Search

Background As a well-recognized Traditional Chinese medicine (TCM), immature fruits of Citrus grandis ‘Tomentosa’ serve to cure chronic cough in humans, to which flavonoids may contribute. Knowledge about the molecular mechanisms underlying flavonoid biosynthesis in ‘Tomentosa’ fruits would, therefore, favor the breeding of varieties with improved health supporting properties. Results To this end, we determined the transcriptomes, using RNA-seq, and metabolic profiles of two contrasting C. grandis varieties, namely ‘Tomentosa’ (‘ZM’) used for TCM production, and a locally cultivated pomelo (‘GQ’), at four fruit development stages. A total of 39 flavonoids, including 14 flavanone/flavone, five isoflavonoids, 12 flavonols, and six anthocyanins, were identified and 16 of them were quantitatively determined in fruits of the two varieties. We found that ‘ZM’ fruits contain more flavonoids than ‘GQ’, whereby rhoifolin accumulated to significantly higher levels in ‘ZM’. Transcriptome sequencing of ‘ZM’ and ‘GQ’ allowed the annotation of 31,510 genes, including 1,387 previously unknown ones. A total of 646 genes were found to be differentially expressed between ‘ZM’ and ‘GQ’ throughout all four fruit developmental stages, indicating them as robust expression markers in future breeding programs. Weighted gene co-expression network analysis (WGCNA) identified 18 modules. Conclusions Combined transcriptional and metabolic analysis revealed 25 flavonoid biosynthesis structural genes and 16 transcriptional regulators (MYBs, bHLHs, WD40) to be likely involved in the biosynthesis of flavonoids in C. grandis ‘Tomentosa’ fruits.

Citron (Citrus. medica L.) fruits are commonly utilized in the production of essential oil, therefore, the fruits residues turn out to be industrial byproducts. In the present study, a crude polysaccharide was extracted from citron fruit residues by hot water extraction and precipitation of ethanol (95%), after deproteinization, a major polysaccharide component (CMLP-2) was obtained by gradient ethanol precipitation (20%-80%). The physicochemical properties of CMLP-2 such as surface morphology, functional groups, and thermostability were examined by FT-IR spectroscopy, SEM, and thermogravimetric analysis. Moreover, the chemical structure of CMLP-2 was elucidated that CMLP-2 is an acidic pectic polysaccharide consisting of arabinose (Ara), galacturonic acid (GalA), and rhamnose (Rha) in a molar ratio of 4:2:1 with a molecular weight of 202.18 kDa. CMLP-2 is a novel pectic polysaccharide rich in rhamnogalacturonan I (RG-I). Moreover, rheological tests revealed that CMLP-2 solution is pseudoplastic and temperature resistant. The result could be a good basis for the utilization of Citrus medica L. fruits residues as plant-derived food additive.

Huanglongbing (HLB), also known as citrus greening, is the most severe pandemics in citrus in more than 50 countries in Asia, Africa and America, causing serious economic losses worldwide (Gottwald, 2020; Wang, 2019). The disease is associated with a phloem-limited and fastidious member of the α-proteobacteriacea. 'Candidatus Liberibacter asiaticus' (CLas) is the most prevalent strain. It was frequently detected in leaves, stems and roots based on PCR detection which recognized both living and dead cells.

Citrus grandis “Tomentosa” (“Huajuhong”) is a famous traditional Chinese medicine. The aim of the present study is to provide a comprehensive characterization of the m6A regulatory genes from C. grandis, and examine their expression patterns in fruits of C. grandis “Tomentosa” during various developmental stages. A total of 26 N6-methyladenosine (m6A) regulatory proteins were identified from the genome of C. grandis, which were distributed across nine chromosomes in C. grandis. Phylogenetic relationships revealed that all m6A regulatory genes were divided into groups of m6A writers, erasers, and readers. The m6A writer groups included CgMTA, CgMTB, and CgMTC three MTs (methyltransferases), one CgVIR (virilizer), one CgHAKAI (E3 ubiquitin ligase HAKAI), and one CgFIP37 (FKBP interacting protein 37). Moreover, 10 CgALKBH (α-ketoglutarate-dependent dioxygenase homolog) members (numbered from CgALKBH1 to CgALKBH10) and 10 CgECT (C-terminal region) members (numbered from CgECT1 to CgECT10) in C. grandis were identified as m6A erasers and readers, respectively. The domain structures and motif architectures among the groups of m6A writers, erasers, and readers were diverse. Cis-acting elements in the promoters of the 26 m6A regulatory genes predicted that the abscisic acid-responsive (ABA) element (ABRE) was present on the promoters of 19 genes. In addition, the expression profiles of all m6A regulatory genes were examined in the fruits of two varieties of C. grandis “Tomentosa” during different growth stages to give basic hints for further investigation of the function of the N6-methyladenosine regulatory genes in C. grandis “Tomentosa”.

