ACC Synthase Genes Related to Cold-dependent Ripening in Pear Fruit

The differential regulation of ACC synthase genes has been studied in pear cultivars that either require a long chilling treatment before they are capable of ripening (‘Passe-Crassane’, PC) or not (‘Old-Home’, OH) and in OH x PC hybrids having no (A16) or intermediate (A50) cold requirement. Among the seven Pc-ACS cDNAs isolated, four of them (Pc-ACS1a/b and Pc-ACS2a/b) showed differential expression in relation with cold requirement. Pc-ACS1a transcripts accumulated specifically during chilling and ripening of cold-dependent cultivars while Pc-ACS1b transcripts were detected only during ripening of cold-independent genotypes. Pc- ACS2a mRNA was expressed specifically in cold-dependent genotypes and negatively regulated by ethylene while Pc-ACS2b transcripts accumulated only in cold-independent genotypes and positively regulated by ethylene. Pc-ACS3, 4 and 5 transcripts accumulation was similar in all genotypes, independently of coldrequirements

Transcriptome analysis of an apple (Malus × domestica) yellow fruit somatic mutation identifies a gene network module highly associated with anthocyanin and epigenetic regulation

An apple loss-of-colour fruit somatic mutation is likely the consequence of collective repression of a co-expression gene network module highly associated with anthocyanin and methylation in the promoter of MdMYB1

Molecular Control of Fruit Ripening and Sensory Quality of Charentais Melon

Traditional Charentais melons have a typical climacteric behavior with ethylene playing a major role in the regulation of the ripening process. Genetic studies using climacteric and non-climacteric types of Cucumis melo demonstrated that the climacteric character is dominant and conferred by 2 duplicated loci only which are of great importance for the regulation of storability and sensory quality. Commercial varieties of Charentais melon with long shelf-life have been generated, some of them by crossing with a non-ripening Charentais genotype (Vauclusien). The introduction of the long shelf-life character resulted in undesirable loss of aroma volatiles production. The inhibition of ethylene synthesis by knocking-down ACC oxidase gene expression has been achieved in Charentais melon. It results is a strong inhibition of the synthesis of aroma volatiles while the accumulation of sugars is not affected or is even improved and the softening of the flesh is strongly affected but not abolished. It was also demonstrated that ethylene-inhibited fruit exhibited better resistance to chilling injury. Due to the importance of aroma volatiles in sensory quality and to the strong negative correlation between aroma production and ethylene synthesis, we have developed a research program aimed at isolating genes involved in the synthesis of volatile esters, compounds that are essential for the flavor of Cantaloupe melons. We report here on the recent advances in the field with special emphasis on the characterization of two families of genes encoding aldehyde reductases and alcohol acyl transferases

Characterization of Genes Involved in the Formation of Aroma Volatiles in "Charentais" Melon Fruit

Volatiles esters impart distinct characteristics to the fruit quality. "Charentais" cantaloupe melon (Cucumis melo "cantalupensis") is characterized by abundant sweetness and aromatic flavour. Plant alcohol acyl transferase (AAT) genes have been identified and shown to be involved in aromas production. Recently,two cDNAs (Cm-AAT1 and Cm-AAT2) putatively involved in the formation of aroma volatile esters have been isolated from melon fruit. Cm-AAT1 protein exhibit alcohol acyl transferase activity while no such activity could be detected for Cm-AAT2. Two new cDNAs (Cm-AAT3 and Cm-AAT4) have been isolated from melon fruit that showed 69% and 36% similarity, respectively, with Cm-AAT1. The percentage similarity over the whole amino acid sequence between them is 34%. Cm-AAT3 and Cm-AAT4 show the highest similarity to the tobacco Nt-HSR201 protein and a rose alcohol acyltransferase Rh-AAT1, respectively. All Cm-AATs genes, share three conserved regions common to the BAHD acyltransferase gene superfamily. Heterologous expression in yeast revealed that some of the encoded proteins have a wide range of specificity while others are specific to a narrow range of substrates

