Identification of Mendel's White Flower Character

BACKGROUND: The genetic regulation of flower color has been widely studied, notably as a character used by Mendel and his predecessors in the study of inheritance in pea. METHODOLOGY/PRINCIPAL FINDINGS: We used the genome sequence of model legumes, together with their known synteny to the pea genome to identify candidate genes for the A and A2 loci in pea. We then used a combination of genetic mapping, fast neutron mutant analysis, allelic diversity, transcript quantification and transient expression complementation studies to confirm the identity of the candidates. CONCLUSIONS/SIGNIFICANCE: We have identified the pea genes A and A2. A is the factor determining anthocyanin pigmentation in pea that was used by Gregor Mendel 150 years ago in his study of inheritance. The A gene encodes a bHLH transcription factor. The white flowered mutant allele most likely used by Mendel is a simple G to A transition in a splice donor site that leads to a mis-spliced mRNA with a premature stop codon, and we have identified a second rare mutant allele. The A2 gene encodes a WD40 protein that is part of an evolutionarily conserved regulatory complex

Stock price reaction to profit warnings: The role of time-varying betas

This study investigates the role of time-varying betas, event-induced variance and conditional heteroskedasticity in the estimation of abnormal returns around important news announcements. Our analysis is based on the stock price reaction to profit warnings issued by a sample of firms listed on the Hong Kong Stock Exchange. The standard event study methodology indicates the presence of price reversal patterns following both positive and negative warnings. However, incorporating time-varying betas, event-induced variance and conditional heteroskedasticity in the modelling process results in post-negative-warning price patterns that are consistent with the predictions of the efficient market hypothesis. These adjustments also cause the statistical significance of some post-positive-warning cumulative abnormal returns to disappear and their magnitude to drop to an extent that minor transaction costs would eliminate the profitability of the contrarian strategy

Analysis of expressed sequence tags generated from full-length enriched cDNA libraries of melon

Abstract Background Melon (Cucumis melo), an economically important vegetable crop, belongs to the Cucurbitaceae family which includes several other important crops such as watermelon, cucumber, and pumpkin. It has served as a model system for sex determination and vascular biology studies. However, genomic resources currently available for melon are limited. Result We constructed eleven full-length enriched and four standard cDNA libraries from fruits, flowers, leaves, roots, cotyledons, and calluses of four different melon genotypes, and generated 71,577 and 22,179 ESTs from full-length enriched and standard cDNA libraries, respectively. These ESTs, together with ~35,000 ESTs available in public domains, were assembled into 24,444 unigenes, which were extensively annotated by comparing their sequences to different protein and functional domain databases, assigning them Gene Ontology (GO) terms, and mapping them onto metabolic pathways. Comparative analysis of melon unigenes and other plant genomes revealed that 75% to 85% of melon unigenes had homologs in other dicot plants, while approximately 70% had homologs in monocot plants. The analysis also identified 6,972 gene families that were conserved across dicot and monocot plants, and 181, 1,192, and 220 gene families specific to fleshy fruit-bearing plants, the Cucurbitaceae family, and melon, respectively. Digital expression analysis identified a total of 175 tissue-specific genes, which provides a valuable gene sequence resource for future genomics and functional studies. Furthermore, we identified 4,068 simple sequence repeats (SSRs) and 3,073 single nucleotide polymorphisms (SNPs) in the melon EST collection. Finally, we obtained a total of 1,382 melon full-length transcripts through the analysis of full-length enriched cDNA clones that were sequenced from both ends. Analysis of these full-length transcripts indicated that sizes of melon 5' and 3' UTRs were similar to those of tomato, but longer than many other dicot plants. Codon usages of melon full-length transcripts were largely similar to those of Arabidopsis coding sequences. Conclusion The collection of melon ESTs generated from full-length enriched and standard cDNA libraries is expected to play significant roles in annotating the melon genome. The ESTs and associated analysis results will be useful resources for gene discovery, functional analysis, marker-assisted breeding of melon and closely related species, comparative genomic studies and for gaining insights into gene expression patterns.This work was supported by Research Grant Award No. IS-4223-09C from BARD, the United States-Israel Binational Agricultural Research and Development Fund, and by SNC Laboratoire ASL, de Ruiter Seeds B.V., Enza Zaden B.V., Gautier Semences S.A., Nunhems B.V., Rijk Zwaan B.V., Sakata Seed Inc, Semillas Fitó S.A., Seminis Vegetable Seeds Inc, Syngenta Seeds B.V., Takii and Company Ltd, Vilmorin and Cie S.A. and Zeraim Gedera Ltd (all of them as part of the support to ICuGI). CC was supported by CNRS ERL 8196.Peer Reviewe

