Carotenoids play an important role in many physiological processes in plants and the phytoene desaturase gene (PDS3) encodes one of the important enzymes in the carotenoid biosynthesis pathway. Here we report the identification and analysis of a T-DNA insertion mutant of PDS3 gene. Functional complementation confirmed that both the albino and dwarf phenotypes of the pds3 mutant resulted from functional disruption of the PDS3 gene. Chloroplast development was arrested at the proplastid stage in the pds3 mutant. Further analysis showed that high level of phytoene was accumulated in the pds3 mutant. Addition of exogenous GA3 could partially rescue the dwarf phenotype, suggesting that the dwarf phenotype of the pds3 mutant might be due to GA deficiency. Microarray and RT-PCR analysis showed that disrupting PDS3 gene resulted in gene expression changes involved in at least 20 metabolic pathways, including the inhibition of many genes in carotenoid, chlorophyll, and GA biosynthesis pathways. Our data suggest that the accumulated phytoene in the pds3 mutant might play an important role in certain negative feedbacks to affect gene expression of diverse cellular pathways.

Disruption of Phytoene Desaturase Gene Results in Albino and Dwarf Phenotypes in Arabidopsis by Impairing ChlorophyII,Carotenoid,and Gibberellin Biosynthesis

Free amino acids, including theanine, glutamine and glutamate, contribute greatly to the pleasant taste and multiple health benefits of tea. Amino acids in tea plants are mainly synthesized in roots and transported to new shoots, which are significantly affected by nitrogen (N) level and forms. However, the regulatory amino acid metabolism genes have not been systemically identified in tea plants. Here, we investigated the dynamic changes of free amino acid contents in response to N deficiency and forms in tea plant roots, and systemically identified the genes associated amino acid contents in individual metabolism pathways. Our results showed that glutamate-derived amino acids are the most dynamic in response to various forms of N and N deficiency. We then performed transcriptomic analyses of roots treated with N deficiency and various forms of N, and differentially expressed amino acid metabolic genes in each pathway were identified. The analyses on expression patterns and transcriptional responses of metabolic genes to N treatments provided novel insights for the molecular basis of high accumulation of theanine in tea plant root. These analyses also identified potential regulatory genes in dynamic amino acid metabolism in tea plant root. Furthermore, our findings indicated that the dynamic expression levels of CsGDH, CsAlaDC, CsAspAT, CsSDH, CsPAL, CsSHMT were highly correlated with changes of amino acid contents in their corresponding pathways. Herein, this study provides comprehensive insights into transcriptional regulation of amino acid metabolism in response to nitrogen deficiency and nitrogen forms in tea plant root.

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Transcriptional regulation of amino acid metabolism in response to nitrogen deficiency and nitrogen forms in tea plant root (Camellia sinensis L.).

Chloroplast development and chlorophyll metabolism have been well described in model plants but not in perennial woody crops. Of particular interest is the interplay between light and hormones under shade conditions. We report that the shade induced accumulation of chlorophylls in Camellia sinensis cv. Shuchazao leaves is at least as a result of (a) positive changes in chloroplast development and (b) light/hormonal regulation of genes and transcription factors involved in the chlorophyll biosynthesis pathway. Under shade conditions, leaves developed an abundance of enlarged chloroplasts encapsulating more prominent thylakoid membranes. Four major metabolites in the chlorophyll biosynthesis pathway namely Chl a, Chl b, DPP, and Mg-Proto IX increased under shade conditions while PBG decreased significantly. Significant changes were found at the transcription level of regulators of chloroplast biogenesis (GLK1 and LHCB), the structural genes in the chlorophyll biosynthesis pathway (HEMA1, CLH1, PORA, and CAO) and potential components involved in light signaling (PHYA, CRY1, HY5, and DELLAs). Two central signal integrators (GLK1 and LHCB) between the nucleus and chloroplast showed clear responses to shade, suggesting a crucial role of light in regulating chloroplast development in tea leaves. Concurrent with the changes in gene expression, the concentrations of endogenous phytohormones (auxin, cytokinin, and gibberellins) increased significantly in the later stages of shade conditions. Two key integrators involved in the hormone signal pathways, EIN3 and EBF1/2, increased under shade conditions suggesting that shade induced changes to hormone levels may play some role in modulating chlorophyll biosynthesis in the tea leaves. Overall, this data suggests that the light and hormone influence over chloroplast development and chlorophyll biosynthesis in Camellia is similar to that of Arabidopsis. This study provides new insights into the molecular mechanisms that regulate chlorophyll biosynthesis in response to light and hormones in a commercially important woody plant such as Camellia, which may facilitate the breeding of high-chlorophyll tea cultivars for the improvement of sensory features of the green tea product.

/pdf/from-chloroplast-biogenesis-to-chlorophyll-accumulation-the-2txe94vk73.pdf

From Chloroplast Biogenesis to Chlorophyll Accumulation: The Interplay of Light and Hormones on Gene Expression in Camellia sinensis cv. Shuchazao Leaves.

