1.Characteristics of the chloroplast genome of Camellia insularis.
Jin ZHANG ; Yongbiao DENG ; Bo ZHAO
Chinese Journal of Biotechnology 2024;40(1):280-291
In this study, the chloroplast genome of Camellia insularis Orel & Curry was sequenced using high-throughput sequencing technology. The results showed that the chloroplast genome of C. insularis was 156 882 bp in length with a typical tetrad structure, encoding 132 genes, including 88 protein-coding genes, 36 tRNA genes, and 8 rRNA genes. Codon preference analysis revealed that the highest number of codons coded for leucine, with a high A/U preference in the third codon position. Additionally, 67 simple sequence repeats (SSR) loci were identified, with a preference for A and T bases. The inverted repeat (IR) boundary regions of the chloroplast genome of C. insularis were relatively conserved, except for a few variable regions. Phylogenetic analysis indicated that C. insularis was most closely related to C. fascicularis. Yellow camellia is a valuable material for genetic engineering breeding. This study provides fundamental genetic information on chloroplast engineering and offers valuable resources for conducting in-depth research on the evolution, species identification, and genomic breeding of yellow Camellia.
Genome, Chloroplast/genetics*
;
Phylogeny
;
Plant Breeding
;
Camellia/genetics*
;
Chloroplasts/genetics*
2.Transcriptome analysis reveals the role of withering treatment in flavor formation of oolong tea (Camellia sinensis).
Chen ZHU ; Shuting ZHANG ; Chengzhe ZHOU ; Biying SHI ; Linjie HUANG ; Yuling LIN ; Zhongxiong LAI ; Yuqiong GUO
Chinese Journal of Biotechnology 2022;38(1):303-327
Oolong tea is a semi-fermented tea with strong flavor, which is widely favored by consumers because of its floral and fruity aroma as well as fresh and mellow taste. During the processing of oolong tea, withering is the first indispensable process for improving flavor formation. However, the molecular mechanism that affects the flavor formation of oolong tea during withering remains unclear. Transcriptome sequencing was used to analyze the difference among the fresh leaves, indoor-withered leaves and solar-withered leaves of oolong tea. A total of 10 793 differentially expressed genes were identified from the three samples. KEGG enrichment analysis showed that the differentially expressed genes were mainly involved in flavonoid synthesis, terpenoid synthesis, plant hormone signal transduction and spliceosome pathways. Subsequently, twelve differentially expressed genes and four differential splicing genes were identified from the four enrichment pathways for fluorescence quantitative PCR analysis. The results showed that the expression patterns of the selected genes during withering were consistent with the results in the transcriptome datasets. Further analysis revealed that the transcriptional inhibition of flavonoid biosynthesis-related genes, the transcriptional enhancement of terpenoid biosynthesis-related genes, as well as the jasmonic acid signal transduction and the alternative splicing mechanism jointly contributed to the flavor formation of high floral and fruity aroma and low bitterness in solar-withered leaves. The results may facilitate better understanding the molecular mechanisms of solar-withering treatment in flavor formation of oolong tea.
Camellia sinensis/genetics*
;
Gene Expression Profiling
;
Plant Leaves
;
Plant Proteins/metabolism*
;
Taste
;
Tea
;
Transcriptome/genetics*
3.Genome-wide identification of CsCCD gene family in tea plant (Camellia sinensis) and expression analysis of the oolong tea processing with supplementary LED light.