In this study, the non‐targeted large‐scale plant metabolomics (UPLC‐Q‐Orbitrap‐MS) was performed for the comparison of chemical profiling of the leaves, barks, flowers, peels, pulps, and seeds of Clausena lansium (Lour.) Skeels (called “wampee”). A total of 364 metabolites were identified, and 62 potential biomarkers were selected by the multivariate statistical analysis. Hierarchical cluster analysis suggested that the selected biomarkers were significant differential metabolites among various parts of wampee. Metabolic pathway analysis showed a significant enrichment of the “Flavone and flavonol synthesis” and “Isoquinoline alkaloid biosynthesis” pathway. This study provides important information for the isolation and identification of functional components from different tissues of wampee and the metabolic biosynthesis pathway elucidation in detail.

Electrocatalysis of bioflavonoids in carbon nanomaterials plays an important role in electrochemical sensors for the detection of their content in fruits. In this study, three types of carbon nanomaterials with 1D, 2D, and 3D structures, namely carbon nanotubes (CNTs), graphene oxide (GO), and Ketjen black (KB), were modified onto glassy carbon electrodes for the electrocatalysis of hesperidin and naringin, which are two important bioflavonoids in fruits. As a result, the CNT-modified electrodes showed the highest electrocatalytic activity for both hesperidin and naringin compared to GO and KB. The morphology and surface chemistry of the carbon nanomaterials were characterized. The structural defects and carbon status of carbon nanomaterials are proposed to be the most important factors affecting the electrocatalysis of hesperidin and naringin. Finally, a CNT-based electrochemical sensor was fabricated to simultaneously detect hesperidin and naringin. Real sample tests on the fruit extract of

Background Mandarin ‘Shatangju’ is susceptible to Huanglongbing (HLB) and the HLB-infected fruits are small, off-flavor, and stay-green at the maturity period. To understand the relationship between pericarp color and HLB pathogen and the effect mechanism of HLB on fruit pericarp coloration, quantitative analyses of HLB bacterial pathogens and carotenoids and also the integrative analysis of metabolome and transcriptome profiles were performed in the mandarin ‘Shatangju’ variety with four different color fruits, whole green fruits (WGF), top-yellow and base-green fruits (TYBGF), whole light-yellow fruits (WLYF), and whole dark-yellow fruits (WDYF) that were infected with HLB. Results the HLB bacterial population followed the order WGF > TYBGF > WLYF > WDYF. And there were significant differences between each group of samples. Regarding the accumulation of chlorophyll and carotenoid, the chlorophyll-a content in WGF was the highest and in WDYF was the lowest. The content of chlorophyll-b in WGF was significantly higher than that in other three pericarps. There were significant differences in the total content of carotenoid between each group. WGF and TYBGF pericarps were low in phytoene, γ-carotene, β-cryptoxanthin and apocarotenal, while other kinds of carotenoids were significantly higher than those in WDYF. And WLYF was only short of apocarotenal. We comprehensively compared the transcriptome and metabolite profiles of abnormal (WGF, TYBGF and WLYF) and normal (WDYF, control) pericarps. In total, 2,880, 2,782 and 1,053 differentially expressed genes (DEGs), including 121, 117 and 43 transcription factors were identified in the three comparisons, respectively. The qRT-PCR confirmed the expression levels of genes selected from transcriptome. Additionally, a total of 77 flavonoids and other phenylpropanoid-derived metabolites were identified in the three comparisons. Most (76.65 %) showed markedly lower abundances in the three comparisons. The phenylpropanoid biosynthesis pathway was the major enrichment pathway in the integrative analysis of metabolome and transcriptome profiles. Conclusions Synthesizing the above analytical results, this study indicated that different color pericarps were associated with the reduced levels of some carotenoids and phenylpropanoids derivatives products and the down-regulation of proteins in flavonoids, phenylpropanoids derivatives biosynthesis pathway and the photosynthesis-antenna proteins.