Endophytic fungi and bacteria are the most ubiquitous and representative commensal members that have been studied so far in various higher plants. Within colonization and interaction with their host plants, endophytic microbiota are reportedly to modulate not only the host's growth but also holobiont resilience to abiotic and biotic stresses, providing a natural reservoir and a promising solution for sustainable agricultural development challenged by global climate change. Moreover, possessing the talent to produce a wide array of high-value natural products, plant endophytic microbiota also serve as an alternative way for novel drug discovery. In this review, tea, one of the world's three largest nonalcoholic beverages and a worldwide economic woody crop, was highlighted in the context of endophytic microbiota. We explore the recent studies regarding isolation approaches, distribution characteristics and diversity, and also biological functions of endophytic microbiota in Camellia sinensis (L.) O. Kuntze. Profoundly, the future insight into interaction mechanism between endophytic microbiota and tea plants will shed light on in-depth exploration of tea microbial resources.

https://www.tandfonline.com/doi/pdf/10.1080/21655979.2020.1816788

Implications of endophytic microbiota in Camellia sinensis: a review on current understanding and future insights

Multi-omics research in albino tea plants: Past, present, and future

A normal tea plant with one albino branch was discovered. RNA sequencing, albinism phenotype and ultrastructural observations provided a valuable understanding of the albino mechanism in tea plants. Tea plants with a specific color (white or yellow) have been studied extensively. A normal tea plant (Camellia sinensis cv. quntizhong) with one albino branch was discovered in a local tea plantation in Huangshan, Anhui, China. The pure albino leaves on this special branch had accumulated a fairly high content of amino acids, especially theanine (45.31 mg/g DW), and had a low concentration of polyphenols and an extremely low chlorophyll (Chl) content compared with control leaves. Ultrastructural observation of an albino leaf revealed no chloroplasts, whereas it was viable in the control leaf. RNA sequencing and differentially expressed gene (DEG) analysis were performed on the albino leaves and on control leaves from a normal green branch. The related genes involved in theanine and polyphenol biosynthesis were also investigated in this study. DEG expression patterns in Chl biosynthesis or degradation, carotenoid biosynthesis or degradation, chloroplast development, and biosynthesis were influenced in the albino leaves. Chloroplast deletion in albino leaves had probably destroyed the balance of carbon and nitrogen metabolism, leading to a high accumulation of free amino acids and a low concentration of polyphenols in the albino leaves. The obtained results can provide insight into the mechanism underlying this special albino branch phenotype, and are a valuable contribution toward understanding the albino mechanism in tea plants.

Significantly increased amino acid accumulation in a novel albino branch of the tea plant (Camellia sinensis)

Methyl jasmonate (MeJA), a volatile organic compound, is a principal flowery aromatic compound in tea. During the processing of black tea, MeJA is produced by jasmonic acid carboxyl methyltransferase (JMT) of the jasmonic acid (JA) substrate, forming a specific floral fragrance. CsJMT was cloned from tea leaves; the three-dimensional structure of CsJMT was predicted. Enzyme activity was identified, and protein purification was investigated. Site-directed deletions revealed that N-10, S-22, and Q-25 residues in the beginning amino acids played a key functional role in enzyme activity. The expression patterns of CsJMT in tea organs differed; the highest expression of CsJMT was observed in the fermentation process of black tea. These results aid in further understanding the synthesis of MeJA during black tea processing, which is crucial for improving black tea quality using specific fragrances and could be applied to the aromatic compound regulation and tea breeding improvement in further studies.

Molecular and Biochemical Characterization of Jasmonic Acid Carboxyl Methyltransferase Involved in Aroma Compound Production of Methyl Jasmonate during Black Tea Processing

Bacteria survived and thrived inside any part of plant without damaging its host are called endophytic bacteria. Many studies have focused on plant–endophytic bacterium interactions and their Plant-growth-promoting (PGP) abilities. Albino tea plants are widely investigated because of their high amino acid concentrations. Previous studies mainly elucidated specific metabolite mechanisms in albino tea plants, while the present study identified the variation in diversity and endophytic bacterial abundance in albino and non-albino tea leaves using culture-dependent and culture-independent methods. With different culture media, we identified a total of 110 isolates, of which 58 endophytic bacteria belonged to three phyla (Proteobacteria, Firmicutes, and Actinobacteria). Moreover, we identified a total of 1856 operational taxonomic units as 513 endophytic bacteria belonging to 27 phyla including Proteobacteria, Firmicutes, and Actinobacteria. In addition, comparing to non-albino tea leaves, endophytic bacteria obtained from albino tea leaves showed higher diversity and abundance. Endophytic bacteria isolated from albino leaves also exhibited higher PGP properties than those from non-albino ones, and their PGP effects on rice and tea cuttings were examined. The results indicated differences in endophytic bacteria between albino and non-albino leaves. Specific endophytic bacteria isolated from albino leaves may have a relationship with the albino phenotype. Our study indicated that PGP bacteria isolated from albino leaves are expected to become a plant growth promoter in the future.

Huiyan Jia

Papers

Significantly increased amino acid accumulation in a novel albino branch of the tea plant (Camellia sinensis)

Molecular and Biochemical Characterization of Jasmonic Acid Carboxyl Methyltransferase Involved in Aroma Compound Production of Methyl Jasmonate during Black Tea Processing

Endophytic bacteria from the leaves of two types of albino tea plants, indicating the plant growth promoting properties