A Rapid and Efficient Method for Purifying High Quality Total RNA from Peaches (Prunus persica) for Functional Genomics Analyses

http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602005000100010&lng=es&nrm=isoPrunus persica has been proposed as a genomic model for deciduous trees and the Rosaceae family. Optimized protocols for RNA isolation are necessary to further advance studies in this model species such that functional genomics analyses may be performed. Here we present an optimized protocol to rapidly and efficiently purify high quality total RNA from peach fruits (Prunus persica). Isolating high-quality RNA from fruit tissue is often difficult due to large quantities of polysaccharides and polyphenolic compounds that accumulate in this tissue and co-purify with the RNA. Here we demonstrate that a modified version of the method used to isolate RNA from pine trees and the woody plant Cinnamomun tenuipilum is ideal for isolating high quality RNA from the fruits of Prunus persica. This RNA may be used for many functional genomic based experiments such as RT-PCR and the construction of large-insert cDNA libraries

アントシアニジン合成系遺伝子DFRの系統解析

Plants have various color pigments. Many plants have fruits and flowers of vivid colors such as orange, red, purple and blue. Anthocyanins with anthocyanidins as aglycone are one group of plant pigments. Each type of anthocyanidins is determined by the substrate specificity of dihydroflavonol 4-reductase (DFR) in its biosynthetic pathway and has a major influence on the determination of plant color. We performed a phylogenetic analysis of DFR genes, which are key enzymes of this anthocyanidin biosynthesis pathway. We collected 170 DFR clones in various plants by browsing database (NCBI). The collected DFRs were classified into ten groups according tosequence similarity. The consensus sequences within each group were aligned, and amino acids involved in the responsible region for determination of the substrate specificity of DFR have been shown. In addition, DFR's intron sites were highly conserved. These results would be useful to unveil the relationship between DFR and plant pigments

Plant Cellular and Molecular Biotechnology: Following Mariotti's Steps

This review is dedicated to the memory of Prof. Domenico Mariotti, who significantly contributed to establishing the Italian research community in Agricultural Genetics and carried out the first experiments of Agrobacterium-mediated plant genetic transformation and regeneration in Italy during the 1980s. Following his scientific interests as guiding principles, this review summarizes the recent advances obtained in plant biotechnology and fundamental research aiming to: (i) Exploit in vitro plant cell and tissue cultures to induce genetic variability and to produce useful metabolites; (ii) gain new insights into the biochemical function of Agrobacterium rhizogenes rol genes and their application to metabolite production, fruit tree transformation, and reverse genetics; (iii) improve genetic transformation in legume species, most of them recalcitrant to regeneration; (iv) untangle the potential of KNOTTED1-like homeobox (KNOX) transcription factors in plant morphogenesis as key regulators of hormonal homeostasis; and (v) elucidate the molecular mechanisms of the transition from juvenility to the adult phase in Prunus tree species

Transcriptional regulatory networks controlling woolliness in peach in response to preharvest gibberellin application and cold storage

BACKGROUND: Postharvest fruit conservation relies on low temperatures and manipulations of hormone metabolism to maintain sensory properties. Peaches are susceptible to chilling injuries, such as ‘woolliness’ that is caused by juice loss leading to a ‘wooly’ fruit texture. Application of gibberellic acid at the initial stages of pit hardening impairs woolliness incidence, however the mechanisms controlling the response remain unknown. We have employed genome wide transcriptional profiling to investigate the effects of gibberellic acid application and cold storage on harvested peaches. RESULTS: Approximately half of the investigated genes exhibited significant differential expression in response to the treatments. Cellular and developmental process gene ontologies were overrepresented among the differentially regulated genes, whereas sequences in cell death and immune response categories were underrepresented. Gene set enrichment demonstrated a predominant role of cold storage in repressing the transcription of genes associated to cell wall metabolism. In contrast, genes involved in hormone responses exhibited a more complex transcriptional response, indicating an extensive network of crosstalk between hormone signaling and low temperatures. Time course transcriptional analyses demonstrate the large contribution of gene expression regulation on the biochemical changes leading to woolliness in peach. CONCLUSION: Overall, our results provide insights on the mechanisms controlling the complex phenotypes associated to postharvest textural changes in peach and suggest that hormone mediated reprogramming previous to pit hardening affects the onset of chilling injuries. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12870-015-0659-2) contains supplementary material, which is available to authorized users

A survey of sequences of KT-HAK-KUP transporters in green algae and basal land plants

