Molecular regulation of anthocyanin discoloration under water stress and high solar irradiance in pluckable shoots of purple tea cultivar.

MAIN CONCLUSION: During water-deficit stress, antioxidant enzymes use anthocyanin molecules as co-substrates to scavenge for reactive oxygen species leading to reduced anthocyanin content and ultimately loss of purple leaf pigmentation in tea. Anthocyanins are an important class of flavonoids responsible for liquor color and market acceptability of processed tea from the anthocyanin-rich purple tea cultivar 'TRFK 306'. However, the color in pluckable shoots fade and turn green during the dry and hot season, before rapidly reverting back to purple when weather is favorably wet and cool/cold. Our study revealed that loss of purple leaf pigmentation correlated well with reduced precipitation, high soil water-deficit, increased intensity and duration of sunlight and temperature. Richly purple pigmented leaves harvested during the cool, wet conditions recorded significantly higher anthocyanin content compared to faded samples harvested during the dry season. Similarly, individual anthocyanins were affected by seasonal changes with malvidin being the most abundant. Comparative transcriptomics of two RNA-seq libraries, dry/discolored and wet/colored seasons, revealed depression of most metabolic processes related to anthocyanin accumulation in dry conditions. Specifically, transcripts encoding pathway regulators, MYB-bHLH-WD40 (MBW) complex, were repressed possibly contributing to the suppression of late biosynthetic genes of the pathway. Further, suppression of anthocyanin transport genes could be linked to reduced accumulation of anthocyanin in the vacuole during the dry season. However, slight increase in expression of some transporter and reactive oxygen species (ROS) antioxidant genes in the discolored leaf suggests non-enzymatic degradation of anthocyanin, ultimately leading to loss of purple color during the dry season. Based on increased expression of ROS antioxidant genes (especially catalase and superoxide dismutase) in the discolored leaf, we speculate that anthocyanins are used as co-substrates by antioxidant enzymes to scavenge for ROS (especially hydrogen peroxide) that escape from organelles, leading to reduced anthocyanins and loss of pigmentation during the dry season.

Underpinning the molecular programming attributing heat stress associated thermotolerance in tea (Camellia sinensis (L.) O. Kuntze).

The most daunting issue of global climate change is the deleterious impact of extreme temperatures on tea productivity and quality, which has resulted in a quest among researchers and growers. The current study aims to unravel molecular programming underpinning thermotolerance by characterizing heat tolerance and sensitivity response in 20 tea cultivars. The significantly higher negative influence of heat stress was recorded in a sensitive cultivar with reduced water retention (47%), chlorophyll content (33.79%), oxidation potential (32.48%), and increase in membrane damage (76.4%). Transcriptional profiling of most tolerant and sensitive cultivars identified 78 differentially expressed unigenes with chaperon domains, including low and high molecular weight heat shock protein (HSP) and heat shock transcription factors (HSFs) involved in heat shock response (HSR). Further, predicted transcriptional interactome network revealed their key role in thermotolerance via well-co-ordinated transcriptional regulation of aquaporins, starch metabolism, chlorophyll biosynthesis, calcium, and ethylene mediated plant signaling system. The study identified the key role of HSPs (CsHSP90) in regulating HSR in tea, wherein, structure-based molecular docking revealed the inhibitory role of geldanamycin (GDA) on CsHSP90 by blocking ATP binding site at N-terminal domain of predicted structure. Subsequently, GDA mediated leaf disc inhibitor assay further affirmed enhanced HSR with higher expression of CsHSP17.6, CsHSP70, HSP101, and CsHSFA2 genes in tea. Through the current study, efforts were made to extrapolate a deeper understanding of chaperons mediated regulation of HSR attributing thermotolerance in tea.

Transcriptional analysis reveals key insights into seasonal induced anthocyanin degradation and leaf color transition in purple tea (Camellia sinensis (L.) O. Kuntze).

