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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.

Aim: To investigate the potential hepato-protective effect of Cassia auriculata leaf extract against ethanol-induced oxidative stress in albino rats, by exploring the antioxidant properties of Cassia auriculata and its ability to mitigate oxidative damage caused by ethanol consumption, and the underlying mechanisms involved. Methodology: In the present study, Group – I served as Control, Group – II (Ethanol induced group), where 50% ethanol was orally administered to the rats, Group – III (treatment group), where 250 mg kg-1 of Cassia auriculata leaf extract was orally administered to the hepato-toxic albino rats and in Group – IV, rats were treated with standard reference drug Silymarin (100 mg kg-1) followed by estimating the parameters related to oxidative stress such as TBARS and antioxidants enzyme levels such as GST, GPx, SOD and CAT in the blood and liver tissue samples. Phytochemical analysis of Cassia auriculata leaf powder was conducted to determine the active phytoconstituents. Results: Treatment with Cassia auriculata extract significantly (p<0.001) reduced the damage caused by oxidative stress by decreasing the levels of TBARS and significantly (p<0.001) elevated the levels of antioxidant enzymes such as GST, GPx, SOD and CAT in the serum and liver samples. Interpretation: The present study provides an opportunity to explore the potential therapeutic properties of Cassia auriculata in combating ethanol-induced oxidative stress. Key words: Albino rats, Cassia auriculata, Ethanol, Oxidative stress

Summary Hemp (Cannabis sativa L.) is an annual and typically dioecious crop. Due to the therapeutic potential for human diseases, phytocannabinoids as a medical therapy is getting more attention recently. Several candidate genes involved in cannabinoid biosynthesis have been elucidated using omics analysis. However, the gene function was not fully validated due to few reports of stable transformation for Cannabis tissues. In this study, we firstly report the successful generation of gene‐edited plants using an Agrobacterium‐mediated transformation method in C. sativa. DMG278 achieved the highest shoot induction rate, which was selected as the model strain for transformation. By overexpressing the cannabis developmental regulator chimera in the embryo hypocotyls of immature grains, the shoot regeneration efficiency was substantially increased. We used CRISPR/Cas9 technology to edit the phytoene desaturase gene and finally generated four edited cannabis seedlings with albino phenotype. Moreover, we propagated the transgenic plants and validated the stable integration of T‐DNA in cannabis genome.

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.

Summary Plant protoplasts are useful for assessing the efficiency of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐associated protein 9 (Cas9) mutagenesis. We improved the process of protoplast isolation and transfection of several plant species. We also developed a method to isolate and regenerate single mutagenized Nicotianna tabacum protoplasts into mature plants. Following transfection of protoplasts with constructs encoding Cas9 and sgRNAs, target gene DNA could be amplified for further analysis to determine mutagenesis efficiency. We investigated N. tabacum protoplasts and derived regenerated plants for targeted mutagenesis of the phytoene desaturase (NtPDS) gene. Genotyping of albino regenerants indicated that all four NtPDS alleles were mutated in amphidiploid tobacco, and no Cas9 DNA could be detected in most regenerated plants.

Cancer is a global health problem and the second-leading cause of death worldwide. Gold nanoparticles (GNPs) are used in a wide range of medical fields, particularly in cancer diagnosis and treatment, therapy, and anti-inflammation. However, the question that always arises is whether GNPs affect normal, healthy cells when used as treatment. Therefore, GNPs were used in this research to treat muscle and brain cancer and to demonstrate its effect on tissues in living organs in healthy mice. The objectives were to determine the efficiency of GNPs in killing cancer cells through cytotoxicity assay, examine the effect of GNPs on different healthy organ tissues, and show the influence of different GNP concentrations, cell types, and incubation times through in vitro experiments.  Two cell lines, AMGM (brain cancer cell line) and RD (muscle cancer cell line), at different concentrations (4.5×1012 – 6×1010 particles mL-1) with two incubation periods (24 and 72h) were used in this study. Ten adult male albino mice were randomly distributed into two groups. The experiment was extended for 10 days to study the effect of a single dose of 250 µl of GNPs (4.5×1012 particles mL-1) administered through intraperitoneal injection by examining the histological changes of the liver, kidney, and spleen of the mice. In the cytotoxicity experiment, high GNP concentrations had the greatest effect at 24h, and the RD cell line was more sensitive than the AMGM cell line at 72h. Histological tests of the mouse organs revealed different changes such as severe hepatic granulomas, renal apoptosis, amyloidosis of spleen tissues, and inflammation with fibrosis of intestinal tissues. GNPs are an influential factor in cancer therapy, and their effects vary with cell line type, exposure time, and dose. They also cause histological harm (fibrosis, inflammation, and apoptosis) on vital organs.

