Mature Citrus Fruit Research Articles

Effects of light and plant hormones on regreening in flavedos of Ponkan mandarin in vitro

In this study, the regulation of regreening in the flavedos of citrus fruit was investigated in vitro. The results showed blue light was much more effective than red light for inducing regreening in citrus fruit. In the blue light treatment, the up-regulation of chlorophyll biosynthetic genes and the down-regulation of chlorophyll degradation genes led to the re-accumulation of chlorophyll in the flavedos of mature fruit. In addition, we found that blue light was a necessary factor for GA-induced regreening in citrus fruit. Under blue light, GA treatment accelerated the re-accumulation of chlorophyll in the flavedos. In contrast to GA, ABA treatment did not induce regreening under blue light, and the flavedos kept in orange color. Compared with the blue light treatment alone, the contents of chlorophyll a, chlorophyll b, and total chlorophyll were decreased by the combination of ABA and blue light treatments. The results presented in this study suggested that blue light and GA are positive regulators for inducing regreening, while ABA is a negative regulator of regreening in the flavedo of mature citrus fruit.

Transcriptomic and physiological comparison of Shatangju (Citrus reticulata) and its late-maturing mutant provides insights into auxin regulation of citrus fruit maturation.

Previous studies have shown that abscisic acid (ABA) and ethylene are involved in pulp maturation and peel coloration in the nonclimacteric citrus fruits. There are also signs indicating that other plant hormones may play some roles in citrus fruit ripening. In this study, we compared profiles of genome-wide gene expression and changes in hormones and peel pigments between fruits of Shatangju mandarin (Citrus reticulata Blanco, designated WT) and its natural mutant, Yuenongwanju (designated MT). The MT fruit matures ~2 months later than the WT fruit. Significant differences in fruit diameter, total soluble solids, titratable acid content, chlorophylls and carotenoids were detected between the fruits of the two genotypes at the sampled time points. Genome-wide transcriptome profiling showed that many genes involved in auxin and ABA metabolism and/or signaling pathways were differentially expressed between the MT and the WT fruits. Importantly, the expression of CrYUCCA8 was significantly lower and the expression of CrNCED5 was significantly higher in WT than in MT fruits at 230 and 250 DPA, respectively. In addition, the indole-3-acetic acid (IAA) level in the MT fruit was significantly higher than that in the WT counterpart, whereas a significantly lower level of ABA was detected in the mutant. Treatment of the WT fruit with exogenous IAA significantly delayed fruit maturation. Our results provide experimental evidence supporting the notion that auxin is a negative regulator of fruit maturation in citrus.

The Surface Defects Detection of Citrus on Trees Based on a Support Vector Machine

Machine learning and image processing have been combined to identify and detect defects in mature citrus fruit at night, which has great research and development significance. First, a multi-light vision system was used to collect citrus UV images, and from these, 1500 samples were obtained, 80% of which were training and 20% were experimental sets. For a support vector machine (SVM) model with “2*Cb-Cr”, “4*a-b-l”, and “H” as the training features, the accuracy of the final training model in the experimental set is 99.67%. Then, the SVM model was used to identify mature citrus regions, detect defects, and output the defective citrus regions label. The average running time of the detection algorithm was 0.84097 s, the accuracy of citrus region detection was 95.32%, the accuracy of citrus defect detection was 96.32%, the precision was 95.24%, and the recall rate was 87.91%. The results show that the algorithm had suitable accuracy and real-time performance in recognition and defect detection in citrus in a natural environment at night.

Essential Oils in Citrus Fruit Ripening and Postharvest Quality

Citrus essential oils (EOs) are widely used as flavoring agents in food, pharmaceutical, cosmetical and chemical industries. For this reason, their demand is constantly increasing all over the world. Besides industrial applications, the abundance of EOs in the epicarp is particularly relevant for the quality of citrus fruit. In fact, these compounds represent a natural protection against postharvest deteriorations due to their remarkable antimicrobial, insecticidal and antioxidant activities. Several factors, including genotype, climatic conditions and cultural practices, can influence the assortment and accumulation of EOs in citrus peels. This review is focused on factors influencing variation of the EOs’ composition during ripening and on the implications on postharvest quality of the fruit.