Involvement of LaeA in various biological processes of filamentous fungi has been demonstrated. However, its role in Penicillium digitatum, the causal agent of citrus postharvest green mold, remains unclear. In this study, a ΔPdLaeA mutant was constructed using homologous recombination. The production of conidia by the ΔPdLaeA mutant was reduced by half compared with that of the wild-type strain. The sensitivity of the ΔPdLaeA mutant increased under alkaline conditions. The virulence assay revealed that PdLaeA was dispensable for the virulence of P. digitatum. Comparative transcriptome analysis revealed that the function loss of PdLaeA resulted in the reduced expression of several secondary metabolite gene clusters. In addition, expression of several key regulators of conidiation (BrlA, FlbA, FlbC, FlbD, and FluG) was also downregulated in the ΔPdLaeA mutant. In summary, the present work demonstrated that PdLaeA was involved in the regulation of SM biosynthesis, as well as the development and environmental adaptation of P. digitatum.

Penicillium digitatum is a destructive post-harvest pathogen of citrus fruit during storage and marketing. The aim of this study is to evaluate the activity of the natural polyphenolic compounds, tannins, on P. digitatum. Tannins acted against P. digitatum by inhibiting its mycelial growth and spore germination. In vivo tests showed that tannins significantly (P

The total phenolic, flavonoid, coumarin, and volatile contents and antioxidant activities in three tissues of six Citrus grandis ‘Tomentosa’ (CGT) fruit varieties were investigated. Two flavones, 3 coumarins and 147 volatiles were identified. The exocarp contained 3 coumarins, however, no coumarins were identified in the mesocarp, and only isoimperatorin was found in the pulp. In the exocarp, mesocarp and pulp, 104, 79 and 71 volatiles were identified, respectively. FuMao (FM) contained the highest total phenolic, naringin and rhoifolin amounts; HouYeZhengMao (HY-ZM) contained the highest meranzin hydrate and bergapten amounts; and XiYangZhengMao (XY-ZM) contained the highest isoimperatorin amount. The ABTS radical scavenging power of HuangLongZhengMao (HL-ZM) was the strongest, and FM had the strongest DPPH radical scavenging effect and ferric ion reducing antioxidant power. According to overall performance, FM and HY-ZM are the top two varieties with the highest total phenolic, flavonoid, and coumarin contents and antioxidant capacities.

Color formation in Hylocereus spp. (pitayas) has been ascribed to the accumulation of betalains. However, several studies have reported the presence of anthocyanins in pitaya fruit and their potential role in color formation has not yet been explored. In this study, we profiled metabolome and transcriptome in fruit of three cultivars with contrasting flesh colors (red, pink and white) to investigate their nutritional quality and the mechanism of color formation involving anthocyanins. Results revealed that pitaya fruit is enriched in amino acid, lipid, carbohydrate, polyphenols, vitamin and other bioactive components with significant variation among the three cultivars. Anthocyanins were detected in the fruit flesh and accumulation levels of Cyanidin 3-glucoside, Cyanidin 3-rutinoside, Delphinidin 3-O-(6-O-malonyl)-beta-glucoside-3-O-beta-glucoside and Delphinidin 3-O-beta-D-glucoside 5-O-(6-coumaroyl-beta-D-glucoside) positively correlated with the reddish coloration. Transcriptome data showed that the white cultivar tends to repress the anthocyanin biosynthetic pathway and divert substrates to other competing pathways. This perfectly contrasted with observations in the red cultivar. The pink cultivar however seems to keep a balance between the anthocyanin biosynthetic pathway and the competing pathways. We identified several active transcription factors of the MYB and bHLH families which can be further investigated as potential regulators of the anthocyanin biosynthetic genes. Collectively, our results suggest that anthocyanins partly contribute to color formation in pitaya fruit. Future studies aiming at manipulating the biosynthetic pathways of anthocyanins and betalains will better clarify the exact contribution of each pathway in color formation in pitayas. This will facilitate efforts to improve pitaya fruit quality and appeal.

Interfacial electron transfer between redox enzymes and electrodes is a key step for enzymatic bioelectrocatalysis in various bioelectrochemical devices. Although the use of carbon nanomaterials enables an increasing number of redox enzymes to carry out bioelectrocatalysis involving direct electron transfer (DET), the role of carbon nanomaterials in interfacial electron transfer remains unclear. Based on the recent progress reported in the literature, in this mini review, the significance of carbon nanomaterials on DET-type bioelectrocatalysis is discussed. Strategies for the oriented immobilization of redox enzymes in rationally modified carbon nanomaterials are also summarized and discussed. Furthermore, techniques to probe redox enzymes in carbon nanomaterials are introduced.

