Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) is a rapid and sensitive method for analyzing microRNA (miRNA) expression. However, accurate qRT-PCR results depend on the selection of reliable reference genes as internal positive controls. To date, few studies have identified reliable reference genes for differential expression analysis of miRNAs among tissues, and among experimental conditions in plants. In this study, three miRNAs and four non-coding small RNAs (ncRNA) were selected as reference candidates, and the stability of their expression was evaluated among different tissues and under different experimental conditions in the tea plant (Camellia sinensis) using the geNorm and NormFinder programs. It was shown that miR159a was the best single reference gene in the bud to the fifth leaf, 5S rRNA was the most suitable gene in different organs, miR6149 was the most stable gene when the leaves were attacked by Ectropis oblique and U4, miR5368n and miR159a were the best genes when the leaves were treated by methyl jasmonate (MeJA), salicylic acid (SA) and abscisic acid (ABA), respectively. Our results provide suitable reference genes for future investigations on miRNA functions in tea plants.

Climate change and extreme weather affect tea growing. A competitive tea market needs quick, short-term solutions. This study evaluates the effects of various shade nets under mild and extreme cold stress on tea leaf physiology, photosynthetic alterations, antioxidant activities, and physiochemical characteristics. Tea plants were treated with SD0 (0% non-shading), SD1 (30% shading), SD2 (60% shading), and SD3 (75% shading). The 30%, 60%, and 75% shade nets shielded tea leaves from cold damage and reduced leaf injury during mild and extreme cold conditions compared with SD0% non-shading. Shading regulates photochemical capacity and efficiency and optimizes chlorophyll a and b, chlorophyll, and carotenoid contents. Moreover, carbon and nitrogen increased during mild cold and decreased in extreme cold conditions. Shading promoted antioxidant activity and physiochemical attributes. In fact, under 60% of shade, superoxide dismutase, peroxidase, catalase, and ω-3 alpha-linolenic acid were improved compared with SD0% non-shading during both mild and extreme cold conditions. From these findings, we hypothesized that the effect of different shades played an important role in the protection of tea leaves and alleviated the defense mechanism for “Zhong Cha 102” during exposure to a cold environment.

Intercropping soybean in tea plantations is a sustainable cultivation system that can improve the growing environment of tea plants compared to monoculture tea. However, the effects of this system on the photosynthesis activity of tea seedlings have yet to be reported. Therefore, we used tea cultivar ‘Zhongcha108’ as experimental materials to investigate the effects of intercropping soybean on the canopy spectral parameters and photosynthesis activity of tea seedlings. Canopy spectral reflectance data showed that soybean–tea intercropping (STS) improved the reflectance of 720, 750 and 840 nm bands in tea seedlings’ canopy. The vegetation indexes (VIs) value related to photosynthetic pigments in STS was obviously higher than monoculture tea (T). In addition, the F_v/F_m and SPAD value in STS were also clearly higher. Transcriptome analysis data indicated that STS induced the expression of light-harvesting complex (LHC) genes, photosystem subunit (Psbs and Psas) genes and dark reaction biological process genes (FBP1, RPE, Calvin cycle protein CP12-1 and transketolase). These results indicate that STS enhanced the photosynthesis activity. The metabolome analysis showed that STS promoted the accumulation of carbohydrate metabolites, which further provided evidence for the enhancement of photosynthesis in the leaves of tea seedlings. This study enhanced our understanding of how intercropping soybeans in a young tea plantation improves the photosynthesis activity to promote tea seedlings’ growth and development.

Doxorubicin (Dox)-induced cardiotoxicity presents a significant challenge to fully harnessing its chemotherapeutic potential. Crocetin (Cro), a dicarboxylic acid found in the crocus flower and gardenia fruit, has shown remarkable antioxidant and anti-inflammatory activities. However, its poor aqueous solubility and limited cellular uptake severely constrain its further application in treating diseases. In this study, we developed Arg-Gly-Asp (RGD) peptide-decorated nanomicelles delivering Cro to alleviate Dox-induced cardiac injury. The RGD@M(Cro) nanomicelles exhibited excellent aqueous solubility and a drug-loading efficiency of 93.3 %. RGD-decorated micelles could enhance the cellular uptake of Cro in cardiomyocytes and inhibit approximately 60 % of HL-1 cell apoptosis through efficient reactive oxygen species (ROS) scavenging. In a cardiomyopathy mouse model, RGD@M(Cro) substantially reduced cardiac damage and improved cardiac indicators. This study highlights the great potential of RGD-decorated micelles in treating cardiac injury and other diseases.

Fungal attacks have become a major obstacle in tea plantations. Colletotrichum gloeosporioides is one of the most devastating fungal pathogens in tea plantations that can severely affect tea yield and quality. However, the molecular mechanism of resistance genes involved in anthracnose is still largely unknown in tea plants. Here, we found that the laccase gene CsLAC37 was involved in the response to fungal infection based on a transcriptome analysis. The full-length CDS of CsLAC37 was cloned, and its protein sequence had the closest relationship with the Arabidopsis AtLAC15 protein compared to other AtLACs. Tissue-specific expression analysis showed that CsLAC37 had higher expression levels in mature leaves and stems than in the other tissues. Subcellular localization showed that the CsLAC37 protein was predominantly localized in the cell membrane. The expression levels of CsLAC37 were upregulated at different time points under cold, salt, SA, and ABA treatments. qRT-PCR confirmed that CsLAC37 responded to both Pestalotiopsis-like species and C. gloeosporioides infections. Functional validation showed that the hydrogen peroxide (H₂O₂) content increased significantly, and POD activity decreased in leaves after antisense oligonucleotide (AsODN) treatment compared to the controls. The results demonstrated that CsLAC37 may play an important role in resistance to anthracnose, and the findings provide a theoretical foundation for molecular breeding of tea varieties with resistance to fungal diseases.