Towards a TILLING platform for functional genomics in Piel de Sapo melons

Background The availability of genetic and genomic resources for melon has increased significantly, but functional genomics resources are still limited for this crop. TILLING is a powerful reverse genetics approach that can be utilized to generate novel mutations in candidate genes. A TILLING resource is available for cantalupensis melons, but not for inodorus melons, the other main commercial group. Results A new ethyl methanesulfonate-mutagenized (EMS) melon population was generated for the first time in an andromonoecious non-climacteric inodorus Piel de Sapo genetic background. Diverse mutant phenotypes in seedlings, vines and fruits were observed, some of which were of possible commercial interest. The population was first screened for mutations in three target genes involved in disease resistance and fruit quality (Cm-PDS, Cm-eIF4E and Cm-eIFI(iso)4E). The same genes were also tilled in the available monoecious and climacteric cantalupensis EMS melon population. The overall mutation density in this first Piel de Sapo TILLING platform was estimated to be 1 mutation/1.5 Mb by screening four additional genes (Cm-ACO1, Cm-NOR, Cm-DET1 and Cm-DHS). Thirty-three point mutations were found for the seven gene targets, six of which were predicted to have an impact on the function of the protein. The genotype/phenotype correlation was demonstrated for a loss-of-function mutation in the Phytoene desaturase gene, which is involved in carotenoid biosynthesis. Conclusions The TILLING approach was successful at providing new mutations in the genetic background of Piel de Sapo in most of the analyzed genes, even in genes for which natural variation is extremely low. This new resource will facilitate reverse genetics studies in non-climacteric melons, contributing materially to future genomic and breeding studies.González, M.; Xu, M.; Esteras Gómez, C.; Roig Montaner, MC.; Monforte Gilabert, AJ.; Troadec, C.; Pujol, M.... (2011). Towards a TILLING platform for functional genomics in Piel de sapo melons. BMC Research Notes. 4(289):289-299. doi:10.1186/1756-0500-4-289S2892994289The International Cucurbit Genomics Initiative (ICuGI). [ http://www.icugi.org ]González-Ibeas D, Blanca J, Roig C, González-To M, Picó B, Truniger V, Gómez P, Deleu W, Caño-Delgado A, Arús P, Nuez F, García-Mas J, Puigdomènech P, Aranda MA: MELOGEN: an EST database for melon functional genomics. BMC Genomics. 2007, 8: 306-10.1186/1471-2164-8-306.Fita A, Picó B, Monforte A, Nuez F: Genetics of Root System Architecture Using Near-isogenic Lines of Melon. J Am Soc Hortic Sci. 2008, 133: 448-458.Fernandez-Silva I, Eduardo I, Blanca J, Esteras C, Picó B, Nuez F, Arús P, Garcia-Mas J, Monforte AJ: Bin mapping of genomic and EST-derived SSRs in melon (Cucumis melo L.). Theor Appl Genet. 2008, 118: 139-150. 10.1007/s00122-008-0883-3.Deleu W, Esteras C, Roig C, González-To M, Fernández-Silva I, Blanca J, Aranda MA, Arús P, Nuez F, Monforte AJ, Picó MB, Garcia-Mas J: A set of EST-SNPs for map saturation and cultivar identification in melon. BMC Plant Biol. 2009, 9: 90-10.1186/1471-2229-9-90.Mascarell-Creus A, Cañizares J, Vilarrasa J, Mora-García S, Blanca J, González-Ibeas D, Saladié M, Roig C, Deleu W, Picó B, López-Bigas N, Aranda MA, Garcia-Mas J, Nuez F, Puigdomènech P, Caño-Delgado A: An oligo-based microarray offers novel transcriptomic approaches for the analysis of pathogen resistance and fruit quality traits in melon (Cucumis melo L.). BMC Genomics. 2009, 10: 467-10.1186/1471-2164-10-467.Blanca JM, Cañizares J, Ziarsolo P, Esteras C, Mir G, Nuez F, Garcia-Mas J, Pico B: Melon transcriptome characterization. SSRs and SNPs discovery for high throughput genotyping across the species. Plant Genome. 2011, 4 (2): 118-131. 10.3835/plantgenome2011.01.0003.González VM, Benjak A, Hénaff EM, Mir G, Casacuberta JM, Garcia-Mas J, Puigdomènech P: Sequencing of 6.7 Mb of the melon genome using a BAC pooling strategy. BMC Plant Biology. 2010, 10: 246-10.1186/1471-2229-10-246.Moreno E, Obando JM, Dos-Santos N, Fernández-Trujillo JP, Monforte AJ, Garcia-Mas J: Candidate genes and QTLs for fruit ripening and softening in melon. Theor Appl Genet. 2007, 116: 589-602.Essafi A, Díaz-Pendón JA, Moriones E, Monforte AJ, Garcia-Mas J, Martín-Hernández AM: Dissection of the oligogenic resistance to Cucumber mosaic virus in the melon accession PI 161375. Theor Appl Genet. 2009, 118: 275-284. 