Differential Transcriptomic Regulation in Sweet Orange Fruit (Citrus sinensis L. Osbeck) Following Dehydration and Rehydration Conditions Leading to Peel Damage.

Water stress is the most important environmental agent that contributes to the crop productivity and quality losses globally. In citrus, water stress is the main driver of the fruit peel disorders that impact the quality and market ability. An increasingly present post-harvest peel disorder is non-chilling peel pitting (NCPP). Non-chilling peel pitting is manifested as collapsed areas of flavedo randomly scattered on the fruit and its incidence increases due to abrupt increases in the environmental relative humidity (RH) during post-harvest fruit manipulation. In this study, we have used a custom-made cDNA microarray containing 44k unigenes from Citrus sinensis (L. Osbeck), covering for the first time the whole genome from this species, to study transcriptomic responses of mature citrus fruit to water stress. In the study, the global gene expression profiles of flavedo from Navelate oranges subjected to severe water stress are compared with those fruits subjected to rehydration stress provoked by changes in the RH during post-harvest, which enhances the development of NCPP. The study results show that NCPP is a complex physiological process that shares molecular responses with those from prolonged dehydration in fruit, but the damage associated with NCPP may be explained by unique features of rehydration stress at the molecular level, such as membrane disorganization, cell wall modification, and proteolysis.

On how auxin, ethylene and IDA-peptide relate during mature Citrus fruit abscission

While the ethylene-auxin interactions are well documented in model dicots such as tomato (climacteric fruit) during mature fruit abscission, the process is not clearly understood in citrus (non-climacteric fruit). The mature fruit produces very little ethylene but is sensitive to ethylene treatments to induce abscission. By contrast, auxin treatments delay fruit abscission, but the particular role of auxin in the process is unknown. Since the IDA-HAE/HSL2 ethylene-independent pathway seem to regulate organ abscission in both model and crop species, we proposed that auxin treatment delays citrus fruit abscission by reactivating the basipetal auxin flux and reducing CitIDA3 expression, without modifying ethylene synthesis.Comparing orange (C. sinensis) genotypes which differ in their abscission rate, ‘Navelate’ vs. ‘Valencia Late’, we found that the force needed to detach the fruit from the tree (FDF) declines in parallel with 1) an increase of ethylene synthesis and CitIDA3 gene expression, and 2) a reduction of PIN1-like (auxin transporter) gene expresion. Further, auxin (2,4-D) treatment maintains a higher force in the abscission zone upregulating PIN1-like and AUX1-like (auxin transporter) gene expression, and downregulating CitIDA3 gene expression, but without modifying ethylene production. We conclude that the 2,4-D treatment delays citrus mature fruit abscission through an ethylene-independent pathway.

Comparative transcriptome analyses of a late-maturing mandarin mutant and its original cultivar reveals gene expression profiling associated with citrus fruit maturation.

Characteristics of late maturity in fruit are good agronomic traits for extending the harvest period and marketing time. However, underlying molecular basis of the late-maturing mechanism in fruit is largely unknown. In this study, RNA sequencing (RNA-Seq) technology was used to identify differentially expressed genes (DEGs) related to late-maturing characteristics from a late-maturing mutant ‘Huawan Wuzishatangju’ (HWWZSTJ) (Citrus reticulata Blanco) and its original line ‘Wuzishatangju’ (WZSTJ). A total of approximately 17.0 Gb and 84.2 M paried-end reads were obtained. DEGs were significantly enriched in the pathway of photosynthesis, phenylpropanoid biosynthesis, carotenoid biosynthesis, chlorophyll and abscisic acid (ABA) metabolism. Thirteen candidate transcripts related to chlorophyll metabolism, carotenoid biosynthesis and ABA metabolism were analyzed using real-time quantitative PCR (qPCR) at all fruit maturing stages of HWWZSTJ and WZSTJ. Chlorophyllase (CLH) and divinyl reductase (DVR) from chlorophyll metabolism, phytoene synthase (PSY) and capsanthin/capsorubin synthase (CCS) from carotenoid biosynthesis, and abscisic acid 8′-hydroxylase (AB1) and 9-cis-epoxycarotenoid dioxygenase (NCED1) from ABA metabolism were cloned and analyzed. The expression pattern of NCED1 indicated its role in the late-maturing characteristics of HWWZSTJ. There were 270 consecutive bases missing in HWWZSTJ in comparison with full-length sequences of NCED1 cDNA from WZSTJ. Those results suggested that NCED1 might play an important role in the late maturity of HWWZSTJ. This study provides new information on complex process that results in the late maturity of Citrus fruit at the transcriptional level.

