scholarly journals Isolation and molecular identification of four culturable endophytes from TV22 clone of Camellia sinensis (L.)

2018 ◽  
Vol 7 (4) ◽  
pp. 2119 ◽  
Author(s):  
Pranjal Pratim Das ◽  
Tapas Medhi

Endophytes are microorganisms presents within plant in asymptomatic manner and often act as a reservoir of novel bioactive secondary metabolites having antimicrobial, anti-insect and other beneficial properties. In absence of any reports on the presence of endophyte and their possible metabolic role in the process of infestation of tea plants (Camellia sinensis L.) by tea mosquito bug (Halopeltis theivora Waterhouse), the present study was undertaken to trace and isolate them from tea leaves for their molecular characterisation based on 16S rRNA sequencing. In this process, four endophytic bacteria have been identified as Brachybacterium sp. strain TMCS1, Bacillus pumilus strain TMCS2, Moraxella osloensis strain TMCS3 and Moraxella osloensis strain TMCS4 from the 2nd leaf of a young flash of C.   sinensis (TV22 clone) which will enable us to study their culturable properties and role as biocontrol agents.

2015 ◽  
Vol 10 (5) ◽  
pp. 1934578X1501000 ◽  
Author(s):  
Wei-Wei Deng ◽  
Min Li ◽  
Chen-Chen Gu ◽  
Da-Xiang Li ◽  
Lin-Long Ma ◽  
...  

Caffeine, a purine alkaloid, is a major secondary metabolite in tea leaves. The demand for low caffeine tea is increasing in recent years, especially for health reasons. We report a novel grafted tea material with low caffeine content. The grafted tea plant had Camellia sinensis as scions and C. oleifera as stocks. The content of purine alkaloids was determined in the leaves of one-year-old grafted tea plants by HPLC. We also characterized caffeine synthase (CS), a key enzyme involved in caffeine biosynthesis in tea plants, at the expression level. The expression patterns of CS were examined in grafted and control leaves by Western blot, using a self-prepared polyclonal antibody with high specificity and sensitivity. The expression of related genes ( TCS1, tea caffeine synthase gene, GenBank accession No. AB031280; sAMS, SAM synthetase gene, AJ277206; TIDH, IMP dehydrogenase gene, EU106658) in the caffeine biosynthetic pathway was investigated by qRT-PCR. HPLC showed that the caffeine content was only 38% as compared with the non-grafted tea leaves. Immunoblotting analysis showed that CS protein decreased by half in the leaves of grafted tea plants. qRT-PCR revealed no significant changes in the expression of two genes in the upstream pathway ( sAMS and TIDH), while the expression of TCS1 was greatly decreased (50%). Taken together, these data revealed that the low caffeine content in the grafted tea leaves is due to low TCS1 expression and CS protein accumulation.


2020 ◽  
Vol 21 (16) ◽  
pp. 5684 ◽  
Author(s):  
Xiaochen Zhou ◽  
Lanting Zeng ◽  
Yingjuan Chen ◽  
Xuewen Wang ◽  
Yinyin Liao ◽  
...  

In tea (Camellia sinensis) plants, polyphenols are the representative metabolites and play important roles during their growth. Among tea polyphenols, catechins are extensively studied, while very little attention has been paid to other polyphenols such as gallic acid (GA) that occur in tea leaves with relatively high content. In this study, GA was able to be transformed into methyl gallate (MG), suggesting that GA is not only a precursor of catechins, but also can be transformed into other metabolites in tea plants. GA content in tea leaves was higher than MG content—regardless of the cultivar, plucking month or leaf position. These two metabolites occurred with higher amounts in tender leaves. Using nonaqueous fractionation techniques, it was found that GA and MG were abundantly accumulated in peroxisome. In addition, GA and MG were found to have strong antifungal activity against two main tea plant diseases, Colletotrichum camelliae and Pseudopestalotiopsis camelliae-sinensis. The information will advance our understanding on formation and biologic functions of polyphenols in tea plants and also provide a good reference for studying in vivo occurrence of specialized metabolites in economic plants.


2019 ◽  
Vol 20 (17) ◽  
pp. 4151 ◽  
Author(s):  
Xuewen Wang ◽  
Lanting Zeng ◽  
Yinyin Liao ◽  
Jianlong Li ◽  
Jinchi Tang ◽  
...  