10.1007/s00122-008-0897-x.Comai L, Henikoff S: TILLING: practical single-nucleotide mutation discovery. Plant J. 2006, 45: 684-94. 10.1111/j.1365-313X.2006.02670.x.Cooper JL, Till BJ, Laport RG, Darlow MC, Kleffner JM, Jamai A, El-Mellouki T, Liu S, Ritchie R, Nielsen N, et al: TILLING to detect induced mutations in soybean. BMC Plant Biol. 2008, 8 (1): 9-10.1186/1471-2229-8-9.Dalmais M, Schmidt J, Le Signor C, Moussy F, Burstin J, Savois V, Aubert G, de Oliveira Y, Guichard C, Thompson R, Bendahmane A: UTILLdb, a Pisum sativum in silico forward and reverse genetics tool. Genome Biol. 2008, 9: R43-10.1186/gb-2008-9-2-r43.Dierking EC, Bilyeu KD: New sources of soybean meal and oil composition traits identified through TILLING. BMC Plant Biol. 2009, 9: 89-10.1186/1471-2229-9-89.Perry J, Brachmann A, Welham T, Binder A, Charpentier M, Groth M, Haage K, Markmann K, Wang TL, Parniske M: TILLING in Lotus japonicus identified large allelic series for symbiosis genes and revealed a bias in functionally defective ethyl methanesulfonate alleles toward glycine replacements. Plant Physiol. 2009, 151 (3): 1281-1291. 10.1104/pp.109.142190.Caldwell DG, McCallum N, Shaw P, Muehlbauer GJ, Marshall DF, Waugh R: A structured mutant population for forward and reverse genetics in Barley (Hordeum vulgare L.). Plant J. 2004, 40 (1): 143-150. 10.1111/j.1365-313X.2004.02190.x.Henikoff S, Bradley JT, Comai L: TILLING. Traditional mutagenesis meets functional genomics. Plant Physiol. 2004, 135: 630-636. 10.1104/pp.104.041061.Wu JL, Wu C, Lei C, Baraoidan M, Bordeos A, Madamba MR, Ramos-Pamplona M, Mauleon R, Portugal A, Ulat VJ, et al: Chemical- and irradiation-induced mutants of indica rice IR64 for forward and reverse genetics. Plant Mol Biol. 2005, 59 (1): 85-97. 10.1007/s11103-004-5112-0.Slade AJ, Fuerstenberg SI, Loeffler D, Steine MN, Facciotti D: A reverse genetic, nontransgenic approach to wheat crop improvement by TILLING. Nat Biotechnol. 2005, 23: 75-81. 10.1038/nbt1043.Till BJ, Cooper J, Tai TH, Colowit P, Greene EA, Henikoff S, Comai L: Discovery of chemically induced mutations in rice by TILLING. BMC Plant Biol. 2007, 7: 19-10.1186/1471-2229-7-19.Xin Z, Wang ML, Barkley NA, Burow G, Franks C, Pederson G, Burke J: Applying genotyping (TILLING) and phenotyping analyses to elucidate gene function in a chemically induced sorghum mutant population. BMC Plant Biol. 2008, 8: 103-10.1186/1471-2229-8-103.Dong C, Dalton-Morgan J, Vincent K, Sharp P: A modified TILLING method for wheat breeding. Plant Genome. 2009, 2: 39-47. 10.3835/plantgenome2008.10.0012.Sestili F, Botticella E, Bedo Z, Phillips A, Lafiandra D: Production of novel allelic variation for genes involved in starch biosynthesis through mutagenesis. Mol Breeding. 2010, 25: 145-154. 10.1007/s11032-009-9314-7.Watanabe S, Mizoguchi T, Aoki K, Kubo Y, Mori H, Imanishi S, Yamazaki Y, Shibata D, Ezura H: Ethylmethanesulfonate (EMS) mutagenesis of Solanum lycopersicum cv. Micro-Tom for large-scale mutant screens. Plant Biotech. 2007, 24: 33-38. 10.5511/plantbiotechnology.24.33.Elias R, Till BJ, Mba Ch, Al-Safadi B: Optimizing TILLING and Ecotilling techniques for potato (Solanum tuberosum L). BMC Res Notes. 2009, 2: 141-10.1186/1756-0500-2-141.Piron F, Nicolaı M, Minoıa S, Piednoir E, Moretti A, Salgues A, Zamir D, Caranta C, Bendahmane A: An induced mutation in tomato eIF4E leads to immunity to two Potyviruses. PLoS ONE. 2010, 5 (6): e11313-10.1371/journal.pone.0011313.Himelblau E, Gilchrist EJ, Buono K, Bizell C, Mentzer L, Vogelzang R, Osborn T, Amasino RM, Parkin IAP, Haughn : Forward and reverse genetics of papid cycling Brassica oleracea. Theor Appl Genet. 2009, 118: 953-961. 10.1007/s00122-008-0952-7.Stephenson P, Baker D, Girin T, Perez A, Amoah S, King GJ, Østergaard L: A rich TILLING resource for studying gene function in Brassica rapa. BMC Plant Biol. 2010, 10: 62-10.1186/1471-2229-10-62.Pitrat M: Melon (Cucumis melo L.). Handbook of Crop Breeding Vol I. Vegetables. Edited by: Prohens J, Nuez F. 2008, New York:Springer, 283-315.Dahmani-Mardas F, Troadec Ch, Boualem A, Leveque S, Alsadon AA, Aldoss AA, Dogimont C, Bendahman A: Engineering Melon Plants with Improved Fruit Shelf Life Using the TILLING Approach. PLoS ONE. 2010, 5: e15776-10.1371/journal.pone.0015776.Nieto C, Piron F, Dalmais M, Marco CF, Moriones E, Gómez-Guillamón ML, Truniger V, Gómez P, Garcia-Mas J, Aranda MA, Bendahmane A: EcoTILLING for the identification of allelic variants of melon eIF4E, a factor that controls virus susceptibility. BMC Plant Biol. 2007, 7: 34-10.1186/1471-2229-7-34.Qin G, Gu H, Ma L, Peng Y, Deng XW, Chen Z, Qu LJ: Disruption of phytoene desaturase gene results in albino and dwarf phenotypes in Arabidopsis by impairing chlorophyll, carotenoid, and gibberellin biosynthesis. Cell Res. 2007, 17: 471-482. 