Prunus domestica Pathogenesis-Related Protein-5 Activates the Defense Response Pathway and Enhances the Resistance to Fungal Infection

Pathogenesis-related protein-5 (PR-5) has been implicated in plant disease resistance and its antifungal activity has been demonstrated in some fruit species. However, their roles, especially their interactions with the other defense responses in plant cells, are still not fully understood. In this study, we have cloned and characterized a new PR-5 cDNA named PdPR5-1 from the European plum (Prunus domestica). Expression of PdPR5-1 was studied in different cultivars varying in resistance to the brown rot disease caused by the necrotrophic fungus Monilinia fructicola. In addition transgenic Arabidopsis, ectopically expressing PdPR5-1 was used to study its role in other plant defense responses after fungal infection. We show that the resistant cultivars exhibited much higher levels of transcripts than the susceptible cultivars during fruit ripening. However, significant rise in the transcript levels after infection with M. fructicola was observed in the susceptible cultivars too. Transgenic Arabidopsis plants exhibited more resistance to Alternaria brassicicola. Further, there was a significant increase in the transcripts of genes involved in the phenylpropanoid biosynthesis pathway such as phenylalanine ammonia-lyase (PAL) and phytoalexin (camalexin) pathway leading to an increase in camalexin content after fungal infection. Our results show that PdPR5-1 gene, in addition to its anti-fungal properties, has a possible role in activating other defense pathways, including phytoalexin production

Recent Developments on the Role of Ethylene in the Ripening of Climacteric Fruit

It has long been recognised that ethylene plays a major role in the ripening process of climacteric fruit. A more thorough analysis, however, has revealed that a number of biochemical and molecular processes associated with climacteric fruit ripening are ethylene-independent. One of the crucial steps of the onset of ripening is the induction of autocatalytic ethylene production. In ethylene-suppressed melons, ACC synthase activity is induced at the same time as in control melons, indicating that ACC biosynthesis during the early stages of ripening seems to be a developmentally-regulated (ethylene-independent) process. The various ripening events exhibit differential sensitivity to ethylene. For instance, the threshold level for degreening of the rind is 1ppm, while 2.5 ppm are required to trigger some components of the softening process. The saturating level of ethylene producing maximum effects is less than 5 ppm, which is by far lower than the internal ethylene concentrations found in the fruit at the climacteric peak (over 100 ppm). In many fruit chilling temperatures hasten ethylene production and ripening and in some late season pear varieties, exposure to chilling temperatures is even absolutely required for the attainment of the capacity to synthesize autocatalytic ethylene. This is correlated with the stimulation of expression of ACC oxidase and of members of the ACC synthase gene family. Ethylene operates via a perception and transduction pathway to induce the expression of genes responsible for the biochemical and physiological changes observed during ripening. However, only a few genes induced via the ethylene transduction pathway have been described so far. We have used a differential display method to isolate novel ethylene-reponsive (ER) cDNA clones of tomato that potentially play a role in propagating the ethylene response and in regulating fruit ripening. Collectively, these data permit a general scheme of the molecular mechanisms of fruit ripening to be proposed

Identification and characterization of genes involved in the fruit color development of european plum

