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Analysis of Natural Allelic Variation Controlling Arabidopsis thaliana Seed Germinability in Response to Cold and Dark： Identification of Three Major Quantitative Trait Loci

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Molecular Plant 2008年第1期1卷 145-154页

作者： Ping-Hong Meng Audrey Macquet Olivier Loudet Annie Marion-Poll Helen M. North Laboratoire de Biologie des Semences UMR 204 INRA AgroParisTech Institut Jean-Pierre Bourgin F-78026 Versailles Cedex France Station de Genetique et Amelioration des Plantes INRA Institut Jean-Pierre Bourgin F-78026 Versailles Cedex France Present address: Institute of Horticulture Guizhou Academy of Agricultural Sciences GuiYang Guizhou Province 550006 RR. China

Light and temperature are key external factors in the control of Arabidopsis thaliana seed germination and dormancy mechanisms. Perception and response to these stimuli have to ensure that seedling emergence and growth occur at the most advantageous time for correct establishment. Analysis of over 300 Arabidopsis accessions identified 14, from 12 different geographical locations, that were able to germinate to greater than 20% at 6℃ in the dark. This natural variation was exploited to identify genetic loci responsible for cold-tolerant, dark germination. A quantitative trait loci approach was used on recombinant inbred line progeny of a cross between Bay-0 and Shahdara. Six distinct quantitative trait loci were identified, three of which were major loci, each responsible for 17-25% of the phenotypic variability in this trait. Parental phenotypes indicated that the majority of the cold-tolerant, dark-germination characteristics are related to light responses. Validation of the three major loci using heterogeneous inbred families confirmed the feasibility of fine mapping and cloning the genes at the quantitative trait loci responsible for cold-tolerant, dark germination.

关键词： Three Major Quantitative Cold and Dark Natural Allelic Variation

Dynamics of Strigolactone Function and Shoot Branching Responses in Pisum sativum

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Molecular Plant 2013年第1期6卷 128-140页

作者： Elizabeth A. Dun Alexandre de Saint Germain Catherine Rameau Christine A. Beveridge The University of Queensland School of Biological Sciences St Lucia QLD 4072 Australia Institut jean-pierre bourgin INRA UMR1318 INRA-AgroParisTech F-78000 Versailles France

Strigolactones （SLs）, or their metabolites, were recently identified as endogenous inhibitors of shoot branch- ing. However, certain key features and dynamics of SL action remained to be physiologically characterized. Here we show that successive direct application of SL to axillary buds at every node along the stem can fully inhibit branching. The SL inhibition of early outgrowth did not require inhibitory signals from other growing buds or the shoot tip. In add- ition to this very early or initial suppression of outgrowth, we also found SL to be effective, up to a point, at moderating the continuing growth of axillary branches. The effectiveness of SL at affecting bud and branch growth correlated with the ability of SL to regulate expression of PsBRC1. PsBRC1 is a transcription factor that is expressed strongly in axillary buds and is required for SL inhibition of shoot branching. Consistent with a dynamic role of the hormone, SL inhibition of bud growth did not prevent buds from later responding to a decapitation treatment, even though SL treatment immediately after decapitation inhibits the outgrowth response. Also, as expected from the hypothesized branching control network in plants, treatment of exogenous SL caused feedback down-regulation of SL biosynthesis genes within 2 h. Altogether, these results reveal new insights into the dynamics of SL function and support the premise that SLs or SL-derived metabolites function dynamically as a shoot branching hormone and that they act directly in axillary buds.

关键词： hormonal regulation dormancy shoot branching.

The ABA-Deficiency Suppressor Locus HAS2 Encodes the PPR Protein LOI1/MEF11 Involved in Mitochondrial RNA Editing

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Molecular Plant 2015年第4期8卷 644-656页

作者： Julien Sechet Camille Roux Anne Plessis Delphine Effroy Anne Frey Francois Perreau Catherine Biniek Anja Krieger-Liszkay David Macherel Helen M. North Hakim Mireau Annie Marion-Poll INRA Institut Jean-Pierre Bourgin UMR 1318 ERL CNRS 3559 Saclay Plant Sciences RD10 F-78026 Versailles France AgroParisTech Institut Jean-Pierre Bourgin UMR 1318 ERL CNRS 3559 Saclay Plant Sciences RD10 F-78026 Versailles France CEA Saclay IBiTec-S CNRS UMR 8221 Serv Bioenerget Biol Struct & Mecanisme F-91191 Gif Sur Yvette France Universite d'Angers UMR IRHS 1345 INRA Agrocampus Ouest F-49045 Angers France Institut jean-pierre bourgin INRA Bat 2 F-78026 Versailles Cedex France Institut jean-pierre bourgin INRA Bat 7 F-78026 Versailles Cedex France Department of Biology New York University 1009 Main Building 100 Washington Square East New York NY 10003-6688 USA