Background Color formation in Hylocereus spp. (pitayas) has been ascribed to the accumulation of betalains. However, several studies have reported the presence of anthocyanins in pitaya fruit and their potential role in color formation has not yet been explored. In this study, we profiled metabolome and transcriptome in fruit of three cultivars with contrasting flesh colors (red, pink and white) to investigate their nutritional quality and the mechanism of color formation involving anthocyanins. Results Results revealed that pitaya fruit is enriched in amino acid, lipid, carbohydrate, polyphenols, vitamin and other bioactive components with significant variation among the three cultivars. Anthocyanins were detected in the fruit flesh and accumulation levels of Cyanidin 3-glucoside, Cyanidin 3-rutinoside, Delphinidin 3-O-(6-O-malonyl)-beta-glucoside-3-O-beta-glucoside and Delphinidin 3-O-beta-D-glucoside 5-O-(6-coumaroyl-beta-D-glucoside) positively correlated with the reddish coloration. Transcriptome data showed that the white cultivar tends to repress the anthocyanin biosynthetic pathway and divert substrates to other competing pathways. This perfectly contrasted with observations in the red cultivar. The pink cultivar however seems to keep a balance between the anthocyanin biosynthetic pathway and the competing pathways. We identified several active transcription factors of the MYB and bHLH families which can be further investigated as potential regulators of the anthocyanin biosynthetic genes. Conclusions Collectively, our results suggest that anthocyanins partly contribute to color formation in pitaya fruit. Future studies aiming at manipulating the biosynthetic pathways of anthocyanins and betalains will better clarify the exact contribution of each pathway in color formation in pitayas. This will facilitate efforts to improve pitaya fruit quality and appeal.

In this study, the non‐targeted large‐scale plant metabolomics (UPLC‐Q‐Orbitrap‐MS) was performed for the comparison of chemical profiling of the leaves, barks, flowers, peels, pulps, and seeds of Clausena lansium (Lour.) Skeels (called “wampee”). A total of 364 metabolites were identified, and 62 potential biomarkers were selected by the multivariate statistical analysis. Hierarchical cluster analysis suggested that the selected biomarkers were significant differential metabolites among various parts of wampee. Metabolic pathway analysis showed a significant enrichment of the “Flavone and flavonol synthesis” and “Isoquinoline alkaloid biosynthesis” pathway. This study provides important information for the isolation and identification of functional components from different tissues of wampee and the metabolic biosynthesis pathway elucidation in detail., The comparison of chemical profiling of the leaves, barks, flowers, peels, pulps, and seeds of wampee was investigated. This study provides important information for the isolation and identification of functional components from different tissues of wampee and the metabolic biosynthesis pathway elucidation in detail.

Mandarin (Citrus reticulata) is one of the most important citrus crops worldwide. Its domestication is believed to have occurred in South China, which has been one of the centers of mandarin cultivation for four millennia. We collected natural wild populations of mandarin around the Nanling region and cultivated landraces in the vicinity. We found that the citric acid level was dramatically reduced in cultivated mandarins. To understand genetic basis of mandarin domestication, we de novo assembled a draft genome of wild mandarin and analyzed a set of 104 citrus genomes. We found that the Mangshan mandarin is a primitive type and that two independent domestication events have occurred, resulting in two groups of cultivated mandarins (MD1 and MD2) in the North and South Nanling Mountains, respectively. Two bottlenecks and two expansions of effective population size were identified for the MD1 group of cultivated mandarins. However, in the MD2 group there was a long and continuous decrease in the population size. MD1 and MD2 mandarins showed different patterns of interspecific introgression from cultivated pummelo species. We identified a region of high divergence in an aconitate hydratase (ACO) gene involved in the regulation of citrate content, which was possibly under selection during the domestication of mandarin. This study provides concrete genetic evidence for the geographical origin of extant wild mandarin populations and sheds light on the domestication and evolutionary history of mandarin.

Citrus is a globally consumed fruit with great popularity. Mandarin (Citrus reticulata cv. ‘Shatangju’) is a local variety, and its planting area and yield are the greatest regarding fruit tree planting in Guangdong Province, China. However, its resistance to Huanglongbing (HLB) is weak. After infection by HLB, the fruits cannot develop normally. In this study, four kinds of fruits were classified as HBG, XQG, ZQG, and DHG, according to the color of their peels. The metabolomes of the three abnormally colored groups (HBG, XQG, and ZQG) and the normally colored group (DHG) were compared using a UPLC-QQQ-MS-based metabolomics approach. In total, 913 metabolites were identified and classified into 23 different categories, including phenylpropanoids and flavonoids; among them, 215 (HBG, 177; XQG, 124; and ZQG, 62) metabolites showed differential accumulation in the three comparison groups (HBG/XQG/ZQG versus DHG). A total of 2 unique metabolites, O-caffeoyl maltotriose and myricetin were detected only in DHG samples. When comparing HBG with DHG, there were 109 decreased and 68 increased metabolites; comparing XQG with DHG, there were 88 decreased and 36 increased metabolites; comparing ZQG with DHG, 41 metabolites were decreased, and 21 metabolites were increased. Metabolic pathway enrichment analysis of these differential metabolites showed significant enrichment of the “phenylpropanoid biosynthesis” pathway in all comparison groups. The hierarchical cluster analysis of the differential metabolites of the four groups showed a clear grouping patterns. The relative contents of three phenylpropanoids, four flavonoids, two alkaloids, one anthocyanin, and two other metabolites were significantly different between each comparison group. This study might provide fundamental insight for the isolation and identification of functional compounds from the peels of citrus fruit infected with HLB and for in-depth research on the effect of HLB on the formation of fruits pigment and the development of HLB-resistant citrus varieties.