European plum fruit (Prunus domestica) are normally blue-black to dark purple. However, some genotypes remain green/yellow after ripening. We hypothesized that in such genotypes anthocyanin biosynthesis is genetically disturbed. To examine this hypothesis, six european plum genotypes with diverse fruit colors were investigated for the expression pattern of several anthocyanin biosynthetic genes (ABGs)—e.g., phenylalanine ammonia-lyase, chalcone synthase (CHS), dihydroflavonol 4-reductase (DFR), anthocyanin synthase (ANS), and UDP-glucose:flavonoid 3-O-glucosyltransferase 1 and 2 (UFGT 1 and 2). Expression profiles indicated that ABGs, especially Pd-CHS and UFGT 2, were significantly downregulated in the green/yellow fruit compared with the dark-purple fruit. Furthermore, the quantification of total polyphenols and individual flavonoid compounds showed substantial differences between the off-colored and the purple genotype. To further examine the contribution of each of the ABGs in color development, the open reading frame (ORP) of Pd-CHS, Pd-DFR, Pd-ANS, and Pd-UFGT 2 was ectopically expressed in tobacco (Nicotiana tabacum). The characterization of transgenic plants showed that the petals of plants expressing Pd-CHS were darker in color and had higher anthocyanin content than control or even other transgenic types, suggesting the significant contribution of CHS in determining anthocyanin production levels and hence fruit coloration. The results of this study provides better understanding of color development in european plum, which can be rewarding in developing european plum cultivars with desired colors through classical or modern breeding tools

Differential regulation of ACC synthase genes in cold-dependent and -independent ripening in pear fruit

Late pear cultivars such as Passe-Crassane (PC) require a long chilling treatment before they are capable of ripening. Early cultivars such as Old-Home (OH) have no cold prerequisite. The regulation of 1-aminocyclopropane-1-carboxylic acid synthase (ACS) genes was studied in OH, PC and in OH x PC hybrids in order to determine the role of this gene family in the cold requirement. Of the seven Pc-ACS cDNAs isolated, four (Pc-ACSla/b and Pc-ACS2a/b) showed differential expression associated with the cold requirement. Pc-ACS1a transcripts accumulated throughout the cold treatment and, with Pc-ACS2a, during ripening of cold-dependent cultivars. Pc-ACS1b and Pc-ACS2b were detected only during ripening of cold-independent genotypes. Furthermore, Pc-ACS2a transcript accumulation was negatively regulated by ethylene, whereas Pc-ACS2b was positively regulated by the hormone. Pc-ACS3, 4 and 5 transcript accumulation was similar in all genotypes. Genetic analyses of OH, PC, and 22 OH x PC progenies demonstrated that late, cold-dependent cultivars were homozygous for Pc-ACS1a and 2a whereas early, cold-independent cultivars were heterozygous for Pc-ACS1(a/b) and homozygous for Pc-ACS2b. A model is presented in which differences in Pc-ACS alleles and gene expression between cold- and non-cold-requiring pears are critical in determining the ripening behaviour of the cultivars

Two highly divergent alcohol dehydrogenases of melon exhibit fruit ripening-specific expression and distinct biochemical characteristics

Alcohol dehydrogenases (ADH) participate in the biosynthetic pathway of aroma volatiles in fruit by interconverting aldehydes to alcohols and providing substrates for the formation of esters. Two highly divergent ADH genes (15% identity at the amino acid level) of Cantaloupe Charentais melon (Cucumis melo var. Cantalupensis) have been isolated. Cm-ADH1 belongs to the medium-chain zinc-binding type of ADHs and is highly similar to all ADH genes expressed in fruit isolated so far. Cm-ADH2 belongs to the short-chain type of ADHs. The two encoded proteins are enzymatically active upon expression in yeast. Cm-ADH1 has strong preference for NAPDH as a co-factor, whereas Cm-ADH2 preferentially uses NADH. Both Cm-ADH proteins are much more active as reductases with Kms 10–20 times lower for the conversion of aldehydes to alcohols than for the dehydrogenation of alcohols to aldehydes. They both show strong preference for aliphatic aldehydes but Cm-ADH1 is capable of reducing branched aldehydes such as 3-methylbutyraldehyde, whereas Cm-ADH2 cannot. Both Cm-ADH genes are expressed specifically in fruit and up-regulated during ripening. Gene expression as well as total ADH activity are strongly inhibited in antisense ACC oxidase melons and in melon fruit treated with the ethylene antagonist 1-methylcyclopropene (1-MCP), indicating a positive regulation by ethylene. These data suggest that each of the Cm-ADH protein plays a specific role in the regulation of aroma biosynthesis in melon fruit