Zixin NI ; Qingyang WU ; Yun YANG ; Huili DENG ; Ziwei ZHOU ; Zhongxiong LAI ; Yun SUN
Chinese Journal of Biotechnology 2022;38(1):359-373
Carotenoid cleavage dioxygenase (CCD) family is important for production of volatile aromatic compounds and synthesis of plant hormones. To explore the biological functions and gene expression patterns of CsCCD gene family in tea plant, genome-wide identification of CsCCD gene family was performed. The gene structures, conserved motifs, chromosome locations, protein physicochemical properties, evolutionary characteristics, interaction network and cis-acting regulatory elements were predicted and analyzed. Real time-quantitative reverse transcription PCR (RT-qPCR) was used to detect the relative expression level of CsCCD gene family members under different leaf positions and light treatments during processing. A total of 11 CsCCD gene family members, each containing exons ranging from 1 to 11 and introns ranging from 0 to 10, were identified. The average number of amino acids and molecular weight were 519 aa and 57 643.35 Da, respectively. Phylogenetic analysis showed the CsCCD gene family was clustered into 5 major groups (CCD1, CCD4, CCD7, CCD8 and NCED). The CsCCD gene family mainly contained stress response elements, hormone response elements, light response elements and multi-factor response elements, and light response elements was the most abundant (142 elements). Expression analysis showed that the expression levels of CsCCD1 and CsCCD4 in elder leaves were higher than those in younger leaves and stems. With the increase of turning over times, the expression levels of CsCCD1 and CsCCD4 decreased, while supplementary LED light strongly promoted their expression levels in the early stage. The expression level of NCED in younger leaves was higher than that in elder leaves and stems on average, and the expression trend varied in the process of turning over. NCED3 first increased and then decreased, with an expression level 15 times higher than that in fresh leaves. In the late stage of turning over, supplementary LED light significantly promoted its gene expression. In conclusion, CsCCD gene family member expressions were regulated by mechanical force and light. These understandings may help to optimize tea processing techniques and improve tea quality.
Camellia sinensis/genetics*
;
Gene Expression Regulation, Plant
;
Phylogeny
;
Plant Leaves/genetics*
;
Plant Proteins/metabolism*
;
Tea
4.Effect of Lactobacillus coryniformis FZU63 on the flavor quality of black tea beverage.
Ruili LI ; Yifeng LIU ; Weibo LUO ; Huilin HUANG ; Meiting HUANG ; Chi CHEN ; Ronghui XIAO ; Jinzhi HAN ; Xucong LÜ
Chinese Journal of Biotechnology 2022;38(12):4731-4743
The tea beverages will be endowed with distinct aroma and taste, as well as various biologically active compounds including probiotic factors, when fermented with lactic acid bacteria (LAB). However, at present, few studies on the dynamics of flavors in tea soup at different fermentation stages were conducted. In this study, the composition of monosaccharides, aromatic components, free amino acids, and organic acids were measured, when the black tea beverages were fermented with Lactobacillus coryniformis FZU63 which was isolated from Chinese traditional kimchi. The results indicated that monosaccharides including glucose, fructose, mannose and xylose in black tea beverages are the main carbon sources for fermentation. In addition, the abundance of aromatic compounds in black tea soup are increased significantly at different fermentation stages, which endow the fermented black tea soup with fruit aroma on the basis of flowery and nutty aroma. Moreover, some bitter amino acids are reduced, whereas the content of sweet and tasty amino acids is elevated. Furthermore, the levels of lactic acid, malic acid, citric acid and other organic acids are accumulated during the fermentation. Additionally, sensory evaluation displays that black tea beverage is acquired with comprehensive high-quality after being fermented for 48 h. This study provides a theoretical basis to steer and control the flavor formation and quality of the fermented tea beverages during LAB fermentation.
Tea/chemistry*
;
Beverages/microbiology*
;
Camellia sinensis
;
Fermentation
;
Acids
;
Amino Acids
;
Glucose
5.The albino mechanism of a new theanine-rich tea cultivar 'Fuhuang 2'.
