Análisis transcriptómico del daño por frío en fruto de melocotón

En la industria hortofrutícola las bajas temperaturas se utilizan para retrasar la maduración y el decaimiento de los frutos. A pesar de tener un uso extendido, los frutos de melocotón sometidos a almacenamiento prolongado en frío (CS) a menudo desarrollan una forma de daño por frío (CI) genéticamente controlada llamada harinosidad (mealiness/woolliness [WLT]): un desorden textural de la pulpa del fruto caracterizado por la perdida de suculencia Aunque durante el frío se han observado algunas alteraciones microscópicas, los síntomas visibles o macroscópicos de daño se desarrollan, no obstante, cuando los frutos se transfieren a temperatura ambiente (temperaturas de simulación de vida útil, SLR) para inducir la maduración después del almacenamiento. Con el objetivo de analizar los mecanismos moleculares subyacentes al desarrollo de la harinosidad durante el almacenamiento en frío (estadio pre-sintomático) y maduración en condiciones de simulación (estadio sintomático), diversos estudios transcriptómicos y proteómicos se han realizado. Estos estudios han resultado en la identificación de diferentes funciones celulares probablemente importantes para el desarrollo de los síntomas de la harinosidad, aunque no profundizan en el análisis y algunos resultados son aparentemente contradictorios (ya que en ellos se usan diferentes variedades de melocotón y protocolos de CS/SLR). Para proporcionar una visión más comprehensiva del desorden de la harinosidad en esta tesis se ha utilizado una aproximación genómica-genética que consiste en la combinación de grupos de líneas hermanas de la población de melocotón Pop-DG (con sensibilidad contrastante para desarrollar harinosidad) con una micromatriz de cDNA (Chillpeach microarray) dedicada al daño por frío para perfilar la expresión génica. Esta estrategia nos ha permitido identificar un gran número de genes asociados a la sensibilidad/tolerancia a desarrollar la harinosidad tanto a en estadios pre-sintomático (fruto maduro y CS) como sintomático, mientras que se compensan otras diferencias fenotípicas que segregan al azar en la población. Mediante la el análisis Fluidigm RT PCR, que se podría definir como qRT-PCR de media capacidad, hemos confirmado la validez de esos marcadores de expresión en los grupos de líneas hermanas, pero más importante, estos marcadores de expresión han resultado ser validos y reproducibles en lineas hermanas individuales de la población Pop-DG y en otros genotipos de melocotón (Oded y Hermoza) con diferentes grados de tolerancia al daño por frío. Además los perfiles de expresión de Od, Hz y los dos grupos de la población Pop-DG se han analizado también con la herramienta bioinformática ROSMETER, que proporciona información de la especificidad de la respuesta transcriptómica al estrés oxidativo. La anotación funcional de los genes diferencialmente expresados nos ha permitido proponer algunas hipótesis sobre los procesos moleculares que ocurren en el fruto de melocotón mientras está almacenado a 5ºC y cuando, posteriormente se deja madurar a temperatura ambiente, que extienden nuestra hipótesis de trabajo sobre el desorden de la harinosidad más allá de los cambios en la pared celular.Low temperatures are commonly used in the horticultural industry to delay peach ripening s and fruit decay. Despite widespread use, peach fruits subjected to prolonged cold storage (CS) periods often develop a genetically controlled form of chilling injury (CI) called mealiness/woolliness (WLT): a flesh textural disorder characterized by a lack of juiciness. Although some microscopic alterations had been observed during CS, visual symptoms (or macroscopic) of WLT appeared however upon transferring the fruits to the room temperature conditions (shelf life ripening [SLR] temperatures,) that are used for inducing ripening after CS. To dissect the molecular mechanisms underlying the WLT development during CS (pre-symptomatic stage) and SLR (symptomatic stage), several transcriptomic and proteomic studies have been reported. These studies have resulted in the identification of different cellular functions as important for the development of the WLT but they did not go deep in the analysis and some results were apparently contradictory (as they used different peach varieties and CS/SLR protocols). To give a more comprehensive view of WLT disorder we have used a genetical genomic approach consisting in the combination of pools of siblings of the Pop-DG population, with contrasting sensitivity to develop WLT, with a CI-dedicated Chillpeach microarray to profile gene expression. This strategy have enable us to identify a large number of genes associated to the sensitivity /tolerance to develop WLT both at pre-symptomatic (mature and CS) and symptomatic (SLR) stages, while compensating for other phenotypic differences randomly segregating in the population. By medium throughput qRT PCR analysis we confirmed the validity of these gene expression markers in the pools, but most important, there were proved to be reliable in individual siblings of the Pop-DG population and other peach genotypes (Oded and Hermoza) with different tolerance degree to chilling injury. Further the expression profiles of Oded, Hermoza and of the two pools from the Pop‐DG population were also analyzed with the bioinformatic tool ROSMETER that provides information on the specificity of the transcriptomic response to oxidative stress. Functional annotation of genes differentially expressed has enabled us to propose some hypotheses of the molecular processes occurring in the peach fruit while stored at 5ºC and later when let to ripen at room temperature that extends our working hypothesis about WLT disorder beyond the cell wall change