Herbivore-induced plant volatiles (HIPVs) play important ecological roles in defense against stresses. In contrast to model plants, reports on HIPV formation and function in crops are limited. Tea (Camellia sinensis) is an important crop in China. α-Farnesene is a common HIPV produced in tea plants in response to different herbivore attacks. In this study, a C. sinensis α-farnesene synthase (CsAFS) was isolated, cloned, sequenced, and functionally characterized. The CsAFS recombinant protein produced in Escherichia coli was able to transform farnesyl diphosphate (FPP) into α-farnesene and also convert geranyl diphosphate (GPP) to β-ocimene in vitro. Furthermore, transient expression analysis in Nicotiana benthamiana plants indicated that CsAFS was located in the cytoplasm and could convert FPP to α-farnesene in plants. Wounding, to simulate herbivore damage, activated jasmonic acid (JA) formation, which significantly enhanced the CsAFS expression level and α-farnesene content. This suggested that herbivore-derived wounding induced α-farnesene formation in tea leaves. Furthermore, the emitted α-farnesene might act as a signal to activate antibacterial-related factors in neighboring undamaged tea leaves. This research advances our understanding of the formation and signaling roles of common HIPVs in crops such as tea plants.


Horticulturae ◽  
2021 ◽  
Vol 7 (10) ◽  
pp. 410
Author(s):  
Yingzi Wang ◽  
Qin Yu ◽  
Yinhua Li ◽  
Juan Li ◽  
Jinhua Chen ◽  
...  

Tea [Camellia sinensis (L.)] plants are important economic crop in China. Chilling stress and freezing damages have seriously affected the quality of tea products that have been already regarded as the main restricting factors to industry’s development. Nitric oxide (NO) plays a crucial role in resistance of abiotic stresses. An experiment was conducted in an artificial climate chamber to study the effect of NO on tea plants grown under chilling stress (–2 °C) for 0, 6, 24, 48, and 72 h. Foliar application of sodium nitroprusside (SNP) at a rate of 500 μmol·L−1 was used as NO donor. The experiment contained two factors: the first was the foliar application with SNP or distilled water, and the scond one was the chilling (–2 °C) exposure time (0, 6, 24, 48, and 72 h). The effects of NO on membrane lipid peroxidation, osmotic adjustment substances, and antioxidant activity under cold stress were studied. In addition, the gene expression of CsICE1 and CsCBF1 in respond to NO addition were also investigated using real-time polymerase chain reaction (RT-PCR). The results show that foliar addition of NO (500 μmol·L−1 of SNP) reduce the relative conductivity of tea leaves, inhibits the elevated malondialdehyde content, promotes the accumulation of proline, soluble protein and sugar, and increases the superoxide dismutase, catalase activities, thereby alleviates the damage of cold stress on tea leaves. The CsICE1 expression in 500 μM SNP treatment was peaked at 24 h of low temperature stress, while it did not express at normal temperature. Therefore, the current study is considered a good scientific material in understanding how tea plants sense and defense the chilling stress and that plays an important role to improve the level of production and economic benefits. It is also provided significant theory bas to control chilling stress in tea plants.


2009 ◽  
Vol 66 (5) ◽  
pp. 583-592 ◽  
Author(s):  
Flávia Mandolesi Pereira de Melo ◽  
Marli Fátima Fiore ◽  
Luiz Alberto Beraldo de Moraes ◽  
Maria Estela Silva-Stenico ◽  
Shirlei Scramin ◽  
...  

In the search for new organisms and new secondary metabolites, a study was conducted to evaluate the diversity of endophytic bacteria from ethnovarieties of cassava cultivated by Brazilian Amazon Indian tribes and also to study the secondary metabolites produced by a Bacillus pumilus strain. Sixty seven cassava endophytic bacteria were subjected to 16S rRNA sequencing and FAME analysis. The bacterial profile revealed that 25% of all endophytic isolates belonged to the genus Bacillus. The isolate B. pumilus MAIIIM4a showed a strong inhibitory activity against the fungi Rhizoctonia solani, Pythium aphanidermatum and Sclerotium rolfsii. Secondary metabolites of this strain were extracted using hexane, dichloromethane and ethyl acetate. Extracts were subjected to bioautography and LC/MS analysis, which allowed the identification of pumilacidin, an antifungal compound produced by B. pumilus MAIIIM4a. The bacterial endophytic localization was confirmed by cassava cell tissue examination using scanning electron microscopy.


Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 282
Author(s):  
Jianlong Li ◽  
Yangyang Xiao ◽  
Qian Fan ◽  
Yinyin Liao ◽  
Xuewen Wang ◽  
...  

Salicylic acid (SA) is a well-known immune-related hormone that has been well studied in model plants. However, less attention has been paid to the presence of SA and its derivatives in economic plants, such as tea plants (Camellia sinensis). This study showed that tea plants were rich in SA and responded differently to different pathogens. Feeding experiments in tea tissues further confirmed the transformation of SA into salicylic acid 2-O-β-glucoside (SAG) and methyl salicylate. Nonaqueous fractionation techniques confirmed that SA and SAG were mostly distributed in the cytosol of tea leaves, consistent with distributions in other plant species. Furthermore, the stem epidermis contained more SA than the stem core both in C. sinensis cv. “Jinxuan” (small-leaf species) and “Yinghong No. 9” (large-leaf species). Compared with cv. “Yinghong No. 9”, cv. “Jinxuan” contained more SAG in the stem epidermis, which might explain its lower incidence rate of wilt disease. This information will improve understanding of SA occurrence in tea plants and provide a basis for investigating the relationship between SA and disease resistance in tea plants.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Peixian Bai ◽  
Liyuan Wang ◽  
Kang Wei ◽  
Li Ruan ◽  
Liyun Wu ◽  
...  

Abstract Background Alanine decarboxylase (AlaDC), specifically present in tea plants, is crucial for theanine biosynthesis. Serine decarboxylase (SDC), found in many plants, is a protein most closely related to AlaDC. To investigate whether the new gene AlaDC originate from gene SDC and to determine the biochemical properties of the two proteins from Camellia sinensis, the sequences of CsAlaDC and CsSDC were analyzed and the two proteins were over-expressed, purified, and characterized. Results The results showed that exon-intron structures of AlaDC and SDC were quite similar and the protein sequences, encoded by the two genes, shared a high similarity of 85.1%, revealing that new gene AlaDC originated from SDC by gene duplication. CsAlaDC and CsSDC catalyzed the decarboxylation of alanine and serine, respectively. CsAlaDC and CsSDC exhibited the optimal activities at 45 °C (pH 8.0) and 40 °C (pH 7.0), respectively. CsAlaDC was stable under 30 °C (pH 7.0) and CsSDC was stable under 40 °C (pH 6.0–8.0). The activities of the two enzymes were greatly enhanced by the presence of pyridoxal-5′-phosphate. The specific activity of CsSDC (30,488 IU/mg) was 8.8-fold higher than that of CsAlaDC (3467 IU/mg). Conclusions Comparing to CsAlaDC, its ancestral enzyme CsSDC exhibited a higher specific activity and a better thermal and pH stability, indicating that CsSDC acquired the optimized function after a longer evolutionary period. The biochemical properties of CsAlaDC might offer reference for theanine industrial production.


Plants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1434
Author(s):  
Hiran A. Ariyawansa ◽  
Ichen Tsai ◽  
Jian-Yuan Wang ◽  
Patchareeya Withee ◽  
Medsaii Tanjira ◽  
...  

Camellia sinensis is one of the major crops grown in Taiwan and has been widely cultivated around the island. Tea leaves are prone to various fungal infections, and leaf spot is considered one of the major diseases in Taiwan tea fields. As part of a survey on fungal species causing leaf spots on tea leaves in Taiwan, 19 fungal strains morphologically similar to the genus Diaporthe were collected. ITS (internal transcribed spacer), tef1-α (translation elongation factor 1-α), tub2 (beta-tubulin), and cal (calmodulin) gene regions were used to construct phylogenetic trees and determine the evolutionary relationships among the collected strains. In total, six Diaporthe species, including one new species, Diaporthe hsinchuensis, were identified as linked with leaf spot of C. sinensis in Taiwan based on both phenotypic characters and phylogeny. These species were further characterized in terms of their pathogenicity, temperature, and pH requirements under laboratory conditions. Diaporthe tulliensis, D. passiflorae, and D. perseae were isolated from C. sinensis for the first time. Furthermore, pathogenicity tests revealed that, with wound inoculation, only D. hongkongensis was pathogenic on tea leaves. This investigation delivers the first assessment of Diaporthe taxa related to leaf spots on tea in Taiwan.


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