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西藏野生拟南芥开花时间变异的遗传基础

植物学报 2024年第3期59卷 373-382页

作者：杨继轩王雪霏顾红雅北京大学生命科学学院蛋白质与植物基因研究国家重点实验室北京100871

开花时间是被子植物生活史中的关键节点。十字花科植物拟南芥(Arabidopsisthaliana)广布于世界各地,在海拔4 000m以上的青藏高原也发现了该物种的自然居群,高原独特的环境塑造了其生活史的独特表型,在开花时间上表现为中等程度早花。该... 详细信息

开花时间是被子植物生活史中的关键节点。十字花科植物拟南芥(Arabidopsisthaliana)广布于世界各地,在海拔4 000m以上的青藏高原也发现了该物种的自然居群,高原独特的环境塑造了其生活史的独特表型,在开花时间上表现为中等程度早花。该研究构建了西藏拟南芥Lhasa居群的F2代作图群体,基于全基因组测序的QTL-seq定位分析,在该居群中定位到主效基因FLC,并且鉴定到其第1个内含子中存在2307bp的缺失,这种单倍型只存在于西藏拟南芥居群。利用CRISPR-Cas9技术构建了Lhasa背景的flc^(-/-)突变体,表现为开花时间显著提前。研究结果表明,西藏拟南芥开花时间改变的主要原因是FLC第1个内含子缺失,该变异并未使其丧失全部功能,这可能有利于西藏拟南芥适应青藏高原独特的气候环境。

关键词：西藏拟南芥开花时间 Flowering Locus C 适应性演化

Duplication and divergent evolution of the CHS and CHS-like genes in the chalcone synthase(CHS)superfamily

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Chinese Science Bulletin 2006年第5期51卷 505-509页

作者： YANG Ji gu hongya College of Life Sciences Peking University Beijing 100871 China

The enzymes of the CHS-superfamily are responsible for biosynthesis of a wide range of natural products in plants. They are important for flower pigmentation, protection against UV light and defense against phytopathogens. Many plants were found to contain multiple copies of CHS genes. This review summarizes the recent progress in the studies of the CHS-superfamily, focusing on the duplication and divergent evolution of the CHS and CHS-like genes. Comparative analyses of gene structure, ex- pression patterns and catalytic properties revealed extensive differentiation in both regulation and func- tion among duplicate CHS genes. It is also proposed that the CHS-like enzymes in the CHS-superfamily evolved from CHS at different times in various or- ganisms. The CHS-superfamily thus offers a valuable model to study the rates and patterns of sequence divergence between duplicate genes.

关键词：苯基苯乙烯酮合酶基因复制分裂演进 CHS基因植物生物化学

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Journal of Integrative Plant Biology 2012年第10期54卷 814-827页

作者： guo Luo hongya gu Jingjing Liu Li-Jia Qu State Key Laboratory for Protein and Plant Gene Research College of Life SciencesPeking-Tsinghua Center of Life SciencesPeking UniversityBeijing 100871China The National Plant Gene Research Center Beijing 100101China

Ubiquitination of proteins is one of the critical regulatory mechanisms in eukaryotes. In higher plants, protein ubiquitination plays an essential role in many biological processes, including hormone signaling, photomorphogenesis, and pathogen defense. However, the roles of protein ubiquitination in the repro- ductive process are not clear. In this study, we identified four plant-specific RING-finger genes designated Aberrant_Pollen Development 1_ （APD1） to APD4, as regulators of pollen mitosis II （PMII） in Arabidopsis thaliana （L.）. The apdl apd2 double mutant showed a significantly increased percentage of bicellular-like pollen at the mature pollen stage. Further downregulation of the APD3 and APD4 transcripts in apdl apd2 by RNA interference （RNAi） resulted in more severe abnormal bicellular-like pollen phenotypes than in apdl apd2, suggesting that cell division was defective in male gametogenesis. All of the four genes were expressed in multiple stages at different levels during male gametophyte development. Confocal analysis using green florescence fusion proteins （GFP） GFP-APD1 and GFP-APD2 showed that APDs are associated with intracellular membranes. Furthermore, APD2 had E2-dependent E3 ligase activity in vitro, and five APD2-interacting proteins were identified. Our results suggest that these four genes may be involved, redundantly, in regulating the PMII process during male gametogenesis.

关键词： Male gametogenesis pollen mitosis II RING-finger E3 ligase Arabidopsis.

Active role of small peptides in Arabidopsis reproduction: Expression evidence

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Journal of Integrative Plant Biology 2015年第6期57卷 518-521页

作者： Qingpei Huang Thomas Dresselhaus hongya gu Li-Jia Qu State Key Laboratory of Protein and Plant Gene Research Peking-Tsinghua Center for Life SciencesSchool of Life SciencesPeking University Cell Biology and Plant Biochemistry Biochemie-Zentrum RegensburgUniversity of Regensburg

Dear Editor, Successful sexual reproduction in plants requires exten- sive communication between male and female gametophytes, their gametes, and with the surrounding sporophytic tissues （Dresselhaus and Franklin-Tong 2013）. Over the past two decades, many peptides including small cysteine-rich peptides （***） and non-CRPs were found to play key roles in plant reproduction, mainly acting as signaling cues in this male- female communication （Marshall et al.2011）.