Interaction of two MADS-box genes leads to growth phenotype divergence of all-flesh type of tomatoes

[EN] All-flesh tomato cultivars are devoid of locular gel and exhibit enhanced firmness and improved postharvest storage. Here, we show that SlMBP3 is a master regulator of locular tissue in tomato fruit and that a deletion at the gene locus underpins the All-flesh trait. Intriguingly, All-flesh varieties lack the deleterious phenotypes reported previously for SlMBP3 under-expressing lines and which preclude any potential commercial use. We resolve the causal factor for this phenotypic divergence through the discovery of a natural mutation at the SlAGL11 locus, a close homolog of SlMBP3. Misexpressing SlMBP3 impairs locular gel formation through massive transcriptomic reprogramming at initial phases of fruit development. SlMBP3 influences locule gel formation by controlling cell cycle and cell expansion genes, indicating that important components of fruit softening are determined at early pre-ripening stages. Our findings define potential breeding targets for improved texture in tomato and possibly other fleshy fruits. The all-flesh type of tomato fruits is caused by mutation of the MBP3 gene, however, knocking down MBP3 in certain genotypes also affect plant and fruit development. Here, the authors show that a natural mutation of AGL11, a close homolog of MBP3, is responsible for the phenotypic divergence.The authors are grateful to L. Lemonnier and D. Saint-Martin for transformation and cultivation of tomato plants and GeT-PlaGe core facility (INRAe Toulouse) for ChIP deep sequencing. The authors also want to thank Dr. Christian Chevalier (INRAE et Univsersite de Bordeaux) for helping in analyzing genes related to cell cycle, cell division, and endoreduplication in tomato. This research was supported by the EU H2020 TomGEM 679796 and HARNESSTOM 101000716 projects.Huang, B.; Hu, G.; Wang, K.; Frasse, P.; Maza, E.; Djari, A.; Deng, W.... (2021). Interaction of two MADS-box genes leads to growth phenotype divergence of all-flesh type of tomatoes. Nature Communications. 12(1):1-14. https://doi.org/10.1038/s41467-021-27117-711412

La tomata ‘Valenciana’ del Perelló: comparació de les seues característiques genètiques i fenotípiques amb el conjunt de tomates tradicionals europees.