Citrus grandis ‘Tomentosa’ which is a special Citrus cultivar, has been employed as cough suppressant and expectorant in traditional Chinese medicine for thousands of years. The aim of this study is to investigate the immunomodulatory role of an acidic polysaccharide (designated as CGTP-AP) purified from C. grandis ‘Tomentosa’. CGTP-AP showed effective immune activation in RAW264.7 macrophages at the concentration of 1–100 μg/mL. CGTP-AP could promote the release of NO in dose- and time-dependent manners. Enzyme-Linked Immunosorbent Assay (ELISA) and RT-PCR analysis demonstrated that CGTP-AP could stimulate the secretion of TNF-α and IL-6 in a dosage-dependent way. Western blot analysis and RT-PCR analysis indicated that CGTP-AP treatment could induce the iNOS and COX-2 expression in RAW264.7 macrophages. By conducting the inhibitors experiments, the activation of NF-κB and MAPK signaling pathways by CGTP-AP treatment was confirmed. Therefore, the present results declared that CGTP-AP could be a promising candidate as a potent immunomodulator for the application in future pharmaceutical development.

Citrus grandis ‘Tomentosa’ (CGT) which is a region-famous medicinal and edible plant contains plentiful bioactive polysaccharides, however, its chemical structures and specific bioactivities still need to be further explored. In the present study, an acidic polysaccharide (CGTP-AP) was extracted and purified from the pulps of CGT. The structure elucidation and anti-cancer activity of CGTP-AP were investigated. Structure characterization indicated that CGTP-AP was a homogeneous heteropolysaccharide composed of arabinose, galactose and galacturonic acid in a molar ratio of 2.45:1:2.77, with an average molecular weight of 2721.68 kDa. Partial acid hydrolysis, methylation and NMR spectrometry revealed that the backbone of CGTP-AP mainly composed of (1 → 4)-α-D-galacturonan, while the branch principally consisted of (1 → 5)-α-L-Araf. In addition, CGTP-AP exhibited effective anti-proliferation against colon cancer cells LOVO and SW620 cells in dose- and time-dependent manners, with IC50 values of 5.55, 4.35 and 3.52 mg mL−1 after 24, 48 and 72 h, and 5.33, 3.63 and 2.97 mg mL−1 after 24, 48 and 72 h, respectively. This study indicated that CGTP-AP might be utilized as a promising food supplement for the patients of colon disorders.

Citrus is a globally consumed fruit with great popularity. Mandarin (Citrus reticulata cv. &lsquo, Shatangju&rsquo, ) is a local variety, and its planting area and yield are the greatest regarding fruit tree planting in Guangdong Province, China. However, its resistance to Huanglongbing (HLB) is weak. After infection by HLB, the fruits cannot develop normally. In this study, four kinds of fruits were classified as HBG, XQG, ZQG, and DHG, according to the color of their peels. The metabolomes of the three abnormally colored groups (HBG, XQG, and ZQG) and the normally colored group (DHG) were compared using a UPLC-QQQ-MS-based metabolomics approach. In total, 913 metabolites were identified and classified into 23 different categories, including phenylpropanoids and flavonoids, among them, 215 (HBG, 177, XQG, 124, and ZQG, 62) metabolites showed differential accumulation in the three comparison groups (HBG/XQG/ZQG versus DHG). A total of 2 unique metabolites, O-caffeoyl maltotriose and myricetin were detected only in DHG samples. When comparing HBG with DHG, there were 109 decreased and 68 increased metabolites, comparing XQG with DHG, there were 88 decreased and 36 increased metabolites, comparing ZQG with DHG, 41 metabolites were decreased, and 21 metabolites were increased. Metabolic pathway enrichment analysis of these differential metabolites showed significant enrichment of the &ldquo, phenylpropanoid biosynthesis&rdquo, pathway in all comparison groups. The hierarchical cluster analysis of the differential metabolites of the four groups showed a clear grouping patterns. The relative contents of three phenylpropanoids, four flavonoids, two alkaloids, one anthocyanin, and two other metabolites were significantly different between each comparison group. This study might provide fundamental insight for the isolation and identification of functional compounds from the peels of citrus fruit infected with HLB and for in-depth research on the effect of HLB on the formation of fruits pigment and the development of HLB-resistant citrus varieties.