10.1038/cr.2007.40.Codons Optimized to Deliver Deleterious Lesions (CODDLe). [ http://www.proweb.org/input ]Lasserre E, Bouquin T, Hernández JA, Bull J, Pech JC, Balague C: Structure and expression of three genes encoding ACC oxidase homologs from melon (Cucumis melo L.). Mol Gen Genet. 1996, 251 (1): 81-90.Giovannoni JJ: Fruit ripening mutants yield insights into ripening control. Curr Opin Plant Biol. 2007, 10: 1-7. 10.1016/j.pbi.2006.11.012.Davuluri GR, van Tuinen A, Mustilli AC, Manfredonia A, Newman R, Burgess D, Brummell DA, King SR, Palys J, Uhlig J, Pennings HMJ, Bowler C: Manipulation of DET1 expression in tomato results in photomorphogenic phenotypes caused by post-transcriptional gene silencing. Plant J. 2004, 40: 344-354. 10.1111/j.1365-313X.2004.02218.x.Wei S, Li X, Gruber MI, Li R, Zhou R, Zebarjadi A, Hannoufa A: RNAi-mediated suppression of DET1 alters the levels of carotenoids and sinapate esters in seeds of Brassica napus. J Agric Food Chem. 2009, 57 (12): 5326-5333. 10.1021/jf803983w.Wang TW, Zhang CG, Wu W, Nowack LM, Madey E, Thompson JE: Antisense suppression of deoxyhypusine synthase in tomato delays fruit softening and alters growth and development DHS mediates the first of two sequential enzymatic reactions that activate eukaryotic translation initiation factor-5A. Plant Physiol. 2005, 138: 1372-1382. 10.1104/pp.105.060194.Ng PC, Henikoff S: SIFT: predicting amino acid changes that affect protein function. Nucleic Acids Res. 2003, 31 (13): 3812-3814. 10.1093/nar/gkg509.Guzman P, Ecker JR: Exploiting the triple response of Arabidopsis to identify ethylene-related mutants. The Plant Cell. 1990, 2: 513-523.Henikoff S, Comai L: Single-nucleotide mutations for plant functional genomics. Ann Rev Plant Biol. 2003, 54: 375-401. 10.1146/annurev.arplant.54.031902.135009.Greene EA, Codomo CA, Taylor NE, Henikoff JG, Till BJ, Reynolds SH, Enns LC, Burtner C, Johnson JE, Odden AR, et al: Spectrum of chemically induced mutations from a large-scale reverse genetic screen in Arabidopsis. Genetics. 2003, 164 (2): 731-740.Britt AB: DNA damage and repair in plants. Annu Rev Plant Physiol Plant Mol Biol. 1996, 47: 75-100. 10.1146/annurev.arplant.47.1.75.Truniger V, Nieto C, González-Ibeas D, Aranda M: Mechanism of plant eIF4E-mediated resistance against a Carmovirus (Tombusviridae): cap-independent translation of a viral RNA controlled in cis by an (a)virulence determinant. Plant J. 2008, 56 (5): 716-727. 10.1111/j.1365-313X.2008.03630.x.Gao Z, Johansen E, Eyers S, Thomas CL, Ellis THN, Maule AJ: The potyvirus recessive resistance gene, sbm1, identifies a novel role for translation initiation factor eIF4E in cell-to-cell trafficking. Plant J. 2004, 40 (3): 376-385. 10.1111/j.1365-313X.2004.02215.x.Kang BC, Yeam I, Frantz JD, Murphy JF, Jahn MM: The pvr1 locus in Capsicum encodes a translation initiation factor eIF4E that interacts with Tobacco etch virus VPg. Plant J. 2005, 42 (3): 392-405. 10.1111/j.1365-313X.2005.02381.x.Ruffel S, Gallois J, Lesage M, Caranta C: The recessive potyvirus resistance gene pot-1 is the tomato orthologue of the pepper pvr2-eiF4 genes. Mol Genet Genom. 2005, 274 (4): 346-353. 10.1007/s00438-005-0003-x.Nicaise V, German-Retana S, Sanjuán R, Dubrana MP, Mazier M, Maisonneuve B, Candresse T, Caranta C, LeGall O: The Eukaryotic Translation Initiation Factor 4E Controls Lettuce Susceptibility to the Potyvirus Lettuce mosaic virus1. Plant Physiol. 2003, 132: 1272-1282. 10.1104/pp.102.017855.Esteras C, Pascual L, Saladie M, Dogimont C, Garcia-Mas J, Nuez F, Picó B: Use of Ecotilling to identify natural allelic variants of melon candidate genes involved in fruit ripening. Proceedings Plant GEM8 Lisbon. 2009Levin I, Frankel P, Gilboa N, Tanny S, Lalazar A: The tomato dark green mutation is a novel allele of the tomato homolog of the DEETIOLATED1 gene. Theor Appl Genet. 2003, 106: 454-460.Kolotilin I, Koltai H, Tadmor Y, Bar-Or C, Reuveni M, Meir A, Nahon S, Shlomo S, Chen L, I Levin: Transcriptional profiling of high pigment-2dg tomato mutant links early fruit plastid biogenesis with its overproduction of phytonutrients. Plant Physiol. 2007, 145: 389-401. 10.1104/pp.107.102962