In this data article, information is provided on sequences of KT-HAK-KUP transporters from green algae and basal land plants. A data set is offered containing sequences corresponding to the chlorophyte algae Chlamydomonas eustigma, Gonium pectorale and Coccomyxa subellipsoidea, the charophyte algae Coleochaete orbicularis and Klebsormidium flaccidum, the bryophyte Sphagnum fallax, the marchantophyte Marchantia polymorpha and the gymnosperm Pinus taeda, which have been not formerly analyzed. In addition, an analysis of similarity scores among representatives of the clusters recognized in photosynthetic green organisms (namely, chlorophyte algae, charophyte algae, basal embryophytes and higher embryophytes) is performed as well as an analysis of membrane topology for them.Fil: Santa Maria, Guillermo Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Universidad Nacional de San Martín; ArgentinaFil: Oliferuk, Sonia. Universidad Nacional de San Martín; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); ArgentinaFil: Moriconi, Jorge Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Universidad Nacional de San Martín; Argentin

Diversity and Abundance of Hymenopterous Parasitoids Associated with Anastrepha fraterculus (Diptera: Tephritidae) in Native and Exotic Host Plants in Misiones, Northeastern Argentina

Some Major host species used by the tephritid fruit flies Anastrepha fraterculus (Wiede-mann) and Ceratitis capitata (Wiedemann), including Acca sellowiana (O. Berg) Burret, Campomanesia xanthocarpa O. Berg, Psidium guajava L., Prunus persica (L.) Batsch, Eriobotrya japonica (Thunb.) Lindl., Citrus reticulata Blanco var. Murcott, C. aurantium L., C. paradisi Macfadyen var. Dalan Dan, and C. paradisi var. Sudashi, were sampled for fruit fly larvae between Feb and Dec 2000 in the northernmost section of the Paranaense forest, in the Province of Misiones, NE Argentina. Both A. fraterculus and C. capitata were obtained from these host plant species, with A. fraterculus accounting for 93% of all tephritid puparia identified. Ten species of larval-pupal parasitoids were recovered from A. fraterculus; Doryctobracon areolatus (Szépligeti), D. brasiliensis (Szépligeti), Utetes anastrephae (Viereck), Opius bellus (Gahan), Diachasmimorpha longicaudata (Ashmead) (Opiinae, raconidae), Odontosema anastrephae Borgmeier, Lopheucoila anastrephae (Rohwer), Aganaspis pelleranoi (Brèthes) (Eucoilinae, Figitidae), Asobara anastrephae (Muessebeck) (Alyssinae, Braconidae), and Aceratoneuromyia indica (Silvestri) (Tetrastichinae, Eulophidae). All these parasitoids, with the exception of D. longicaudata and A. indica, are native to the Neotropical region. No parasitoids were recovered from C. capitata puparia. Asobara anastrephae and O. anastrephae are newly recorded in Argentina, whereas D. brasiliensis, U. anastrephae, and L. anastrephae are newly reported in Misiones. The eucoiline A. pelleranoi wasthe most abundant parasitoid species. Acca sellowiana and P. guajava harbored the highest parasitoid abundance and diversity.Fil: Schliserman, Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Catamarca. Universidad Nacional de Catamarca. Centro de Investigaciones y Transferencia de Catamarca; ArgentinaFil: Ovruski Alderete, Sergio Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Decoll, Olga. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Misiones; ArgentinaFil: Wharton, Robert. Texas A&M University; Estados Unido

Physiological characterization of drought stress response and expression of two transcription factors and two LEA genes in three Prunus genotypes

Global warming has led to a progressive decrease in rainfall, which is reflected by a reduction of water resources in the soil and a negative effect on crop production in Mediterranean areas. Under drought stress, many plants react by inducing a different series of responses at both physiological and molecular levels, allowing them to survive for a variable period of time. Therefore, in order to understand the response of roots to drought conditions, the genotypes peach × almond ‘Garnem’ [P. amygdalus Batsch × P. persica (L.) Batsch] and their progeny, the hybrid ‘P.2175’ × ‘Garnem’-3 and OP-‘P.2175’ (P. cerasifera Ehrh.) were subjected to a period of water deficit. Drought conditions with a subsequent re-watering period were tested for potted plants for one month. Stomatal conductance and leaf water potential were measured to monitor the plant physiological responses. Significant differences among the drought stress and drought stress recovery treatments and among the genotypes were observed. In addition, four genes related to the ABA biosynthesis pathway were studied for their expression by RT-qPCR: an AN20/AN1 zinc finger protein (ppa012373m); a bZIP transcription factor (ppa013046m); a dehydrin (ppa005514m) and a LEA protein (ppa008651m). Their expression profiles correlated with our physiological results of drought response, being higher in roots than in phloem tissue. In general, the expression of the four studied genes was higher after 15 days under drought conditions. Under drought and recovery conditions, the zinc finger and bZIP transcription factors showed significant differences in their relative expression levels from LEA and dehydrin. These results suggest the role of LEA and dehydrin in the regulatory response to drought stress in Prunus genotypes. Therefore, the dehydrin and the protein LEA might be potential biomarkers to select rootstocks for tolerance to drought conditions.We wish to thank Michael Glenn for helpful comments and ideas on the ash content discussion. This work was supported by RTA2014-00062 from the Instituto Nacional de Investigación y Tecnología Agraria (INIA) and by the Research Group A12 of Aragon, Spain. We kindly appreciate the FPI-INIA 2012 grant for B. Bielsa