The hotABA-deficiency suppressor2 （has2） mutation increases drought tolerance and the ABA sensitivity of stomata closure and seed germination. Here we report that the HAS2 locus encodes the MITOCHONDRIAL EDITING FACTOR11 （MEF11）, also known as LOVASTATIN INSENSITIVE1. has21mef11 mutants exhibited phenotypes very similar to the ABA-hypersensitive mutant, hail-1 pp2ca-1 hab 1-1 abil-2, which is impaired in four genes encoding type 2C protein phosphatases （PP2C） that act as upstream negative regulators of the ABA signaling cascade. Like pp2c, mef11 plants were more resistant to progressive water stress and seed germination was more sensitive to paclobutrazol （a gibberellin biosynthesis inhibitor） as well as mannitol and NaCI, compared with the wild-type plants. Phenotypic alterations in mef11 were associated with the lack of editing of transcripts for the mitochondrial cytochrome c maturation FN2 （cCmFN2） gene, which encodes a cytochrome c-heme lyase subunit involved in cytochrome c biogenesis. Although the abundance of electron transfer chain complexes was not affected, their dysfunction could be deduced from increased respiration and altered production of hydrogen peroxide and nitric oxide in reef11 seeds. As minor defects in mitochondrial respiration affect ABA signaling, this suggests an essential role for ABA in mitochondrial retrograde regulation.

关键词： abscisic acid mitochondria pentatricopeptide repeat protein RNA editing drought tolerance germination

The miR166-SIHB15A regulatory module controls ovule development and parthenocarpic fruit set under adverse temperatures in tomato

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Molecular Plant 2021年第7期14卷 1185-1198页

作者： Christian Clepet Ravi Sureshbhai Devani Rachid Boumlik Yanwei Hao Halima Morin Fabien Marcel Marion Verdenaud Brahim Mania Gwilherm Brisou Sylvie Citerne Gregory Mouille jean-Christophe Lepeltier Shai Koussevitzky Adnane Boualem Abdelhafid Bendahmane Institute of Plant Sciences Paris-Saclay INRAECNRSUniversite Paris-SaclayOrsay 91405France HM Clause Mas Saint-PierreQuartier La GalineSaint-Remy de Provence 13210France Hazera Seeds Ltd Berurim M.P.Shikmim 7983700Israel lnstitut jean-pierre bourgin INRAEVersaillesFrance Present address:Key Laboratory of Horticultural Crop Biology and Germplasm Innovation in South China College of HorticultureSouth China Agricultural UniversityGuangzhou 510642China

Fruit set is inhibited by adverse temperatures,with consequences on yield.We isolated a tomato mutant producing fruits under non-permissive hot temperatures and identified the causal gene as SIHB15A,belonging to classⅢhomeodomain leucine-zipper transcription factors.SIHB15A,loss-of-function mu-tants display aberrant ovule development that mimics transcriptional changes occurring in fertilized ovules and leads to parthenocarpic fruit set under optimal and non-permissive temperatures,in field and greenhouse conditions.Under cold growing conditions,SIHB15A is subjected to conditional haploinsufficiency and recessive dosage sensitivity controlled by microRNA 166(miR166).Knockdown of SIHB15A alleles by miR166 leads to a continuum of aberrant ovules correlating with parthenocarpic fruit set.Consistent with this,plants harboring an Slhb15a-miRNA166-resisiant allele developed normal ovules and were unable to set parthenocarpic fruit under cold conditions.DNA affinity purification sequencing and RNA-sequencing analyses revealed that SIHB15A is a bifunctional transcription factor expressed in the ovule integument.SIHB15A binds to the promoters of auxin-related genes to repress auxin signaling and to the promoters of ethylene-related genes to activate their expression.A survey of tomato genetic biodiversity identified pat and pat-1,two historical parthenocarpic mutants,as alleles of SIHB15A.Taken together,our findings demonstrate the role of SIHB15A as a sentinel to prevent fruit set in the absence of fertilization and provide a mean to enhance fruiting under extreme temperatures.

