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BACKGROUND: Nitrogen (N) fertilization influences plant growth and yield, and may also affect fruit quality. For two consecutive seasons, we examined the effects of various N fertilization levels – 5 to 200 mg L−1 – on pomegranate fruit, aril and juice quality. Evaluations included fruit and aril weight, size and color, appearance of peel blemishes, internal black rot and nutritional composition of extracted juices. RESULTS: Nitrogen fertilization affected pomegranate fruit, aril and juice quality. The most pronounced effects were observed in trees grown under the lowest N fertilization level, which bore smaller fruit and arils, the latter with lighter color; the fruit suffered from sunburn, and the juice had lower total soluble solid, acidity and anthocyanin contents. The proportion of edible aril weight per total fruit weight gradually increased with an increase in N fertilization concentration. In contrast, N fertilization did not affect peel color, roughness or cracking incidence. Black rot incidence increased with increasing N concentration. CONCLUSIONS: Nitrogen fertilization affected pomegranate fruit, aril and juice quality, and the total number of marketable fruits per tree. The optimal N fertilization levels, which were most beneficial for achieving high‐quality marketable fruit, were between 40 and 100 mg L−1. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Cold storage of pomegranates is essential for prolonging postharvest storage and for the implementation of cold‐quarantine insect disinfestation treatments required for international trading. However, pomegranates are chilling sensitive; they may develop chilling injuries upon exposure to unfavorable low temperatures. In this mini‐review, we summarize molecular data obtained from three different RNA Seq transcriptome analyses of responses of pomegranate fruits to cold storage. These experiments included comparisons among the transcriptomic responses following a 2‐week exposure to 1 °C in three different model systems: 1) unconditioned chilling‐sensitive fruits versus relatively chilling‐tolerant low‐temperature‐conditioned fruits; 2) chilling‐sensitive early harvested fruits versus relatively chilling‐tolerant late‐harvested ones; and 3) chilling‐sensitive 'Ganesh' variety versus the relatively chilling‐tolerant 'Wonderful' variety. Comparisons among differentially expressed transcripts that were exclusively and significantly up‐regulated in the relatively chilling‐tolerant fruits in all three model systems enabled identification of 573 common chilling tolerance‐associated genes in pomegranates. Functional categorization and classification of the differentially expressed transcripts revealed several regulatory, metabolic, and stress‐adaptation pathways that were uniquely activated in response to cold storage in relatively chilling‐tolerant fruits. More specifically, we identified common up‐regulation of transcripts involved in activation of jasmonic acid and ethylene hormone biosynthesis and signaling, stress‐related transcription factors, calcium and MAPK signaling, starch degradation and galactinol and raffinose biosynthesis, phenol biosynthesis, lipid metabolism, and heat‐shock proteins. We hypothesized these pathways to be involved in imparting chilling tolerance to pomegranate fruits. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Summary: We recently developed a postharvest low‐temperature‐conditioning (LTC) treatment for ‘Wonderful’ pomegranate fruits that involve prestorage exposure to 15 °C for 10 days, and it markedly enhanced chilling tolerance. To elucidate the molecular mechanisms involved in the fruit responses to LTC, we conducted RNA‐Seq analysis of ‘Wonderful’ pomegranate inner membrane tissue at four different time points: (i) immediately after harvest; (ii) after the LTC treatment; (iii) after 2 weeks of cold storage at 1 °C; and (iv) after LTC + 2 weeks at 1 °C. Pairwise comparisons revealed that 7798 transcripts were significantly (P ≤ 0.05) induced or repressed by a factor of at least four in the chilling treatment (2422 upregulated and 5376 downregulated) as compared with just 1997 transcripts in the LTC + chilling treatment (974 upregulated and 1023 downregulated). Thus, application of LTC mitigated the overall changes in transcript levels associated with cold storage. Furthermore, the LTC + chilling affected gene expression patterns related to regulatory and stress mechanisms, including modification of specific transcription factor, signalling and stress‐related genes. [ABSTRACT FROM AUTHOR]