A preliminary transcriptomic approach to elucidate post harvest ripening of plum fruit

The aim of the present study was to dissect common and/or diverse mechanisms regulating plum (Prunus salicina) fruit ripening in genotypes characterized by different patterns of ethylene production. Fruit of an ethylenesuppressed cultivar ('Shiro') and a cultivar characterized by the typical increase of ethylene production during fruit ripening ('Santa Rosa') were harvested at commercial maturity stage and allowed to further ripen at room temperature (23°C) up to 4 days. While non-detectable amounts of ethylene were recorded in 'Shiro' fruit, a typical climacteric behavior was observed in 'Santa Rosa' plums. For comparative purposes, the peach microarray μPEACH 1.0 containing 4,806 oligonucleotides corresponding to an equal number of genes expressed in peach fruit was employed for transcript profiling during postharvest ripening of both cultivars. Intriguingly, transcript levels of genes involved in ethylene biosynthesis, primarily 1-aminocyclopropane-1-carboxylate synthase, appeared to increase during the progress of 'Shiro' fruit ripening, following the same pattern as in 'Santa Rosa' plums. These data suggest that an induction of the ethylene biosynthetic pathway is present also in plum cultivars in which the burst of ethylene is not detectable. Expression levels of other genes implicated in auxin metabolism, antioxidant system and stress response followed the same pattern in both cultivars. Overall, this preliminary transcriptomic approach tried to elucidate the flow of events that accompany postharvest ripening of plum cultivars with diverse properties in relation to ethylene evolutio

The Peach RGF/GLV Signaling Peptide pCTG134 Is Involved in a Regulatory Circuit That Sustains Auxin and Ethylene Actions

In vascular plants the cell-to-cell interactions coordinating morphogenetic and physiological processes are mediated, among others, by the action of hormones, among which also short mobile peptides were recognized to have roles as signals. Such peptide hormones (PHs) are involved in defense responses, shoot and root growth, meristem homeostasis, organ abscission, nutrient signaling, hormone crosstalk and other developmental processes and act as both short and long distant ligands. In this work, the function of CTG134, a peach gene encoding a ROOT GROWTH FACTOR/GOLVEN-like PH expressed in mesocarp at the onset of ripening, was investigated for its role in mediating an auxin-ethylene crosstalk. In peach fruit, where an auxin-ethylene crosstalk mechanism is necessary to support climacteric ethylene synthesis, CTG134 expression peaked before that of ACS1 and was induced by auxin and 1-methylcyclopropene (1-MCP) treatments, whereas it was minimally affected by ethylene. In addition, the promoter of CTG134 fused with the GUS reporter highlighted activity in plant parts in which the auxin-ethylene interplay is known to occur. Arabidopsis and tobacco plants overexpressing CTG134 showed abnormal root hair growth, similar to wild-type plants treated with a synthetic form of the sulfated peptide. Moreover, in tobacco, lateral root emergence and capsule size were also affected. In Arabidopsis overexpressing lines, molecular surveys demonstrated an impaired hormonal crosstalk, resulting in a re-modulated expression of a set of genes involved in both ethylene and auxin synthesis, transport and perception. These data support the role of pCTG134 as a mediator in an auxin-ethylene regulatory circuit and open the possibility to exploit this class of ligands for the rational design of new and environmental friendly agrochemicals able to cope with a rapidly changing environment

Espectroscopía NIR como método no destructivo de determinación de parámetros de calidad del fruto en melocotón tardío de Calandra y en Miraflores

1 copia .pdf (a-3) de póster original presentado. 3 Figs.El melocotonero tardío de Calanda (Prunus persica (L.) Batsch.) es una variedad población de melocotonero (Prunus persica (L.) Batsch.) de gran importancia económica por su alto valor añadido. La espectroscopía NIR es una técnica cuyo principal interés radica en la posibilidad de estimación, por métodos no destructivos, de parámetros de calidad del fruto.Trabajo financiado por los proyectos “PET2007-09-COS-02 (INIA) y PM005/2006 (Gobierno de Aragón). El equipo de Investigación de Aula Dei está integrado en el Grupo Consolidado “Alimentos de Origen Vegetal”, reconocido por el Gobierno de Aragón.Peer reviewe