关键词： parthenocarpy dosage sensitivity HD-Ziplll miRNA166 heat stress cold stress

Alternative splicing enhances transcriptome complexity in desiccating seeds

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Journal of Integrative Plant Biology 2016年第12期58卷 947-958页

作者： Arunkumar Srinivasan Jose M. Jimenez-Gomez Fabio Fornara Wim J. J. Soppe Vittoria Brambilla Department of Plant Breeding and Genetics Max Planck Institute for Plant Breeding Research Cologne Germany Open Analytics Antwerp Belgium Institut jean-pierre bourgin Institut National de la Recherche Agronomique Versailles France University of Milan Department of Biosciences Milano 20133 Italy University of Milan Department of Agricultural and Environmental Sciences via Celoria 220133 Milano Italy

Before being dispersed in the environment, mature seeds need to be dehydrated. The survival of seeds after dispersal depends on their low hydration in combination with high desiccation tolerance. These characteristics are estab- lished during seed maturation. Some key seed maturation genes have been reported to be regulated by alternative splicing （AS）. However, so far AS was described only for single genes and a comprehensive analysis of AS during seed maturation has been lacking. We investigated gene expression and AS during Arabidopsis thaliana seed development at a global level, before and after desiccation. Bioinformatics tools were developed to identify differentially spliced regions within genes. Our data suggest the importance and shows the peculiar features of AS during seed desiccation. We identified AS in 347o of genes that are expressed at both timepoints before and after desiccation. Most of these AS transcript variants had not been found before in other tissues. Among the AS genes some seed master regulators could be found. Interestingly, 6% of all expressed transcripts were not transcriptionally regulated during desiccation, but only modified by AS. We propose that AS should be more routinely taken into account in the analysis of transcriptomic data to prevent overlooking potentially important regulators.

关键词： Alternative splicing Arabidopsis thaliana seed desiccation

Precision Breeding Made Real with CRISPR: Illustration through Genetic Resistance to Pathogens

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Plant Communications 2020年第5期1卷 1-20页

作者： Florian Veillet Mickael Durand Thomas Kroj Stella Cesari jean-Luc Gallois IGEPP INRAEInstitut AgroUniv RennesPloudaniel 29260France Germicopa Breeding KerguivarchChateauneuf Du Faou 29520France Institut jean-pierre bourgin INRAEAgroParisTechUniversitéParis-SaclayVersailles 78000France INRAE BGPIBiology and Genetics of Plant-Pathogen InteractionsCampus International de BaillarguetMontpellierFrance INRAE GAFLMontfavet F-84143France

Since its discovery as a bacterial adaptive immune system and its development for genome editing in eukaryotes,the CRISPR technology has revolutionized plant research and precision crop breeding.The CRISPR toolbox holds great promise in the production of crops with genetic disease resistance to increase agriculture resilience and reduce chemical crop protection with a strong impact on the environment and public health.In this review,we provide an extensive overviewon recent breakthroughs in CRISPR technology,including the newly developed prime editing system that allows precision gene editing in plants.We present how each CRISPR tool can be selected for optimal use in accordance with its specific strengths and limitations,and illustrate how the CRISPR toolbox can foster the development of genetically pathogen-resistant crops for sustainable agriculture.

关键词： CRISPR-Cas9 gene targeting base editing prime editing plant/pathogen interactions precision crop breeding

Homogalacturonan MethyI-Esterification and Plant Development

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Molecular Plant 2009年第5期2卷 851-860页

作者： Sebastian Wolf Gregory Mouille Jerome Pelloux Heidelberg Institute for Plant Science Im Neuenheimer Feld 360 69120 Heidelberg Germany Laboratoire de Biologie Cellulaire Institut Jean-Pierre Bourgin INRA Centre de Versailles-Grignon Route de St Cyr 78026 Versailles France EA3900-BioPI Biologie des Plantes et contrble des Insectes ravageurs Universite de Picardie 33 Rue St Leu 80039 Amiens France

The ability of a plant cell to expand is largely defined by the physical constraints imposed by its cell wall. Accordingly, cell wall properties have to be regulated during development. The pectic polysaccharide homogalacturonan is a major component of the plant primary walls. Biosynthesis and in muro modification of homogalacturonan have recently emerged as key determinants of plant development, controlling cell adhesion, organ development, and phyllotactic patterning. This review will focus on recent findings regarding impact of homogalacturonan content and methylesterification status of this polymer on plant life. Demethyl-esterification of homogalacturonan occurs through the action of the ubiquitous enzyme ＇pectin methyl-esterase＇. We here describe various strategies developed by the plant to finely tune the methyl-esterification status of homogalacturonan along key events of the plant lifecycle.

关键词： Carbohydrate metabolism cell walls Arabidopsis Pectin.

