Identification of genes associated with soluble sugar and organic acid accumulation in 'Huapi' kumquat (Fortunella crassifolia Swingle) via transcriptome analysis.

BACKGROUND: The levels and ratios of sugar and acid are important contributors to fruit taste. Kumquat is one of the most economically important citrus crops, but information on the soluble sugar and organic acid metabolism in kumquat is limited. Here, two kumquat varieties - 'Rongan' (RA) and its mutant 'Huapi' (HP) - were used to assess soluble sugar and organic acid accumulation and the related genes. RESULTS: Soluble sugars include sucrose, glucose and fructose, while malate, quinic acid and citrate are the dominant organic acids in the fruits of both kumquat varieties. HP accumulated more sugars but fewer organic acids than did RA. Transcriptome analysis revealed 63 and 40 differentially expressed genes involved in soluble sugar and organic acid accumulation, respectively. The genes associated with sugar synthesis and transport, including SUS, SPS, TST, STP and ERD6L, were up-regulated, whereas INVs, FRK and HXK genes related to sugar degradation were down-regulated in HP kumquat. For organic acids, the up-regulation of PEPC and NAD-MDH could accelerate malate accumulation. In contrast, high expression of NAD-IDH and GS resulted in citric acid degradation during HP fruit development. Additionally, the PK, PDH, PEPCK and FBPase genes responsible for the interconversion of soluble sugars and organic acids were also significantly altered in the early development stages in HP. CONCLUSION: The high sugar accumulation in HP fruit was associated with up-regulation of SUS, SPS, TST, STP and ERD6L genes. The PEPCK, PEPC, NAD-MDH, NADP-IDH, GS and FBPase genes played important roles in acid synthesis and degradation in HP kumquat. These findings provide further insight into understanding the mechanisms underlying metabolism of sugars and organic acids in citrus. © 2021 Society of Chemical Industry.

Genome-Wide Identification of Long Non-coding RNA in Trifoliate Orange (Poncirus trifoliata (L.) Raf) Leaves in Response to Boron Deficiency.

Long non-coding RNAs (lncRNAs) play important roles in plant growth and stress responses. As a dominant abiotic stress factor in soil, boron (B) deficiency stress has impacted the growth and development of citrus in the red soil region of southern China. In the present work, we performed a genome-wide identification and characterization of lncRNAs in response to B deficiency stress in the leaves of trifoliate orange (Poncirus trifoliata), an important rootstock of citrus. A total of 2101 unique lncRNAs and 24,534 mRNAs were predicted. Quantitative real-time polymerase chain reaction (qRT-PCR) experiments were performed for a total of 16 random mRNAs and lncRNAs to validate their existence and expression patterns. Expression profiling of the leaves of trifoliate orange under B deficiency stress identified 729 up-regulated and 721 down-regulated lncRNAs, and 8419 up-regulated and 8395 down-regulated mRNAs. Further analysis showed that a total of 84 differentially expressed lncRNAs (DELs) were up-regulated and 31 were down-regulated, where the number of up-regulated DELs was 2.71-fold that of down-regulated. A similar trend was also observed in differentially expressed mRNAs (DEMs, 4.21-fold). Functional annotation of these DEMs was performed using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, and the results demonstrated an enrichment of the categories of the biosynthesis of secondary metabolites (including phenylpropanoid biosynthesis/lignin biosynthesis), plant hormone signal transduction and the calcium signaling pathway. LncRNA target gene enrichment identified several target genes that were involved in plant hormones, and the expression of lncRNAs and their target genes was significantly influenced. Therefore, our results suggest that lncRNAs can regulate the metabolism and signal transduction of plant hormones, which play an important role in the responses of citrus plants to B deficiency stress. Co-expression network analysis indicated that 468 significantly differentially expressed genes may be potential targets of 90 lncRNAs, and a total of 838 matched lncRNA-mRNA pairs were identified. In summary, our data provides a rich resource of candidate lncRNAs and mRNAs, as well as their related pathways, thereby improving our understanding of the role of lncRNAs in response to B deficiency stress, and in symptom formation caused by B deficiency in the leaves of trifoliate orange.

Vitamin D level in patients with menstrually related migraine / 上海交通大学学报（医学版）

Objective · To compare vitamin D levels in the patients with menstrually related migraine and those with non-menstrual migraine.Methods · A total of 100 patients with menstrually related migraine were enrolled as case group, and 100 patients with non-menstrual migraine were matched as control group in Renji Hospital, Shanghai Jiao Tong University School of Medicine, from June 2017 to June 2018. The patients' information was collected and serum vitamin D levels were measured. Univariate analysis was performed to analyze the differences between the groups, and Logistic regression model was performed to analyze the relationship between menstrually related migraine and vitamin D levels. Results · Vitamin D levels were lower in case group than those in control group [ (13.48±5.02) ng/mL vs (14.93±4.94) ng/mL, P=0.042]. There were no statistically significant differences in other indexes between the groups (P＞0.05). Logistic regression analysis showed that the patients with vitamin D less than 10 ng/mL were more likely to have menstrually related migraine than those with vitamin D more than 20 ng/ml (OR=3.656, 95％ CI 1.382-9.672, P=0.009). In addition, there were no statistically significant differences in clinical characteristics and vitamin D levels between the patients with migraine occurring in premenstrual period and menstrual period (P＞0.05). Conclusion · Low vitamin D levels may be associated with menstrually related migraine, but not with its occurrence in premenstrual or menstrual period.