[CA] La tomata ‘Valenciana del Perelló’, originaria de l’horta de València, representa una de les més importants i apreciades varietats tradicional valencianes de consum en fresc. A pesar d’açò manquem d’una tipificació i caracterització en profunditat, el que dificulta identificar de manera especifica i objectiva característiques distintives d’aquest tipus de tomata tant especial. En aquest treball comparem les característiques genètiques i fenotípiques de la tomata la ‘Valenciana’ i, concretament la ‘Valenciana del Perelló’ amb un conjunt de mes de 1000 varietats de tomates tradicionals Europees amb la finalitat de proporcionar les bases per distingir la tomata ‘Valenciana’ del Perelló’ d’aquelles pertanyents a altres varietats tradicionals anomenades també valencianes o aquelles que són similars dins del conjunt tradicional europeu. La caracterització morfològica, basada en 10 caràcters morfològics i qualitatius ens indica que encara que hi ha diferències poblacionals, no són suficients per discriminar entre tipus varietals. El genotipat de les varietats tradicionals europees revela una estructura genètica ben definida per a la varietat de tomata ‘Valenciana del Perelló’ i respecte a altres tomates valencianes o tomates europees de característiques similars. En particular, 18 variants de seqüència SNPs en llocs específics del genoma de la tomata són suficients per distingir clarament la tomata ‘Valenciana’ tipus ‘ el Perelló’ de les altres varietats de tomata ‘Valenciana’ i la tomata ‘Valenciana’ del conjunt de tomata tradicional europeu. Tenint en compte açò, els nostres resultats proporcionen l’empremta genètica de la tomata ‘Valenciana del Perelló’, punt de partida per a la valorització d’aquesta varietat local i per a la seua utilització en programes de millora o el seu us certificat en mercats de productes d’alta qualitat.[EN] The ‘Valenciana d’El Perelló’ tomato, originating from the Spanish region l’Horta de València, represents one of the most important and appreciated Valencian landraces for fresh market. Despite this, we still lack a detailed typification and characterization of this variety which is a prerequisite for identifying specific and objective distinctive characteristics of this type of tomato. In this work we compared genetic and phenotypic traits of the tomato variety ‘Valenciana’, in particular the ‘Valenciana d’El Perelló’, against more than 1000 traditional varieties in order to provide the basis to distinguish the tomato ‘Valenciana d’El Perelló’ from a number of other landraces named ‘Valenciana’ or to those similar within the traditional European tomato collection. Morphological characterization, based in 10 fruit morphological and qualitative traits indicated that despite differences between populations, these traits do not discriminate varietal types. The genotyping of European traditional varieties reveal a well defined genetic structure of ‘Valenciana d’El Perelló’ with respect to other Valencian or European tomatoes with similar characteristics. In particular, 18 sequence SNP variants are sufficient to distinguish clearly the tomato ‘Valenciana d’El Perelló’ type from other ‘Valenciana’ varieties and the ‘Valenciana’ variety from the rest of European traditional tomato. Taking this into account, our results provide the ‘Valenciana d’El Perelló’ tomato genetic fingerprint, starting point for the valorization of this landrace and for its use in breeding programs or its certified use in high quality markets.Aquest projecte ha rebut financiació del programa Horizon 2020 de la Unió Europea a través del projecte No 634561 [This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 634561].Pons Puig, C.; Monforte Gilabert, AJ.; Figás Moreno, MDR.; Soler Aleixandre, S.; Blanca Postigo, JM.; Ziarsolo Areitioaurtena, P.; Cañizares Sales, J.... (2020). La tomata ‘Valenciana’ del Perelló: comparació de les seues característiques genètiques i fenotípiques amb el conjunt de tomates tradicionals europees. En I Congrés de la Tomaca Valenciana: La Tomaca Valenciana d'El Perelló. Editorial Universitat Politècnica de València. 141-152. https://doi.org/10.4995/TOMAVAL2017.2017.6196OCS14115

Atlas of phenotypic, genotypic and geographical diversity present in the European traditional tomato