Comparative Analysis of miRNAs and Their Target Transcripts between a Spontaneous Late-Ripening Sweet Orange Mutant and Its Wild-Type Using Small RNA and Degradome Sequencing.

Fruit ripening in citrus is not well-understood at the molecular level. Knowledge of the regulatory mechanism of citrus fruit ripening at the post-transcriptional level in particular is lacking. Here, we comparatively analyzed the miRNAs and their target genes in a spontaneous late-ripening mutant, “Fengwan” sweet orange (MT) (Citrus sinensis L. Osbeck), and its wild-type counterpart (“Fengjie 72-1,” WT). Using high-throughput sequencing of small RNAs and RNA degradome tags, we identified 107 known and 21 novel miRNAs, as well as 225 target genes. A total of 24 miRNAs (16 known miRNAs and 8 novel miRNAs) were shown to be differentially expressed between MT and WT. The expression pattern of several key miRNAs and their target genes during citrus fruit development and ripening stages was examined. Csi-miR156k, csi-miR159, and csi-miR166d suppressed specific transcription factors (GAMYBs, SPLs, and ATHBs) that are supposed to be important regulators involved in citrus fruit development and ripening. In the present study, miRNA-mediated silencing of target genes was found under complicated and sensitive regulation in citrus fruit. The identification of miRNAs and their target genes provide new clues for future investigation of mechanisms that regulate citrus fruit ripening.

Effects of exogenous abscisic acid on the expression of citrus fruit ripening-related genes and fruit ripening

Despite exogenous abscisic acid (ABA) treatment is a widely-used effective method to prove its role in plants, this method has been limitedly explored in citrus fruits. Here, we examined the effects of exogenous ABA and nordihydroguaiaretic acid (NDGA) on the color index H value and the contents of soluble sugars, organic acids and endogenous hormones in the fruits of Citrus reticulata Blanco cv. Ponkan. Additionally, the expression profiles of genes involved in the ABA, sugar and organic acid metabolism and signal transduction pathway in fruits were also investigated. The results indicated that exogenous ABA could accelerate fruit coloring, significantly decrease the organic acid content, and affect the ripening of citrus fruit, while treatment with NDGA could restrain fruit coloring and acid degradation. Exogenous ABA treatment produced minor effect on the accumulation of sugars but largely regulated the expression of most sugar- and acid-related genes, indicating a coordinate interaction between the ABA signaling pathway and sucrose signaling pathway to regulate citrus fruit ripening. Especially, CsSUC 3 plays a vital role in accumulation of sugars during the last ripening stage of citrus fruit, which could be induced by ABA treatment. The transcript levels of CsACO1 and CsNADP-IDH in ABA-treated fruits were significantly higher than in the control fruits during the late ripening stages, which was conducive to the degradation of organic acids. Moreover, ABA could restrict its own accumulation at 14 DAY not only by inducing the expression of ABA 8′-hydroxylase 1 but also by reducing the expression of CsNCED1, and the transcriptional levels of other ABA-related genes were largely triggered by exogenous ABA. Overall, ABA is a positive regulator of ripening and could be used to regulate citrus fruit ripening effectively.

