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Disruption of phytoene desaturase gene results in albino and dwarf phenotypes in Arabidopsis by impairing chlorophyll, carotenoid, and gibberellin biosynthesis

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Cell Research 2007年第5期17卷 471-482页

作者： Genji Qin1 hongya gu1,2 Ligeng Ma1,3 Yiben Peng1 Xing Wang Deng1,3 Zhangliang Chen1 Li-Jia Qu1,2 1 Peking-Yale Joint Center for Plant Molecular Genetics and Agrobiotechnology, National Laboratory for Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China 2 The National Plant Gene Research Center (Beijing), Beijing 100101, China 3 Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven Connecticut 62520-8104, USA Peking-Yale Joint Center for Plant Molecular Genetics and Agrobiotechnology National Laboratory for Protein Engineering and Plant Genetic Engineering College of Life Sciences Peking University Beijing China The National Plant Gene Research Center (Beijing) Beijing China Department of Molecular Cellular and Developmental Biology Yale University New Haven Connecticut USA

Carotenoids play an important role in many physiological processes in plants and the phytoene desaturase gene （PDS3）encodes one of the important enzymes in the carotenoid biosynthesis pathway. Here we report the identification andanalysis of a T-DNA insertion mutant of PDS3 gene. Functional complementation confirmed that both the albino anddwarf phenotypes of the pds3 mutant resulted from functional disruption of the PDS3 gene. Chloroplast development wasarrested at the proplastid stage in the pds3 mutant. Further analysis showed that high level of phytoene was accumulatedin the pds3 mutant. Addition of exogenous GA3could partially rescue the dwarf phenotype, suggesting that the dwarfphenotype of the pds3 mutant might be due to GA deficiency. Microarray and RT-PCR analysis showed that disruptingPDS3 gene resulted in gene expression changes involved in at least 20 metabolic pathways, including the inhibition ofmany genes in carotenoid, chlorophyll, and GA biosynthesis pathways. Our data suggest that the accumulated phytoenein the pds3 mutant might play an important role in certain negative feedbacks to affect gene expression of diverse cel-lular pathways.

关键词： Arabidopsis thaliana phytoene desaturase albino dwarf microarray analysis

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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.

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Characterization of Arabidopsis MYB transcription factor gene AtMYB17 and its possible regulation by LEAFY and AGL15

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Journal of Genetics and Genomics 2009年第2期36卷 99-107页

作者： Yunfei Zhang guangyu Cao Li-Jia Qu hongya gu National Laboratory of Protein Engineering and Plant Genetic Engineering College of Life Sciences Peking University Beijing 100871 China

MYB transcription factors compose one of the largest transcription factor families in Arabidopsis, which play important roles in vari- ous developmental processes as well as defense responses against environmental stresses. In this study, we report the characterization of AtMYB17 gene, a putative R2R3 type MYB gene family member in Arabidopsis. AtMYB17 was found exclusively localized in nuclear, with an activation domain at its C-terminus. AtMYBI7 was highly expressed in inflorescences and siliques, especially at early flower developmental stages. The level of AtMYB17 transcripts was also found to increase after imbibition during seed germination and gradually concentrate to the shoot apex. Bioinformatics analysis identified several binding sites of LEAFY （LFY） and AGL15 in the promoter re- gion of AtMYB17. Promoter-guS fusion analysis showed that the LFY binding sites were important in fine-tuning regulation of the spatio-temporal expression ofAtMYB17 in transgenic plants. Moreover, AtMYB17 was up-regulated in 35S：：AGL15 plants. Taken together, our data suggest that LFY may be involved in the regulation of AtMYB17, possibly together with AGL 15, and thereafter in early inflores- cence development and seed germination.

关键词： AtMYB17 transcription factor Arabidopsis LEAFY AGL 15

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Expression and functional analysis of the rice plasma-membrane intrinsic protein gene family

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Cell Research 2006年第3期16卷 277-286页

作者： Lei guo~1 Zi Yi Wang~1 Hong Lin~1 Wei Er Cui~1 Jun Chen~1 Meihua Liu~1 Zhang Liang Chen~(1,2) Li Jia Qu~(1,2) hongya gu~(1,2)1 Peking-Yale Joint Research Center for Plant Molecular Genetics and AgroBiotechnology,National Laboratoryfor Protein Engineering and Plant Genetic Engineering,College of Life Sciences,Peking University,Beijing 100871,China 2 The National Plant Gene Research Center,Beijing 100101,China Peking-Yale Joint Research Center for Plant Molecular Genetics and AgroBiotechnology National Laboratory for Protein Engineering and Plant Genetic Engineering College of Life Sciences Peking University Beijing China The National Plant Gene Research Center Beijing China

Plasma membrane intrinsic proteins (PIPs) are a subfamily of aquaporins that enable fast and controlled translocationof water across the *** this study,we systematically identified and cloned ten PIP genes from *** on thesimilarity of the amino acid sequences they encoded,these rice PIP genes were classified into two groups and designatedas OsPIPI-1 to OsPIP1-3 and OsPIP2-1 to OsPIP2-7 following the nomenclature of PIP genes in ***-PCR analysis identified three root-specific and one leaf-specific OsPIP ***,the expression profile ofeach OsPIP gene in response to salt,drought and ABA treatment was examined in *** on transgenic plantsover-expressing of either OsPIP1 (OsPIPI-1) or OsPIP2 (OsPIP2-2) in wild-type Arabidopsis,showed enhanced toler-ance to salt (100 mM of NaCl) and drought (200 mM of mannitol),but not to salt treatment of higher concentration (150mM of NaCl).Taken together,these data suggest a distinct role of each OsPIP gene in response to different stresses,andshould add a new layer to the understanding of the physiological function of rice PIP genes.

关键词： aquaporins PIP quantitative RT-PCR Oryza sativa organ-specific

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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

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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.

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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

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Arabidopsis AtBECLIN 1/AtAtg6/AtVps30 is essential for pollen germination and plant development

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Cell Research 2007年第3期17卷 249-263页

作者： Genji Qin~1 Zhiqiang Ma~1 Li Zhang~1 Shufan Xing~1 Xianhui Hou~1 Jie Deng~1 Jingjing Liu~1 Zhangliang Chen~(1,2) Li-Jia Qu~(1,2) hongya gu~(1,2) ~1National 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 ~2The National Plant Gene Research Center (Beijing),Beijing 100101,China 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

Pollen germination on the surtace ot compatible stigmatic tissues is an essential step tor plant *** wereport that the Arabidopsis mutant bcl1 is male sterile as a result of the failure of pollen *** show that the bcl1mutant allele cannot be transmitted by male gametophytes and no homozygous bcl1 mutants were *** ofpollen developmental stages indicates that the bcl1 mutation affects pollen germination but not pollen ***-lar analysis demonstrates that the failure of pollen germination was caused by the disruption of AtBECLIN ***1 is expressed predominantly in mature pollen and encodes a protein with significant homology to Beclin1/Atg6/Vps30required for the processes of autophagy and vacuolar protein sorting(VPS)in *** also show that AtBECLIN 1 isrequired for normal plant development,and that genes related to autophagy,VPS and the glycosylphosphatidylinositolanchor system,were affected by the deficiency of AtBECLIN 1.

关键词： Arabidopsis thaliana AtBECLIN 1 pollen germination plant normal development

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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

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