[EN] The Mediterranean basin countries are considered secondary centres of tomato diversification. However, information on phenotypic and allelic variation of local tomato materials is still limited. Here we report on the evaluation of the largest traditional tomato collection, which includes 1499 accessions from Southern Europe. Analyses of 70 traits revealed a broad range of phenotypic variability with different distributions among countries, with the culinary end use within each country being the main driver of tomato diversification. Furthermore, eight main tomato types (phenoclusters) were defined by integrating phenotypic data, country of origin, and end use. Genome-wide association study (GWAS) meta-analyses identified associations in 211 loci, 159 of which were novel. The multidimensional integration of phenoclusters and the GWAS meta-analysis identified the molecular signatures for each traditional tomato type and indicated that signatures originated from differential combinations of loci, which in some cases converged in the same tomato phenotype. Our results provide a roadmap for studying and exploiting this untapped tomato diversity.We thank Universitat Illes Balears, the Greek Gene Bank (GGB-NAGREF), Universita degli Studi Mediterranea Reggio Calabria, the CRB-Leg (INRA-GAFL)", the Genebank of CNR-IBBR (Bari, Italy) and ARCA 2010 for seed sharing. CNR-IBBR also acknowledges the seed donors, the Leibniz Institute of Plant Genetics and Crop Plant Research, Maria Cristina Patane (CNR-IBE, Catania, Italy) and La Semiorto Sementi SRL, as well as Mrs. Roberta Nurcato for technical assistance. IBMCP-UPV acknowledges Maurizio Calduch (ALCALAX) for technical assistance and Mario Fon for English grammar editing. This work was supported by European Commission H2020 research and innovation program through TRADITOM grant agreement No.634561, G2P-SOL, grant agreement No. 677379, and HARNESSTOM grant agreement No. 101000716. Clara Pons and Mariola Plazas are grateful to Spanish Ministerio de Ciencia e Innovacion for postdoctoral grants FJCI-2016-29118 and IJC2019-039091I/AEI/10.13039/501100011033; Joan Casals to a Serra Hunter Fellow at Universitat Politècnica de Catalunya.Pons Puig, C.; Casals, J.; Palombieri, S.; Fontanet, L.; Riccini, A.; Rambla Nebot, JL.; Ruggiero, A.... (2022). Atlas of phenotypic, genotypic and geographical diversity present in the European traditional tomato. Horticulture Research. 9:1-16. https://doi.org/10.1093/hr/uhac112116

Prospective individual patient data meta-analysis of two randomized trials on convalescent plasma for COVID-19 outpatients

Data on convalescent plasma (CP) treatment in COVID-19 outpatients are scarce. We aimed to assess whether CP administered during the first week of symptoms reduced the disease progression or risk of hospitalization of outpatients. Two multicenter, double-blind randomized trials (NCT04621123, NCT04589949) were merged with data pooling starting when = 50 years and symptomatic for <= 7days were included. The intervention consisted of 200-300mL of CP with a predefined minimum level of antibodies. Primary endpoints were a 5-point disease severity scale and a composite of hospitalization or death by 28 days. Amongst the 797 patients included, 390 received CP and 392 placebo; they had a median age of 58 years, 1 comorbidity, 5 days symptoms and 93% had negative IgG antibody-test. Seventy-four patients were hospitalized, 6 required mechanical ventilation and 3 died. The odds ratio (OR) of CP for improved disease severity scale was 0.936 (credible interval (CI) 0.667-1.311); OR for hospitalization or death was 0.919 (CI 0.592-1.416). CP effect on hospital admission or death was largest in patients with <= 5 days of symptoms (OR 0.658, 95%CI 0.394-1.085). CP did not decrease the time to full symptom resolution

Pre-symptomatic transcriptome changes during cold storage of chilling sensitive and resistant peach cultivars to elucidate chilling injury mechanisms