An Induced Mutation in Tomato eIF4E Leads to Immunity to Two Potyviruses

BACKGROUND: The characterization of natural recessive resistance genes and Arabidopsis virus-resistant mutants have implicated translation initiation factors of the eIF4E and eIF4G families as susceptibility factors required for virus infection and resistance function. METHODOLOGY/PRINCIPAL FINDINGS: To investigate further the role of translation initiation factors in virus resistance we set up a TILLING platform in tomato, cloned genes encoding for translation initiation factors eIF4E and eIF4G and screened for induced mutations that lead to virus resistance. A splicing mutant of the eukaryotic translation initiation factor, S.l_eIF4E1 G1485A, was identified and characterized with respect to cap binding activity and resistance spectrum. Molecular analysis of the transcript of the mutant form showed that both the second and the third exons were miss-spliced, leading to a truncated mRNA. The resulting truncated eIF4E1 protein is also impaired in cap-binding activity. The mutant line had no growth defect, likely because of functional redundancy with others eIF4E isoforms. When infected with different potyviruses, the mutant line was immune to two strains of Potato virus Y and Pepper mottle virus and susceptible to Tobacco each virus. CONCLUSIONS/SIGNIFICANCE: Mutation analysis of translation initiation factors shows that translation initiation factors of the eIF4E family are determinants of plant susceptibility to RNA viruses and viruses have adopted strategies to use different isoforms. This work also demonstrates the effectiveness of TILLING as a reverse genetics tool to improve crop species. We have also developed a complete tool that can be used for both forward and reverse genetics in tomato, for both basic science and crop improvement. By opening it to the community, we hope to fulfill the expectations of both crop breeders and scientists who are using tomato as their model of study

Engineering Melon Plants with Improved Fruit Shelf Life Using the TILLING Approach