Overexpression of a citrus basic helix-loop-helix transcription factor (CubHLH1), which is homologous to Arabidopsis activation-tagged bri1 suppressor 1 interacting factor genes, modulates carotenoid metabolism in transgenic tomato

To explore the transcription factors associated with carotenoid metabolism in citrus fruit, one transcription factor (CubHLH1) was selected through microarray screening in Satsuma mandarin (Citrus unshiu Marc.) fruit, which was treated with exogenous ethylene or gibberellin (GA), accelerating or retarding carotenoid accumulation in peel, respectively. The amino acid sequence of CubHLH1 has homology to Arabidopsis activation-tagged bri1 suppressor 1 (ATBS1) interacting factor (AIF), which is functionally characterized as a negative regulator of the brassinolide (BR) signalling pathway. Yeast two-hybrid analysis revealed that protein for CubHLH1 could interact with Arabidopsis and tomato ATBS1. Overexpression of CubHLH1 caused a dwarf phenotype in transgenic tomato (Solanum lycopersicum L.), suggesting that CubHLH1 has a similar function to Arabidopsis AIF. In the transgenic tomato fruit at ripening stage, the lycopene content was reduced along with the changes in carotenoid biosynthetic gene expression. The abscisic acid (ABA) content of all the transgenic tomato fruit was higher than that of the wild type. These results implied that CubHLH1 is considered to have a similar function to Arabidopsis AIFs and might be directly involved in carotenoid metabolism in mature citrus fruit.

Profiling gene expression in citrus fruit calyx abscission zone (AZ-C) treated with ethylene.

On-tree storage and harvesting of mature fruit account for a large proportion of cost in the production of citrus, and a reduction of the cost would not be achieved without a thorough understanding of the mechani sm of the mature fruit abscission. Genome-wide gene expression changes in ethylene-treated fruit calyx abscission zone (AZ-C) of Citrus sinensis cv. Olinda were therefore investigated using a citrus genome array representing up to 33,879 citrus transcripts. In total, 1313 and 1044 differentially regulated genes were identified in AZ-C treated with ethylene for 4 and 24h, respectively. The results showed that mature citrus fruit abscission commenced with the activation of ethylene signal transduction pathway that led to the activation of ethylene responsive transcription factors and the subsequent transcriptional regulation of a large set of ethylene responsive genes. Significantly down-regulated genes included those of starch/sugar biosynthesis, transportation of water and growth promoting hormone synthesis and signaling, whereas significantly up-regulated genes were those involved in defense, cell wall degradation, and secondary metabolism. Our data unraveled the underlying mechanisms of some known important biochemical events occurring at AZ-C and should provide informative suggestions for future manipulation of the events to achieve a controllable abscission for mature citrus fruit.

Oxidative metabolism in ‘Valencia’ sweet orange [Citrus sinensis (L.) Osbeck] flavedo tissue treated with the abscission agent 5-chloro-3- methyl-4-nitro-1H-pyrazole (CMNP)

Summary5-Chloro-3-methyl-4-nitro-1H-pyrazole (CMNP) is a pyrazole derivative that promotes the abscission of mature citrus fruit and is being developed to facilitate mechanical commercial harvesting of mature fruit. Understanding the mode of action of CMNP in promoting abscission will assist in decisions on its commercial use and in the development of alternative, but superior compounds. Fully-mature ‘Valencia’ sweet orange trees were sprayed with 2.0 mM CMNP. Fruit detachment force (FDF) values, hydrogen peroxide (H2O2) concentrations, and malondialdehyde (MDA) concentrations, as well as superoxide dismutase (SOD), ascorbate peroxidase (APOD), glutathione reductase (GR), and peroxidase (POD) activities were measured 0, 1, 2, and 3 d after CMNP treatment (DACT). FDF values declined linearly from 65 N to 28 N after CMNP treatment. H2O2 and MDA concentrations were higher in CMNP-treated flavedo tissue (FT) compared to the untreated controls. SOD activity was higher in the FT of CMNP-treated fruit than in untreated controls and was largely responsible for the accumulation of H2O2. The activities of enzymes in the APOD/GR cycle (i.e., ascorbate peroxidase and glutathione reductase) declined significantly throughout the abscission process. CMNP-induced higher SOD and lower APOD and GR activities, which maintained elevated levels of H2O2. H2O2 may serve as a signal, in association with previously reported lipid signals, in the CMNP-mediated abscission of citrus fruit. POD activities increased up to 1 DACT, then declined to control values. POD may be involved in reducing free indole-acetic acid (IAA) concentrations in the FT, thus enhancing the sensitivity of the FT to ethylene, which is known to promote abscission in citrus fruit.