Background: Cold storage induces chilling injury (CI) disorders in peach fruit (woolliness/mealiness, flesh browning and reddening/bleeding) manifested when ripened at shelf life. To gain insight into the mechanisms underlying CI, we analyzed the transcriptome of 'Oded' (high tolerant) and 'Hermoza' (relatively tolerant to woolliness, but sensitive to browning and bleeding) peach cultivars at pre-symptomatic stages. The expression profiles were compared and validated with two previously analyzed pools (high and low sensitive to woolliness) from the Pop-DG population. The four fruit types cover a wide range of sensitivity to CI. The four fruit types were also investigated with the ROSMETER that provides information on the specificity of the transcriptomic response to oxidative stress. Results: We identified quantitative differences in a subset of core cold responsive genes that correlated with sensitivity or tolerance to CI at harvest and during cold storage, and also subsets of genes correlating specifically with high sensitivity to woolliness and browning. Functional analysis indicated that elevated levels, at harvest and during cold storage, of genes related to antioxidant systems and the biosynthesis of metabolites with antioxidant activity correlates with tolerance. Consistent with these results, ROSMETER analysis revealed oxidative stress in 'Hermoza' and the progeny pools, but not in the cold resistant 'Oded'. By contrast, cold storage induced, in sensitivity to woolliness dependant manner, a gene expression program involving the biosynthesis of secondary cell wall and pectins. Furthermore, our results indicated that while ethylene is related to CI tolerance, differential auxin subcellular accumulation and signaling may play a role in determining chilling sensitivity/tolerance. In addition, sugar partitioning and demand during cold storage may also play a role in the tolerance/sensitive mechanism. The analysis also indicates that vesicle trafficking, membrane dynamics and cytoskeleton organization could have a role in the tolerance/sensitive mechanism. In the case of browning, our results suggest that elevated acetaldehyde related genes together with the core cold responses may increase sensitivity to browning in shelf life. Conclusions: Our data suggest that in sensitive fruit a cold response program is activated and regulated by auxin distribution and ethylene and these hormones have a role in sensitivity to CI even before fruit are cold stored.This research was funded by US-Israel Binational Agriculture Research and Development Fund (BARD) Grant no. 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A genetic genomics-expression approach reveals components of the molecular mechanisms beyond the cell wall that underlie peach fruit woolliness due to cold storage

[EN] Peach fruits subjected to prolonged cold storage (CS) to delay decay and over-ripening often develop a form of chilling injury (CI) called mealiness/woolliness (WLT), a flesh textural disorder characterized by lack of juiciness. Transcript profiles were analyzed after different lengths of CS and subsequent shelf life ripening (SLR) in pools of fruits from siblings of the Pop-DG population with contrasting sensitivity to develop WLT. This was followed by quantitative PCR on pools and individual lines of the Pop-DG population to validate and extend the microarray results. Relative tolerance to WLT development during SLR was related to the fruit's ability to recover from cold and the reactivation of normal ripening, processes that are probably regulated by transcription factors involved in stress protection, stress recovery and induction of ripening. 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AUXIN RESPONSE FACTOR 2 Intersects Hormonal Signals in the Regulation of Tomato Fruit Ripening.

The involvement of ethylene in fruit ripening is well documented, though knowledge regarding the crosstalk between ethylene and other hormones in ripening is lacking. We discovered that AUXIN RESPONSE FACTOR 2A (ARF2A), a recognized auxin signaling component, functions in the control of ripening. ARF2A expression is ripening regulated and reduced in the rin, nor and nr ripening mutants. It is also responsive to exogenous application of ethylene, auxin and abscisic acid (ABA). Over-expressing ARF2A in tomato resulted in blotchy ripening in which certain fruit regions turn red and possess accelerated ripening. ARF2A over-expressing fruit displayed early ethylene emission and ethylene signaling inhibition delayed their ripening phenotype, suggesting ethylene dependency. Both green and red fruit regions showed the induction of ethylene signaling components and master regulators of ripening. Comprehensive hormone profiling revealed that altered ARF2A expression in fruit significantly modified abscisates, cytokinins and salicylic acid while gibberellic acid and auxin metabolites were unaffected. Silencing of ARF2A further validated these observations as reducing ARF2A expression let to retarded fruit ripening, parthenocarpy and a disturbed hormonal profile. Finally, we show that ARF2A both homodimerizes and interacts with the ABA STRESS RIPENING (ASR1) protein, suggesting that ASR1 might be linking ABA and ethylene-dependent ripening. These results revealed that ARF2A interconnects signals of ethylene and additional hormones to co-ordinate the capacity of fruit tissue to initiate the complex ripening process