Polystyrene microplastics (PSMPs) have emerged as a ubiquitous environmental toxicant that affects different organs including testes. Ginkgetin (GNG) is a biflavonoid that shows antioxidant properties. The current research was undertaken to evaluate the ameliorative potential of GNG against PSMPs-instigated testicular damages. Forty-eight albino rats (male) were randomly divided into 4 equal groups: control, PSMPs-treated group (0.01 mgkg −1 ), GNG + PSMPs–exposed group (25 mgkg −1  + 0.01 mgkg −1 ), and only GNG-supplemented group (25 mgkg −1 ). After 56 days of treatment, it was revealed that PSMPs significantly reduced the activity of glutathione peroxidase (GPx), catalase (CAT), superoxide dismutase (SOD), and glutathione reductase (GSR), while concurrently augmented the levels of lipid peroxidation marker, i.e., malondialdehyde (MDA) along with reactive oxygen species (ROS). Rats administered with PSMPs showed a significant reduction in the spermatogenic indices (sperm count, viability, and motility), HOS coiled tail sperm along with increased sperm structural deformities, i.e., tail, head, and mid-piece. Additionally, PSMPs exposure decreased the levels of testosterone, luteinizing (LH), and follicle-stimulating hormones (FSH). Besides, administration of PSMPs reduced the steroidogenic enzymes (13β-HSD, StAR, and 17β-HSD) and Bcl-2 expression, while augmented the caspase-3 and Bax expression. PSMPs also elevated the levels of inflammatory markers (IL-6, IL-1β, TNF-α, and NF-κB) and activity of COX-2 in the testes. Furthermore, PSMPs treatment induced various histopathological damages in the testes of rats. Therefore, findings of the current study suggested that GNG effectively mitigated the PSMPs-induced testicular toxicity owing to its chemoprotective potential possibly through its anti-inflammatory, antioxidant, anti-apoptotic, and androgenic properties.

The present study identified the probable mechanism behind the anti-cancer activity of the hexane fraction of Pleurotus osteratus (HFPO) using network pharmacology and experimental validation. HFPO myco-metabolites targets and targets related to the cancer were mined from the online web server, and overlapping targets were screened. Out of the 74 overlapping targets, 33 targets were identified in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway of cancer. Furthermore, the main active myco-metabolites and hub targets were identified by network analysis of the compound-targets network and protein–protein interaction (PPI), respectively. Molecular docking results showed good binding affinity of the hub targets with their respective myco-metabolites. HFPO induced in-vitro anti-cancer activity by affecting the PI3K-AKT-mTOR pathway, besides time-dependent cell cytotoxicity and apoptotic cell body formation. Additionally, tumor volume reduction was observed in HFPO-treated Ehrlich ascites carcinoma (EAC) bearing Swiss albino mice. Overall, HFPO induces anti-cancer potential by modulating the PI3K-AKT-mTOR signaling pathway.

Summary In recent years, the type II CRISPR system has become a widely used and robust technique to implement site‐directed mutagenesis in a variety of species including model and crop plants. However, few studies manipulated metabolic pathways in plants using the CRISPR system. Here, we introduced the pYLCRISPR/Cas9 system with one or two single‐site guide RNAs to target the tomato phytoene desaturase gene. An obvious albino phenotype was observed in T0 regenerated plants, and more than 61% of the desired target sites were edited. Furthermore, we manipulated the γ‐aminobutyric acid (GABA) shunt in tomatoes using a multiplex pYLCRISPR/Cas9 system that targeted five key genes. Fifty‐three genome‐edited plants were obtained following single plant transformation, and these samples represented single to quadruple mutants. The GABA accumulation in both the leaves and fruits of genomically edited lines was significantly enhanced, and the GABA content in the leaves of quadruple mutants was 19‐fold higher than that in wild‐type plants. Our data demonstrate that the multiplex CRISPR/Cas9 system can be exploited to precisely edit tomato genomic sequences and effectively create multisite knockout mutations, which could shed new light on plant metabolic engineering regulations.

A rice low temperature-induced albino variant was determined by the recessive ltia1 and ltia2 genes. LTIA1 and LTIA2 encode highly conserved mini-ribonucleases III located in chloroplasts and expressed in aerial parts of the plant. At low temperature, LTIA1 and LTIA2 redundantly affect chlorophyll levels, non-photochemical quenching, photosynthetic quantum yield of PS II and seedling growth. LTIA1 and LTIA2 proteins are involved in splicing of atpF and the biogenesis of 16S and 23S rRNA in chloroplasts. Presence/absence variation of LTIA1, the ancestral copy, was found only in japonica but that of LTIA2 in all rice subgroups. Accessions with LTIA2 presence tended to be distributed more remote from the equator compared to those with LTIA2 absence. LTIA2 duplicated from LTIA1 at the early stage of divergence of the AA genome Oryza species but deleted againin O. nivara. In cultivated rice, absence of LTIA2 is derived from O. nivara. LTIA1 absence occurred more recently in japonica.