Localization of QTLs for in vitro plant regeneration in tomato

Background Low regeneration ability limits biotechnological breeding approaches. The influence of genotype in the regeneration response is high in both tomato and other important crops. Despite the various studies that have been carried out on regeneration genetics, little is known about the key genes involved in this process. The aim of this study was to localize the genetic factors affecting regeneration in tomato. Results We developed two mapping populations (F2 and BC1) derived from a previously selected tomato cultivar (cv. Anl27) with low regeneration ability and a high regeneration accession of the wild species Solanum pennellii (PE-47). The phenotypic assay indicated dominance for bud induction and additive effects for both the percentage of explants with shoots and the number of regenerated shoots per explant. Two linkage maps were developed and six QTLs were identified on five chromosomes (1, 3, 4, 7 and 8) in the BC1 population by means of the Interval Mapping and restricted Multiple QTL Mapping methods. These QTLs came from S. pennellii, with the exception of the minor QTL located on chromosome 8, which was provided by cv. Anl27. The main QTLs correspond to those detected on chromosomes 1 and 7. In the F2 population, a QTL on chromosome 7 was identified on a similar region as that detected in the BC1 population. Marker segregation distortion was observed in this population in those areas where the QTLs of BC1 were detected. Furthermore, we located two tomato candidate genes using a marker linked to the high regeneration gene: Rg-2 (a putative allele of Rg-1) and LESK1, which encodes a serine/threonine kinase and was proposed as a marker for regeneration competence. As a result, we located a putative allele of Rg-2 in the QTL detected on chromosome 3 that we named Rg-3. LESK1, which is also situated on chromosome 3, is outside Rg-3. In a preliminary exploration of the detected QTL peaks, we found several genes that may be related to regeneration. Conclusions In this study we have identified new QTLs related to the complex process of regeneration from tissue culture. We have also located two candidate genes, discovering a putative allele of the high regeneration gene Rg-1 in the QTL on chromosome 3. The identified QTLs could represent a significant step toward the understanding of this process and the identification of other related candidate genes. It will also most likely facilitate the development of molecular markers for use in gene isolation.Trujillo Moya, C.; Gisbert Domenech, MC.; Vilanova Navarro, S.; Nuez Viñals, F. (2011). Localization of QTLs for in vitro plant regeneration in tomato (Solanum lycopersicum L.). BMC Plant Biology. 11:140-152. doi:10.1186/1471-2229-11-140S1401521

Recovering Ancient Grapevine Varieties: From Genetic Variability to In Vitro Conservation, A Case Study

A great number of varieties have been described in grapevine; however, few of them are currently in use. The increasing concern on varietal diversity loss has encouraged actions for recovering and preserving grapevine germplasm, which represents valuable resources for breeding as well as for diversification in grapevine-derived products. On the other hand, it is expected that this important crop, which is distributed in warm areas worldwide, will suffer the climate changes. Therefore, it is also convenient the identification of intravarietal variability and the recovery of accessions well adapted to particular environments. In this chapter, we will contribute to highlight the importance of recovering ancient materials, the usefulness of SSR markers to determine their molecular profile, the importance to analyze their virus status, and the possibilities that offer biotechnological tools for virus sanitation and in vitro storage as a complement of field preservation. In this context, we have evaluated different grapevine accessions and developed in vitro culture protocols for micropropagation, sanitation, and storage grapevine cultivars. In this work, we report the results obtained for the historic variety “Valencí Blanc” (or “Beba”) and the historic and endangered variety “Esclafagerres” (“Esclafacherres” or “Esclafacherris”)

New findings and actions in the recovery of old Mediterranean grapevine varieties

In this work, we report new findings related to the recovery of grapevine diversity in the Comunitat Valenciana (Eastern Spain): accessions of old varieties at risk of disappearance, new genotypes, synonymies, homonymies, and some foreign old varieties. In addition, the in vitro establishment of some rescued varieties has been carried out as a complement to ex-situ conservation, as well as to provide material for biotechnological applications

Overexpression of BvHb2, a Class 2 Non-Symbiotic Hemoglobin from Sugar Beet, Confers Drought-Induced Withering Resistance and Alters Iron Content in Tomato