Pectin May Hinder the Unfolding of Xyloglucan Chains during Cell Deformation： Implications of the Mechanical Performance of Arabidopsis Hypocotyls with Pectin Alterations

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Molecular Plant 2009年第5期2卷 990-999页

作者： Willie Abasolo Michaela Eder Kazuchika Yamauchi Nicolai Obel Antje Reinecke Lutz Neumetzler John W.C. Dunlop Gregory Mouille Markus Pauly Herman Hofte Ingo Burgert Max-Planck-lnstitute of Colloids and Interfaces Department of Biomaterials Potsdam Germany College of Forestry and Natural Resources University of the Philippines Los Banos Philippines Department of Socio-Environmental Energy Science Kyoto University Japan Max-Planck-lnstitute for Molecular Plant Physiology Potsdam Germany Laboratoire de Biologie Cellulaire UR501 Institute Jean-Pierre Bourgin INRA Versailles France Michigan State University Plant Research Laboratory East Lansing Michigan USA

Plant cell walls, like a multitude of other biological materials, are natural fiber-reinforced composite materials. Their mechanical properties are highly dependent on the interplay of the stiff fibrous phase and the soft matrix phase and on the matrix deformation itself. Using specific Arabidopsis thaliana mutants, we studied the mechanical role of the matrix assembly in primary cell walls of hypocotyls with altered xyloglucan and pectin composition. Standard microtensile tests and cyclic loading protocols were performed on tour1 hypocotyls with affected RGII borate diester cross-links and a hindered xyloglucan fucosylation as well as qua2 exhibiting 50% less homogalacturonan in comparison to wild-type. As a control, wild-type plants （Col-0） and mur2 exhibiting a specific xyloglucan fucosylation and no differences in the pectin network were utilized. In the standard tensile tests, the ultimate stress levels （-tensile strength） of the hypocotyls of the mutants with pectin alterations （mur1, qua2） were rather unaffected, whereas their tensile stiffness was noticeably reduced in comparison to Col-0. The cyclic loading tests indicated a stiffening of all hypocotyls after the first cycle and a plastic deformation during the first straining, the degree of which, however, was much higher for tour1 and qua2 hypocotyls. Based on the mechanical data and current cell wall models, it is assumed that folded xyloglucan chains between cellulose fibrils may tend to unfold during straining of the hypocotyls. This response is probably hindered by geometrical constraints due to pectin rigidity.

关键词： Arabidopsis thaliana mutants cellulose xyloglucan pectin cyclic loading tests.

An atypical NLR gene confers bacterial wilt susceptibility in Arabidopsis

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Plant Communications 2023年第5期4卷 277-294页

作者： Choghag Demirjian Narjes Razavi Gang Yu Baptiste Mayjonade Lu Zhang Fabien Lonjon Fabien Chardon Se bastien Carrere Je rome Gouzy Ste phane Genin Alberto PMacho Fabrice Roux Richard Berthome Fabienne Vailleau LIPME Universite de ToulouseINRAECNRSCastanet-TolosanFrance Shanghai Center for Plant Stress Biology CAS Center for Excellence in Molecular Plant SciencesChinese Academy of SciencesShanghaiChina Universite Paris-Saclay INRAEAgroParisTechInstitut Jean-Pierre Bourgin(IJPB)78000 VersaillesFrance Present address:Department of Cell&Systems Biology University of TorontoTorontoONCanada Present address:Department of Biology NYU12 Waverly PlaceNew YorkNY 10003USA

Quantitative disease resistance(QDR)remains the most prevalent form of plant resistance in crop fields and wild habitats.Genome-wide association studies(GWAS)have proved to be successful in deciphering the quantitative genetic basis of complex traits such as QDR.To unravel the genetics of QDR to the devastating worldwide bacterial pathogen Ralstonia solanacearum,we performed a GWAS by challenging a highly polymorphic local mapping population of Arabidopsis thaliana with four R.solanacearum type III effector(T3E)mutants,identified as key pathogenicity determinants after a first screen on an A.thaliana core collection of 25 accessions.Although most quantitative trait loci(QTLs)were highly specific to the identity of the T3E mutant(ripAC,ripAG,ripAQ,and ripU),we finely mapped a common QTL located on a cluster of nucleotide-binding domain and leucine-rich repeat(NLR)genes that exhibited structural variation.We functionally validated one of these NLRs as a susceptibility factor in response to R.solanacearum,named it Bacterial Wilt Susceptibility 1(BWS1),and cloned two alleles that conferred contrasting levels of QDR.Further characterization indicated that expression of BWS1 leads to suppression of immunity triggered by different R.solanacearum effectors.In addition,we showed a direct interaction between BWS1 and RipAC T3E,and BWS1 and SUPPRESSOR OF G2 ALLELE OF skp1(SGT1b),the latter interaction being suppressed by RipAC.Together,our results highlight a putative role for BWS1 as a quantitative susceptibility factor directly targeted by the T3E RipAC,mediating negative regulation of the SGT1-dependent immune response.

关键词： Ralstonia solanacearum typeⅢeffectors Arabidopsis thaliana GWAS R gene susceptibility