Intercropping, a green and sustainable planting pattern, has demonstrated positive effects on plant growth and the soil environment. However, there is currently little research on the influence of intercropping leguminous plants and using them as green manure on the soil environment and tea quality. During the profuse flowering period of Chinese milkvetch, the contents of tea amino acids and soluble sugar in intercropping tea plants with soybean increased by 6.89 and 54.58%. Moreover, there was 27.42% increase in soil ammonium nitrogen and 21.63% increase in available nitrogen. When Chinese milkvetch was returned to soil for 1 month during its profuse flowering period, the soybean and Chinese milkvetch as green manure enhanced tea amino acids and soluble sugar by 9.11 and 33.96%, and soil ammonium nitrogen, nitrate nitrogen and available nitrogen increased by 25.04, 77.84, and 48.90%. Intercropping systems also have positive effects on tea quality components, soil fertility, and soil microbial communities during the profuse flowering period of soybeans and when soybeans with this period were returned to the field for 1 month. Furthermore, the soil fertility index was significantly increased, especially in the intercropping system of tea–soybean–Chinese milkvetch. The soil bacterial community complexity and fungal community interactions were significantly increased. Soil pH, nitrate nitrogen, and available phosphorus were found to be crucial influencing factors on soil microbial communities, specifically bacterial communities. These results highlight the significance of optimizing intercropping systems to improve the soil environment and tea quality components. They also provide a theoretical foundation for promoting the sustainable development of tea plantations.

Low-temperature stress is an increasing problem for the cultivation of tea (Camellia sinensis), with adverse effects on plant growth and development and subsequent negative impacts on the tea industry. Methyl jasmonate (MeJA), as a plant inducer, can improve the cold-stress tolerance in tea plants. R2R3-MYB transcription factors (TFs) are considered potentially important regulators in the resistance to cold stress in plants. However, the molecular mechanisms, by which MYB TFs via the jasmonic acid pathway respond to cold stress in the tea plant, remain unknown. In this study, physiological and biochemical assays showed that exogenous MeJA application could effectively promote ROS scavenging in the tea plant under cold stress, maintaining the stability of the cell membrane. Sixteen R2R3-MYB TFs genes were identified from the tea plant genome database. Quantitative RT-PCR analysis showed that three CsMYB genes were strongly induced under a combination of MeJA and cold-stress treatment. Subcellular localization assays suggest CsMYB45, CsMYB46, and CsMYB105 localized in the nucleus. Exogenous MeJA treatment enhanced the overexpression of CsMYB45, CsMYB46, and CsMYB105 in E. coli and improved the growth and survival rates of recombinant cells compared to an empty vector under cold stress. Yeast two-hybrid and bimolecular fluorescence complementation experiments confirmed that CsMYB46 and CsMYB105 interacted with CsJAZ3, CsJAZ10, and CsJAZ11 in the nucleus. Taken together, these results highlight that CsMYB45, CsMYB46, and CsMYB105 are not only key components in the cold-stress signal response pathway but also may serve as points of confluence for cold stress and JA signaling pathways. Furthermore, our findings provide new insight into how MYB TFs influence cold tolerance via the jasmonic acid pathway in tea and provide candidate genes for future functional studies and breeding.

Microorganisms play essential roles in soil-ecosystem multifunctionality. However, the contribution of their community assembly processes, composition, diversity, and keystone species to ecosystem multifunctionality is unclear, especially in tea-plantation ecosystems. In order to assess the effects of various intercropping patterns (tea-plant monoculture and tea plants, respectively, intercropped with soybean, soybean—milk vetch, soybean—red clover, and soybean—smooth vetch) on soil rare and abundant taxa, a field experiment was carried out. We found that tea plantation intercropping with legumes improved the soil-ecosystem multifunctionality by altering the soil environment, and ultimately benefited nutrient absorption and quality improvement of tea leaves. Whether it was in bacteria or fungi, rare taxa had a higher proportion of deterministic processes in community assembly than abundant taxa. Additionally, intercropping practices changed the soil environment, and rare bacterial taxa were assembled and shifted from variable selection to homogeneous dispersal. Intercropping practices significantly changed the bacterial and fungal communities’ composition, and rare taxa had higher α-diversity than abundant taxa. Increasing legume species in intercropping practice enhanced community dissimilarity to the tea monoculture by affecting soil pH, ammonium nitrogen, and nitrate nitrogen. Rare bacterial and fungal β-diversity exhibited stronger positive relationships with ecosystem multifunctionality (both average and multi-threshold approaches) compared to the corresponding abundant taxa. Furthermore, ecosystem multifunctionality under different intercropping practices was closely related to the keystone rare operational taxonomic units, especially rare bacterial species of Chloroflexi. Our results emphasize the disparate feedbacks of rare and abundant taxa to diverse intercropping practices, as well as the important connection between rare bacterial taxa and ecosystem multifunctionality.

Plant resistance inducers represent an alternative strategy that mitigate stress-induced damage in plants. Previously, 2-amino-3-methylhexanoic acid (AMHA), a novel natural plant resistance inducer, was shown to significantly bolster cold tolerance, thermotolerance, and pathogen resistance in plants. However, the intricate mechanisms underlying AMHA’s response to cold stress remain elusive. Thus, we investigated the physiological and transcriptomic analyses of AMHA pretreatment on tea plant to determine its substantial role of AMHA under cold stress. The results showed that pretreatment with 100 nM AMHA effectively mitigated the detrimental effects of cold stress on photosynthesis and growth. Furthermore, differentially expressed genes were identified through RNA-seq during pretreatment, cold stress, and 2 days of recovery. These genes were mainly enriched in pathways related to flavonoid/anthocyanin, carotenoid, and ascorbic acid-glutathione (AsA-GSH) cycle, including GST (encoding glutathione S-transferase). Potential regulatory relationships between the identified genes and transcription factors were also established. Antisense oligodeoxynucleotide-silencing and overexpression experiments revealed that CsGSTU7 enhances cold resistance by maintaining redox homeostasis. In conclusion, our study suggests that antioxidant-related signaling molecules play a critical role in the signaling cascades and transcriptional regulation mediating AMHA-induced cold-stress resistance in tea plant.

The elite germplasm resources are key to the beautiful appearance and pleasant flavor of Biluochun tea. We collected and measured the agronomic traits of 95 tea plants to reveal the trait diversity and breeding value of Biluochun tea plant populations. The results revealed that the agronomic traits of Biluochun tea plant populations were diverse and had high breeding value. Additionally, we resequenced these tea plant populations to reveal genetic diversity, population structure, and selection pressure. The Biluochun tea plant populations contained two groups and were least affected by natural selection based on the results of population structure and selection pressure. More importantly, four non-synonymous single nucleotide polymorphisms (nsSNPs) and candidate genes associated with (−)-gallocatechin gallate (GCG), (−)-gallocatechin (GC), and caffeine (CAF) were detected using at least two GWAS models. The results will promote the development and application of molecular markers and the utilization of elite germplasm from Biluochun populations.