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the only natural vector of Candidatus Liberibacter asiaticus that causes citrus huanglongbing (HLB), a most destructive disease of citrus. Currently, no remedial therapy exists for the disease, and so effective control of ACP is very important in curbing the transmission of the disease. The push-pull strategy should be thoroughly explored as an approach to ACP management. This mini-review summarises the current progress towards more effective repellent and attractant chemicals through investigating known repellent and attractive plants. Interactions between ACP and its host plants are also addressed, with emphasis on the possible involvement of the host biochemicals in attracting the insect. Potential ways to increase the effectiveness of the pull-push strategy are briefly discussed. It is expected that the pull-push strategy will be gradually developed following more extensive research.

Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is the insect vector of Candidatus Liberibacter asiaticus (Las), the causal bacteria of the devastating citrus disease Huanglongbing (HLB). Many citrus orchards in China have been destroyed by HLB infection. The survival time of ACPs without food was related to the temperature they experienced. The longest survival time of adult ACPs was 5.93 ± 0.12 days when the mean temperature was approximately 12 °C. The shortest lifespan of ACP adults was 2.33 ± 0.12 days when the mean temperature on the test day was approximately 26 °C. Surprisingly, ACP could feed on herbaceous plants in the absence of preferred hosts. The most suitable weed species for the ACP was Solanum nigrum, with the longest survival time of 41 days for all individuals tested. The survival time of ACPs feeding on herbaceous plants was also related to temperature. The surviving 20% and 10% of ACP adults in the Solanum nigrum treatment showed no decline but slight improvement as compared with control in the fecundity. In conclusion, some ACPs may be able to live for a month or even longer in an orchard if suitable plants and temperatures are available. The Las-infected ACPs that can survive by feeding on herbaceous plants may be sources of Las infection in replanted orchards. The suggested time to replant citrus in groves that were previously infected with Las is summer to autumn, or it is recommended to leave the land idle for more than three months.

Mingliutianju (Citrus reticuiata Blanco cv.Mingliutianju, MP) is a novel late ripening cultivar selected from Chuntianju (Citrus reticuiata Blanco cv.Chuntianju, CP) through spontaneous bud mutation. Large-scale analysis of gene expression differences between of peels of MP and CP is studied during the course of 12 weeks and 23 weeks after flowering (WAF) using Affymetrix GeneChip® citrus genome array to provide new insights into the mechanisms behind this bud mutation. Altogether, 395 differentially expressed genes are identified including 132 up-regulated and 263 down-regulated genes. Singular enrichment analysis (SEA) and parametric analysis of gene set enrichment (PAGE) are performed and 7 statistically significant differentially expressed genes are selected finally for further study. Results from semiquantitative RT-PCR performed on these 7 genes show that one gene's expression situation is totally correlated with that observed with the microarray. These seven genes include three UDP-glucosyltransferase genes, two disease resistance-responsive protein-related genes, one chitinase gene and one protein kinase family protein gene, all of which are involved in 18 different biological processes according to the hierarchical tree of annotated gene ontology (GO) terms. The candidate biological process and its relation to bud mutation are discussed.

Litchi and longan are famous southern subtropical fruits in China. Besides closely related to each other taxonomically, litchi and longan share many similarities in origin, cultivation history and environmental requirements; furthermore, the developments of research on litchi and longan germplasm resources are somewhat similar and comparable. This paper reviews the literature on origin and distribution, collection and conservation, classification and identification, selection and breeding of litchi and longan germplasm resources in China. The classification of litchi and longan can be divided into three stages: classification based on morphological traits, classification by isozyme analysis and DNA markers. The expression of morphological traits is often affected by environmental conditions; the number of loci that isozyme analysis can examine is small, cultivars, which are closely related often have identical isozyme patterns. The limitations of classification based on morphological traits and isozyme analysis are then overcome by the utilization of molecular markers. At present, most new litchi and longan cultivars are selected from superior seedlings obtained after open-pollination or sport; germplasm with desirable characteristics such as aborted seeds, rather big-fruit and rather late-bearing have not been fully exploited. In order to achieve the goal of improving fruit quality and agronomic characteristics, it is suggested that additional studies of genetic diversity on litchi and longan germplasm should be carried out with more powerful markers such as SSR and ISSR; marker assisted selection (MAS) should be employed to improve litchi and longan's cross breeding efficiency.

