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.

From the Wild to the Cup: Tracking Footprints of the Tea Species in Time and Space.

Tea is one of the world's most popular beverages, known for its cultural significance and numerous health benefits. A clear understanding of the origin and history of domestication of the tea species is a fundamental pre-requisite for effective germplasm conservation and improvement. Though there is a general consensus about the center of origin of the tea plant, the evolutionary origin and expansion history of the species remain shrouded in controversy, with studies often reporting conflicting findings. This mini review provides a concise summary of the current state of knowledge regarding the origin, domestication, and dissemination of the species around the world. We note that tea was domesticated around 3000 B.C. either from non-tea wild relatives (probably Camellia grandibracteata and/or C. leptophylla) or intra-specifically from the wild Camellia sinensis var. assamica trees, and that the genetic origins of the various tea varieties may need further inquiry. Moreover, we found that lineage divergence within the tea family was apparently largely driven by a combination of orogenic, climatic, and human-related forces, a fact that could have important implications for conservation of the contemporary tea germplasm. Finally, we demonstrate the robustness of an integrative approach involving linguistics, historical records, and genetics to identify the center of origin of the tea species, and to infer its history of expansion. Throughout the review, we identify areas of debate, and highlight potential research gaps, which lay a foundation for future explorations of the topic.

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.

Functional annotation of putative QTL associated with black tea quality and drought tolerance traits.

The understanding of black tea quality and percent relative water content (%RWC) traits in tea (Camellia sinensis) by a quantitative trait loci (QTL) approach can be useful in elucidation and identification of candidate genes underlying the QTL which has remained to be difficult. The objective of the study was to identify putative QTL controlling black tea quality and percent relative water traits in two tea populations and their F1 progeny. A total of 1,421 DArTseq markers derived from the linkage map identified 53 DArTseq markers to be linked to black tea quality and %RWC. All 53 DArTseq markers with unique best hits were identified in the tea genome. A total of 5,592 unigenes were assigned gene ontology (GO) terms, 56% comprised biological processes, cellular component (29%) and molecular functions (15%), respectively. A total of 84 unigenes in 15 LGs were assigned to 25 different Kyoto Encyclopedia of Genes and Genomes (KEGG) database pathways based on categories of secondary metabolite biosynthesis. The three major enzymes identified were transferases (38.9%), hydrolases (29%) and oxidoreductases (18.3%). The putative candidate proteins identified were involved in flavonoid biosynthesis, alkaloid biosynthesis, ATPase family proteins related to abiotic/biotic stress response. The functional annotation of putative QTL identified in this current study will shed more light on the proteins associated with caffeine and catechins biosynthesis and % RWC. This study may help breeders in selection of parents with desirable DArTseq markers for development of new tea cultivars with desirable traits.

Multiple origins and a narrow genepool characterise the African tea germplasm: concordant patterns revealed by nuclear and plastid DNA markers.

Despite the highly economic value of tea in Africa, its genetic and geographic origins remain largely unexplored. Here we address this by collecting 439 samples across 11 countries in Africa and Asia to investigate the origin and genepool composition of African tea based on 23 nuclear microsatellites loci (nSSRs) and three cpDNA intergenic spacer regions. Our results indicated that the African tea represents a potpourri originating from multiple introductions over time. The nSSR analysis revealed that the majority (79%) of tea accessions collected in Africa belong to Indian Assam tea which have likely originated from India and/or Sri Lanka. The patterns of nSSR variation also showed that Chinese Assam tea is genetically distinct from Indian Assam tea, and has rarely been used in African tea breeding efforts since only 4% of the African tea accessions possessed this genotype. We found a total of 22 cpDNA haplotypes, which grouped into three main geographic clades that were concordant with the distribution of microsatellite genotypes. Several private cpDNA haplotypes were identified in Chinese Assam tea in Southern Yunnan province of China. Therefore Chinese Assam tea will be important for the enrichment of African tea gene pools. Our results is a useful guide in future tea breeding programmes in Africa.

Insights into the Genetic Relationships and Breeding Patterns of the African Tea Germplasm Based on nSSR Markers and cpDNA Sequences.

Africa is one of the key centers of global tea production. Understanding the genetic diversity and relationships of cultivars of African tea is important for future targeted breeding efforts for new crop cultivars, specialty tea processing, and to guide germplasm conservation efforts. Despite the economic importance of tea in Africa, no research work has been done so far on its genetic diversity at a continental scale. Twenty-three nSSRs and three plastid DNA regions were used to investigate the genetic diversity, relationships, and breeding patterns of tea accessions collected from eight countries of Africa. A total of 280 African tea accessions generated 297 alleles with a mean of 12.91 alleles per locus and a genetic diversity (H S) estimate of 0.652. A STRUCTURE analysis suggested two main genetic groups of African tea accessions which corresponded well with the two tea types Camellia sinensis var. sinensis and C. sinensis var. assamica, respectively, as well as an admixed "mosaic" group whose individuals were defined as hybrids of F2 and BC generation with a high proportion of C. sinensis var. assamica being maternal parents. Accessions known to be C. sinensis var. assamica further separated into two groups representing the two major tea breeding centers corresponding to southern Africa (Tea Research Foundation of Central Africa, TRFCA), and East Africa (Tea Research Foundation of Kenya, TRFK). Tea accessions were shared among countries. African tea has relatively lower genetic diversity. C. sinensis var. assamica is the main tea type under cultivation and contributes more in tea breeding improvements in Africa. International germplasm exchange and movement among countries within Africa was confirmed. The clustering into two main breeding centers, TRFCA, and TRFK, suggested that some traits of C. sinensis var. assamica and their associated genes possibly underwent selection during geographic differentiation or local breeding preferences. This study represents the first step toward effective utilization of differently inherited molecular markers for exploring the breeding status of African tea. The findings here will be important for planning the exploration, utilization, and conservation of tea germplasm for future breeding efforts in Africa.