Drought stress is one of the major threats to agriculture and concomitantly to food production. Tomato is one of the most important industrial crops, but its tolerance to water scarcity is very low. Traditional plant breeding has a limited margin to minimize this water requirement. In order to design novel biotechnological approaches to cope with this problem, we have screened a plant cDNA library from the halotolerant crop sugar beet (Beta vulgaris L.) for genes able to confer drought/osmotic stress tolerance to the yeast model system upon overexpression. We have identified the gene that encodes BvHb2, a class 2 non-symbiotic hemoglobin, which is present as a single copy in the sugar beet genome, expressed mainly in leaves and regulated by light and abiotic stress. We have evaluated its biotechnological potential in the model plant Arabidopsis thaliana and found that BvHb2 is able to confer drought and osmotic stress tolerance. We also generated transgenic lines of tomato (Solanum lycopersicum) overexpressing BvHb2 and found that the resulting plants are more resistant to drought-induce withering. In addition, transgenic lines overexpressing BvHb2 exhibit increased levels of iron content in leaves. Here, we show that class 2 non-symbiotic plant hemoglobins are targets to generate novel biotechnological crops tolerant to abiotic stress. The fact that these proteins are conserved in plants opens the possibility for using Non-GMO approaches, such as classical breeding, molecular breeding, or novel breeding techniques to increase drought tolerance using this protein as a target.</jats:p

Comportamiento de líneas de Cucumis metuliferus, Citrullus lanatus var. citroides y C. colocynthis frente a Meloidogyne spp. como potenciales portainjertos de melón, pepino y sandía

La resistencia vegetal para el control de Meloidogyne es efectiva y económicamente rentable, pero su uso continuado puede seleccionar poblaciones virulentas, como se ha citado en tomate y pimiento. En cucurbitáceas existe germoplasma resistente que podría utilizarse como patrón en rotación con solanáceas para mejorar el control del nematodo y la durabilidad de los genes implicados. Se llevaron a cabo una serie de ensayos para identificar resistencia a M. incognita y M. javanica en dos líneas de Cucumis metuliferusPostprint (published version

Desarrollo de un protocolo de embriogénesis somática para el saneamiento de las variedades de vid infectadas con GLRaV-3 y GFkV

La embriogénesis somática es una técnica de cultivo in vitro que permite el saneamiento de cultivares de vid (Vitis vinifera L.). Se ha analizado la presencia de los virus ArMvV, GELY, GFkV, GLRaV-1 y GLRaV-3 mediante real-time multiplex RT-PCR en hojas de plantas adultas de los cultivares “Valencí Blanc” y “Botó de Gall”. Además, se ha establecido un protocolo de embriogénesis somática para ambas variedades. La adición de 2 mgL-1 de 2,4D en el medio de cultivo incrementa el porcentaje de embriones cultivados así como el número de embriones por callos

Cucumis metuliferus es resistente a poblaciones de Meloidogyne spp. incluso virulentas al gen MI

Se realizaron diversos ensayos para determinar la respuesta de C. metuliferus frente diversas poblaciones de Meloidogyne arenaria, M. incognita y M. javanica, algunas de las cuales habían sido ya caracterizadas como virulentas al gen de resistencia Mien tomatePostprint (published version

The influence of ethylene and ethylene modulators on shoot organogenesis in tomato

