The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) has been reported for precise genome modification in many plants. In the current study, we demonstrate a successful mutation in phytoene desaturase (RAS-PDS) of banana cv. Rasthali using the CRISPR/Cas9 system. Two PDS genes were isolated from Rasthali (RAS-PDS1 and RAS-PDS2), and their protein sequence analysis confirmed that both PDS comprises conserved motifs for enzyme activity. Phylogenetic analysis of RAS-PDS1 and RAS-PDS2 revealed a close evolutionary relationship with other monocot species. The tissue-specific expression profile of RAS-PDS1 and RAS-PDS2 in Rasthali suggested differential regulation of the genes. A single 19-bp guide RNA (gRNA) was designed to target the conserved region of these two RAS-PDS and transformed with Cas9 in embryogenic cell suspension (ECS) cultures of cv. Rasthali. Complete albino and variegated phenotype were observed among regenerated plantlets. DNA sequencing of 13 plants confirmed the indels with 59% mutation frequency in RAS-PDS, suggesting activation of the non-homologous end-joining (NHEJ) pathway. The majority of mutations were either insertion (1-5) or deletion (1-4) of nucleotides near to protospacer adjacent motif (PAM). These mutations have created stop codons in RAS-PDS sequences which suggest premature termination of RAS-PDS protein synthesis. The decreased chlorophyll and total carotenoid contents were detected in mutant lines that revealed the functional disruption of both RAS-PDS genes. Our results demonstrate that genome editing through CRISPR/Cas9 can be applied as an efficient tool for banana genome modification.

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BackgroundGlobally, the routinely used case-based reporting and IgG serosurveys underestimate the actual prevalence of COVID-19. Simultaneous estimation of IgG antibodies and active SARS-CoV-2 markers can provide a more accurate estimation.MethodsA cross-sectional survey of 16416 people covering all risk groups was done between 3-16 September 2020 using the state of Karnataka’s infrastructure of 290 hospitals across all 30 districts. All participants were subjected to simultaneous detection of SARS-CoV-2 IgG using a commercial ELISA kit, SARS-CoV-2 antigen using a rapid antigen detection test (RAT), and reverse transcription-polymerase chain reaction (RT-PCR) for RNA detection. Maximum-likelihood estimation was used for joint estimation of the adjusted IgG, active, and total prevalence, while multinomial regression identified predictors.FindingsThe overall adjusted prevalence of COVID-19 in Karnataka was 27 ·3% (95% CI: 25 ·7-28 ·9), including IgG 16 ·4% (95% CI: 15 ·1 - 17 ·7) and active infection 12 ·7% (95% CI: 11 ·5-13 ·9). The case-to-infection ratio was 1:40, and the infection fatality rate was 0 ·05%. Influenza-like symptoms or contact with a COVID-19 positive patient are good predictors of active infection. The RAT kits had higher sensitivity (68%) in symptomatic participants compared to 47% in asymptomatic.InterpretationThis is the first comprehensive survey providing accurate estimates of the COVID-19 burden anywhere in the world. Further, our findings provide a reasonable approximation of population immunity threshold levels. Using the RAT kits and following the syndromic approach can be useful in screening and monitoring COVID-19. Leveraging existing surveillance platforms, coupled with appropriate methods and sampling framework, renders our model replicable in other settings.

Background:Free radicals or highly reactive oxygen species are capable of inducing oxidative damage to human body. Antioxidants are the compounds which terminate the attack of reactive species and reduce the risk of diseases. Both Baccopa monnieri and Centella asiatica are used in treatment of brain disorders in humans and have almost similar effects.Objective:The study was conducted to determine the antioxidant properties of two well-known memory enhancer medicinal plants Baccopa monnieri and Centella asiatica.Results:The antioxidant activity of these two medicinal plants was evaluated by measuring reducing ability, free radical scavenging activity by DPPH and hydrogen peroxide methods. The antioxidants compounds like ascorbic acid, total phenols and tannins were also evaluated in these plants. Baccopa monnieri and Centella asiatica exhibited significant differences (P<0.05) in their antioxidant values. The methanolic extract of whole leaf powder of Baccopa monnieri exhibited significantly higher antioxidant activity than the Centella asiatica. The antioxidant components viz. ascorbic acid, total phenols and tannins were also found in a higher concentration in Baccopa monnieri as compared to Centella asiatica.Conclusion:It can be concluded from the study that regular use of Baccopa monnieri as a supplement could be more helpful compared to Centella asiatica in treatment of neurological disorders caused by free radical damage.