BACKGROUND: Pear psylla is amajor obstacle to efficient integrated pestmanagement in pear orchards in Israel and around the world.We used two accessions with natural resistance to pear psylla Cacopsylla bidens (Šulc) – Py.760-261 (760) and Py.701-202 (701), both apparently of Pyrus communis L. origin – as interstock grafts to confer psylla resistance to the commercially important 'Spadona Estiva' (Pyrus communis) scion (Spadona) cultivar. The interaction of the interstocks with quince (Cydonia oblong Mill.) and Pyrus betulifolia Bunge rootstocks was also tested. RESULTS: Usage of Py.760-261 (760) and Py.701-202 (701) as interstocks for the psylla-sensitive Spadona resulted in a five-fold decrease in the C. bidens population, apparently as a consequence of antibiosis affecting nymph survival. Additionally, psylla survival was negatively correlated with the interstock length and amount of foliage. The yield and fruit quality of Spadona grafted on the '701' interstock equaled or even exceeded those of the control in fruit quantity, fruit size and soluble solids content, especially on P. betulifolia rootstock. CONCLUSION: Susceptibility to pear psylla decreased significantly following grafting of commercial Spadona on resistant interstock. This is the first demonstration of increased resistance to pear psylla conferred by the use of resistant interstock in pear trees and among the few examples demonstrating transfer of resistance to insects from the interstock in fruit trees. [ABSTRACT FROM AUTHOR]

BACKGROUND The pear psylla, Cacopsylla bidens (Šulc), is one of the most damaging pests of commercial pear orchards in Israel. Psylla control is a major obstacle to efficient integrated pest management, necessitating research on cultivars with natural resistance to pear psylla. Recently, two pear accessions (Py.760-261 and Py.701-202) from the local Newe Ya'ar fruit tree live collection were identified as having apparent resistance to pear psylla. Our goal was to evaluate the resistance of these two accessions relative to the commercial cultivar Spadona Estiva, and to identify whether the resistance mechanisms in the former interfere with insect colonisation of the plant (antixenosis) or inhibit insect growth, development, reproduction and survival (antibiosis). RESULTS Settlement and development of C. bidens was evaluated under natural conditions (pear orchard), semi-natural conditions (potted plants), and on detached branches and leaves (laboratory). Our results indicate that the selection Py.760-261 is 10 times more resistant than Spadona while Py.701-202 is five times more resistant. CONCLUSIONS The resistance mechanism in both accessions appears to be antibiosis affecting nymph survival. These resistant accessions may be used as rootstock or as a source of resistant genes in breeding programmes. © 2013 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

•Chilling requirement, together with heat requirement, determines the bloom date, which has an impact on the climatic distribution of the genotypes of tree species. The molecular basis of floral bud chilling requirement is poorly understood, despite its importance to the adaptation and production of fruit trees. In addition, the genetic nature of heat requirement and the genetic interrelationships among chilling requirement, heat requirement and bloom date remain unclear. •A peach ( Prunus persica) F2 population of 378 genotypes developed from two genotypes with contrasting chilling requirements was used for linkage map construction and quantitative trait loci (QTL) mapping. The floral bud chilling and heat requirements of each genotype were evaluated over 2 yr and the bloom date was scored over 4 yr. •Twenty QTLs with additive effects were identified for three traits, including one major QTL for chilling requirement and two major QTLs for bloom date. The majority of QTLs colocalized with QTLs for other trait(s). In particular, one genomic region of 2 cM, pleiotropic for the three traits, overlapped with the sequenced peach EVG region. •This first report on the QTL mapping of floral bud chilling requirement will facilitate marker-assisted breeding for low chilling requirement cultivars and the map-based cloning of genes controlling chilling requirement. The extensive colocalization of QTLs suggests that there may be one unified temperature sensing and action system regulating chilling requirement, heat requirement and bloom date together [ABSTRACT FROM AUTHOR]

Citrus fruits possess unique aromas rarely found in other fruit species. While fruit flavor is composed of complex combinations of soluble and volatile compounds, several low-abundance sesquiterpenes, such as valencene, nootkatone, alpha-sinensal, and beta-sinensal, stand out in citrus as important flavor and aroma compounds. The profile of terpenoid volatiles in various citrus species and their importance as aroma compounds have been studied in detail, but much is still lacking in our understanding of the physiological, biochemical, and genetic regulation of their production. Here, we report on the isolation, functional expression, and developmental regulation of Cstps1, a sesquiterpene synthase-encoding gene, involved in citrus aroma formation. The recombinant enzyme encoded by Cstps1 was shown to convert farnesyl diphosphate to a single sesquiterpene product identified as valencene by gas chromatography–mass spectrometry (GC–MS). Phylogenetic analysis of plant terpene synthase genes localized Cstps1 to the group of angiosperm sesquiterpene synthases. Within this group, Cstps1 belongs to a subgroup of citrus sesquiterpene synthases. Cstps1 was found to be developmentally regulated: transcript was found to accumulate only towards fruit maturation, corresponding well with the timing of valencene accumulation in fruit. Although citrus fruits are non-climacteric, valencene accumulation and Cstps1 expression were found to be responsive to ethylene, providing further evidence for the role of ethylene in the final stages of citrus fruit ripening. Isolation of the gene encoding valencene synthase provides a tool for an in-depth study of the regulation of aroma compound biosynthesis in citrus and for metabolic engineering for fruit flavor characteristics. [ABSTRACT FROM AUTHOR]