Purple-tea, an anthocyanin rich cultivar has recently gained popularity due to its health benefits and captivating leaf appearance. However, the sustainability of purple pigmentation and anthocyanin content during production period is hampered by seasonal variation. To understand seasonal dependent anthocyanin pigmentation in purple tea, global transcriptional and anthocyanin profiling was carried out in tea shoots with two leaves and a bud harvested during in early (reddish purple: S1_RP), main (dark gray purple: S2_GP) and backend flush (moderately olive green: S3_G) seasons. Of the three seasons, maximum accumulation of total anthocyanin content was recorded in S2_GP, while least amount was recorded during S3_G. Reference based transcriptome assembly of 412 million quality reads resulted into 71,349 non-redundant transcripts with 6081 significant differentially expressed genes. Interestingly, key DEGs involved in anthocyanin biosynthesis [PAL, 4CL, F3H, DFR and UGT/UFGT], vacuolar trafficking [ABC, MATE and GST] transcriptional regulation [MYB, NAC, bHLH, WRKY and HMG] and Abscisic acid signaling pathway [PYL and PP2C] were significantly upregulated in S2_GP. Conversely, DEGs associated with anthocyanin degradation [Prx and lac], repressor TFs and key components of auxin and ethylene signaling pathways [ARF, AUX/IAA/SAUR, ETR, ERF, EBF1/2] exhibited significant upregulation in S3_G, correlating positively with reduced anthocyanin content and purple coloration. The present study for the first-time elucidated genome-wide transcriptional insights and hypothesized the involvement of anthocyanin biosynthesis activators/repressor and anthocyanin degrading genes via peroxidases and laccases during seasonal induced leaf color transition in purple tea. Futuristically, key candidate gene(s) identified here can be used for genetic engineering and molecular breeding of seasonal independent anthocyanin-rich tea cultivars.

Multiple-genotypes transcriptional analysis revealed candidates genes and nucleotide variants for improvement of quality characteristics in tea (Camellia sinensis (L.) O. Kuntze).

Tea quality is a polygenic trait that exhibits tremendous genetic variability due to accumulation of array of secondary metabolites. To elucidate global molecular insights controlling quality attributes, metabolite profiling and transcriptome sequencing of twelve diverse tea cultivars was performed in tea shoots harvested during quality season. RP-HPLC-DAD analysis of quality parameters revealed significant difference in catechins, theanine and caffeine contents. Transcriptome sequencing resulted into 50,107 non-redundant transcripts with functional annotations of 81.6% (40,847) of the transcripts. Interestingly, 2872 differentially expressed transcripts exhibited significant enrichment in 38 pathways (FDR ≤ 0.05) including secondary metabolism, amino acid and carbon metabolism. Thirty-eight key candidates reportedly involved in biosynthesis of fatty acid derived volatiles, volatile terpenes, glycoside hydrolysis and key quality related pathways (flavonoid, caffeine and theanine-biosynthesis) were highly expressed in catechins-rich tea cultivars. Furthermore, enrichment of candidates involved in flavonoid biosynthesis, transcriptional regulation, volatile terpene and biosynthesis of fatty acid derived volatile in Protein-Protein Interactome network revealed well-coordinated regulation of quality characteristics in tea. Additionally, ascertainment of 23,649 non-synonymous SNPs and validation of candidate SNPs present in quality related genes suggests their potential utility in genome-wide mapping and marker development for expediting breeding of elite compound-rich tea cultivars.

Combating Climate Change in the Kenyan Tea Industry.

Climate change triggered by global warming poses a major threat to agricultural systems globally. This phenomenon is characterized by emergence of pests and diseases, extreme weather events, such as prolonged drought, high intensity rains, hailstones and frosts, which are becoming more frequent ultimately impacting negatively to agricultural production including rain-fed tea cultivation. Kenya is predominantly an agricultural based economy, with the tea sector generating about 26% of the total export earnings and about 4% gross domestic product (GDP). In the recent years, however, the country has witnessed unstable trends in tea production associated with climate driven stresses. Toward mitigation and adaptation of climate change, multiple approaches for impact assessment, intensity prediction and adaptation have been advanced in the Kenyan tea sub-sector. Further, pressure on tea breeders to release improved climate-compatible cultivars for the rapidly deteriorating environment has resulted in the adoption of a multi-targeted approach seeking to understand the complex molecular regulatory networks associated with biotic and abiotic stresses adaptation and tolerance in tea. Genetic modeling, a powerful tool that assists in breeding process, has also been adopted for selection of tea cultivars for optimal performance under varying climatic conditions. A range of physiological and biochemical responses known to counteract the effects of environmental stresses in most plants that include lowering the rates of cellular growth and net photosynthesis, stomatal closure, and the accumulation of organic solutes such as sugar alcohols, or osmolytes have been used to support breeding programs through screening of new tea cultivars suitable for changing environment. This review describes simulation models combined with high resolution climate change scenarios required to quantify the relative importance of climate change on tea production. In addition, both biodiversity and ecosystem based approaches are described as a part of an overall adaptation strategy to mitigate adverse effects of climate change on tea in Kenya and gaps highlighted for urgent investigations.