Background: Fruit ripening and softening are key traits that have an effect on food supply, fruit nutritional value and consequently, human health. Since ethylene induces ripening of climacteric fruit, it is one of the main targets to control fruit over ripening that leads to fruit softening and deterioration. The characterization of the ethylene pathway in Arabidopsis and tomato identified key genes that control fruit ripening. [br/] Methodology/Principal Findings: To engineer melon fruit with improved shelf-life, we conducted a translational research experiment. We set up a TILLING platform in a monoecious and climacteric melon line, cloned genes that control ethylene production and screened for induced mutations that lead to fruits with enhanced shelf life. Two missense mutations, L124F and G194D, of the ethylene biosynthetic enzyme, ACC oxidase 1, were identified and the mutant plants were characterized with respect to fruit maturation. The L124F mutation is a conservative mutation occurring away from the enzyme active site and thus was predicted to not affect ethylene production and thus fruit ripening. In contrast, G194D modification occurs in a highly conserved amino acid position predicted, by crystallographic analysis, to affect the enzymatic activity. Phenotypic analysis of the G194D mutant fruit showed complete delayed ripening and yellowing with improved shelf life and, as predicted, the L124F mutation did not have an effect. [br/] Conclusions/Significance: We constructed a mutant collection of 4023 melon M2 families. Based on the TILLING of 11 genes, we calculated the overall mutation rate of one mutation every 573 kb and identified 8 alleles per tilled kilobase. We also identified a TILLING mutant with enhanced fruit shelf life. This work demonstrates the effectiveness of TILLING as a reverse genetics tool to improve crop species. As cucurbits are model species in different areas of plant biology, we anticipate that the developed tool will be widely exploited by the scientific community

First TILLING Platform in Cucurbita pepo: A New Mutant Resource for Gene Function and Crop Improvement