Comparative transcriptome analyses between a spontaneous late-ripening sweet orange mutant and its wild type suggest the functions of ABA, sucrose and JA during citrus fruit ripening.

A spontaneous late-ripening mutant of ‘Jincheng’ (C. sinensis L. Osbeck) sweet orange exhibited a delay of fruit pigmentation and harvesting. In this work, we studied the processes of orange fruit ripening through the comparative analysis between the Jincheng mutant and its wild type. This study revealed that the fruit quality began to differ on 166th days after anthesis. At this stage, fruits were subjected to transcriptome analysis by RNA sequencing. 13,412 differentially expressed unigenes (DEGs) were found. Of these unigenes, 75.8% were down-regulated in the wild type, suggesting that the transcription level of wild type was lower than that of the mutant during this stage. These DEGs were mainly clustered into five pathways: metabolic pathways, plant-pathogen interaction, spliceosome, biosynthesis of plant hormones and biosynthesis of phenylpropanoids. Therefore, the expression profiles of the genes that are involved in abscisic acid, sucrose, and jasmonic acid metabolism and signal transduction pathways were analyzed during the six fruit ripening stages. The results revealed the regulation mechanism of sweet orange fruit ripening metabolism in the following four aspects: First, the more mature orange fruits were, the lower the transcription levels were. Second, the expression level of PME boosted with the maturity of the citrus fruit. Therefore, the expression level of PME might represent the degree of the orange fruit ripeness. Third, the interaction of PP2C, PYR/PYL, and SnRK2 was peculiar to the orange fruit ripening process. Fourth, abscisic acid, sucrose, and jasmonic acid all took part in orange fruit ripening process and might interact with each other. These findings provide an insight into the intricate process of sweet orange fruit ripening.

Identification of fruit and branch in natural scenes for citrus harvesting robot using machine vision and support vector machine

Abstract: With the decrease of agricultural labor and the increase of production cost, the researches on citrus harvesting robot (CHR) have received more and more attention in recent years. For the success of robotic harvesting and the safety of robot, the identification of mature citrus fruit and obstacle is the priority of robotic harvesting. In this work, a machine vision system, which consisted of a color CCD camera and a computer, was developed to achieve these tasks. Images of citrus trees were captured under sunny and cloudy conditions. Due to varying degrees of lightness and position randomness of fruits and branches, red, green, and blue values of objects in these images are changed dramatically. The traditional threshold segmentation is not efficient to solve these problems. Multi-class support vector machine (SVM), which succeeds by morphological operation, was used to simultaneously segment the fruits and branches in this study. The recognition rate of citrus fruit was 92.4%, and the branch of which diameter was more than 5 pixels, could be recognized. The results showed that the algorithm could be used to detect the fruits and branches for CHR. Keywords: citrus, machine vision, citrus harvesting robot (CHR), branch, identification, multi-class support vector machine (SVM) DOI: 10.3965/j.ijabe.20140702.014 Citation: Lu Q, Cai J R, Liu B, Deng L, Zhang Y J. Identification of fruit and branch in natural scenes for citrus harvesting robot using machine vision and support vector machine. Int J Agric & Biol Eng, 2014; 7(2): 115－121.

Mechanism of Carotenoid Accumulation in Citrus Fruit

Citrus is a complex source of carotenoids with the largest number of carotenoids found in any fruit. Carotenoid concentration and composition vary greatly among citrus varieties. Satsuma mandarin (Citrus unshiu Marc.) predominantly accumulates β-cryptoxanthin in the juice sacs. Valencia orange (Citrus sinensis Osbeck) predominantly accumulates violaxanthin isomers in the juice sacs. Lisbon lemon (Citrus limon Burm.f.) accumulates low level of carotenoids in the juice sacs. To elucidate the carotenoid accumulation in citrus fruit maturation, the expression of genes related to carotenoid biosynthesis and catabolism was investigated in the three citrus varieties exhibited different carotenoid profile. The results showed that the carotenoid accumulation during citrus fruit maturation is highly regulated by the coordination of the expression for the genes related to carotenoid biosynthesis and catabolism in both flavedo and juice sacs. ‘Tamami’ is a hybrid between ‘Kiyomi’ tangor (Citrus unshiu Marc. × Citrus sinensis Osbeck) and ‘Wilking’ mandarin (Citrus nobilis Lour. × Citrus deliciosa Ten.). To elucidate the mechanism of the accumulation of β-cryptoxanthin in ‘Tamami’, a variety accumulating higher β-cryptoxanthin than Satsuma mandarin, the expression of genes related to carotenoid biosynthesis and catabolism was investigated in the juice sacs of ‘Tamami’. The results showed that the mechanism of β-cryptoxanthin accumulation in ‘Tamami’ was similar to that in Satsuma mandarin. Furthermore, in the recent studies, possible factors, which regulate carotenoid concentration and composition in citrus juice sacs were investigated in vitro.

