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

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.

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SERK Family Receptor-like Kinases Function as Co-receptors with PXY for Plant Vascular Development

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Molecular Plant 2016年第10期9卷 1406-1414页

作者： Heqiao Zhang Xiaoya lin Zhifu Han Jizong Wang li-jia qu Jijie Chai Ministry of Education Key Laboratory of Protein Science Center for Structural Biology School of Life Sciences Tsinghua-Peking Joint Center for Life SciencesTsinghua University Beijing 100084 China State Key Laboratory of Protein and Plant Gene Research School of Life Sciences Peking-Tsinghua Joint Center for Life Sciences Peking UniversityBeijing 100871 China

In Arabidopsis, the CLAVATA3/EMBRYO SURROUNDING REGION-RELATED （CLE） peptides play important roles in regulating proliferation and differentiation of plant-specific stem cells. Although receptors of CLEs are reported to be leucine-rich repeat receptor kinases, the mechanisms underlying CLE-induced receptor activation remain largely unknown. Here we show that SOMATIC EMBRYOGENESIS RECEPTOR KINASEs （SERKs） serve as co-receptors in CLE41/TDIF-PXY signaling to regulate plant vascular development. TDIF induces interaction of its receptor PXY with SERKs in vitro and in vivo. Furthermore, the serk1-1 serk2-1 bakl-5 mutant plants are less sensitive to TDIF, phenocopying the pxy mutant with a compromised promotion of procambial cell proliferation. Crystal structure of the PXY-TDIF-SERK2 complex reveals that the last amino acid of TDIF conserved among CLEs and other evolutionary-related peptides is important for the interaction between SERK2 and PXY. Taken together, our current study identifies SERKs as signaling components of the TDIF-PXY pathway and suggests a conserved activation mechanism of CLE receptors.

关键词： receptor-like kinase SERK PXY-TDIF vascular development co-receptor

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A nuclear-encoded mitochondrial gene AtCIB22 is essential for plant development in Arabidopsis

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Journal of Genetics and Genomics 2010年第10期37卷 667-683页

作者： lihua Han Genji Qin Dingming Kang Zhangliang Chen Hongya Gu li-jia qu College of Agronomy and Biotechnology China Agricultural University Beijing 100094 China National Laboratory of Protein Engineering and Plant Genetic Engineering Peking University Beijing 100871 China The National Plant Gene Research Center (Beijing) Beijing 100101 China

Complex I （the NADH：ubiquinone oxidoreductase） of the mitochondrial respiratory chain is a complicated, multi-subunit, membrane- bound assembly and contains more than 40 different proteins in higher plants. In this paper, we characterize the Arabidopsis homologue （designated as AtCIB22） of the B22 subunit of eukaryotic mitochondriai Complex I. AtCIB22 is a single-copy gene and is highly con- served throughout eukaryotes. AtCIB22 protein is located in mitochondria and the AtC1B22 gene is widely expressed in different tissues. Mutant Arabidopsis plants with a disrupted AtC1B22 gene display pleiotropic phenotypes including shorter roots, smaller plants and de- layed flowering. Stress analysis indicates that the AtC1B22 mutants＇ seed germination and early seedling growth are severely inhibited by sucrose deprivation stress but more tolerant to ethanol stress. Molecular analysis reveals that in moderate knockdown AtCIB22 mutants, genes including cell redox proteins and stress related proteins are significantly up-regulated, and that in severe knockdown AtCIB22 mu- tants, the alternative respiratory pathways including NDA1, NDB2, AOXla and AtPUMP1 are remarkably elevated. These data demon- strate that AtCIB22 is essential for plant development and mitochondrial electron transport chains in Arabidopsis. Our findings also en- hance our understanding about the physiological role of Complex I in plants.

关键词： mitochondria Complex I B22 subunit ethanol treatment alternative oxidase uncoupling protein

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Allelic Analyses of the Arabidopsis YUC1 Locus Reveal Residues and Domains Essential for the Functions of YUC Family of Flavin Monooxygenases

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Journal of Integrative Plant Biology 2011年第1期53卷 54-62页

作者： Xianhui Hou Sainan liu Florencia Pierri Xinhua Dai li-jia qu Yunde Zhao 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 Section of Cell and Developmental Biology University of California at San Diego La Jolla California 92093-0116 USA