Although the availability of genetic and genomic resources for Cucurbita pepo has increased significantly, functional genomic resources are still limited for this crop. In this direction, we have developed a high throughput reverse genetic tool: the first TILLING (Targeting Induced Local Lesions IN Genomes) resource for this species. Additionally, we have used this resource to demonstrate that the previous EMS mutant population we developed has the highest mutation density compared with other cucurbits mutant populations. The overall mutation density in this first C. pepo TILLING platform was estimated to be 1/133 Kb by screening five additional genes. In total, 58 mutations confirmed by sequencing were identified in the five targeted genes, thirteen of which were predicted to have an impact on the function of the protein. The genotype/phenotype correlation was studied in a peroxidase gene, revealing that the phenotype of seedling homozygous for one of the isolated mutant alleles was albino. These results indicate that the TILLING approach in this species was successful at providing new mutations and can address the major challenge of linking sequence information to biological function and also the identification of novel variation for crop breeding.Financial support was provided by the Spanish Project INIA (Instituto Nacional de Investigacion y Tecnologia Agraria y Almentaria) RTA2011-00044C02-01, the ANR MELODY (ANR-11-BSV7-0024), the European Research Council (ERCSEXYPARTH), FEDER, and FSE funds. NVD has been awarded a grant by the Andalusian Institute of Agronomy Research IFAPA. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Vicente-Dolera, N.; Troadec, C.; Moya, M.; Río-Celestino, MD.; Pomares-Viciana, T.; Bendahmane, A.; Picó Sirvent, MB.... (2014). First TILLING Platform in Cucurbita pepo: A New Mutant Resource for Gene Function and Crop Improvement. PLoS ONE. 9(11):112743-112743. https://doi.org/10.1371/journal.pone.0112743S112743112743911Paris, H. S., Yonash, N., Portnoy, V., Mozes-Daube, N., Tzuri, G., & Katzir, N. (2002). Assessment of genetic relationships in Cucurbita pepo (Cucurbitaceae) using DNA markers. Theoretical and Applied Genetics, 106(6), 971-978. doi:10.1007/s00122-002-1157-0Parry, M. A. J., Madgwick, P. J., Bayon, C., Tearall, K., Hernandez-Lopez, A., Baudo, M., … Phillips, A. L. (2009). Mutation discovery for crop improvement. Journal of Experimental Botany, 60(10), 2817-2825. doi:10.1093/jxb/erp189Gilchrist, E., & Haughn, G. (2010). Reverse genetics techniques: engineering loss and gain of gene function in plants. Briefings in Functional Genomics, 9(2), 103-110. doi:10.1093/bfgp/elp059McCallum, C. M., Comai, L., Greene, E. A., & Henikoff, S. (2000). Targeting Induced LocalLesions IN Genomes (TILLING) for Plant Functional Genomics. Plant Physiology, 123(2), 439-442. doi:10.1104/pp.123.2.439Colbert, T., Till, B. J., Tompa, R., Reynolds, S., Steine, M. N., Yeung, A. T., … Henikoff, S. (2001). High-Throughput Screening for Induced Point Mutations. Plant Physiology, 126(2), 480-484. doi:10.1104/pp.126.2.480Wang, T. L., Uauy, C., Robson, F., & Till, B. (2012). TILLINGin extremis. Plant Biotechnology Journal, 10(7), 761-772. doi:10.1111/j.1467-7652.2012.00708.xDong, C., Dalton-Morgan, J., Vincent, K., & Sharp, P. (2009). A Modified TILLING Method for Wheat Breeding. The Plant Genome Journal, 2(1), 39. doi:10.3835/plantgenome2008.10.0012Uauy, C., Paraiso, F., Colasuonno, P., Tran, R. K., Tsai, H., Berardi, S., … Dubcovsky, J. (2009). A modified TILLING approach to detect induced mutations in tetraploid and hexaploid wheat. BMC Plant Biology, 9(1), 115. doi:10.1186/1471-2229-9-115Kumar, A. P., Boualem, A., Bhattacharya, A., Parikh, S., Desai, N., Zambelli, A., … Bendahmane, A. (2013). SMART -- Sunflower Mutant population And Reverse genetic Tool for crop improvement. BMC Plant Biology, 13(1), 38. doi:10.1186/1471-2229-13-38Kurowska, M., Daszkowska-Golec, A., Gruszka, D., Marzec, M., Szurman, M., Szarejko, I., & Maluszynski, M. (2011). TILLING - a shortcut in functional genomics. Journal of Applied Genetics, 52(4), 371-390. doi:10.1007/s13353-011-0061-1Rigola, D., van Oeveren, J., Janssen, A., Bonné, A., Schneiders, H., van der Poel, H. J. A., … van Eijk, M. J. T. (2009). High-Throughput Detection of Induced Mutations and Natural Variation Using KeyPoint™ Technology. PLoS ONE, 4(3), e4761. doi:10.1371/journal.pone.0004761González, M., Xu, M., Esteras, C., Roig, C., Monforte, A. J., Troadec, C., … Picó, B. (2011). Towards a TILLING platform for functional genomics in Piel de Sapo melons. BMC Research Notes, 4(1). doi:10.1186/1756-0500-4-289Elias, R., Till, B. J., Mba, C., & Al-Safadi, B. (2009). Optimizing TILLING and Ecotilling techniques for potato (Solanum tuberosum L). BMC Research Notes, 2(1), 141. doi:10.1186/1756-0500-2-141Dahmani-Mardas, F., Troadec, C., Boualem, A., Lévêque, S., Alsadon, A. A., Aldoss, A. A., … Bendahmane, A. (2010). Engineering Melon Plants with Improved Fruit Shelf Life Using the TILLING Approach. PLoS ONE, 5(12), e15776. doi:10.1371/journal.pone.0015776Boualem, A., Fleurier, S., Troadec, C., Audigier, P., Kumar, A. P. K., Chatterjee, M., … Bendahmane, A. (2014). Development of a Cucumis sativus TILLinG Platform for Forward and Reverse Genetics. PLoS ONE, 9(5), e97963. doi:10.1371/journal.pone.0097963Blanca, J., Cañizares, J., Roig, C., Ziarsolo, P., Nuez, F., & Picó, B. (2011). Transcriptome characterization and high