Tomato PYR/PYL/RCAR ABA receptors show high expression in root, differential sensitivity to the ABA-agonist quinabactin and capability to enhance plant drought resistance

This article contains supplementary data at: http://jxb.oxfordjournals.org/content/65/15/4451/suppl/DC1 © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology[EN] Abscisic acid (ABA) plays a crucial role in the plant’s response to both biotic and abiotic stress. Sustainable production of food faces several key challenges, particularly the generation of new varieties with improved water use efciency and drought tolerance. Different studies have shown the potential applications of Arabidopsis PYR/PYL/ RCAR ABA receptors to enhance plant drought resistance. Consequently the functional characterization of orthologous genes in crops holds promise for agriculture. The full set of tomato (Solanum lycopersicum) PYR/PYL/RCAR ABA receptors have been identied here. From the 15 putative tomato ABA receptors, 14 of them could be grouped in three subfamilies that correlated well with corresponding Arabidopsis subfamilies. High levels of expression of PYR/ PYL/RCAR genes was found in tomato root, and some genes showed predominant expression in leaf and fruit tissues. Functional characterization of tomato receptors was performed through interaction assays with Arabidopsis and tomato clade A protein phosphatase type 2Cs (PP2Cs) as well as phosphatase inhibition studies. Tomato receptors were able to inhibit the activity of clade A PP2Cs differentially in an ABA-dependent manner, and at least three receptors were sensitive to the ABA agonist quinabactin, which inhibited tomato seed germination. Indeed, the chemical activation of ABA signalling induced by quinabactin was able to activate stress-responsive genes. Both dimeric and monomeric tomato receptors were functional in Arabidopsis plant cells, but only overexpression of monomeric-type receptors conferred enhanced drought resistance. In summary, gene expression analyses, and chemical and transgenic approaches revealed distinct properties of tomato PYR/PYL/RCAR ABA receptors that might have biotechnological implications.This work was supported by the Ministerio de Ciencia e Innovacion, Fondo Europeo de Desarrollo Regional and Consejo Superior de Investigaciones Cientificas (grants BIO2011-23446 to PLR, BFU2011-25384 to AA, and Fontagro and COST1106 for networking activities to AG); fellowships to LR and MP; and a Juan de la Cierva contract to MGG. Cristina Martinez-Andujar and the Bioinformatics Core Service of the IBMCP are acknowledged for the SlEF1a marker and help in bioinformatics analyses, respectively.González Guzmán, M.; Rodriguez, L.; Lorenzo Orts, L.; Pons Puig, C.; Sarrion-Perdigones, A.; Fernandez, MA.; Peirats Llobet, M.... (2014). Tomato PYR/PYL/RCAR ABA receptors show high expression in root, differential sensitivity to the ABA-agonist quinabactin and capability to enhance plant drought resistance. Journal of Experimental Botany. 65(15):4451-4464. https://doi.org/10.1093/jxb/eru219S44514464651

Uniform ripening Encodes a Golden 2-like Transcription Factor Regulating Tomato Fruit Chloroplast Development