A simple new method, competitive hybridization, for identification of differentially regulated genes was used to isolate novel genes induced by ethylene in citrus (Citrus sinensis [L.] Osbeck cv. Shamouti) leaves. One of the isolated genes, an ethylene-induced esterase gene (EIE), was further characterized. The deduced protein sequence of this gene shows a similarity to those of several plant α/β hydrolase gene family members, which are known to be involved in secondary metabolism. Northern blot analysis demonstrated that EIE mRNA was induced by ethylene within 4 h and accumulated to a very high level 24 h after the initiation of ethylene treatment. Induction of EIE by ethylene could be counteracted by 1-methylcyclopropene, a potent ethylene perception inhibitor, indicating that the expression of EIE is ethylene-dependent. The bacterially expressed protein of EIE was recognized by antiserum against Pir7b, a naphthol AS esterase induced in rice by the non-host pathogen, Pseudomonas syringae pv. syringae. The EIE protein was identified in ethylene-treated leaves using anti-Pir7b antibodies. An α-naphthyl acetate esterase accumulated concomitantly with the increase in EIE protein in ethylene-treated citrus leaves. An enzyme activity assay followed by western analysis confirmed that the esterase was EIE. [ABSTRACT FROM AUTHOR]

Pre-treatment of citrus leaves and leaf explants (Citrus sinensis [L.] Osbeck cv. Shamouti), with 1-methylcyclopropene (1-MCP), induced endogenous ethylene production when leaves were further incubated in air. The induction of ethylene production was 1-MCP concentration-dependent. Abscission was concomitantly delayed. In leaves pre-treated with 1-MCP followed by exposure to ethylene, abscission was significantly delayed in comparison with those without 1-MCP pre-treatment. When leaf explants were co-treated for 24 h with ethylene and 1-MCP, abscission was delayed quite efficiently. The Lineweaver-Burke plot yielded a half-maximal value of 0.234 μl l-1 for the effect of ethylene on abscission. 1-MCP-1 competed kinetically with ethylene with a Ki value of approximately 1.4-5.5 nl l-1 1-MCP. Under these experimental conditions there was some competition between 1-MCP and ethylene. However, ethylene was not able to completely counteract the inhibitory effect of 1-MCP. Pre-treatment with 1-MCP, followed by exogenous ethylene treatment, suppressed the induction of endo-β-glucanase (EG) activity at the laminar abscission zone. The ethylene-dependent accumulation of the hydrolyse gene was demonstrated by blocking the accumulation of CsCel a1 mRNA by 1-MCP. Six hours of exposure of leaves to 1-MCP at various times during a total of 24 h ethylene treatment efficiently reversed ethylene induction of CsCel a1 gene at mRNA level up to 18 h. The results demonstrate that the induction of abscission by ethylene is controlled at mRNA level at the abscission zone. [ABSTRACT FROM AUTHOR]

Summary: We report on the isolation, functional expression and characterization of a cDNA encoding chlorophyllase, the enzyme catalyzing the first step in the chlorophyll breakdown pathway. The Chlase1 cDNA from Valencia Orange (Citrus sinensis cv. Valencia) was obtained by RT–PCR using degenerate primers based on the amino acid sequence of the previously purified protein. Chlase1 encodes a protein of 329 amino acids, including a sequence domain characterizing serine-lipases and a putative chloroplast-directing transit peptide. The Chlase1 gene encodes an active chlorophyllase enzyme which catalyzes the dephytylation of chlorophyll as shown by in vitro recombinant enzyme assays. Chlorophyllase expression at the transcript level in Valencia orange peel was found to be low and constitutive during natural fruit development without significant increase towards color-break and ripening. However, ethylene treatment induced an increase in chlorophyllase transcript at all stages of development. An enhanced response to ethylene treatment was observed during the months of October and November, corresponding to the time of natural color-break. The senescence-delaying regulator gibberellin-A[sub 3] (GA[sub 3]) inhibited the effect of ethylene on chlorophyllase transcript accumulation. The data presented suggest that chlorophyllase may not be the regulator of chlorophyll breakdown during natural fruit ripening but is consistent with the notion that chlorophyll is gradually degraded during ripening due to a negative balance between synthesis and breakdown. According to this model, exogenous application of ethylene accelerates chlorophyll breakdown due to increased de novo synthesis of chlorophyllase. Further experi- mentation on the regulation and role of chlorophyllase in planta will be facilitated by the gene tools established in this work. [ABSTRACT FROM AUTHOR]