throughput SSRs and SNPs discovery in Cucurbita pepo (Cucurbitaceae). BMC Genomics, 12(1). doi:10.1186/1471-2164-12-104Esteras, C., Gomez, P., Monforte, A. J., Blanca, J., Vicente-Dolera, N., Roig, C., … Pico, B. (2012). High-throughput SNP genotyping in Cucurbita pepo for map construction and quantitative trait loci mapping. BMC Genomics, 13(1), 80. doi:10.1186/1471-2164-13-80Vicente-Dólera, N., Pinillos, V., Moya, M., Del Río-Celestino, M., Pomares-Viciana, T., Román, B., & Gómez, P. (2014). An improved method to obtain novel mutants in Cucurbita pepo by pollen viability. Scientia Horticulturae, 169, 14-19. doi:10.1016/j.scienta.2014.01.045Martín, B., Ramiro, M., Martínez-Zapater, J. M., & Alonso-Blanco, C. (2009). A high-density collection of EMS-induced mutations for TILLING in Landsberg erecta genetic background of Arabidopsis. BMC Plant Biology, 9(1), 147. doi:10.1186/1471-2229-9-147Wienholds, E. (2003). Efficient Target-Selected Mutagenesis in Zebrafish. Genome Research, 13(12), 2700-2707. doi:10.1101/gr.1725103Dalmais, M., Schmidt, J., Le Signor, C., Moussy, F., Burstin, J., Savois, V., … Bendahmane, A. (2008). UTILLdb, a Pisum sativum in silico forward and reverse genetics tool. Genome Biology, 9(2), R43. doi:10.1186/gb-2008-9-2-r43Triques, K., Sturbois, B., Gallais, S., Dalmais, M., Chauvin, S., Clepet, C., … Bendahmane, A. (2007). Characterization of Arabidopsis thaliana mismatch specific endonucleases: application to mutation discovery by TILLING in pea. The Plant Journal, 51(6), 1116-1125. doi:10.1111/j.1365-313x.2007.03201.xTaylor, N. E. (2003). PARSESNP: a tool for the analysis of nucleotide polymorphisms. Nucleic Acids Research, 31(13), 3808-3811. doi:10.1093/nar/gkg574Ng, P. C. (2003). SIFT: predicting amino acid changes that affect protein function. Nucleic Acids Research, 31(13), 3812-3814. doi:10.1093/nar/gkg509Obrero, Á., González-Verdejo, C. I., Die, J. V., Gómez, P., Del Río-Celestino, M., & Román, B. (2013). Carotenogenic Gene Expression and Carotenoid Accumulation in Three Varieties of Cucurbita pepo during Fruit Development. Journal of Agricultural and Food Chemistry, 61(26), 6393-6403. doi:10.1021/jf4004576Cosio, C., Vuillemin, L., De Meyer, M., Kevers, C., Penel, C., & Dunand, C. (2009). An anionic class III peroxidase from zucchini may regulate hypocotyl elongation through its auxin oxidase activity. Planta, 229(4), 823-836. doi:10.1007/s00425-008-0876-0Sisko, M. (2003). Genome size analysis in the genus Cucurbita and its use for determination of interspecific hybrids obtained using the embryo-rescue technique. Plant Science, 165(3), 663-669. doi:10.1016/s0168-9452(03)00256-5Campa A (1991) Biological roles of plant peroxidases: known and potential function. In Peroxidases in Chemistry and Biology Vol. II. (Everse, J., Everse, K.E. & Grisham, M.B., eds), pp. 25–50, CRC Press, Boca Raton, FL.Schuller, D. J., Ban, N., van Huystee, R. B., McPherson, A., & Poulos, T. L. (1996). The crystal structure of peanut peroxidase. Structure, 4(3), 311-321. doi:10.1016/s0969-2126(96)00035-4Stephenson, P., Baker, D., Girin, T., Perez, A., Amoah, S., King, G. J., & Østergaard, L. (2010). A rich TILLING resource for studying gene function in Brassica rapa. BMC Plant Biology, 10(1), 62. doi:10.1186/1471-2229-10-62Suzuki, T., Eiguchi, M., Kumamaru, T., Satoh, H., Matsusaka, H., Moriguchi, K., … Kurata, N. (2007). MNU-induced mutant pools and high performance TILLING enable finding of any gene mutation in rice. Molecular Genetics and Genomics, 279(3), 213-223. doi:10.1007/s00438-007-0293-2Chantreau, M., Grec, S., Gutierrez, L., Dalmais, M., Pineau, C., Demailly, H., … Hawkins, S. (2013). PT-Flax (phenotyping and TILLinG of flax): development of a flax (Linum usitatissimum L.) mutant population and TILLinG platform for forward and reverse genetics. BMC Plant Biology, 13(1), 159. doi:10.1186/1471-2229-13-159Rawat, N., Sehgal, S. K., Joshi, A., Rothe, N., Wilson, D. L., McGraw, N., … Gill, B. S. (2012). A diploid wheat TILLING resource for wheat functional genomics. BMC Plant Biology, 12(1), 205. doi:10.1186/1471-2229-12-205Minoia, S., Petrozza, A., D’Onofrio, O., Piron, F., Mosca, G., Sozio, G., … Carriero, F. (2010). A new mutant genetic resource for tomato crop improvement by TILLING technology. BMC Research Notes, 3(1). doi:10.1186/1756-0500-3-69Markiewicz, P., Kleina, L. G., Cruz, C., Ehret, S., & Miller, J. H. (1994). Genetic Studies of the lac Repressor. XIV. Analysis of 4000 Altered Escherichia coli lac Repressors Reveals Essential and Non-essential Residues, as well as «Spacers» which do not Require a Specific Sequence. Journal of Molecular Biology, 240(5), 421-433. doi:10.1006/jmbi.1994.1458Carpin, S., Crèvecoeur, M., Greppin, H., & Penel, C. (1999). Molecular Cloning and Tissue-Specific Expression of an Anionic Peroxidase in Zucchini. Plant Physiology, 120(3), 799-810. doi:10.1104/pp.120.3.799Welinder, K. G., Justesen, A. F., Kjaersgård, I. V. H., Jensen, R. B., Rasmussen, S. K., Jespersen, H. M., & Duroux, L. (2002). Structural diversity and transcription of class III peroxidases from Arabidopsis thaliana. European Journal of Biochemistry, 269(24), 6063-6081. doi:10.1046/j.1432-1033.2002.03311.

Minéralisation accélérée de l'atrazine dans les sols: conditions de mise en place, caractérisation et influence de la disponibilité en carbone et en azote

*INRA, Laboratoire de science du sol Diffusion du document : INRA, Laboratoire de science du sol Diplôme : Dr. Ing