关键词： gene Figure Expression evidence CRP

Molecular evolution of the exon 2 of CHS genes and the possibility of its application to plant phylogenetic analysis

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Chinese Science Bulletin 2000年第19期45卷 1735-1742页

作者： WANG Jinling QU Lijia CHEN Jun gu hongya CHEN Zhangliang The National Laboratory of Protein Engineering and Plant Genetic Engineering Peking UniversityBeijing 100871China De Montfort University The GatewayLeicesterLEI 9BHUK

The exon 2 of chalcone synthase (CHS) gene is relatively conserved during evolution. In this study, three exon 2 fragments from two species in gymnosperm (Cycas panzhihuaensis, Ginkgo biloba ) and seven from four species in angiosperm (Magnolia denudata, Salix babylonica, Nymphaea tetragona, Camellia japonica) have been amplified by PCR from genomic DNA and sequenced. Together with other 73 sequences of CHS collected from EMBL database and literature, these sequences, which embrace 19 families of gymnosperm and angiosperm, have been analyzed for their phylogenetic relations by parsimony method. The result indicated that sequences from the same systematic family usually grouped together except those from Theaceae, Magnoliaceae and Nymphaeaceae. The relative rate test revealed the rate heterogeneity of CHS genes among the families. For the nucleotide substitution the sequences from Asteraceae and Solanaceae evolve faster than those from the other families analyzed while the sequences from Poaceae,

关键词： chalcone synthase (CHS) PCR phylogeny molecular evolution.

GAMT2 Encodes a Methyltransferase of Gibberellic Acid That is Involved in Seed Maturation and Germination in Arabidopsis

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Journal of Integrative Plant Biology 2007年第3期49卷 368-381页

作者： Shufan Xing Genji Qin Yan Shi Zhangliang Chen hongya gu Li-Jia Qu National Laboratory for Protein Engineering and Plant Genetic Engineering Peking-Yale Joint Research Center for Plant Molecular Genetics and AgroBiotechnology College of Life Sciences Peking University Beijing 100871 China National Plant Gene Research Center (Beijing) Beijing 100101 China

Salicylic acid methyltransferase （SAMT）, benzoic acid methyltransferase （BAMT） and theobromine methyltransferase （TH）（henceforth, SABATH） family proteins belong to a unique class of mehtyltransferase that can methylate small molecular compounds Including indole-3-acidic acid （IAA）, salicylic acid （SA） and jasmonic acid （JA）, in plants. Here we report that the GAMT2 protein, which has 34.2% similarity with IAMT1 in the amino acid sequence, can methylate gibberellic acid （GA）. Biolnformatics analysis suggests that GAMT2 may be able to methylate one molecule larger than SA. GAMT2 is predominantly expressed in the developing seed embryo and endosperm in Arabidopsis. During seed germination, the expression of GAMT2 decreases until the cotyledons expand out of the seed coat. Overexpression of GAMT2 in Arabidopsis resulted in multiple phenotypes, including dwarfism, retarded growth, late flowering, and reduced fertility, which are similar to the phenotypes of GA-deficient mutants. Seed germination assay showed that GAMT2 overexpression in plants was hypersensitive to GA biosynthesis inhibitor （ancymidol） and abscisic acid （ABA） treatments, whereas the GAMT2 null mutant （SALK_075450） was slightly Insensitive to such treatments, suggesting that GAMT2 may methylate GA or ABA. Enzyme activity analysis indicated that GAMT2 was able to methylate GA3 into Methyi-GA3 in vitro, but could not methylate ABA. Microarray analysis on GAMT2 overexpression plants suggested that Methyl-GA may be an Inactive form of GA in Arabidopsis. These data suggest that GAMT2 Is Involved in seed maturation and germination by modulating GA activity.

关键词： GAMT2 GA-deficient seed germination seed maturation Me-GA methyl acceptors SABATH methyltransferase

Over-expression of WOX1 Leads to Defects in Meristem Development and Polyamine Homeostasis in Arabidopsis

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Journal of Integrative Plant Biology 2011年第6期53卷 493-506页

作者： Yanxia Zhang Renhong Wu Genji Qin Zhangliang Chen hongya gu Li-Jia Qu State KeyLaboratory for Protein- and Plant 'Gene-I~esearch 'Pe--king-Yale Joi~ Research Center iorl~la-nt Molecular Genetics andAgroBiotechnology College of Life Sciences Peking University Beijing 100871 China The National Plant Gene Research Center (Beijingi Beijing 100101 China