Huanglongbing (HLB) is an economical and destructive disease of citrus in South China, such as in Guangdong, Guangxi that is caused by the bacterium Candidatus Liberibacter asiaticus. The interaction at mRNA level between pathogen and citrus (Ponkan, Citrus reticulata Blanco ) was primarily researched by Digital Gene Expression Tag Profiling. Ponkan leaves at 13 weeks and 26 weeks after HLB inoculation were used for analysis. The numbers of up-regulated genes were increased from 37% in 13 wpi (weeks post inoculation) to 64% in 26 wpi. The differentially expressed genes (DEGs) fold change increased more than 8 times from 16.7% to 87.3%. Gene ontology (GO) process molecular function enrichment analysis showed that the DEGs with oxidation reduction function increased from 4.41% to 8.48% and that DEGs responsive to stresses increased from 1.10% to 2.08%, but those related to defense responses decreased from 0.74% to 0.64%. However, those related to defense responses of down-regulated genes increased from 0.55% to 0.79%. Apparently, the expression level of resistance genes strengthened, while the defense ability of host declined along with enhanced stresses caused by HLB infection. Photosynthesis-related genes were down-regulated at both 13 wpi and 26 wpi, which indicated that HLB infection greatly reduced the citrus photosynthesis, perhaps via feedback regulation of the accumulated starches resulted from blockage of sieve tubes by the bacteria in the phloem tissue. RIN4,a negative regulator of plant immunity, was also found up-regulated by approximately 9-fold.

Solexa sequencing was used to reveal the changes in small RNAome profile in roots of mock-inoculated (CK) and Huanglongbing bacteria-inoculated (HLB) ‘Sanhu’ tangerine (Citrus reticulata Blanco) trees. Results showed that the number of reads of both unique and total sRNAs decreased apparently in roots following infection with HLB. Distribution in length of sRNAs changed also remarkably, showing an increase in 22 nt and 21 nt small RNAs (sRNAs) and a decrease in 24 nt sRNAs in HLB-infected samples. A total of 42 known micro RNAs (miRNAs) belonging to 27 highly conserved miRNA families were identified. Comparisons showed that 33 known miRNAs exhibited a significant expression difference between CK and HLB-inoculated samples. In addition, 34 novel miRNAs, among which 24 were differentially expressed, were also identified, and their expression levels were analyzed by qRT-PCR. Three hundred and eighty five potential target genes were predicted for most of the 57 differentially expressed miRNAs. GO and KEGG annotation analysis revealed that most miRNA-target genes were those implicated in developmental process, response to stress and stimulus, transcription and protein metabolism. The characterization of the miRNAomes between the healthy and HLB infected Sanhu tangerine roots provided new insight into the involvement of miRNAs in HLB infection of citrus.

MS is the most commonly used basal medium for the micropropagation of banana (Musa spp.), in which inorganic N source consists of NH4+-N and NO3--N, and NH4+:NO3- = 1:2. In the present study, basal medium for rooting culture was modified by supplying NO3--N as the sole N source at the concentration of 17.80 to 19.78 mmol·L-1. Not only was the percentage of qualified plantlets higher than that of MS or ½ MS, but the cost for medium preparation (per liter) was about $0.6 lower than that of MS based on local retail price, for the concentration of N decreased from about 60 mmol·L-1 to nearly 20 mmol·L-1. The effects of four factors: sucrose (25 to 35 g·L-1), NAA (0.2 to 1.0 mg·L-1), IBA (0.2 to 1.0 mg·L-1), and basal medium with modified N source on the percentage of qualified plantlets were studied by using orthogonal design. The variance analysis of data showed that all the four factors significantly affected the process of rooting culture. Among them, sucrose was the most important factor, followed by revised basal medium, IBA, and NAA. The optimal medium for rooting culture was BM II supplemented with 30 g·L-1 sucrose, 0.5 mg·L-1 NAA, and 0.5 mg·L-1 IBA. During the following acclimation phase, >90% of plantlets survived. Chemical name used: 6-benzylaminopurine (BA); indole-3-butyric acid (IBA); α-naphthaleneacetic acid (NAA).

