Cheng Pan

Saliva plays a crucial role in shaping the compatibility of piercing–sucking insects with their host plants. Understanding the complex composition of leafhopper saliva is important for developing effective and eco-friendly control strategies for the tea green leafhopper, Empoasca flavescens Fabrecius, a major piercing–sucking pest in Chinese tea plantations. This study explored the saliva proteins of tea green leafhopper adults using a custom collection device, consisting of two layers of Parafilm stretched over a sucrose diet. A total of 152 proteins were identified using liquid chromatography–tandem mass spectrometry (LC-MS/MS) following the filter-aided sample preparation (FASP). These proteins were categorized into six groups based on their functions, including enzymes, transport proteins, regulatory proteins, cell structure proteins, other proteins, and unknown proteins. Bioinformatics analyses predicted 16 secreted proteins, which were successfully cloned and transcriptionally analyzed across various tissues and developmental stages. Genes encoding putative salivary secretory proteins, including Efmucin1, EfOBP1, EfOBP2, EfOBP3, Efmucin2, low-density lipoprotein receptor-related protein (EfLRP), EFVg1, and EFVg2, exhibited high expressions in salivary gland (SG) tissues and feeding-associated expressions at different developmental stages. These findings shed light on the potential elicitors or effectors mediating the leafhopper feeding and defense responses in tea plants, providing insights into the coevolution of tea plants and leafhoppers. The study’s conclusions open avenues for the development of innovative leafhopper control technologies that reduce the reliance on pesticides in the tea industry.

The Hangzhou White Chrysanthemum (Chrysanthemum morifolium Ramat) is one of the “Zhejiang eight flavors” in traditional Chinese medicine. The phosphatidylethanolamine-binding protein (PEBP) plays an important role in flowering and floral organ development. Even so, the biological role of PEBPs in the Hangzhou White Chrysanthemum has not been studied, which attracted us. Here, nine CmPEBP genes that contain the PF01161 domain were identified in the Hangzhou White Chrysanthemum for the first time, and their biological role in flowering was preliminarily studied. A phylogenetic analysis classified the CmPEBP genes into three subfamilies: MFT-like, TFL-like, and FT-like genes. The differential expression analysis was performed under different tissues and different stressors using qRT-PCR. It showed that each CmPEBP displayed tissue-specific expression patterns. Expression patterns in response to different temperatures and hormone stressors were investigated. They were finally demonstrated to be differentially expressed. TFL-like gene expression, which delayed reproductive growth, was upregulated under heat stress. Conversely, FT-like gene expression was upregulated under low temperatures. CmFT1 expression could be inhibited by GA (gibberellin), 6-BA (benzylaminopurine), ET (ethylene), and MeSA (methyl salicylate) but could be activated by IAA (indole-3-aceticacid), ABA (abscisic acid), and SA (salicylic acid) in the dark, whereas CmFT2 and CmFT3 expression levels were upregulated by ET, MeJA (methyl jasmonate), and ABA but were downregulated by 6-BA, SA, and MeSA. GA, IAA, SA, and MeSA inhibited CmTFL gene expression under light and dark treatments. Further research on CmPEBP genes in the Hangzhou White Chrysanthemum could better determine their roles in flowering and floral organ development, especially in response to the prolonged spraying of exogenous hormones.

Gibberellins (GAs), a class of diterpenoid phytohormones, play a key role in regulating diverse processes throughout the life cycle of plants. Bioactive GA levels are rapidly regulated by Gibberellin-dioxygenases (GAox), which are involved in the biosynthesis and deactivation of gibberellin. In this manuscript, a comprehensive genome-wide analysis was carried out to find all GAox in Camellia sinensis. For the first time in a tea plant, 14 CsGAox genes, containing two domains, DIOX_N (PF14226) and 2OG-FeII_Oxy, were identified (PF03171). These genes all belong to 2-oxoglutarate-dependent dioxygenases (2-ODD), including four CsGA20ox (EC: 1.14.11.12), three CsGA3ox (EC: 1.14.11.15), and seven CsGA2ox (EC: 1.14.11.13). According to the phylogenetic classification as in Arabidopsis, the CsGAox genes spanned five subgroups. Each CsGAox shows tissue-specific expression patterns, although these vary greatly. Some candidate genes, which may play an important role in response to external abiotic stresses, have been identified with regards to patterns, such as CsGA20ox2, CsGA3ox2, CsGA3ox3, CsGA2ox1, CsGA2ox2, and CsGA2ox4. The bioactive GA levels may be closely related to the GA20ox, GA3ox and GA2ox genes. In addition, the candidate genes could be used as marker genes for abiotic stress resistance breeding in tea plants.

As mobile genetic elements, the diversity and activity of the Tyl-copia retrotransposons are key contributors to genome organisation and evolution, which have been investigated in many plants but little in the tea plant, Camellia sinensis. We selected a total of 12 varieties of tea plant distributed across a large geographical area and sequenced the reverse transcriptase (RT) sequences of Ty1-copia retrotransposons using degenerate primers. The sequences that were widespread among tea varieties, were approximately 260 bp in length and exhibited high heterogeneity among tea plants. The RT sequences from tea plants were similar to other known Ty1-copia retrotransposon RT sequences. Phylogenetic analysis showed that tea RT sequences were closely related to those from woody plants, such as pear, poplar and apple. In contrast, they were more distantly related to RT sequences from herbaceous plants, such as tomato and rice. Activity assay revealed that Ty1-copia retrotransposons are transcriptionally active during the normal development of tea plants. These results will support further research on Ty1-copia retrotransposons in C. sinensis.