[EN] The influence of ethylene and ethylene modulators on the in vitro organogenesis of tomato was studied using a highly regenerating accession of the wild tomato Solanum pennellii and an F1 plant resulting from a cross between Solanum pennellii and Solanum lycopersicum cv. Anl27, which is known to have a low regeneration frequency. Four ethylene-modulating compounds, each at four levels, were used, namely: cobalt chloride (CoCl 2), which inhibits the production of ethylene; AgNO 3 (SN), which inhibits ethylene action; and Ethephon and the precursor 1-aminocyclopropane-1-carboxylic acid (ACC), which both promote ethylene synthesis. Leaf explants of each genotype were incubated on shoot induction medium supplemented with each of these compounds at 0, 10 or 15 days following bud induction. The results obtained in our assays indicate that ethylene has a significant influence on tomato organogenesis. Concentrations of ethylene lower than the optimum (according to genotype) at the beginning of the culture may decrease the percentage of explants with buds (B), produce a delay in their appearance, or indeed inhibit bud formation. This was observed in S. pennellii and the F1 explants cultured on media with SN (5.8-58.0 ¿M) as well as in the F1 explants cultured on medium with 21.0 ¿M CoCl 2. The percentage of explants with shoots (R) and the mean number of shoots per explant with shoots (PR) also diminished in media that contained SN. Shoots isolated from these explants were less developed compared to those isolated from control explants. On the other hand, ethylene supplementation may contribute to enhancing shoot development. The number of isolable shoots from S. pennellii explants doubled in media with ACC (9.8-98.0 ¿M). Shoots isolated from explants treated with ethylene releasing compounds showed a higher number of nodes when ACC and Ethephon were added at 10 days (in F1 explants) or at 15 days (in S. pennellii) after the beginning of culture. Thus, the importance of studying not only the concentration but also the timing of the application of regulators when developing regeneration protocols has been made manifest. An excess of ethylene supplementation may produce an inhibitory effect, as was observed when using Ethephon (17.2-69.0 ¿M). These results show the involvement of ethylene in tomato organogenesis and lead us to believe that ethylene supplementation may contribute to enhancing regeneration and shoot development in tomato. © 2012 Springer Science+Business Media B.V.Carlos Trujillo has a predoctoral fellowship from the Spanish 'Ministerio de Educacion y Ciencia'. This work has been funded by Universitat Politecnica de Valencia (PAID 05-10). The technical assistance of N. Palacios and the revision of the manuscript's English by J. Bergen are gratefully acknowledged.Trujillo Moya, C.; Gisbert Domenech, MC. (2012). The influence of ethylene and ethylene modulators on shoot organogenesis in tomato. Plant Cell, Tissue and Organ Culture. 111(1):141-148. https://doi.org/10.1007/s11240-012-0168-zS1411481111Abeles FB, Morgan PW, Saltveit ME (1992) Ethylene in plant biology. Academic Press, San DiegoBhatia P, Ashwath N, Senaratna T, David M (2004) Tissue culture studies of tomato (Lycopersicon esculentum). Plant Cell Tiss Org Cult 78:1–21Bhatia P, Ashwath N, Midmore DJ (2005) Effects of genotype, explant orientation, and wounding on shoot regeneration in tomato. In Vitro Cell Dev Biol-Plant 41:457–464Biddington NL (1992) The Influence of ethylene in plant-tissue culture. Plant Growth Regul 11:173–187Brown DC, Thorpe TA (1995) Crop improvement through tissue culture. World J Microbiol Biotechnol 11(4):409–415Chraibi KMB, Latche A, Roustan JP, Fallot J (1991) Stimulation of shoot regeneration from cotyledons of Helianthus annuus by the ethylene inhibitors,silver and cobalt. Plant Cell Rep 10:204–207Devi R, Dhaliwal MS, Kaur A, Gosal SS (2008) Effect of growth regulators on in vitro morphogenic response of tomato. Indian J Biotechnol 7:526–530Dias LLC, Santa-Catarina C, Ribeiro DM, Barros RS, Floh EIS, Otoni WC (2009) Ethylene and polyamine production patterns during in vitro shoot organogenesis of two passion fruit species as affected by polyamines and their inhibitor. Plant Cell Tiss Org Cult 99:199–208Dimasi-Theriou K, Economou AS (1995) Ethylene enhances shoot formation in cultures of the peach rootstock GF-677 (Prunus persica × P. amygdalus). Plant Cell Rep 15:87–90Gisbert C, Arrillaga I, Roig LA, Moreno V (1999) Adquisition of a collection of Lycopersicon pennellii (Corr. D’Arcy) transgenic plants with uidA and nptII marker genes. J Hortic Sci Biotechnol 74:105–109Hughes KW (1981) In vitro ecology: exogenous factors affecting growth and morphogenesis in plant culture systems. Environ Exp Bot 21:281–288Huxter TJ, Thorpe TA, Reid DM (1981) Shoot initiation in light- and darkgrown tobacco callus: the role of ethylene. Physiol Plant 53:319–326Kumar PP, Lakshmanan P, Thorpe TA (1998) Regulation of morphogenesis in plant tissue culture by ethylene. In Vitro Cell Dev Biol Plant 34:94–103Lima JE, Benedito VA, Figueira A, Peres LEP (2009) Callus, shoot and hairy root formation in vitro as affected by the sensitivity to auxin and ethylene in tomato mutants. Plant Cell Rep 28:1169–1177Lu J, Vahala J, Pappinen A (2011) Involvement of ethylene in somatic embryogenesis in Scots pine (Pinus sylvestris L.). Plant Cell Tiss Org Cult 107:25–33Mohiuddin AKM, Chowdhury MKU, Abdullah ZC, Napis S (1997) Influence of silver nitrate (ethylene inhibitor) on cucumber in vitro shoot regeneration. Plant Cell Tiss Org Cult 51:75–78Moshkov IE, Novikova GV, Hall MA, George EF (2008) Plant Growth Regulators III: ethylene. In: George EF, Hall MA, Klerk G-JD (eds) Plant Propaga-tion by Tissue Culture, vol 1. 3rd edn. Springer, The Netherlands, pp 239–248Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497Osman MG, Khalafalla MM (2010) Promotion of in vitro shoot formation from shoot tip of tomato (Lycopersicon esculentum Mill. cv. Omdurman) by ethylene inhibitors. Int J Curr Res 4:82–86Ptak A, El Tahchy A, Wyzgolik G, Henry M, Laurain-Mattar D (2010) Effects of ethylene on somatic embryogenesis and galantamine content in Leucojum aestivum L. cultures. Plant Cell Tiss Org Cult 102:61–67Pua EC, Sim GE, Chi GL, Kong LF (1996) Synergistic effects of ethylene inhibitors and putrescine on shoot regeneration from hypocotyl explants of Chinese radish (Raphanus sativus L. var. longipinnatus Bailey) in vitro. Plant Cell Rep 15:685–690Reid MS (1995) Ethylene in plant growth, development and senescence. In: Davies PJ (ed) Plant hormones: physiology, biochemistry and molecular biology, 2nd edn. Kluwer Acad Publ, The Netherlands, pp 486–508Trujillo-Moya C, Gisbert C, Vilanova S, Nuez F (2011) Localization of QTLs for in vitro plant regeneration in tomato. BMC Plant Biol 11: art.140Tsuchisaka A, Theologis A (2004) Heterodimeric interactions among the 1-amino-cyclopropane-1-carboxylate synthase polypeptides encoded by the Arabidopsis gene family. Proc Natl Acad Sci USA 101:2275–2280Vogel JP, Woeste KE, Theologis A, Kieber JJ (1998) Recessive and dominant mutations in the ethylene biosynthetic gene ACS5 of Arabidopsis confer cytokinin insensitivity and ethylene overproduction, respectively. Proc Natl Acad Sci USA 95:4766–477