Label-free shotgun proteomics and metabolite analysis reveal a significant metabolic shift during citrus fruit development

Label-free LC-MS/MS-based shot-gun proteomics was used to quantify the differential protein synthesis and metabolite profiling in order to assess metabolic changes during the development of citrus fruits. Our results suggested the occurrence of a metabolic change during citrus fruit maturation, where the organic acid and amino acid accumulation seen during the early stages of development shifted into sugar synthesis during the later stage of citrus fruit development. The expression of invertases remained unchanged, while an invertase inhibitor was up-regulated towards maturation. The increased expression of sucrose-phosphate synthase and sucrose-6-phosphate phosphatase and the rapid sugar accumulation suggest that sucrose is also being synthesized in citrus juice sac cells during the later stage of fruit development.

カンキツ果実におけるカロテノイド集積メカニズム

Citrus is a complex source of carotenoids with the largest number of carotenoids found in any fruit. Carotenoid concentration and composition vary greatly among citrus varieties. Satsuma mandarin (Citrus unshiu Marc.) predominantly accumulates β-cryptoxanthin in the juice sacs. Valencia orange (Citrus sinensis Osbeck) predominantly accumulates violaxanthin isomers in the juice sacs. Lisbon lemon (Citrus limon Burm.f.) accumulates low level of carotenoids in the juice sacs. To elucidate the carotenoid accumulation in citrus fruit maturation, the expression of genes related to carotenoid biosynthesis and catabolism was investigated in the three citrus varieties exhibited different carotenoid profile. The results showed that the carotenoid accumulation during citrus fruit maturation is highly regulated by the coordination of the expression for the genes related to carotenoid biosynthesis and catabolism in both flavedo and juice sacs. ‘Tamami’ is a hybrid between ‘Kiyomi’ tangor (Citrus unshiu Marc. × Citrus sinensis Osbeck) and ‘Wilking’ mandarin (Citrus nobilis Lour. × Citrus deliciosa Ten.). To elucidate the mechanism of the accumulation of β-cryptoxanthin in ‘Tamami’, a variety accumulating higher β-cryptoxanthin than Satsuma mandarin, the expression of genes related to carotenoid biosynthesis and catabolism was investigated in the juice sacs of ‘Tamami’. The results showed that the mechanism of β-cryptoxanthin accumulation in ‘Tamami’ was similar to that in Satsuma mandarin. Furthermore, in the recent studies, possible factors, which regulate carotenoid concentration and composition in citrus juice sacs were investigated in vitro.

Identification of a GCC transcription factor responding to fruit colour change events in citrus through the transcriptomic analyses of two mutants