In plants, the meristem has to maintain a separate population of pluripotent cells that serve two main tasks, i.e., self-maintenance and organ initiation, which are separated spatially in meristem. Prior to our study, WUS and *** WOX genes had been reported as essential for the development of the SAM. In this study, the consequences of gain of WOX1 function are described. Here we report the identification of an Arabidopsis gain-of-function mutant woxl-D, in which the expression level of the WOX1 （WUSCHEL HOMEOBOX 1） was elevated and subtle defects in meristem development were observed. The woxl-D mutant phenotype is dwarfed and slightly bushy, with a smaller shoot apex. The woxl-D mutant also produced small and dark green leaves, and exhibited a failure in anther dehiscence and male sterility. Molecular evidences showed that the transcription of the stem cell marker gene CLV3 was down-regulated in the meristem of woxl-D but accumulated in the other regions, i.e., in the root-hypocotyl junction and at the sites for lateral root initiation. The fact that the organ size and cell size in leaves of woxl-D are smaller than those in wild type suggests that cell expansion is possibly affected in order to have partially retarded the development of lateral organs, possibly through alteration of CLV3 expression pattern in the meristem. An S-adenosylmethionine decarboxylase （SAMDC） protein, SAMDC1, was found able to interact with WOX1 by yeast two-hybrid and pull-down assays in vitro. HPLC analysis revealed a significant reduction of polyamine content in woxl-D. Our results suggest that WOX1 plays an important role in meristem development in Arabidopsis, possibly via regulation of SAMDC activity and polyamine homeostasis, and/or by regulating CLV3 expression.

关键词： WOX CLV3 meristems polyamines SAMDC Arabidopsis.

Novel DYW-type pentatricopeptide repeat(PPR) protein BLX controls mitochondrial RNA editing and splicing essential for early seed development of Arabidopsis

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Journal of Genetics and Genomics 2018年第3期45卷 155-168页

作者： Yan Sun Jiaying Huang Sheng Zhong hongya gu Shan He Li-Jia Qu State Key Laboratory for Protein and Plant Gene Research Peking-Tsinghua Center for Life Sciences at College of Life Sciences Peking University Beijing 100871 China The National Plant Gene Research Center (Beijing) Beijing 100101 China

In plants, RNA editing is a post-transcriptional process that changes specific cytidine to uridine in both mitochondria and plastids. Most pentatricopeptide repeat（PPR） proteins are involved in organelle RNA editing by recognizing specific RNA sequences. We here report the functional characterization of a PPR protein from the DYW subclass, Baili Xi（BLX）, which contains five PPR motifs and a DYW domain. BLX is essential for early seed development, as plants lacking the BLX gene was embryo lethal and the endosperm failed to initiate cellularization. BLX was highly expressed in the embryo and endosperm, and the BLX protein was specifically localized in mitochondria, which is essential for BLX function. We found that BLX was required for the efficient editing of 36 editing sites in mitochondria. Moreover, BLX was involved in the splicing regulation of the fourth intron of nad1 and the first intron of nad2. The loss of BLX function impaired the mitochondrial function and increased the reactive oxygen species（ROS） level. Genetic complementation with truncated variants of BLX revealed that, in addition to the DYW domain, only the fifth PPR motif was essential for BLX function. The upstream sequences of the BLX-targeted editing sites are not conserved, suggesting that BLX serves as a novel and major mitochondrial editing factor（MEF） via a new non-RNA-interacting manner. This finding provides new insights into how a DYW-type PPR protein with fewer PPR motifs regulates RNA editing in plants.

关键词： Arabidopsis PPR RNA editing RNA splicing Embryo and endosperm development

Overexpression of the Wounding-Responsive Gene AtMYB15 Activates the Shikimate Pathway in Arabidopsis

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Journal of Integrative Plant Biology 2006年第9期48卷 1084-1095页

作者： Yanhui Chen Xiangbo Zhang Wei Wu Zhangliang Chen hongya gu Li-Jia Qu National Laboratory for Protein Engineering and Plant Genetic Engineering Peking-Yale Joint Research Center for Plant Molecular Genetics and AgroBiotechnology College of Life Sciences Peking University Beijing 100871 China The National Plant Gene Research Center （Beijing） Beijing 100101 China

The MYB transcription factor genes play important roles in many developmental processes and various defense responses of plants. The shikimate pathway is a major biosynthetic pathway for the production of three aromatic amino acids and other aromatic compounds that are involved in multiple responses of plants, including protection against UV and defense. Herein, we describe the characterization of the R2R3-MYB gene AtMYB15as an activator of the shikimate pathway in Arabidopsis. The AtMYB15 protein is nuclear localized and a transcriptional activation domain is found in its C-terminal portion. Northern blots showed that AtMYB15 is an early wounding-inducible gene. Resutls of microarray analysis, confirmed using quantitative real-time polymerase chain reaction, showed that overexpression of AtMYB15 in transgenic plants resulted in elevated expression of almost all the genes involved in the shikimate pathway. Bioinformatics analysis showed that one or more AtMYB15-binding AC elements were detected in the promoters of these upregulated genes. Furthermore, these genes in the shikimate pathway were also found to be induced by wounding. These data suggest an important role of AtMYB15as a possible direct regulator of the Arabidopsis shikimate pathway in response to wounding.

关键词： MYB transcription factor shikimate pathway transcriptional activation domain wounding inducible