PT-Flax (phenotyping and TILLinG of flax): development of a flax (Linum usitatissimumL.) mutant population and TILLinG platform for forward and reverse genetics

BMC Plant Biology

Research Article

Brigitte Thomasset¹  Olivier van Wuytswinkel²  Reynald Tavernier³  Christophe Pineau³  Xavier Guillot⁴  Jean-Paul Trouvé⁵  Christine Paysant-Leroux⁵  Hervé Demailly⁶  Laurent Gutierrez⁶  Brigitte Chabbert⁷  Abdelhafid Bendahmane⁸  Marion Dalmais⁸  Véronique Brunaud⁸  Maxime Chantreau⁹  Simon Hawkins⁹  Sébastien Grec⁹  Manash Chatterjee^1,10 

[1] CNRS-FRE 3580, GEC, Université de Technologie de Compiègne, CS 60319, 60203, Compiègne, cedex, France;EA 3900-BioPI, UFR des Sciences, UPJV, 33 rue Saint Leu, 80039, Amiens, cedex, France;LINEA, 20 Avenue Saget, 60 210, Grandvilliers, France;Laboulet Semences, 80 270, Airaines, France;Terre de Lin, société cooperative agricole, 76 740, Saint-Pierre-Le-Viger, France;UFR des Sciences, CRRBM, UPJV, 33 rue Saint Leu, 80039, Amiens, cedex, France;UMR614 Fractionnement des AgroRessources et Environnement, INRA, F-51100, Reims, France;UMR614 Fractionnement des AgroRessources et Environnement, Université de Reims Champagne-Ardenne, F-51100, Reims, France;URGV, Unité de Recherche en Génomique Végétale, Université d'Evry Val d'Essonne, INRA, 2 rue Gaston, 5708, 91057, Crémieux, Evry, CP, cedex, France;Université Lille Nord de France, Lille 1 UMR 1281, F-59650, Villeneuve d'Ascq, cedex, France;281 Stress Abiotiques et Différenciation des Végétaux Cultivés, INRA UMR, F-59650, Villeneuve d’Ascq, France;c/o Jai Research Foundation, Bench Bio Pvt Ltd., 396195, Vapi, Gujarat, India;National University of Ireland Galway (NUIG), University Road, Galway, Ireland;

关键词: Flax;    TILLinG;    Mutants;    Fiber;    Lignin;    Lignan;    Oil;    Fatty acids;   

DOI  :  10.1186/1471-2229-13-159

 received in 2013-07-05, accepted in 2013-10-09,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundFlax (Linum usitatissimum L.) is an economically important fiber and oil crop that has been grown for thousands of years. The genome has been recently sequenced and transcriptomics are providing information on candidate genes potentially related to agronomically-important traits. In order to accelerate functional characterization of these genes we have generated a flax EMS mutant population that can be used as a TILLinG (Targeting Induced Local Lesions in Genomes) platform for forward and reverse genetics.ResultsA population of 4,894 M2 mutant seed families was generated using 3 different EMS concentrations (0.3%, 0.6% and 0.75%) and used to produce M2 plants for subsequent phenotyping and DNA extraction. 10,839 viable M2 plants (4,033 families) were obtained and 1,552 families (38.5%) showed a visual developmental phenotype (stem size and diameter, plant architecture, flower-related). The majority of these families showed more than one phenotype. Mutant phenotype data are organised in a database and can be accessed and searched at UTILLdb (http://urgv.evry.inra.fr/UTILLdb). Preliminary screens were also performed for atypical fiber and seed phenotypes. Genomic DNA was extracted from 3,515 M2 families and eight-fold pooled for subsequent mutant detection by ENDO1 nuclease mis-match cleavage. In order to validate the collection for reverse genetics, DNA pools were screened for two genes coding enzymes of the lignin biosynthesis pathway: Coumarate-3-Hydroxylase (C3H) and Cinnamyl Alcohol Dehydrogenase (CAD). We identified 79 and 76 mutations in the C3H and CAD genes, respectively. The average mutation rate was calculated as 1/41 Kb giving rise to approximately 9,000 mutations per genome. Thirty-five out of the 52 flax cad mutant families containing missense or codon stop mutations showed the typical orange-brown xylem phenotype observed in CAD down-regulated/mutant plants in other species.ConclusionsWe have developed a flax mutant population that can be used as an efficient forward and reverse genetics tool. The collection has an extremely high mutation rate that enables the detection of large numbers of independant mutant families by screening a comparatively low number of M2 families. The population will prove to be a valuable resource for both fundamental research and the identification of agronomically-important genes for crop improvement in flax.

【 授权许可】

Unknown   
© Chantreau et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.