BackgroundExternal ripening in Citrus fruits is morphologically characterized by a colour shift from green to orange due to the degradation of chlorophylls and the accumulation of carotenoid pigments. Although numerous genes coding for enzymes involved in such biochemical pathways have been identified, the molecular control of this process has been scarcely studied. In this work we used the Citrus clementina mutants 39B3 and 39E7, showing delayed colour break, to isolate genes potentially related to the regulation of peel ripening and its physiological or biochemical effects.ResultsPigment analyses revealed different profiles of carotenoid and chlorophyll modification in 39B3 and 39E7 mutants. Flavedo from 39B3 fruits showed an overall delay in carotenoid accumulation and chlorophyll degradation, while the flavedo of 39E7 was devoid of the apocarotenoid β-citraurin among other carotenoid alterations. A Citrus microarray containing about 20,000 cDNA fragments was used to identify genes that were differentially expressed during colour change in the flavedo of 39B3 and 39E7 mutants respect to the parental variety. The results highlighted 73 and 90 genes that were respectively up- and down-regulated in both mutants. CcGCC1 gene, coding for a GCC type transcriptional factor, was found to be down-regulated. CcGCC1 expression was strongly induced at the onset of colour change in the flavedo of parental clementine fruit. Moreover, treatment of fruits with gibberellins, a retardant of external ripening, delayed both colour break and CcGCC1 overexpression.ConclusionsIn this work, the citrus fruit ripening mutants 39B3 and 39E7 have been characterized at the phenotypic, biochemical and transcriptomic level. A defective synthesis of the apocarotenoid β-citraurin has been proposed to cause the yellowish colour of fully ripe 39E7 flavedo. The analyses of the mutant transcriptomes revealed that colour change during peel ripening was strongly associated with a major mobilization of mineral elements and with other previously known metabolic and photosynthetic changes. The expression of CcGCC1 was associated with peel ripening since CcGCC1 down-regulation correlated with a delay in colour break induced by genetic, developmental and hormonal causes.

Comprehensive Analysis of Expressed Sequence Tags from the Pulp of the Red Mutant ‘Cara Cara’ Navel Orange (Citrus sinensis Osbeck)

Expressed sequence tag (EST) analysis of the pulp of the red-fleshed mutant 'Cara Cara' navel orange provided a starting point for gene discovery and transcriptome survey during citrus fruit maturation. Interpretation of the EST datasets revealed that the mutant pulp transcriptome held a high section of stress responses related genes, such as the type III metallothionein-like gene (6.0%), heat shock protein (2.8%), Cu/Zn superoxide dismutase (0.8%), late embryogenesis abundant protein 5 (0.8%), etc. 133 transcripts were detected to be differentially expressed between the red mutant and its orange-color wild genotype 'Washington' via digital expression analysis. Among them, genes involved in metabolism, defense/stress and signal transduction were statistical overrepresented. Fifteen transcription factors, composed of NAM, ATAF, and CUC transcription factor (NAC); myeloblastosis (MYB); myelocytomatosis (MYC); basic helix-loop-helix (bHLH); basic leucine zipper (bZIP) domain members, were also included. The data reflected the distinct expression profile and the unique regulatory module associated with these two genotypes. Eight differently expressed genes analyzed in digital were validated by quantitative real-time polymerase chain reaction. For structural polymorphism, both simple sequence repeats and single nucleotide polymorphisms (SNP) loci were surveyed; dinucleotide presentation revealed a bias toward AG/GA/TC/CT repeats (52.5%), against GC/CG repeats (0%). SNPs analysis found that transitions (73%) outnumbered transversions (27%). Seventeen potential cultivar-specific and 387 heterozygous SNP loci were detected from 'Cara Cara' and 'Washington' EST pool.

Profiling gibberellin (GA 3)-responsive genes in mature mandarin fruit using a citrus 22K oligoarray

Gibberellin 3 (GA 3)-responsive genes were investigated with a citrus 22K oligoarray 1 1 The information regarding citrus 22K oligoarray is available at the website http://www.fruit.affrc.go.jp/cgat/. to further the understanding of transcriptional regulation by GA 3 treatment in Satsuma mandarin fruit ( Citrus unshiu Marc.). 213 GA 3-responsive genes were identified that showed a 3-fold or greater expression change after 72 h GA 3 treatment, compared to expression after 72 h air treatment. GA 3 treatment induced expression of pathogenesis-related (PR) proteins and genes that function in photosynthesis, chloroplast biogenesis, resistance, defense and stress. Also, GA 3 treatment reduced the transcription of several ethylene-inducible genes, such as carotenoid metabolic genes, which are associated with fruit ripening. Contrasting effects between GA 3 and ethylene were observed on photosynthesis and chloroplast biogenesis, chlorophyll metabolism, and carotenoid metabolism, indicating that the endogenous GA 3 level might be important for the endogenous regulation of maturation and senescence in mature citrus fruit. It was also found that the GA response pathway was likely to take part in cross-talk with the pathogen-related pathway in mature citrus fruit.