[EN] Modern tomato (Solanum lycopersicum) varieties are bred for uniform ripening (u) light green fruit phenotypes to facilitate harvests of evenly ripened fruit. U encodes a Golden 2-like (GLK) transcription factor, SlGLK2, which determines chlorophyll accumulation and distribution in developing fruit. In tomato, two GLKs-SlGLK1 and SlGLK2-are expressed in leaves, but only SlGLK2 is expressed in fruit. Expressing GLKs increased the chlorophyll content of fruit, whereas SlGLK2 suppression recapitulated the u mutant phenotype. GLK overexpression enhanced fruit photosynthesis gene expression and chloroplast development, leading to elevated carbohydrates and carotenoids in ripe fruit. SlGLK2 influences photosynthesis in developing fruit, contributing to mature fruit characteristics and suggesting that selection of u inadvertently compromised ripe fruit quality in exchange for desirable production traits.Minimum Information About a Microarray Experiment (MIAME)-compliant microarray data are available at http://ted.bti.cornell.edu. and at http://www.ebi.ac.uk/arrayexpress/(accession E-MEXP-3652). F. Carrari and A. Fernie provided S. pennellii SlGLK2, and J. Maloof provided S. habrochaites SlGLK2 sequences. The U. S. Department of Agriculture (USDA)/National Institute of Food and Agriculture Solanaceae Coordinated Agricultural Project provided potato data. We are grateful to the Tomato Genome Consortium and the SOL Genomics Network for prepublication access to the tomato genome sequence. The S. pennellii introgression lines were provided by the C. M. Rick Tomato Genetics Resource Center; the S. pimpinellifolium populations were provided by the Instituto de Hortofruticultura Subtropical y Mediterranea "La Mayora," Consejo Superior de Investigaciones Cientificas; and both populations are available by request from the sources. The AtGLK-expressing lines were provided by Mendel Biotechnology and Seminis/Monsanto Vegetable Seeds. SlGLK2, the corresponding lines, and the F2 10-1 IL x M82 population lines and seeds are available from J. J. G. without restriction. Seminis/Monsanto will make available, upon request, and under a material transfer agreement indicating they are to be used for noncommercial purposes, the following lines: LexA:AtGLK1:p35S:LexA-Gal4; LexA:AtGLK1:pLTP:LexA-Gal4; LexA:AtGLK1:pRbcS:LexA-Gal4; LexA:AtGLK1:pPDS:LexA-Gal4; LexA:AtGLK2:p35S:LexA-Gal4; LexA:AtGLK2:pLTP:LexA-Gal4; LexA:AtGLK2:pRbcS:LexA-Gal4; LexA:AtGLK2:pPDS:LexA-Gal4; plus the T63 control line. Other biological materials are available by request from A. L. T. P. or J.J.G. A. L. T. P., T. H., K.L.-C., R.F.-B., and A. B. B. have filed a provisional U. S. patent application UC #2011-841, "Introduction of wild species GLK genes for improved ripe tomato fruit quality," through the University of California. A. L. T. P. and A. B. B. have filed the U. S. patent application #2010/0154078, " Transcription factors that enhance traits in plant organs," through Mendel Biotechnology. Assistance from B. Blanco-Ulate, S. Phothiset, S. Reyes, A. Abraham, L. Gilani, and G. Arellano is gratefully acknowledged. J. Langdale provided helpful advice regarding GLK phylogeny and nomenclature. G. Adamson and P. Kysar, Electron Microscopy (EM) Laboratory, University of California Davis Medical Center did the EM work. University of California Discovery and partners funded the pepper analysis and the initial investigations of the Arabidopsis GLKs. The Vietnam Education Foundation supported C.N. Fundacion Genoma Espana ESPSOL Project provided partial funding to A.G. USDA-Agricultural Research Service, USDA-National Research Initiative (2007-02773), and NSF (Plant Genome Program IOS-0923312) provided support to J.J.G.Powell, AL.; Nguyen, CV.; Hill, T.; Cheng, KL.; Figueroa-Balderas, R.; Aktas, H.; Ashrafi, H.... (2012). Uniform ripening Encodes a Golden 2-like Transcription Factor Regulating Tomato Fruit Chloroplast Development. Science. 336:1711-1715. doi:10.1126/science.1222218S1711171533