[Effects of drought and rewatering on leaf photosynthesis, chlorophyll fluorescence, and root architecture of citrus seedlings.] / å¹²æ—±å¤æ°´å¯¹æŸ‘æ©˜å¹¼è‹—å¶ç‰‡å ‰åˆã€å¶ç»¿ç´ è§å ‰å’Œæ ¹ç³»æž„åž‹çš„å½±å“.

Drought severely affects citrus growth and development. In order to explore the mechanism of drought response of citrus, two cultivars (Sanhuhongju and Sanhuhuahong) that differing in drought tolerance were used as materials. The drought and rewatering treatment was conducted in pot experiments, with leaf photosynthesis, chlorophyll fluorescence, and root architecture being measured. The results showed that drought significantly decreased net photosynthetic rate (Pn), stomatal conductance (gs), transportation rate (Tr), and intercellular CO2 concentration (Ci) of both cultivars, but Sanhuhongju generally showed less reduction. After rewatering, photosynthetic parameters were partly recovered but still lower than that in control. The water use efficiency (WUE) of Sanhuhongju was significantly increased after drought stress for 15 d, but the WUE of Sanhuhuahong was decreased except at the 15 day of drought stress. In addition, the maximum photosynthesis efficiency of PS II (Fv/Fm) was increased in both cultivars, but the photochemical quantum yield of PS II [Y(II)] was increased in Sanhuhuahong under drought. Both the apparent electron transport rate (ETR) and photochemical quenching (qP) were inhibited in the treated seedlings. The non-photochemical quenching (NPQ) was decreased in Sanhuhongju while increased in Sanhuhuahong under drought and rewatering conditions. Drought stress resulted in the decrease of root surface area and volume of both cultivars, and it inhibited root elongation of Sanhuhuahong while improved the root length and root tip number of Sanhuhongju. The length of first lateral roots of Sanhuhongju was increased after drought stress 10 d, but did not change at the drought stress prophase of Sanhuhuahong, and then significantly decreased after 20 d. Furthermore, drought stress inhibited all lateral roots development except the tertiary lateral root of Sanhuhongju, and root growth could not be recovered by rewatering except root tip number. In conclusion, Sanhuhongju showed less reduction in leaf photosynthesis than Sanhuhuahong, with higher WUE and light use efficiency under drought stress. The increases of root tip number and lateral root length would help improve water uptake ability in Sanhuhongju.

Transcription profiles of boron-deficiency-responsive genes in citrus rootstock root by suppression subtractive hybridization and cDNA microarray.

Boron (B) deficiency has seriously negative effect on citrus production. Carrizo citrange (CC) has been reported as a B-deficiency tolerant rootstock. However, the molecular mechanism of its B-deficiency tolerance remained not well-explored. To understand the molecular basis of citrus rootstock to B-deficiency, suppression subtractive hybridization (SSH) and microarray approaches were combined to identify the potential important or novel genes responsive to B-deficiency. Firstly four SSH libraries were constructed for the root tissue of two citrus rootstocks CC and Trifoliate orange (TO) to compare B-deficiency treated and non-treated plants. Then 7680 clones from these SSH libraries were used to construct a cDNA array and microarray analysis was carried out to verify the expression changes of these clones upon B-deficiency treatment at various time points compared to the corresponding controls. A total of 139 unigenes that were differentially expressed upon B-deficiency stress either in CC or TO were identified from microarray analysis, some of these genes have not previously been reported to be associated with B-deficiency stress. In this work, several genes involved in cell wall metabolism and transmembrane transport were identified to be highly regulated under B-deficiency stress, and a total of 23 metabolic pathways were affected by B-deficiency, especially the lignin biosynthesis pathway, nitrogen metabolism, and glycolytic pathway. All these results indicated that CC was more tolerant than TO to B-deficiency stress. The B-deficiency responsive genes identified in this study could provide further information for understanding the mechanisms of B-deficiency tolerance in citrus.

Digital gene expression analysis of corky split vein caused by boron deficiency in 'Newhall' Navel Orange (Citrus sinensis Osbeck) for selecting differentially expressed genes related to vascular hypertrophy.

Corky split vein caused by boron (B) deficiency in 'Newhall' Navel Orange was studied in the present research. The boron-deficient citrus exhibited a symptom of corky split vein in mature leaves. Morphologic and anatomical surveys at four representative phases of corky split veins showed that the symptom was the result of vascular hypertrophy. Digital gene expression (DGE) analysis was performed based on the Illumina HiSeq™ 2000 platform, which was applied to analyze the gene expression profilings of corky split veins at four morphologic phases. Over 5.3 million clean reads per library were successfully mapped to the reference database and more than 22897 mapped genes per library were simultaneously obtained. Analysis of the differentially expressed genes (DEGs) revealed that the expressions of genes associated with cytokinin signal transduction, cell division, vascular development, lignin biosynthesis and photosynthesis in corky split veins were all affected. The expressions of WOL and ARR12 involved in the cytokinin signal transduction pathway were up-regulated at 1(st) phase of corky split vein development. Furthermore, the expressions of some cell cycle genes, CYCs and CDKB, and vascular development genes, WOX4 and VND7, were up-regulated at the following 2(nd) and 3(rd) phases. These findings indicated that the cytokinin signal transduction pathway may play a role in initiating symptom observed in our study.