Flavin monooxygenases （FMOs） play critical roles in plant growth and development by synthesizing auxin and other signaling molecules. However, the structure and function relationship within plant FMOs is not understood. Here we defined the important residues and domains of the Arabidopsis YUC1 FMO, a key enzyme in auxin biosynthesis. We previously showed that simultaneous inactivation of YUC1 and its homologue YUC4 caused severe defects in vascular and floral development. We mutagenized the yuc4 mutant and screened for mutants with phenotypes similar to those of yucl yuc4 double mutants. Among the isolated mutants, five of them contained mutations in the YUC1 gene. Interestingly, the mutations identified in the new yucl alleles were concentrated in the two GXGXXG motifs that are highly conserved among the plant FMOs. One such motif presumably binds to flavin adenine dinucleotide （FAD） cofactor and the other binds to nicotinamide adenine dinucleotide phosphate （NADPH）. We also identified the Ser139 to Phe conversion in yucl, a mutation that is located between the two nucleotide-binding sites. By analyzing a series of yucl mutants, we identified key residues and motifs essential for the functions of YUC1 FMO.

关键词： Auxins Residues Plants Planting Adenine Mutation Mutants

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Arabidopsis AtVPS15 Plays Essential Roles in Pollen Germination Possibly by Interacting with AtVPS34

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Journal of Genetics and Genomics 2012年第2期39卷 81-92页

作者： Wei-Ying Wang li Zhang Shufan Xing Zhiqiang Ma Jingjing liu Hongya Gu Genji Qin li-jia qu State Key Laboratory of Protein and Plant Gene Research College of Life SciencesPeking UniversityBeijing 100871China The National Plant Gene Research Center(Beijing) Beijing 100101China

VPS 15 protein is a component of the phosphatidylinositol 3-kinase complex which plays a pivotal role in the development of yeast and mammalian cells. The knowledge about the function of its homologue in plants remains limited. Here we report that AtVPS15, a homologue of yeast VPS15p in Arabidopsis, plays an essential role in pollen germination. Homozygous T-DNA insertion mutants of AtVPS15 could not be obtained from the progenies of self-pollinated heterozygous mutants. Reciprocal crosses between atvps15 mutants and wild-type Arabidopsis revealed that the T-DNA insertion was not able to be transmitted by male gametophytes. DAPI staining, Alexander＇s stain and scanning electron microscopic analysis showed that atvps15 heterozygous plants produced pollen grains that were morphologically indistinguishable from wild-type pollen, whereas in vitro germination experiments revealed that germination of the pollen grains was defective. GUS staining analysis of transgenic plants expressing the GUS reporter gene driven by the AtVPS15 promoter showed that AtVPS15 was mainly expressed in pollen grains. Finally, DUALmembrane yeast two-hybrid analysis demonstrated that AtVPS15 might interact directly with AtVPS34. These results suggest that AtVPS15 is very important for pollen germination, possibly through modulation of the activity of PI3-kinase.

关键词： Arabidopsis AtVPS15 Pollen germination PI3-kinase

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The Arabidopsis Anaphase-Promoting Complex/Cyclosome Subunit 1 is Critical for Both Female Gametogenesis and Embryogenesis

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Journal of Integrative Plant Biology 2013年第1期55卷 64-74页

作者： Yanbing Wang Yingnan Hou Hongya Gu Dingming Kang Zhang-liang Chen Jingjing liu li-jia qu State Key Laboratory of Protein and Plant Gene Research College of Life Sciences Peking University College of Agronomy and Biotechnology China Agricultural University The National Plant Gene Research Center

Anaphase-promoting complex/cyclosome （APC/C）, a multisubunit E3 ligase, plays a critical role in cell cycle control, but the functional characterization of each subunit has not yet been completed. To investigate the function of APC1 in Arabidopsis, we analyzed four mutant alleles of APC1, and found that mutation in APC1 resulted in significantly reduced plant fertility, accumulation of cyclin B, and disrupted auxin distribution in embryos. The three mutant alleles apcl-1, apcl-2 and apcl-3 shared variable defects in female gametogenesis including degradation, abnormal nuclear number, and disrupted polarity of nuclei in the embryo sac as well as in embryogenesis, in which embryos were arrested at multiple stages. All of these defects are similar to those previously identified in apc4. The mutant apcl-4, in which the T-DNA was inserted after the transmembrane domain at the C-terminus, showed much more severe phenotypes; that is, most of the ovules were arrested at the one-nucleate female gametophyte stage （stage FG1）. In the apcl apc4 double mutants, the fertility was further reduced by one-third in apcl-ll＋ apc4-1/＋, and in some cases no ovules even survived in siliques of apcl-4/＋ apc4-1/＋. Our data thus suggest that APC1, an essential component of APC/C, plays a synergistic role with APC4 both in female gametogenesis and in embryogenesis.

