Assessment of gamma radiation through agro-morphological characters in camellia sinensis L. (O.) kuntze.

PURPOSE: To study the effects of gamma radiation on tea seed germination, morphological changes, and genetic variation by using gamma radiation. MATERIAL AND METHOD: Fresh Tea seed material were irradiated with twenty different doses of gamma radiation such as 0, 2, 4, 6, 8, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80 90, 100, 200, 500 and 1000 Gy from Cobalt 60Co source from Regional Nuclear Agriculture Research Center, Bidhan Chandra Krishi Viswavidyalaya (BCKV), West Bengal, in between 2019 and 2020. RESULT AND CONCLUSION: The growth behavior of tea seedling was recorded under varying levels of gamma radiation and its performance at nursery stages. It was observed seed irradiated with doses from 35 Gy to 100 Gy could germinate but could not survive beyond five (05) months. When treated with higher doses as 200 Gy, 500 Gy and 1000 Gy, no seed germination takes places due to possible damages occur in the DNA structure. Screening of growth characteristics of tea plant generally monitored by the characteristics like plant height, number of leaves, number of primary branches, base diameter, and total leaf area of plants and we found that these characteristics significantly increased with the progress of time and increasing levels of gamma radiation; however, the plant height showed decreasing trend with the increasing levels of gamma radiation, which could be due to the change in chromosomal structure and genetic alteration. After 90 weeks of planting, the plant height, no. of primary branches, the number of leaves, plant base diameter, and total leaf area per plant recorded were 36.42 cm, 1/plant, 7.11/plant, 0.62 c.m, 22.92 cm2/plant respectively under the radiation level 30 Gy, whereas the corresponding figures of the above parameters at the control treatment were 85.32 cm, 1/plant, 18.84/plant, 1.18 c.m and 26.68 cm2/plant, respectively. The total plant height, no. of primary branches, the number of leaves, plant base diameter, and total leaf area per plant were significantly influenced by the rising levels of gamma radiation (up to 100 Gy), finally, after 90 weeks of planting, the maximum no. of branching was observed in the treatment of 8 Gy, 10 Gy and 15 Gy respectively. The study reveals a hitherto open the possibility of using gamma radiation on tea plant for creation of variation in the tea seed planting materials. Further studies on mutation using tea planting materials would give an insight into its mutable gene behavior.

MicroRNAs and their targeted genes associated with phase changes of stem explants during tissue culture of tea plant.

Elucidation of the molecular mechanism related to the dedifferentiation and redifferentiation during tissue culture will be useful for optimizing regeneration system of tea plant. In this study, an integrated sRNAome and transcriptome analyses were carried out during phase changes of the stem explant culture. Among 198 miRNAs and 8001 predicted target genes, 178 differentially expressed miRNAs and 4264 potential targets were screened out from explants, primary calli, as well as regenerated roots and shoots. According to KEGG analysis of the potential targets, pathway of "aminoacyl-tRNA biosynthesis", "proteasome" and "glutathione metabolism" was of great significance during the dedifferentiation, and pathway of "porphyrin and chlorophyll metabolism", "mRNA surveillance pathway", "nucleotide excision repair" was indispensable for redifferentiation of the calli. Expression pattern of 12 miRNAs, including csn-micR390e, csn-miR156b-5p, csn-miR157d-5p, csn-miR156, csn-miR166a-3p, csn-miR166e, csn-miR167d, csn-miR393c-3p, csn-miR394, csn-miR396a-3p, csn-miR396 and csn-miR396e-3p, was validated by qRT-PCR among 57 differentially expressed phase-specific miRNAs. Validation also confirmed that regulatory module of csn-miR167d/ERF3, csn-miR156/SPB1, csn-miR166a-3p/ATHB15, csn-miR396/AIP15A, csn-miR157d-5p/GST and csn-miR393c-3p/ATG18b might play important roles in regulating the phase changes during tissue culture of stem explants.

Analysis of Differentially Expressed Genes in Tissues of Camellia sinensis during Dedifferentiation and Root Redifferentiation.

Tissue culture is very important for identifying the gene function of Camellia sinensis (L.) and exploiting novel germplasm through transgenic technology. Regeneration system of tea plant has been explored but not been well established since the molecular mechanism of tea plant regeneration is not clear yet. In this study, transcriptomic analysis was performed in the initial explants of tea plant and their dedifferentiated and redifferentiated tissues. A total of 93,607 unigenes were obtained through de novo assembly, and 7,193 differentially expressed genes (DEGs) were screened out from the 42,417 annotated unigenes. Much more DEGs were observed during phase transition rather than at growth stages of callus. Our KOG and KEGG analysis, and qPCR results confirmed that phase transition of tea plant was closely related to the mechanism that regulate expression of genes encoding the auxin- and cytokinin-responsive proteins, transcription factor MYB15 and ethylene-responsive transcription factor ERF RAP2-12. These findings provide a reliable foundation for elucidating the mechanism of the phase transition and may help to optimize the regeneration system by regulating the gene expression pattern.

Free Zn(2+) enhances inhibitory effects of EGCG on the growth of PC-3 cells.

Epigallocatechin-3-gallate (EGCG), a major component of green tea, has both preventive and therapeutic beneficial actions in prostate cancer. In the present study, we compared the growth inhibitory effects and the antioxidant and ability to modify cell membrane permeation of zinc-EGCG complex and Zn2+/EGCG mixture on androgen-insensitive prostate cancer (PC-3) cells. It was noted that free Zn2+ enhanced the growth inhibitory effects of EGCG on PC-3 cells at 160 micromol/L concentration,whereas zinc-EGCG complex was ineffective. EGCG showed potent free radical scavenging ability in the presence of Zn2+. EGCG in the presence of Zn2+ was more effective than EGCG alone in enhancing the permeability of the cell membrane, whereas zinc-EGCG complex had no effect on PC-3 cell membrane permeability. These results indicate that though Zn2+ enhanced the action of EGCG on PC-3 cells, zinc-EGCG complex is highly unlikely to be formed in the presence of Zn2+ and EGCG to explain the potentiating action of Zn2+ on the growth inhibitory property of EGCG on PC-3 cells.