Xinying LIN ; Shuxian SHAO ; Pengjie WANG ; Ruxing YANG ; Yucheng ZHENG ; Xiaomin CHEN ; Lei ZHANG ; Naixing YE
Chinese Journal of Biotechnology 2022;38(10):3956-3972
To explore the mechanism of tea albino variation and high theanine formation, 'Fuyun 6' and a new theanine-rich tea cultivar 'Fuhuang 2' were as materials in this study, pigment content, metabolome and transcriptome of the two cultivars were analyzed by ultramicroelectron microscopy, widely targeted metabolomics, targeted metabolomics and transcriptomics. The results showed that five catechins, theobromine, caffeine, and 20 free amino acids, including theanine, glutamine, arginine, etc., were identified by targeted metabolomics. The amino acid content of 'Fuhuang 2' was significantly higher than that of 'Fuyun 6', and the theanine content was as high as 57.37 mg/g in 'Fuhuang 2'. The ultrastructure of leaves showed that the chloroplast cell structure of 'Fuhuang 2' was fuzzy, most of the grana lamellae were arranged in disorder, with large gaps, and the thylakoids were filiform. The determination of pigments showed that compared with 'Fuyun 6', the contents of chlorophyll A and B, carotenoids, flavonoids and other pigments of 'Fuhuang 2' decreased significantly, some important pigment-related-genes, such as chlorophyllase (CLH), 9-cis-epoxycarotenoid dioxygenase (NCED), flavonoid 3β-hydroxylase (F3H) and flavonoid 3', 5'-hydroxylase (F3'5'H) were significantly changed. Compared with 'Fuyun 6', 'Fuhuang 2' identified 138 significantly changed metabolites (SCMs) and 658 differentially expressed genes (DEGs). KEGG enrichment analysis showed that SCMs and DEGs were significantly enriched in amino acid biosynthesis, glutathione metabolism and TCA cycle. In general, the albino phenotype of 'Fuhuang 2' may be caused by a deficiency in photosynthetic proteins, chlorophyll metabolism genes and chlorophyll content. The accumulation of high theanine in 'Fuhuang 2' may be due to the low nitrogen consumption in yellowed leaves and the lack of carbon skeleton, amino and nitrogen resources are stored more effectively, resulting in the up regulation of metabolites and related gene expression in the amino acid synthesis pathway, theanine has become a significant accumulation of nitrogen-containing compounds in yellowed leaves.
Camellia sinensis/genetics*
;
Chlorophyll A/metabolism*
;
Plant Proteins/genetics*
;
Plant Leaves/chemistry*
;
Chlorophyll/metabolism*
;
Transcriptome
;
Flavonoids/metabolism*
;
Amino Acids/genetics*
;
Tea
;
Mixed Function Oxygenases/metabolism*
;
Nitrogen/metabolism*
6.Active fractions of Camellia nitidissima inhibit non-small cell lung cancer via suppressing epidermal growth factor receptor.
Zi-Ling WANG ; Yu-Jie GUO ; Yun-Yun ZHU ; Le CHEN ; Ting WU ; Da-Hui LIU ; Bi-Sheng HUANG ; Hong-Zhi DU
China Journal of Chinese Materia Medica 2021;46(20):5362-5371
The present study explored the effects and its underlying mechanisms of four active fractions of Camellia nitidissima(leaf polyphenols, leaf saponins, flower polyphenols, and flower saponins in C. nitidissima) in inhibiting the proliferation and migration of non-small cell lung cancer(NSCLC) by suppressing the epidermal growth factor receptor(EGFR). MTT assay was used to detect the effect of four active fractions on the proliferation of NCI-H1975 and HCC827 cells. Wound healing assay and Transwell assay were adopted to evaluate the effect of four active fractions on the migration of NSCLC. The effect of four active fractions on the enzyme activity of EGFR was detected. Molecular docking was carried out to explore the direct action capacity and action sites between representative components of the four active fractions and EGPR. Western blot assay was employed to investigate the effect of four active fractions on the protein expression in EGFR downstream signaling pathways. The results of the MTT assay indicated that the cell viability of NCI-H1975 and HCC827 cells was significantly inhibited by four active fractions at 50, 100, 150, and 200 μg·mL~(-1) in a dose-dependent manner. Wound healing assay and Transwell assay revealed that the migration of NCI-H1975 and HCC827 cells was significantly suppressed by four active fractions. In addition, the results of the protein activity assay showed that the enzyme activity of EGFR was significantly inhibited by four active fractions. The molecular docking results confirmed that various components in four active fractions possessed strong binding activity to EGFR enzymes. Western blot assay revealed that four active fractions down-regulated the protein expression of EGFR and its downstream signaling pathways. It is concluded that the four active fractions of C. nitidissima can inhibit NSCLC. The mechanism may be related to EGFR and its downstream signaling pathways. This study provides a new scientific basis for the clinical treatment of NSCLC with active fractions of C. nitidissima, which is of reference significance for further research on the anti-tumor mechanism of C. nitidissima.