关键词： Arabidopsis anaphase-promoting complex/cyclosome (APC/C) APC1 APC4 female gametogenesis embryogenesis synergistic role

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The egg cell is preferentially fertilized in Arabidopsis double fertilization

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Journal of Integrative Plant Biology 2022年第11期64卷 2039-2046页

作者： ling li Saiying Hou Wei Xiang Zihan Song Yuan Wang li Zhang Jing li Hongya Gu Juan Dong Thomas Dresselhaus Sheng Zhong li-jia qu State Key Laboratory for Protein and Plant Gene Research Peking‐Tsinghua Center for Life Sciences at College of Life SciencesPeking UniversityBeijing 100871China Beijing Advanced Innovation Center for Tree Breeding by Molecular Design College of Bioscience and Resources EnvironmentBeijing University of AgricultureBeijing 102206China The National Plant Gene Research Center Beijing 100101China The Waksman Institute of Microbiology Rutgers the State University of New JerseyPiscatawayNJ"2"08854USA Cell Biology and Plant Biochemistry University of RegensburgRegensburg 93053Germany

In flowering plants(angiosperms),fertilization of the egg cell by one sperm cell produces an embryo,whereas fusion of a second sperm cell with the central cell generates the *** most angiosperms like Arabidopsis,a pollen grain contains two isomorphic sperm cells required for this double fertilization process.A long-standing unsolved question is whether the two fertilization events have any preference.A tool to address this question is the usage of the cyclin-dependent kinase a1(cdka;1)mutant pollen,which produces a single sperm-like cell(SLC).Here,we first adopt a complementation-based fluorescence-labeling method to successfully separate and collect cdka;1 mutant pollen containing a single ***-cell RNA-sequencing analysis revealed that cdka;1 SLCs show a gene expression profile highly similar to that of sperm cells and not to the generative cell,precursor of the two sperm *** assays using a limited number of cdka;1 mutant pollen revealed that in 98.2%of the ovules,single fertilization of the egg cell *** of pistils with excessive cdka;1 mutant pollen allowed the delivery of a second SLC via fertilization recovery,which fertilized the central cell,resulting in 20.7%double-fertilized *** indicates that cdka;1 SLCs are able to fertilize both the egg and the central *** together,our findings have answered a long-standing question and support that preferential fertilization of the egg cell is evident in Arabidopsis.

关键词： Arabidopsis double fertilization egg cell preferential fertilization

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Lack of ethylene does not affect reproductive success and synergid cell death in Arabidopsis

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Molecular Plant 2022年第2期15卷 354-362页

作者： Wenhao li Qiyun li Mohan Lyu Zhijuan Wang Zihan Song Shangwei Zhong Hongya Gu Juan Dong Thomas Dresselhaus Sheng Zhong li-jia qu State Key Laboratory for Protein and Plant Gene Research Peking-Tsinghua Center for Life Sciences at College of Life SciencesPeking UniversityBeijing 100871People's Republic of China The National Plant Gene Research Center(Beijing) Beijing 100101People's Republic of China The Waksman Institute of Microbiology Rutgers the State University of New JerseyPiscatawayNJ 08854USA Cell Biology and Plant Biochemistry University of Regensburg93053 RegensburgGermany

The signaling pathway of the gaseous hormone ethylene is involved in plant reproduction,growth,devel-opment,and stress *** reproduction,the two synergid cells of the angiosperm female gametophyte both undergo programmed cell death(PCD)/degeneration but in a different manner:PCD/degeneration of one synergid facilitates pollen tube rupture and thereby the release of sperm cells,while PCD/degeneration of the other synergid blocks supernumerary pollen *** signaling was postu-lated to participate in some of the synergid cell functions,such as pollen tube attraction and the induction of PCD/***,ethylene-mediated induction of synergid PCD/degeneration and the role of ethylene itself have not been firmly ***,we employed the CRISPR/Cas9 technology to knock out the five ethylene-biosynthesis 1-aminocyclopropane-1-carboxylic acid oxidase(ACO)genes and created Arabidopsis mutants free of ethylene *** ethylene-free mutant plants showed normal triple responses when treated with ethylene rather than 1-aminocyclopropane-1-carboxylic acid,but had increased lateral root density and enlarged petal sizes,which are typical phenotypes of mutants defective in ethylene *** these ethylene-free plants,we further demonstrated that production of ethylene is not necessarily required to trigger PCD/degeneration of the two synergid cells,but certain com-ponents of ethylene signaling including transcription factors ETHYLENE-INSENSITIVE 3(EIN3)and EIN3-liKE 1(EIL1)are necessary for the death of the persistent synergid cell.

