ABSTRACT
The GLK gene family plays a crucial role in the regulation of chloroplast development and participates in chlorophyll synthesis. However, the precise mechanism by which GLK contributes to citrus's chlorophyll synthesis remains elusive. The GLK gene family causes variations in the photosynthetic capacity and chlorophyll synthesis of different citrus varieties. In this study, we identified tissue-specific members and the key CcGLKs involved in chlorophyll synthesis. A total of thirty CcGLK transcription factors (TFs) were discovered in the citrus genome, distributed across all nine chromosomes. The low occurrence of gene tandem duplication events and intronic variability suggests that intronic variation may be the primary mode of evolution for CcGLK TFs. Tissue-specific expression patterns were observed for various GLK family members; for instance, CcGLK12 and CcGLK15 were specifically expressed in the skin, while CcGLK30 was specific to the ovary, and CcGLK10, CcGLK6, CcGLK21, CcGLK2, CcGLK18, CcGLK9, CcGLK28, and CcGLK8 were specifically expressed in the leaves. CcGLK4, CcGLK5, CcGLK11, CcGLK23, CcGLKl7, CcGLK26, and CcGLK20 may participate in the regulation of the ALA, prochlorophylate, protoporphyrin IX, Mg-protoporphyrin IX, Chl b, T-Chl, MG-ProtoIX ME, and POR contents in citrus.
ABSTRACT
Sweet orange 'Newhall' (C. sinensis) is a popular fruit in high demand all over the world. Its peel and pulp are rich in a variety of nutrients and are widely used in catering, medicine, food and other industries. Grafting is commonly practiced in citrus production. Different rootstock types directly affect the fruit quality and nutritional flavor of citrus. However, the studies on citrus metabolites by grafting with different rootstocks are very limited, especially for amino acids (AAs). The preliminary test showed that there were significant differences in total amino acid content of two rootstocks (Poncirus trifoliata (CT) and C. junos Siebold ex Tanaka (CJ)) after grafting, and total amino acid content in the peel was higher than flesh. However, the molecular mechanism affecting amino acid differential accumulation remains unclear. Therefore, this study selected peel as the experimental material to reveal the amino acid components and differential accumulation mechanism of sweet orange 'Newhall' grafted with different rootstocks through combined transcriptome and metabolome analysis. Metabolome analysis identified 110 amino acids (AAs) and their derivatives in sweet orange 'Newhall' peels, with L-valine being the most abundant. L-asparagine was observed to be affected by both developmental periods and rootstock grafting. Weighted gene co-expression network analysis (WGCNA) combined with Redundancy Analysis (RDA) revealed eight hub structural genes and 41 transcription factors (TFs) that significantly influenced amino acid biosynthesis in sweet orange 'Newhall' peels. Our findings further highlight the significance of rootstock selection in enhancing the nutritional value of citrus fruits and might contribute to the development of functional citrus foods and nutritional amino acid supplements.
ABSTRACT
Citrus cultivation involves the widespread practice of grafting, which has a significant impact on citrus development and fruit quality and yield. However, understanding the effect of flavonoid compounds after different rootstock grafting have been limited. Flavonoid compounds, found at the highest levels in citrus peels, contribute to improving fruit quality and nutritional value. In this study, scion-rootstock interaction was investigated at various developmental stages when sweet orange 'Newhall' was hetero-grafted with two commonly used rootstocks (Poncirus trifoliate population, C. junos Siebold ex Tanaka). Physiological index detection showed a higher concentration of total flavonoid content in peels of sweet orange 'Newhall' grafted on Poncirus trifoliate population (ct) than C. junos Siebold ex Tanaka (cj). Further metabolomic analysis identified 703 flavonoid compounds, including flavones, flavonols, and flavanones. Out of the 25 flavonoids affected by different rootstock grafting and developmental stages, most were flavones. Transcriptomic analysis identified 8,562 differentially expressed genes (DEGs). Co-expression and Pearson's correlation analysis discovered six hub structure genes and 19 transcription factors (TFs) that affected flavonoid biosynthesis. In addition to increasing the transcript levels of genes that synthesize flavones, flavonols, and flavanones, the scion-rootstock interaction also affected the expression of many TFs. Taken together, our findings suggested that hetero-grafting could promote the accumulation of flavonoid compounds in citrus peels during the development stages. These results offered fresh perspectives on grafting's application usefulness and the enhancement of the accumulation of nutritive flavonoid components by grafting in citrus.
ABSTRACT
Sweet orange 'Newhall' peels (SOPs) are abundant in flavonoids, making them increasingly popular in the realms of nutrition, food, and medicine. However, there is still much unknown about flavonoid components in SOPs and the molecular mechanism of flavonoid biosynthesis when subjected to magnesium stress. The previous experiment conducted by the research group found that the total flavonoid content of Magnesium deficiency (MD) was higher than Magnesium sufficiency (MS) in SOPs. In order to study the metabolic pathway of flavonoids under magnesium stress, an integrative analysis of the metabolome and transcriptome was performed in SOPs at different developmental stages, comparing MS and MD. A comprehensive analysis revealed the identification of 1,533 secondary metabolites in SOPs. Among them, 740 flavonoids were classified into eight categories, with flavones identified as the dominant flavonoid component. The influence of magnesium stress on flavonoid composition was evaluated using a combination of heat map and volcanic map, which indicated significant variations between MS and MD varieties at different growth stages. The transcriptome detected 17,897 differential genes that were significantly enriched in flavonoid pathways. Further analysis was performed using Weighted gene correlation network analysis (WGCNA) in conjunction with flavonoid metabolism profiling and transcriptome analysis to identify six hub structural genes and ten hub transcription factor genes that play a crucial role in regulating flavonoid biosynthesis from yellow and blue modules. The correlation heatmap and Canonical Correspondence Analysis (CCA) results showed that CitCHS had a significant impact on the synthesis of flavones and other flavonoids in SOPs, as it was the backbone gene in the flavonoid biosynthesis pathway. The qPCR results further validated the accuracy of transcriptome data and the reliability of candidate genes. Overall, these results shed light on the composition of flavonoid compounds in SOPs and highlight the changes in flavonoid metabolism that occur under magnesium stress. This research provides valuable insights for improving the cultivation of high-flavonoid plants and enhancing our understanding of the molecular mechanisms underlying flavonoid biosynthesis.
