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

Dominance induction of fruitlet shedding in Malus × domestica (L. Borkh): molecular changes associated with polar auxin transport

BACKGROUND: Apple fruitlet abscission is induced by dominance, a process in which hormones such as auxin, cytokinins and strigolactone play a pivotal role. The response to these hormones is controlled by transcription regulators such as Aux/IAA and ARR, whereas auxin transport is controlled by influx and efflux carriers. RESULTS: Seven partial clones encoding auxin efflux carriers (MdPIN1_A, MdPIN1_B, MdPIN10_A, MdPIN10_B, MdPIN4, MdPIN7_A and MdPIN7_B), three encoding auxin influx carriers (MdLAX1, MdLAX2 and MdLAX3) and three encoding type A ARR cytokinin response regulators (MdARR3, MdARR4 and MdARR6) were isolated by the use of degenerate primers. The organization of the PIN multigene family in apple is closer to Medicago truncatula than to Arabidopsis thaliana. The genes are differentially expressed in diverse plant organs and at different developmental stages. MdPIN1 and MdPIN7 are largely more expressed than MdPIN10 and MdPIN4. During abscission, the transcription of these genes increased in the cortex whereas in the seed a sharp fall was observed. The expression of these genes was found to be at least partially controlled by ethylene and auxin. CONCLUSION: The ethylene burst preceding abscission of fruitlets may be responsible for the decrease in transcript level of MDPIN1, MDARR5 and MDIAA3 in seed. This situation modulates the status of the fruitlet and its fate by hampering the PAT from the seeds down through the abscission zone (AZ) and this brings about the shedding of the fruitlet

Large-scale Gene Ontology analysis of plant transcriptome-derived sequences retrieved by AFLP technology

<p>Abstract</p> <p>Background</p> <p>After 10-year-use of AFLP (Amplified Fragment Length Polymorphism) technology for DNA fingerprinting and mRNA profiling, large repertories of genome- and transcriptome-derived sequences are available in public databases for model, crop and tree species. AFLP marker systems have been and are being extensively exploited for genome scanning and gene mapping, as well as cDNA-AFLP for transcriptome profiling and differentially expressed gene cloning. The evaluation, annotation and classification of genomic markers and expressed transcripts would be of great utility for both functional genomics and systems biology research in plants. This may be achieved by means of the Gene Ontology (GO), consisting in three structured vocabularies (<it>i</it>.<it>e</it>. ontologies) describing genes, transcripts and proteins of any organism in terms of their associated cellular component, biological process and molecular function in a species-independent manner. In this paper, the functional annotation of about 8,000 AFLP-derived ESTs retrieved in the NCBI databases was carried out by using GO terminology.</p> <p>Results</p> <p>Descriptive statistics on the type, size and nature of gene sequences obtained by means of AFLP technology were calculated. The gene products associated with mRNA transcripts were then classified according to the three main GO vocabularies. A comparison of the functional content of cDNA-AFLP records was also performed by splitting the sequence dataset into monocots and dicots and by comparing them to all annotated ESTs of Arabidopsis and rice, respectively. On the whole, the statistical parameters adopted for the <it>in silico </it>AFLP-derived transcriptome-anchored sequence analysis proved to be critical for obtaining reliable GO results. Such an exhaustive annotation may offer a suitable platform for functional genomics, particularly useful in non-model species.</p> <p>Conclusion</p> <p>Reliable GO annotations of AFLP-derived sequences can be gathered through the optimization of the experimental steps and the statistical parameters adopted. The Blast2GO software was shown to represent a comprehensive bioinformatics solution for an annotation-based functional analysis. According to the whole set of GO annotations, the AFLP technology generates thorough information for angiosperm gene products and shares common features across angiosperm species and families. The utility of this technology for structural and functional genomics in plants can be implemented by serial annotation analyses of genome-anchored fragments and organ/tissue-specific repertories of transcriptome-derived fragments.</p