Cotton ovule epidermal cell differentiation into long fibers primarily depends on wall-oriented processes such as loosening, elongation, remodeling, and maturation. Such processes are governed by cell wall bound structural proteins and interacting carbohydrate active enzymes. Glycosylation plays a major role in the structural, functional, and localization aspects of the cell wall and extracellular destined proteins. Elucidating the glycoproteome of fiber cells would reflect its wall composition as well as compartmental requirement, which must be system specific. Following complementary proteomic approaches, we have identified 334 unique proteins comprising structural and regulatory families. Glycopeptide-based enrichment followed by deglycosylation with PNGase F and A revealed 92 unique peptides containing 106 formerly N-linked glycosylated sites from 67 unique proteins. Our results showed that structural proteins like arabinogalactans and carbohydrate active enzymes were relatively more abundant and showed stage- and isoform-specific expression patterns in the differentiating fiber cell. Furthermore, our data also revealed the presence of heterogeneous and novel forms of structural and regulatory glycoproteins. Comparative analysis with other plant glycoproteomes highlighted the unique composition of the fiber glycoproteome. The present study provides the first insight into the identity, abundance, diversity, and composition of the glycoproteome within single celled cotton fibers. The elucidated composition also indirectly provides clues about unicellular compartmental requirements underlying single cell differentiation.

Background India has experienced the second largest outbreak of COVID-19 globally, yet there is a paucity of studies analysing contact tracing data in the region which can optimise public health interventions (PHI’s). Methods We analysed contact tracing data from Karnataka, India between 9 March and 21 July 2020. We estimated metrics of transmission including the reproduction number (R), overdispersion (k), secondary attack rate (SAR), and serial interval. R and k were jointly estimated using a Bayesian Markov Chain Monte Carlo approach. We studied determinants of risk of further transmission and risk of being symptomatic using Poisson regression models. Findings Up to 21 July 2020, we found 111 index cases that crossed the super-spreading threshold of ≥8 secondary cases. Among 956 confirmed traced cases, 8.7% of index cases had 14.4% of contacts but caused 80% of all secondary cases. Among 16715 contacts, overall SAR was 3.6% [95% CI, 3.4–3.9] and symptomatic cases were more infectious than asymptomatic cases (SAR 7.7% vs 2.0%; aRR 3.63 [3.04–4.34]). As compared to infectors aged 19–44 years, children were less infectious (aRR 0.21 [0.07–0.66] for 0–5 years and 0.47 [0.32–0.68] for 6–18 years). Infectors who were confirmed ≥4 days after symptom onset were associated with higher infectiousness (aRR 3.01 [2.11–4.31]). As compared to asymptomatic cases, symptomatic cases were 8.16 [3.29–20.24] times more likely to cause symptomatic infection in their secondary cases. Serial interval had a mean of 5.4 [4.4–6.4] days, and case fatality rate was 2.5% [2.4–2.7] which increased with age. Conclusion We found significant heterogeneity in the individual-level transmissibility of SARS-CoV-2 which could not be explained by the degree of heterogeneity in the underlying number of contacts. To strengthen contact tracing in over-dispersed outbreaks, testing and tracing delays should be minimised and retrospective contact tracing should be implemented. Targeted measures to reduce potential superspreading events should be implemented. Interventions aimed at children might have a relatively small impact on reducing transmission owing to their low symptomaticity and infectivity. We propose that symptomatic cases could cause a snowballing effect on clinical severity and infectiousness across transmission generations; further studies are needed to confirm this finding.

We report elevated biomass and altered growth characteristics of tobacco plants up on transformation with a NAC (NAM, ATAF1/2,CUC2) gene (GenBank Accession FJ754254) isolated from Lepidium latifolium L. (LlaNAC). Transgenic plants showed significant differences in fresh weight, midrib length of longest leaf, leaf area, height of the plant, root and shoot weights, etc. during vegetative phase. On 100th day after sowing (DAS), plants of transgenic lines were 2-3 times taller than the wild type plants, though no significant difference was recorded in moisture contents of any of the plant tissues. Over-expression of NAC gene up to 2,000 fold was recorded in leaves of transgenic plants on 100th DAS. Interestingly, transgenic plants showed significantly shortened (P(t) = 0.02-0.04) life cycle, as they showed a completely altered growth behaviour. Transgenic plants entered reproductive phase earlier by 60 days, with lines NC2 and NC7b entering first, followed by line NC10. However, the time period spent in the reproductive phase by the plant was nearly twice in case of transgenic lines NC2, NC7b and NC10, as compared to the wild type plants. Despite that, these lines completed their life cycle in 45-60 days lesser than the time taken by wild-type tobacco plants. No difference was recorded in fruit and seed yield of transgenic or wild type plants. To the best of our knowledge, this is the first report on over-expression of NAC gene causing altered growth and biomass patterns. We expect this study to become an important reference towards future engineering of plants for fuel and fodder purposes.