The accumulation of various pigments leads to the formation of different flower colors in plants. However, the regulation mechanism of yellow flower formation and flower color differences between Camellia nitidssima C.W.Chi (CN) and its hybrids C. ‘Zhenghuangqi’ (ZHQ), C. ‘Huangxuanlv’ (HXL), and C. ‘Xinshiji’ (XSJ), remains largely unknown. Here, we showed that the content of two flavonols, quercetin-7-O-glucoside (Qu7G) and quercetin-3-O-glucoside (Qu3G), was positively correlated with the yellow degree of petals in CN and its three hybrids. Additionally, we performed a comparative transcriptomic analysis of petals of the four yellow camellia plants, which revealed 322 common upregulated and 866 common downregulated DEGs (differentially expressed genes) in the CN vs. ZHQ, CN vs. HXL, and CN vs. XSJ comparison groups. Their regulatory pathway analysis showed that flavonol biosynthesis genes (FLSs and GTs) and transcriptional regulatory genes MYBs were all expressed higher in CN than its three hybrids, which corresponded to differences in the flavonol content among the four yellow camellias. Further, two ethylene synthesis genes (ACSs, ACO) and three ethylene signaling genes (EIN2s, EIN3, ERFs) were all upregulated in the yellow petals of CN. In conclusion, the expression of flavonol-related genes and flavonols (Qu7G and Qu3G) accumulation could play a key role in the formation of yellow flowers in camellia, and the ethylene pathway might be involved in the regulation of yellow flower formation of camellias. This work describes the possible regulatory pathway of yellow camellia, thereby laying a foundation for future verification of genes linked to flower coloring and the breeding of yellow camellia.

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Camellia japonica is a woody ornamental plant with multiple flower color variations caused by bud sport; however, the molecular mechanism remains unclear. Here, chemical and transcriptomic analyses of C. japonica were performed with white, pink, red, and dark red flowers caused by bud sport. Seven anthocyanins were detected in these samples, except in C. japonica ’YuDan’ (white petals). The total anthocyanin content of C. japonica ’JinBiHuiHuang’ was the highest, and cyanidin 3-O-β-glucoside (Cy3G) was the main anthocyanin affecting the redness of petals. Furthermore, the ratio of Cy3G and cyanidin-3-O-(6-O-(E)-p-coumaroyl)-B-glucoside) was significantly correlated with the red petal phenotype. In total, 5673 genes were identified as differentially expressed genes (DEGs). The potential co-expression modules related to anthocyanin accumulation were established, which featured transcription factors, anthocyanin biosynthesis, and plant hormone signal transduction. Thirteen structural genes in the anthocyanin biosynthetic pathway were identified as DEGs, most of them were upregulated with deepening of flower redness. An integrated promoter and cluster analysis suggested that CjMYB62, CjMYB52, and CjGATA may play important roles in anthocyanin accumulation. These results provide insight and candidate genes for the transcriptional mechanism responsible for the bud sport phenotype.

Leaves are the main vegetative organs of the aboveground part of plants and play an important role in plant morphogenesis. KNOTTED-LIKE HOMEOBOX ( KNOX ) plays a crucial role in regulating leaf cell fate and maintaining leaf development. In this study, we analyzed LtKNOX1 from Lilium tsingtauense and illustrated its function in transgenic plants. Tissue-specific expression analysis indicated that LtKNOX1 was highly expressed in stems, young flower buds, and shoot apical meristems (SAMs). Ectopic overexpression of LtKNOX1 in Nicotiana benthamiana suggested that transformants with mild phenotypes were characterized by foliar wrinkles and mildly curled leaves; transformants with intermediate phenotypes showed severely crimped blades and narrow leaf angles, and the most severe phenotypes lacked normal SAMs and leaves. Moreover, the expression levels of genes involved in the regulation of KNOX in transgenic plants were detected, including ASYMMETRIC LEAVES1, PIN-FORMED 1, GA20-oxidase, CUP-SHAPED COTYLEDON 2, CLAVATA 1 and WUSCHEL ( WUS ), and the expression of other genes were down-regulated except WUS . This study contributes to our understanding of the LtKNOX1 function.

CBFs belong to the ERF subfamily of the AP2 supergene family and often play an important role in the cold acclimation of temperate plants. However, the role of CBFs in Camellia japonica (Naidong), the only Camellia japonica population found in the temperate zones of China, remains unclear. It is very important to study the genetic composition of C. japonica (Naidong) to adapt to low temperature for Camellia species. Using full-length transcriptome data, we identified four CjCBF genes that respond to cold stress and analyzed their evolutionary relationships, domains, and expression patterns. The phylogeny of CBFs of 19 angiosperms divided the genes into three categories, and the four CjCBFs belong to a small subcluster. The strong response of CjCBF1 to cold treatment and its sustained high level of expression indicated that it plays an important role in the process of cold acclimation. A yeast two-hybrid assay revealed an interaction between CjCBF1, CjCBF2, and CjCBF5, and subcellular localization confirmed this finding. The expression of CjCBFs was tissue-specific: CBF1 was mainly expressed in leaves, and CBF3 was mainly expressed in stem. The responses of the four CjCBFs to drought and high temperature and the effect of light were also characterized. Our study provides new insight into the role of CBFs in the cold response in C. japonica (Naidong).