Comparative transcriptomic analysis of plum fruit treated with 1-MCP

Microarray technology has allowed the large scale transcriptomic analysis of fruit ripening. The μPEACH1.0 microarray containing 4,806 probes corresponding to genes expressed in peach fruit tissues has been used in a heterologous fashion in two studies of plums ripening behavior. Gene expression of different cultivars of plums treated with the ethylene antagonist, 1-methylcyclopropene (1-MCP) and stored for short periods at room temperature or for longer periods of cold storage was examined. In the first study, mature fruit of a suppressed ethylene climacteric cultivar 'Shiro' and a cultivar characterized by a typical increase of ethylene production during ripening ('Santa Rosa') were harvested and incubated for 24h in air (control) or 1-MCP and allowed to ripen at room temperature. Different levels of transcripts of genes implicated in cell wall metabolism, hormone (ethylene and auxin) regulation, stress and defense, and in the transcription/translation machinery, as well as others involved with ripening were identified. In the second study, the effects of 1-MCP on gene expression in relation to the development of chilling injury (CI) in the climacteric cultivars 'Ruby Red' (RR) and 'October Sun' (OS) and 'Zee Lady' peaches (ZP) were analyzed. The fruit were treated for 24h at room temperature with 1-MCP prior to storage at 0°C. For RR, there was no significant effect of 1-MCP on the level of CI symptoms, while 1-MCP significantly reduced CI symptoms in OS fruit and an increase of CI in treated ZP fruit. Microarray analysis showed that immediately following treatment, 186, 134 and 56 genes were differentially expressed between the control and 1-MCP-treated fruit of these cultivars, respectively: after 4 weeks cold storage, 311, 52 and 224 genes for RR, OS and ZP, respectively, were differentially expressed between control and treated fruit. Thus, for OS, the number of differentially expressed genes reduced during storage while the number increased in RR and ZP. Comparisons of the data suggest that the transcript profile is altered by 1-MCP more in plums than peaches. These studies, carried out within an international collaborative network, will increase our understanding of the regulation of pathways involved in plum fruit ripening and in metabolic processes related to storage and shelf lif

A microarray approach to identify genes involved in seed-pericarp cross-talk and development in peach

<p>Abstract</p> <p>Background</p> <p>Field observations and a few physiological studies have demonstrated that peach embryogenesis and fruit development are tightly coupled. In fact, attempts to stimulate parthenocarpic fruit development by means of external tools have failed. Moreover, physiological disturbances during early embryo development lead to seed abortion and fruitlet abscission. Later in embryo development, the interactions between seed and fruit development become less strict. As there is limited genetic and molecular information about seed-pericarp cross-talk and development in peach, a massive gene approach based on the use of the μPEACH 1.0 array platform and quantitative real time RT-PCR (qRT-PCR) was used to study this process.</p> <p>Results</p> <p>A comparative analysis of the transcription profiles conducted in seed and mesocarp (cv Fantasia) throughout different developmental stages (S1, S2, S3 and S4) evidenced that 455 genes are differentially expressed in seed and fruit. Among differentially expressed genes some were validated as markers in two subsequent years and in three different genotypes. Seed markers were a LTP1 (lipid transfer protein), a PR (pathogenesis-related) protein, a prunin and LEA (Late Embryogenesis Abundant) protein, for S1, S2, S3 and S4, respectively. Mesocarp markers were a RD22-like protein, a serin-carboxypeptidase, a senescence related protein and an Aux/IAA, for S1, S2, S3 and S4, respectively.</p> <p>The microarray data, analyzed by using the HORMONOMETER platform, allowed the identification of hormone-responsive genes, some of them putatively involved in seed-pericarp crosstalk. Results indicated that auxin, cytokinins, and gibberellins are good candidates, acting either directly (auxin) or indirectly as signals during early development, when the cross-talk is more active and vital for fruit set, whereas abscisic acid and ethylene may be involved later on.</p> <p>Conclusions</p> <p>In this research, genes were identified marking different phases of seed and mesocarp development. The selected genes behaved as good seed markers, while for mesocarp their reliability appeared to be dependent upon developmental and ripening traits. Regarding the cross-talk between seed and pericarp, possible candidate signals were identified among hormones.</p> <p>Further investigations relying upon the availability of whole genome platforms will allow the enrichment of a marker genes repertoire and the elucidation of players other than hormones that are involved in seed-pericarp cross-talk (i.e. hormone peptides and microRNAs).</p