A citrus salt-stress associated protein (Cit-SAP), partially purified from citrus cultured cells, was previously identified as the first plant phospholipid hydroperoxide glutathione peroxidase (PHGPx). The nucleotide sequence of its isolated gene (csa) revealed that a TGT, known as codon for Cys, encodes the presumed catalytic residue 41 in the polypeptide chain of Cit-SAP. In animals, a TGA encodes the rare amino acid selenocysteine (Sec) as the catalytic residue of the analogous enzyme. It is of interest to establish whether the TGT codon for this catalytic residue in the plant enzyme is indeed translated to Cys and not to Sec, and to demonstrate the effect of such a change, if it exists, on the nature of the enzymatic activity of the plant enzyme as compared to that of the animal. In the present study, we have purified for the first time, by affinity chromatography, enzymatically active citrus PHGPx from recombinant Escherichia coli bearing the csa gene. Tryptic digestion of the purified enzyme followed by HPLC afforded the isolation of a peptide which contains residue 41, and its sequence analysis revealed that this residue is indeed a Cys, and not Sec. The enzymatic activity and specificity of the recombinant Cit-SAP was found to be similar to that observed before for the partially purified plant enzyme. However, the rate of this activity was much lower towards phospholipid hydroperoxides, and none towards hydrogen peroxide, as compared to that of the animal analogue. It is therefore suggested that the presence of a Cys, and not Sec, as the catalytic residue in the plant enzyme, affects its enzymatic activity and may determine a different biological role for the plant PHGPx from that of the animal. [ABSTRACT FROM AUTHOR]

Oxidative stress in plants causes the induction of several enzymes, including superoxide dismutase (EC 1.15.1.1), ascorbate peroxidase (EC 1.11.1.11) and glutathione reductase (EC 1.6.4.2). The first two are responsible for converting superoxide to H2O2 and its subsequent reduction to H2O, and the third is involved in recycling of ascorbate. Glutathione peroxidases (GPXs, EC 1.11.1.9) are a family of key enzymes involved in scavenging oxyradicals in animals. Only recently, indications for the existence of this enzyme in plants were reported. Genes with significant sequence homology to one member of the animal GPX family, namely phospholipid hydroperoxide glutathione peroxidase (PHGPX), were isolated from several plants. Cit-SAP, the protein product encoded by the citrus csa gene, which is induced by salt-stress, is so far the only plant PHGPX that has been isolated and characterized. This protein differs from the animal PHGPX in its rate of enzymatic activity and in containing a Cys instead of selenocysteine (Sec) as its presumed catalytic residue. The physiological role of Cit-SAP and its homologs in other plants is not yet known. [ABSTRACT FROM AUTHOR]

The cyanobacterium Anabaena responds to nitrogen deprivation by producing heterocysts, cells specialized for nitrogen fixation, at well-spaced intervals along its filaments. The gene hepA, required for heterocyst maturation, is expressed in response to nitrogen deprivation, prior to visible differentiation. A spatial pattern of hepA expression indistinguishable from the eventual pattern of heterocysts was made visible by fusing the hepA promoter to luxAB, which encodes bacterial luciferase. Because the resulting signal did not greatly exceed instrumental background, T7 RNA polymerase was used to increase luminescence. The hepA promoter was fused to the gene for that polymerase, and a promoter recognized by that polymerase was fused to luxAB. Filaments containing these two fusions showed spaced luminescing cells many hours before differentiation became discernible morphologically. [ABSTRACT FROM AUTHOR]

Summary: Plastid chaperonin 60 (cpn60) is a chloroplast protein, presumed to assist in assembly and folding of plastid proteins. Although molecular chaperones often accumulate significantly in response to stress, this has never been demonstrated for cpn60. In this study, the accumulation of cpn60 in Nicotiana seedlings during their development was followed under different stress conditions. It was found that cpn60 accumulates markedly in developing seedlings in response to tentoxin and several other (but not all) stresses. Cpn60 accumulates only during a narrow period of seedling development. It is proposed that cpn60 accumulation under stress is developmentally regulated. [ABSTRACT FROM AUTHOR]