个主效eQTL区间（包含200 kb侧翼序列）与拟南芥、白菜、甘蓝和芸薹族近缘物种基因组同源区段具有很好的共线性关系。基因注释结果表明检测到的eQTL均为trans-QTL,2个主效eQTL区段共包含78个基因,包括MYB51、MYB52和bZIP5转录因子,可能为 BnTT3基因的上游直接调控因子,对这些基因功能的深入分析将有助于阐明甘蓝型油菜黄籽性状形成的分子调控机制,为黄籽候选基因的克隆筛选奠定基础。

关键词：甘蓝型油菜种子 TT3 重组自交系表达数量性状位点

基于QTL定位和全基因组关联分析筛选甘蓝型油菜株高和一次有效分枝高度的候选基因

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作物学报 2020年第2期46卷 214-227页

作者：霍强杨鸿陈志友荐红举曲存民卢坤李加纳西南大学农学与生物科技学院/油菜工程研究中心重庆400715 西南大学现代农业科学研究院重庆400715

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... 详细信息

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。

关键词：甘蓝型油菜株高一次有效分枝高度基因定位候选基因筛选