Apoptosis
;
Camellia
;
Carcinoma, Non-Small-Cell Lung/drug therapy*
;
Cell Line, Tumor
;
Cell Proliferation
;
ErbB Receptors/genetics*
;
Humans
;
Lung Neoplasms/drug therapy*
;
Molecular Docking Simulation
7.Effectivity of Camellia sinensis Extract with Nano-chitosan to Fibroblast Amounts of Wistar Rats Gingival Wound Healing Process
Indra Bramanti ; Annisa Hidaratri Uningojati ; Dilla Asriyani ; Urfa Tabtila ; Fathul Muin ; Bramanti Nadya Kausara
Archives of Orofacial Sciences 2021;16(SUPP 1):25-32
ABSTRACT
Green tea (Camellia sinensis) has high level of flavonoids which are proven to have anti-inflammatory
activity. Effect of flavonoids can be enhanced by nano-chitosan capsulation as drug carrier. Chitosan
is polysaccharide derived from crustacean shells that mostly used as matrix of various drugs and plant
extracts. The aim of this study was to determine the effectivity of flavonoids in green tea extract in nanochitosan capsulation towards the number of fibroblasts on proliferative phase of gingival wound healing
process. Green tea was extracted, encapsulated with nano-chitosan and then made into gel. Gingiva
labial of 24 male white 3-month-old Wistar rats were wounded by punch biopsy (2 mm diameter), then
were treated two times a day, and were divided randomly into four groups of topical gel applications:
green tea extract gel encapsulated nano-chitosan, green tea extract gel, base gel as negative control,
and NSAIDs gel as positive control, starting at 0 day until 7th day. At 5th and 7th day, three rats from
each group were decapitated and the mandibular gingiva was taken in order to make histology slides
with hematoxylin eosin staining. Under microscope, the number of fibroblasts were examined. The data
were analysed using ANOVA test with 95% confidence level. The results showed that the number of
fibroblasts on proliferative phase was significantly higher than control negative (p < 0.05) and has no
significant differences (p > 0.05) with control positive. In conclusion, topical application of green tea
extract gel encapsulated nano-chitosan was effective to accelerate rats gingival wound healing process by
increasing the fibroblasts.
Camellia sinensis
;
Chitosan
;
Gingiva--injuries
;
Wound Healing
;
Rats, Wistar
8.New observations on the effect of camellia oil on fatty liver disease in rats.
Journal of Zhejiang University. Science. B 2020;21(8):657-667
Camellia oil has become an important plant oil in China in recent years, but its effects on non-alcoholic fatty liver disease (NAFLD) have not been documented. In this study, the effects of camellia oil, soybean oil, and olive oil on NAFLD were evaluated by analyzing the fatty acid profiles of the plant oils, the serum lipids and lipoproteins of rats fed different oils, and by cytological and ultrastructural observation of the rats' hepatocytes. Analysis of fatty acid profiles showed that the polyunsaturated fatty acid (PUFA) n-6/n-3 ratio was 33.33 in camellia oil, 12.50 in olive oil, and 7.69 in soybean oil. Analyses of serum lipids and lipoproteins of rats showed that the levels of total cholesterol and low-density lipoprotein cholesterol in a camellia oil-fed group (COFG) were lower than those in an olive oil-fed group (OOFG) and higher than those in a soybean oil-fed group (SOFG). However, only the difference in total cholesterol between the COFG and SOFG was statistically significant. Cytological observation showed that the degree of lipid droplet (LD) accumulation in the hepatocytes in the COFG was lower than that in the OOFG, but higher than that in the SOFG. Ultrastructural analysis revealed that the size and number of the LDs in the hepatocytes of rats fed each of the three types of oil were related to the degree of damage to organelles, including the positions of nuclei and the integrity of mitochondria and endoplasmic reticulum. The results revealed that the effect of camellia oil on NAFLD in rats was greater than that of soybean oil, but less than that of olive oil. Although the overall trend was that among the three oil diets, those with a lower n-6/n-3 ratio were associated with a lower risk of NAFLD, and the effect of camellia oil on NAFLD was not entirely related to the n-6/n-3 ratio and may have involved other factors. This provides new insights into the effect of oil diets on NAFLD.
