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Plant Sci ; 311: 110996, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34482908


Rubus chingii, is widely distributed in many Asian countries and well known for its medicinal and dietary properties. Diversity of fruit color in raspberry has been attributed to the presence of either anthocyanins or carotenoids. In this study, we investigated anthocyanins and carotenoids, and their biosynthesis by LC-MS/MS. Six anthocyanins mainly consisted of flavanol-anthocyanins while five carotenoids mainly consisted of ß-citraurin esters. Flavanol-anthocyanins were produced from an offshoot of the anthocyanin biosynthesis, which started with biosynthesis of flavanols and anthocyanidin by leucoanthocyanidin reductase (LAR)/anthocyanidin reductase (ANR) and anthocyanidin synthase (ANS/LDOX) respectively. ß-citraurin esters were produced from cleavage of zeaxanthin and esterification by organic acid, which was an offshoot of the carotenoid biosynthesis. The offshoot started with biosynthesis of zeaxanthin and ß-citraurin by carotene ß-hydroxylase (CHYB/LUT5) and carotenoid cleavage dioxygenase (CCD) respectively. During fruit ripening, biosynthesis of flavanols and anthocyanins was down-regulated by genes/proteins involved in phenylpropanoid and flavonoid biosynthesis, while biosynthesis of ß-citraurin esters was up-regulated by imbalanced expression of genes/proteins involved in ß,ß-ring and ß, ε-ring hydroxylation. Thus, ß-citraurin esters, instead of anthocyanins imparted reddish color to the ripe fruit. These pigments and their biosynthesis in R. chingii are totally different from what occurs in other raspberry species.

Antocianinas/metabolismo , Carotenoides/metabolismo , Frutas/crescimento & desenvolvimento , Frutas/metabolismo , Pigmentação/fisiologia , Rubus/crescimento & desenvolvimento , Rubus/metabolismo , Antocianinas/genética , China , Frutas/anatomia & histologia , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Pigmentação/genética , Rubus/anatomia & histologia
Plant Cell Rep ; 40(10): 1923-1946, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34333679


KEY MESSAGE: Combined transcriptomic and metabolic analyses reveal that fruit of Rubus chingii Hu launches biosynthesis of phenolic acids and flavonols at beginning of fruit set and then coordinately accumulated or converted to their derivatives. Rubus chingii Hu (Chinese raspberry) is an important dual functional food with nutraceutical and pharmaceutical values. Comprehensively understanding the mechanisms of fruit development and bioactive components synthesis and regulation could accelerate genetic analysis and molecular breeding for the unique species. Combined transcriptomic and metabolic analyses of R. chingii fruits from different developmental stages, including big green, green-to-yellow, yellow-to-orange, and red stages, were conducted. A total of 89,188 unigenes were generated and 57,545 unigenes (64.52%) were annotated. Differential expression genes (DEGs) and differentially accumulated metabolites (DAMs) were mainly involved in the biosynthesis of secondary metabolites. The fruit launched the biosynthesis of phenolic acids and flavonols at the very beginning of fruit set and then coordinately accumulated or converted to their derivatives. This was tightly regulated by expressions of the related genes and MYB and bHLH transcription factors. The core genes products participated in the biosynthesis of ellagic acid (EA) and kaempferol-3-O-rutinoside (K-3-R), such as DAHPS, DQD/SDH, PAL, 4CL, CHS, CHI, F3H, F3'H, FLS, and UGT78D2, and their corresponding metabolites were elaborately characterized. Our research reveals the molecular and chemical mechanisms of the fruit development of R. chingii. The results provide a solid foundation for the genetic analysis, functional genes isolation, fruit quality improvement and modifiable breeding of R. chingii.