Pummelo cultivars are usually difficult to identify morphologically, especially when fruits are unavailable. The problem was addressed in this study with the use of two methods: high resolution melting analysis of SNPs and sequencing of DNA segments. In the first method, a set of 25 SNPs with high polymorphic information content were selected from SNPs predicted by analyzing ESTs and sequenced DNA segments. High resolution melting analysis was then used to genotype 260 accessions including 55 from Myanmar, and 178 different genotypes were thus identified. A total of 99 cultivars were assigned to 86 different genotypes since the known somatic mutants were identical to their original genotypes at the analyzed SNP loci. The Myanmar samples were genotypically different from each other and from all other samples, indicating they were derived from sexual propagation. Statistical analysis showed that the set of SNPs was powerful enough for identifying at least 1000 pummelo genotypes, though the discrimination power varied in different pummelo groups and populations. In the second method, 12 genomic DNA segments of 24 representative pummelo accessions were sequenced. Analysis of the sequences revealed the existence of a high haplotype polymorphism in pummelo, and statistical analysis showed that the segments could be used as genetic barcodes that should be informative enough to allow reliable identification of 1200 pummelo cultivars. The high level of haplotype diversity and an apparent population structure shown by DNA segments and by SNP genotypes, respectively, were discussed in relation to the origin and domestication of the pummelo species.

Photorespiration is one of the most intensively studied topics in plant biology. While a number of mutants deficient in photorespiratory enzymes have been identified and characterized for their physiological functions, efforts on glycolate oxidase (GLO; EC 1.1.3.15) have not been so successful. This is a report about the generation of transgenic rice (Oryza sativa L.) plants carrying a GLO antisense gene driven by an estradiol-inducible promoter, which allowed for controllable suppressions of GLO and its detailed functional analyses. The GLO-suppressed plants showed typical photorespiration-deficient phenotypes. More intriguingly, it was found that a positive and linear correlation existed between GLO activities and the net photosynthetic rates (P(N)), and photoinhibition subsequently occurred once P(N) reduction surpassed 60%, indicating GLO can exert a strong regulation over photosynthesis. Various expression analyses identified that Rubisco activase was transcriptionally suppressed in the GLO-suppressed plants, consistent with the decreased Rubisco activation states. While the substrate glycolate accumulated substantially, few changes were observed for the product glyoxylate, and for some other downstream metabolites or genes as well in the transgenic plants. Further analyses revealed that isocitrate lyase and malate synthase, two key enzymes in the glyoxylate cycle, were highly up-regulated under GLO deficiency. Taken together, the results suggest that GLO is a typical photorespiratory enzyme and that it can exert a strong regulation over photosynthesis, possibly through a feed-back inhibition on Rubisco activase, and that the glyoxylate cycle may be partially activated to compensate for the photorespiratory glyoxylate when GLO is suppressed in rice.

Root samples of 'Sanhu' red tangerine trees infected with and without Candidatus Liberibacter asiaticus (CLas) were collected at 50 days post inoculation and subjected to RNA-sequencing and isobaric tags for relative and absolute quantification (iTRAQ) to profile the differentially expressed genes (DEGs) and proteins (DEPs), respectively. Quantitative real-time PCR was subsequently used to confirm the expression of 16 selected DEGs. Results showed that a total of 3956 genes and 78 proteins were differentially regulated by HLB-infection. Among the most highly up-regulated DEPs were sperm specific protein 411, copper ion binding protein, germin-like proteins, subtilisin-like proteins and serine carboxypeptidase-like 40 proteins whose transcript levels were concomitantly up-regulated as shown by RNA-seq data. Comparison between our results and those of the previously reported showed that known HLB-modulated biological pathways including cell-wall modification, protease-involved protein degradation, carbohydrate metabolism, hormone synthesis and signaling, transcription activities, and stress responses were similarly regulated by HLB infection but different or root-specific changes did exist. The root unique changes included the down-regulation in genes of ubiquitin-dependent protein degradation pathway, secondary metabolism, cytochrome P450s, UDP-glucosyl transferases and pentatricopeptide repeat containing proteins. Notably, nutrient absorption was impaired by HLB-infection as the expression of the genes involved in Fe, Zn, N and P adsorption and transportation were significantly changed. HLB-infection induced some cellular defense responses but simultaneously reduced the biosynthesis of the three major classes of secondary metabolites, many of which are known to have anti-pathogen activities. Genes involved in callose deposition were up-regulated whereas those involved in callose degradation were also up-regulated, indicating that the sieve tube elements in roots were hanging on the balance of life and death at this stage. In addition, signs of carbohydrate starvation were already eminent in roots at this stage. Other interesting genes and pathways that were changed by HLB-infection were also discussed based on our findings.