Animals
;
Camellia/chemistry*
;
Fatty Acids/analysis*
;
Hepatocytes/ultrastructure*
;
Lipid Droplets/physiology*
;
Lipids/blood*
;
Male
;
Non-alcoholic Fatty Liver Disease/pathology*
;
Plant Oils/administration & dosage*
;
Rats
;
Rats, Sprague-Dawley
9.Exogenous lipoid pneumonia presented by acute eosinophilic pneumonia.
Allergy, Asthma & Respiratory Disease 2019;7(1):57-60
Exogenous lipoid pneumonia is an uncommon medical condition resulting from aspiration or inhalation of oily material. Generally, lipoid pneumonia has nonspecific clinical and radiological presentations, and may be misdiagnosed as bacterial pneumonia or lung cancer. We describe an unusual case of exogenous lipoid pneumonia accompanied by peripheral blood and pulmonary eosinophilia. A 63-year-old man was admitted with progressively worsening exertional dyspnea and productive cough for 5 days. A chest radiograph showed abnormalities in the lower lobe of the right lung, and a diagnosis of community-acquired pneumonia was made; intravenous antibiotics were administered. However, dyspnea and hypoxia gradually worsened and peripheral blood eosinophilia developed. A bronchoscopy was performed and bronchoalveolar lavage fluid analysis showed markedly increased numbers of eosinophils (40%). Subsequently, a comprehensive review of history revealed that he fell asleep with camellia oil in his mouth for 2 weeks to relieve foreign body sensation of the throat. Sputum and bronchoalveolar lavage fluid cytology showed the presence of lipid-laden macrophages. He was diagnosed with lipoid pneumonia and acute eosinophilic pneumonia. Chest radiograph and symptom were rapidly improved after treatment with intravenous methylprednisolone.
Anoxia
;
Anti-Bacterial Agents
;
Bronchoalveolar Lavage Fluid
;
Bronchoscopy
;
Camellia
;
Cough
;
Diagnosis
;
Dyspnea
;
Eosinophilia
;
Eosinophils*
;
Foreign Bodies
;
Humans
;
Inhalation
;
Lung
;
Lung Neoplasms
;
Macrophages
;
Methylprednisolone
;
Middle Aged
;
Mouth
;
Pharynx
;
Pneumonia*
;
Pneumonia, Bacterial
;
Pneumonia, Lipid
;
Pulmonary Eosinophilia*
;
Radiography, Thoracic
;
Respiratory Aspiration
;
Sensation
;
Sputum
10.Diversity and Antifungal Activity of Endophytic Fungi Associated with Camellia oleifera.
Jinxiu YU ; Ying WU ; Zhen HE ; Mi LI ; Kaiming ZHU ; Bida GAO
Mycobiology 2018;46(2):85-91
Endophytic fungi strains (n = 81) were isolated from the leaves, barks, and fruits of Camellia oleifera from Hunan province (China) to delineate their species composition and potential as biological control agents of C. oleifera anthracnose. The fungi were identified by morphological and phylogenetic analyses. Fungal colonization rates of the leaves, barks, and fruits were 58.02, 27.16, and 14.81%, respectively. The isolates were identified as 14 genera, belonging to two subdivisions, Deuteromycotina and Ascomycotina; 87.65% of all isolates belonged to Deuteromycotina. The dominant species, occurring with a high relative frequency, were Pestalotiopsis sp. (14.81%), Penicillium sp. (14.81%), and Fusarium sp. (12.35%). The Simpson’s and Shannon’s diversity indices revealed the highest species diversity in the leaves, followed by the barks and fruits. The similarity index for the leaves versus barks comparison was the highest, indicating that the number of endophytic fungal species shared by the leaves and barks was higher than barks and fruits or leaves and fruits. Based on the results of dual culture experiments, only five strains exhibited antifungal activity against C. oleifera anthracnose pathogen, with isolate ty-64 (Oidium sp.) generating the broadest inhibition zones. Our results indicate that the endophytes associated with C. oleifera could be employed as natural agents controlling C. oleifera anthracnose.
Biological Control Agents
;
Camellia*
;
Colon
;
Endophytes
;
Fruit
;
Fungi*
;
Fusarium
;
Penicillium


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