红苞凤梨(Ananas bracteatus)金边嵌合性状非编码RNA表观调控机制

批准号:
31570698
项目类别:
面上项目
资助金额:
64.0 万元
负责人:
马均
依托单位:
学科分类:
C1612.园林学
结题年份:
2019
批准年份:
2015
项目状态:
已结题
项目参与者:
马明东、姜福星、周宇爝、刘柿良、雷竹、钟小兰、李夏、刘雯
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中文摘要
红苞凤梨嵌合叶由绿色和白色组织形成金边、金心等多种类型,且花果艳丽多彩,观赏期长,是重要的新型观赏植物。但其嵌合性状在繁殖过程中易发生变异甚至消失,大量再生植株因丧失观赏价值而被淘汰。前期测序发现5个叶绿素合成相关基因在白化细胞中下调表达,其中hemC是导致白化的关键基因。植物表观遗传学己成为当今植物科学研究新的焦点,非编码RNA调控是表观遗传调控的重要内容。本项目将在前期研究的基础上,以组培分离的全绿、全白植株为材料,利用高通量测序构建两种细胞类型的miRNA和lncRNA表达谱并进行表达差异分析;筛选调控细胞白化关键基因表达的关键调控非编码RNA;检测关键调控因子及其靶基因的表达模式和靶基因的DNA甲基化水平;采用CRISPR-Cas9基因敲除、超表达验证关键调控因子功能。探明关键调控因子的结构、功能及相互作用网络,为红苞凤梨金边嵌合性状的的分子改良和稳定性调控奠定基础。
英文摘要
Variegated chimera varieties of Ananas bracteatus have been planted as important landscape plants for its ornamental values of color leaf and fruit. The leaves are composed of green normal tissues and abino tissues. However the variegated chimera character of Bromeliad is unstable. Many plants regenerated by in vitro tissue culture lost ornamental value for the losing of leaf color. It is very important and meaningful to study the molecular mechanism of chimera. Our studies showed that the compressing of the gene expression and synthesis of chlorophyll lead to the chlorosis of the cells. The hemC is the key gene playing an important role the cell chlorosis. Study on epigenetic regulation mechanism is becoming the new research hot point in plant science, and the non-coding RNA regulation is the most important content of epigenetic regulation. Based on our preliminary studies, the complete green or white plants derived from in vitro culture were used in the further study of epigenetic regulation mechanism of the key genes leading to the chlorosis of cells in the applied project. The miRNA and lncRNA expression profiles were constructed by Illumina sequence and the variations between the green plant and white plant were analyzed. The key non-coding RNAs working in cell chlorosis were screened by correlation analysis. The expression patterns of the key non-coding RNAs and their target genes were detected by Real-time PCR and western blot. The DNA-methylation of the target genes was analyzed. The biological function of the key non-coding RNAs were verified by CRISPR-Cas9 gene knockout and gene over expression technique. This study could illuminate the structure, function and interaction network of the key non-coding RNA in cell chlorosis. The research results could lay a foundation of the molecular improvement and stability regulation of the chimera character of Ananas bracteatus.