In vitro propagation of Vitis vinifera L. cv. 'Monastrell'

[EN] Background: A protocol for the micropropagation of the grape (Vitis vinifera L.) cultivar 'Monastrell' was developed. Initial plant material was obtained from the sanitary selection of grapevine plants performed by real-time RT-PCR to confirm the absence of Grapevine fanleaf virus, Arabis mosaic virus, Grapevine leafroll-associated virus 1, Grapevine leafroll-associated virus 3, and Grapevine fleck virus. Results: The effects of the salt composition (comparing Lloyd and McCown woody plant medium and Murashige and Skoog medium 1/2 macronutrients) and the growth regulator benzylaminopurine (BAP), at 0 and 8.9 mu M, on plant propagation were evaluated using nodes as explants. The most efficient procedure consisted of bud induction in the medium with Lloyd and McCown woody plant salts and 8.9 mu M BAP for 30 d along with elongation in cytokinin-free medium for 60 d, which gave 22 nodes/explant (174 plants/initial plant). A second cycle of propagation in a medium without BAP for another 60 d could give approximately 10,000 nodes, which can be obtained after an additional 2 months of culture. All plants acclimatized after the second cycle of multiplication were successfully transferred to soil. Conclusion: We developed an optimal protocol for V. vinifera cv. 'Monastrell' micropropagation, the first described for this cultivar. (C) 2017 Pontificia Universidad Catolica de Valparaiso. Production and hosting by Elsevier B. V. All rights reserved.The study was supported by the projects RTA2011-00067-C04, RTA2014-00061-C03, and PRP-CGL2015-70843-R, all co-funded with FEDER Funds. Tania San Pedro has a grant (01/14-FSE-22) supported by the Instituto Valenciano de Investigaciones Agrarias.San Pedro-Galan, T.; Peiró Barber, RM.; Villanova, J.; Olmos Castelló, A.; Gisbert Domenech, MC. (2017). In vitro propagation of Vitis vinifera L. cv. 'Monastrell'. Electronic Journal of Biotechnology. 27:80-83. https://doi.org/10.1016/j.ejbt.2017.03.006S80832