Ácido Elágico , Frutas/crescimento & desenvolvimento , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas , Rubus/crescimento & desenvolvimento , Ácido Elágico/metabolismo , Flavonóis/biossíntese , Flavonóis/genética , Frutas/genética , Perfilação da Expressão Gênica , Hidroxibenzoatos/metabolismo , Quempferóis/genética , Quempferóis/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Controle de Qualidade , Rubus/genética , Rubus/metabolismo , Terpenos/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
Front Plant Sci ; 12: 706667, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34447402


Rubus chingii HU, is a medicinal and nutritious fruit, which is very rich in flavonoids. However, the biosynthesis of its flavonoids is poorly understood. This study examined flavonoids and the genes/proteins at four fruit ripening phases using LC-MS/MS and qPCR. Six major kinds of anthocyanins, primarily consisted of flavanol-anthocyanins, which differed in form or concentration from other Rubus species. In contrast to other known raspberries species, R. chingii had a decline in flavonoids during fruit ripening, which was due to down-regulation of genes and proteins involved in phenylpropanoid and flavonoid biosynthesis. Unexpectedly, anthocyanin also continuously decreased during fruit maturation. This suggests that anthocyanins are not responsible for the fruit's reddish coloration. Flavanol-anthocyanins were derived from the proanthocyanidin pathway, which consumed two flavonoid units both produced through the same upstream pathway. Their presence indicates a reduction in the potential biosynthesis of anthocyanin production. Also, the constantly low expression of RchANS gene resulted in low levels of anthocyanin biosynthesis. The lack of RchF3'5'H gene/protein hindered the production of delphinidin glycosides. Flavonoids primarily comprising of quercetin/kaempferol-glycosides were predominately located at fruit epidermal-hair and placentae. The proportion of receptacle/drupelets changes with the maturity of the fruit and may be related to a decrease in the content of flavonoids per unit mass as the fruit matures. The profile and biosynthesis of R. chingii flavonoids are unique to Rubus. The unique flavonol pathways of R. chingii could be used to broaden the genetic diversity of raspberry cultivars and to improve their fruit quality.

Plant Physiol Biochem ; 163: 76-86, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33819717


Carotenoids are one of the most abundant pigments in raspberries. Rubus chingii Hu, indigenous to China, is traditionally consumed for health benefits. However, the carotenoid composition and pathways of R. chingii have not yet been studied. In this study, the components of carotenoids and genes/proteins involved in their biosynthesis were investigated during four fruit ripening phases via LC-MS/MS. Zeaxanthin, ß-citraurin and its esters, first identified in Rubus, gradually accumulated during fruit maturation. These compounds, rather than anthocyanins, were responsible for the ripe fruit coloration. In carotenoid metabolism, upstream synthesis genes of RcPSY2 (CL1406.Contig2), RcPDS1 (CL7625.Contig2), RcZDS1 (CL590.Contig6) and RcCRTISO1 (CL6919.Contig2) were up-regulated in gene/protein expression to accelerate carotene biosynthesis. Downstream genes of RcLUT5CHYB/CYP97A (CL8884.Contig3) and RcCHYB/BCH (CL7966.Contig1) were up-regulated in gene/protein expression, while RcCHYE/CYP97C (CL9380.Contig1/2) were maintained at low levels. RcLCYE (Unigene19570) was down-regulated while RcLCYB (CL7586.Contig1) was up-regulated and then down-regulated. These differential gene/protein expressions between LCYB and LCYE, and between CHYE and CHYB led to zeaxanthin accumulation by elevating its biosynthetic enzymes and lowering enzymes for lutein biosynthesis. In apocarotenoid biosynthesis, RcCCD (CL1310.Contig3) was up-regulated in gene/protein expression, which raised the content of ß-citraurin and its esters. Additionally, these genes/proteins diverged into different subgroups with distinct pattens of expression, suggesting their difference in function. For example, RcPSY1/3, RcZDS2, and RcCRTISO2/3 genes were expressed at very low levels, suggesting that they may be active in other tissues rather than in fruit. The mechanism of zeaxanthin and ß-citraurin biosynthesis is responsible for fruit coloration, which is completely novel to Rubus.

Rubus , China , Cromatografia Líquida , Frutas/genética , Regulação da Expressão Gênica de Plantas , Espectrometria de Massas em Tandem