金边红苞凤梨( Ananas comosus var. bracteatues) 具有艳丽的果实和冠芽,是一种重要的花叶观赏植物。叶片由绿色正常组织与白化组织相嵌形成金边,低温和花期叶片呈现艳丽红色。但其嵌合性状在繁殖过程中易发生变异甚至消失,导致大量再生植株因丧失观赏价值而被淘汰。因此,深入研究嵌合体中白化细胞形成的分子机理对嵌合品种繁育有着重要意义。本项目在前期转录水平对嵌合体白化细胞失绿突变机理研究基础之上,以组培分离的全绿、全白植株为材料,利用高通量测序构建叶片发育过程中两种组织类型的miRNA和lncRNA表达谱并进行表达差异分析,构建了蛋白表达谱,检测了蛋白修饰组差异。采用 qRT-PCR验证了测序结果的可靠性。将miRNA和lncRNA的靶基因与Nr, Nt, Swiss-Prot, GO, COG, KEGG, KOG 和 Pfam数据库进行比对进行功能注释和富集分析,厘清miRNA和lncRNA参与调控的叶片主要生理代谢过程。筛选出与光合作用、叶绿素合成、叶绿体发育相关的差异表达miRNA和lncRNA。最终筛选到了 7 个红苞凤梨白化失绿关键 miRNA和86个靶向叶绿素代谢相关蛋白编码基因的lncRNA。构建UBi-pCAMBIA1301-pre-AbmiR163 超表达载体,并转化红苞凤梨愈伤组织,筛选得到转基因植株,验证了AbmiR163通过调控DVR基因的转录调控红苞凤梨叶绿素合成的功能。构建了CRISPR/Cas9-lncABCG11基因编辑载体并转化红苞凤梨愈伤组织,筛选得到转基因植株,验证了lncABCG11通过调控POR基因的表达,调控红苞凤梨叶绿素合成,从而影响叶片颜色的功能。通过蛋白质组和蛋白质修饰组分析,发现叶片的白化在转录、转录调控、翻译和翻译后水平都存在显著的调控机制,筛选出了相关的调控因子用于后续深入研究。
期刊论文列表
专著列表
科研奖励列表
会议论文列表
专利列表
Screening and characterization of long noncoding RNAs involved in the albinism of Ananas comosus var. bracteatus leaves
凤梨白化病相关长链非编码 RNA 的筛选和表征
DOI:10.1371/journal.pone.0225602
发表时间:2019-11
期刊:Plos One
影响因子:3.7
作者:Lin Zhen;Xiong Yingyuan;Xue Yanbin;Mao Meiqin;Xiang Yixuan;He Yehua;Rafique Fatima;Hu Hao;Liu Jiawen;Li Xi;Sun Lingxia;Huang Zhuo;Ma Jun
通讯作者:Ma Jun
High-throughput sequencing analysis revealed the regulation patterns of small RNAs on the development of A. comosus var. bracteatus leaves.
高通量测序分析揭示了小RNA对A. comosus var.发育的调控模式。
DOI:10.1038/s41598-018-20261-z
发表时间:2018-01-31
期刊:Scientific reports
影响因子:4.6
作者:Xiong YY;Ma J;He YH;Lin Z;Li X;Yu SM;Li RX;Jiang FX;Li X;Huang Z;Sun LX
通讯作者:Sun LX
DOI:--
发表时间:2016
期刊:热带亚热带植物学报
影响因子:--
作者:李瑞雪;余三淼;李夏;熊颖媛;蔺珍;唐琰琪;马均
通讯作者:马均
SMRT sequencing analysis reveals the full-length transcripts and alternative splicing patterns in Ananas comosus var. bracteatus
SMRT 测序分析揭示了 Ananas comosus var. 的全长转录本和可变剪接模式。
DOI:10.7717/peerj.7062
发表时间:2019-06-21
期刊:PEERJ
影响因子:2.7
作者:Ma, Jun;Xiang, Yixuan;Huang, Zhuo
通讯作者:Huang, Zhuo
Comparative transcriptomic and proteomic analyses of the green and white parts of chimeric leaves in Ananas comosus var. bracteatus
菠萝嵌合叶绿色和白色部分的比较转录组和蛋白质组分析。
DOI:10.7717/peerj.7261
发表时间:2019-07
期刊:Peer J
影响因子:--
作者:Xue Yanbin;Ma Jun;He Yehua;Yu Sanmiao;Lin Zhen;Xiong Yingyuan;Rafique Fatima;Jiang Fuxing;Sun Lingxia;Ma Mingdong;Zhou Yujue;Li Xi;Huang Zhuo
通讯作者:Huang Zhuo
红苞凤梨叶色嵌合体色素协同竞争积累调控机理研究
- 批准号:31971704
- 项目类别:面上项目
- 资助金额:58.0万元
- 批准年份:2019
- 负责人:马均
- 依托单位:
多组学解析红苞凤梨(Ananas comosus var. bracteatus)嵌合性状形成机理
- 批准号:31770743
- 项目类别:面上项目
- 资助金额:61.0万元
- 批准年份:2017
- 负责人:马均
- 依托单位:
红苞凤梨嵌合体中白化细胞失绿分子机理研究
- 批准号:31300585
- 项目类别:青年科学基金项目
- 资助金额:21.0万元
- 批准年份:2013
- 负责人:马均
- 依托单位:
国内基金
海外基金
