Sugars Will Eventually be Exported Transporter (SWEET) and SemiSWEET are recently characterized families of sugar transporters in eukaryotes and prokaryotes, respectively. SemiSWEETs contain 3 transmembrane helices (TMHs), while SWEETs contain 7. Here, we performed sequence-based comprehensive analyses for SWEETs and SemiSWEETs across the biosphere. In total, 3,249 proteins were identified and ≈60% proteins were found in green plants and Oomycota, which include a number of important plant pathogens. Protein sequence similarity networks indicate that proteins from different organisms are significantly clustered. Of note, SemiSWEETs with 3 or 4 TMHs that may fuse to SWEET were identified in plant genomes. 7-TMH SWEETs were found in bacteria, implying that SemiSWEET can be fused directly in prokaryote. 15-TMH extraSWEET and 25-TMH superSWEET were also observed in wild rice and oomycetes, respectively. The transporters can be classified into 4, 2, 2, and 2 clades in plants, Metazoa, unicellular eukaryotes, and prokaryotes, respectively. The consensus and coevolution of amino acids in SWEETs were identified by multiple sequence alignments. The functions of the highly conserved residues were analyzed by molecular dynamics analysis. The 19 most highly conserved residues in the SWEETs were further confirmed by point mutagenesis using SWEET1 from Arabidopsis thaliana. The results proved that the conserved residues located in the extrafacial gate (Y57, G58, G131, and P191), the substrate binding pocket (N73, N192, and W176), and the intrafacial gate (P43, Y83, F87, P145, M161, P162, and Q202) play important roles for substrate recognition and transport processes. Taken together, our analyses provide a foundation for understanding the diversity, classification, and evolution of SWEETs and SemiSWEETs using large-scale sequence analysis and further show that gene duplication and gene fusion are important factors driving the evolution of SWEETs.

γ-Aminobutyric acid (GABA) is an important bioactive component of tea (Camellia sinensis) providing various health benefits. We studied GABA accumulation via the GABA shunt and polyamine degradation pathways under anoxia in tea leaves. Anoxia caused a ∼20-fold increment in GABA concentration, relative to fresh tea leaves. This increment was due to the increase of glutamate decarboxylase and diamine oxidase activities. Genes involved in GABA formation, such as CsGAD1 and CsGAD2, were significantly up-regulated by anoxia. The concentrations of putrescine and spermine, two substrates for GABA production, were also increased by anoxia. Treating tea leaves with aminoguanidine completely inhibited diamine oxidase activity during anoxia, but the concentration of GABA decreased by only ∼25%. We infer that about one-fourth of GABA formed in tea leaves under anoxia comes from the polyamine degradation pathway, opening the possibility of producing GABA tea based through the regulation of metabolism.

To determine the mechanisms in tea plants responding to temperature stresses (heat and cold), we examined the global transcriptomic and metabolomic profiles of the tea plant cultivar ‘Suchazao’ under moderately low temperature stress (ML), severely low temperature stress (SL), moderately high temperature stress (MH) and severely high temperature stress (SH) using RNA-seq and high performance liquid chromatography tandem mass spectrometry/mass spectrometry (HPLC-MS/MS), respectively. The identified differentially expressed genes indicated that the synthesis of stress-resistance protein might be redirected to cope with the temperature stresses. We found that heat shock protein genes Hsp90 and Hsp70 played more critical roles in tea plants in adapting to thermal stress than cold, while late embryogenesis abundant protein genes (LEA) played a greater role under cold than heat stress, more types of zinc finger genes were induced under cold stress as well. In addition, energy metabolisms were inhibited by SH, SL and ML. Furthermore, the mechanisms of anthocyanin synthesis were different under the cold and heat stresses. Indeed, the CsUGT75C1 gene, encoding UDP-glucose:anthocyanin 5-O-glucosyl transferase, was up-regulated in the SL-treated leaves but down-regulated in SH. Metabolomics analysis also showed that anthocyanin monomer levels increased under SL. These results indicate that the tea plants share certain foundational mechanisms to adjust to both cold and heat stresses. They also developed some specific mechanisms for surviving the cold or heat stresses. Our study provides effective information about the different mechanisms tea plants employ in surviving cold and heat stresses, as well as the different mechanisms of anthocyanin synthesis, which could speed up the genetic breeding of heat- and cold-tolerant tea varieties.

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Tea plant is an economically important crop in China, but long-term monoculture and substantial chemical nitrogen fertilizer input cause soil acidification, which in turn affects the nutrient supply and tea quality. Intercropping has drawn more attention in tea gardens because this pattern is expected to improve soil fertility and tea quality and change the soil microbial community composition. However, the roles of some key microorganisms in rhizosphere soils have not been well characterized. Hereby, a “soybean in summer and smooth vetch in winter” mode was selected to investigate the effects of intercropped legumes in a tea garden on soil fertility, tea quality, and the potential changes in beneficial bacteria such as Bacillus. Our data showed that when soybeans were turned into soil, intercropping system exhibited higher soil organic matter (SOM), total nitrogen (TN), tea quality indices and the expression of Camellia sinensis glutamine synthetase gene (CsGS). Notably, intercropping significantly affected the bacterial communities and decreased the relative abundance of Bacillus but increased its absolute abundance. Bacillus amyloliquefaciens BM1 was isolated from intercropped soil and showed outstanding plant growth-promoting (PGP) properties when coinoculated with rhizobia. In winter, intercropping with smooth vetch had a beneficial effect on soil properties and tea quality. Comparably, coinoculation with strain BM1 and Rhizobium leguminosarum Vic5 on smooth vetch (Vicia villosa) showed huge improvements in SOM, TN and quality of tea leaves, accompanied by the highest level of amino acids and lowest levels of polyphenol and caffeine (p < 0.05). According to these results, our findings demonstrate that intercropping with some legumes in the tea garden is a strategy that increases SOM, TN and tea quality, and some PGP Bacillus species are optional to obtain an amplification effect.