关键词： ethylene ACC PCD synergid cells triple response Arabidopsis

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SNAIL1 is essential for female gametogenesis in Arabidopsis thaliana

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Journal of Integrative Plant Biology 2017年第9期59卷 629-641页

作者： lihong Hao Xiaolin Wei Jiulei Zhu jiao Shi Jingjing liu Hongya Gu Tomohiko Tsuge li-jia qu State Key Laboratory for Protein and Plant Gene Research Peking-Tsinghua Center for Life SciencesSchool of Life SciencesPeking UniversityBeijing 100871China National Plant Gene Research Center(Beijing) Bejing 100101China Institute for Chemical Research Kyoto UniversityGokasho UjiKyoto 611-0011Japan

Two yeast Brix family members Ssfl and Ssf2, involved in large ribosomal subunit synthesis, are essential for yeast cell viability and mating efficiency. Their putative homologs exist in the Arabidopsis genome; however, their role in plant development is unknown. Here, we show that Arabidopsis thaliana SNAIL1 （AtSNAIL1）, a protein sharing high sequence identity with yeast Ssfl and Ssf2, is critical to mitosis progression of female gametophyte development. The snail1 homozygous mutant was nonviable and its heterozygous mutant was semi-sterile with shorter siliques. The mutation in SNAIL1 led to absence of female transmission and reduced male transmission. Further phenotypic analysis showed that the synchronic development of female gametophyte in the snail1 heterozygous mutant was greatly impaired and the snaih pollen tube growth, in vivo, was also compromised. Furthermore, SNAIL1 was a nucleolar- localized protein with a putative role in protein synthesis. Our data suggest that SNAIL1 may function in ribosome biogenesis like Ssf1 and Ssf2 and plays an important role during megagametogenesis in Arabidopsis.

关键词： SNAIL1 is essential for female gametogenesis in Arabidopsis thaliana Figure

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Egg cell-secreted aspartic proteases ECS1/2 promote gamete attachment to prioritize the fertilization of egg cells over central cells in Arabidopsis

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Journal of Integrative Plant Biology 2022年第11期64卷 2047-2059页

作者： jiahao jiang Nils Stührwohldt Tianxu liu Qingpei Huang ling li li Zhang Hongya Gu liumin Fan Sheng Zhong Andreas Schaller li-jia qu State Key Laboratory for Protein and Plant Gene Research Peking-Tsinghua Center for Life Sciences at College of Life SciencesPeking UniversityBeijing 100871China Department of Plant Physiology and Biochemistry Institute of BiologyUniversity of HohenheimStuttgart 70593Germany

Double fertilization is an innovative phenomenon in angiosperms,in which one sperm cell first fuses with the egg cell to produce the embryo,and then the other sperm fuses with the central cell to produce the ***,the molecular mechanism of the preferential fertilization of egg cells is poorly *** this study,we report that two egg cell-secreted aspartic proteases,ECS1 and ECS2,play an important role in promoting preferential fertilization of egg cells in *** show that simultaneous loss of ECS1 and ECS2 function resulted in an approximately 20%reduction in fertility,which can be complemented by the full-length ECS1/2 but not by corresponding active site mutants or by secretion-defective versions of ECS1/*** phenotypic analysis revealed that the egg cellsperm cell attachment was compromised in ecs1 ecs2 *** pollination assays with cyclin-dependent kinase a1(cdka;1)pollen showed that preferential egg cell fertilization was impaired in the ecs1 ecs2 *** together,these results demonstrate that egg cells secret two aspartic proteases,ECS1 and ECS2,to facilitate the attachment of sperm cells to egg cells so that preferential fertilization of egg cells is *** study reveals the molecular mechanism of preferential fertilization in Arabidopsis thaliana.

关键词： Arabidopsis thaliana double fertilization ECS1/2 gamete interaction preferential fertilization

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