In vitro regeneration from internodal explants of bitter melon (Momordica charantia L.) via indirect organogenesis

Organogenic callus induction and high frequency shoot regeneration were achieved from internodal explants of bitter melon. About 97.5% of internodal explants derived from 30 day old in vivo grown plants produced green, compact nodular organogenic callus in Murashige and Skoog (MS) plus Gamborg et al. (1968) (B5) medium containing 5.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.0 μM thidiazuron (TDZ) after two successive transfers at 11 days interval. Adventitious shoots were produced from organogenic callus when it was transferred to MS medium supplemented with 4.0 μM TDZ, 1.5 μM 2,4-D and 0.07 mM L-glutamine with shoot induction frequency of 96.5% and regeneration of adventitious shoots from callus (48 shoots per explant). Shoot proliferation occurred when callus with emerging shoots was transferred in the same medium at an interval of 15 days. The regenerated shoots were elongated on the same medium. The elongated shoots were rooted in MS medium supplemented with 3.0 μM indole 3-butyric acid (IBA). Rooted plants were acclimatized in green-house and subsequently established in soil with a survival rate of 95%. This protocol yielded an average of 48 shoots per internodal explant after 80 days of culture.Keywords: Adventitious shoots, growth regulators, hardening, organogenic callus, Momordica charanti

In vitro regeneration from internodal explants of bitter melon (Momordica charantia L.) via indirect organogenesis

Organogenic callus induction and high frequency shoot regeneration were achieved from internodal explants of bitter melon. About 97.5% of internodal explants derived from 30 day old in vivo grown plants produced green, compact nodular organogenic callus in Murashige and Skoog (MS) plus Gamborg et al. (1968) (B5) medium containing 5.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.0 μM thidiazuron (TDZ) after two successive transfers at 11 days interval. Adventitious shoots were produced from organogenic callus when it was transferred to MS medium supplemented with 4.0 μM TDZ, 1.5 μM 2,4-D and 0.07 mM L-glutamine with shoot induction frequency of 96.5% and regeneration of adventitious shoots from callus (48 shoots per explant). Shoot proliferation occurred when callus with emerging shoots was transferred in the same medium at an interval of 15 days. The regenerated shoots were elongated on the same medium. The elongated shoots were rooted in MS medium supplemented with 3.0 μM indole 3-butyric acid (IBA). Rooted plants were acclimatized in green-house and subsequently established in soil with a survival rate of 95%. This protocol yielded an average of 48 shoots per internodal explant after 80 days of culture.Keywords: Adventitious shoots, growth regulators, hardening, organogenic callus, Momordica charanti

An efficient Agrobacterium tumefaciens -mediated genetic transformation of bitter melon (Momordica charantia L.)

Abstract A simple and efficient protocol for Agrobacterium tumefaciens-mediated genetic transformation of bitter melon (Momordica charantia L.) has been developed. Pre-cultured leaf explants were transformed by co-cultivation with Agrobacterium tumefaciens strain LBA4404 harbouring a binary vector pBAL2 carrying the reporter gene β-glucuronidase intron (gus) and the marker gene neomycin phosphotransferase (nptII). After co-cultivation, explants were transferred in to a callus induction medium containing 7.7 μM naphthaleneacetic acid (NAA) with 2.2 μM thidiazuron (TDZ), 100 mg L -1 kanamycin and 300 mg L -1 carbenicillin. Regeneration of adventitious shoots from callus was achieved on MS medium containing 5.5 μM TDZ, 2.2 μM NAA, 3.3 μM silver nitrate (AgNO 3 ), 100 mg L -1 kanamycin and 300 mg L -1 carbenicillin. Transgenic shoots were excised from callus and elongated in MS medium fortified with 3.5 μM, gibberellic acid (GA 3 ), 100 mg L -1 kanamycin and 300 mg L -1 carbenicillin. The transgenic elongated shoots were rooted in MS medium supplemented with 4.0 μM indole 3-butyric acid (IBA) and 100 mg L -1 kanamycin. The putative transgenic plants were acclimatized in the greenhouse and seeds were subsequently collected from mature fruits. Further, the presence of acetosyringone (300 μM) in the co-cultivation medium, infection of explants for 30 min and 3 days of co-cultivation proved to be critical factors for greatly improving the transformation efficiency. Histochemical GUS assay and polymerase chain reaction of field-established transgenic plants and their offspring confirmed the presence of the gus and nptII genes, respectively. Integration of T-DNA into the genome of putative transgenics was further confirmed by Southern blot analysis. The nptII gene expression in transgenic plants was confirmed by RT-PCR. A transformation efficiency of 7% was obtained

Prevalence of pulmonary tuberculosis among the tribal populations in India

IMPORTANCE: There is no concrete evidence on the burden of TB among the tribal populations across India except for few studies mainly conducted in Central India with a pooled estimation of 703/100,000 with a high degree of heterogeneity. OBJECTIVE: To estimate the prevalence of TB among the tribal populations in India. DESIGN, PARTICIPANTS, SETTING: A survey using a multistage cluster sampling design was conducted between April 2015 and March 2020 covering 88 villages (clusters) from districts with over 70% tribal majority populations in 17 States across 6 zones of India. The sample populations included individuals ≥15 years old. MAIN OUTCOME AND MEASURES: Eligible participants who were screened through an interview for symptoms suggestive of pulmonary TB (PTB); Two sputum specimens were examined by smear and culture. Prevalence was estimated after multiple imputations for non-coverage and a correction factor of 1.31 was then applied to account for non-inclusion of X-ray screening. RESULTS: A total of 74532 (81.0%) of the 92038 eligible individuals were screened; 2675 (3.6%) were found to have TB symptoms or h/o ATT. The overall prevalence of PTB was 432 per 100,000 populations. The PTB prevalence per 100,000 populations was highest 625 [95% CI: 496–754] in the central zone and least 153 [95% CI: 24–281] in the west zone. Among the 17 states that were covered in this study, Odisha recorded the highest prevalence of 803 [95% CI: 504–1101] and Jammu and Kashmir the lowest 127 [95% CI: 0–310] per 100,000 populations. Findings from multiple logistic regression analysis reflected that those aged 35 years and above, with BMI <18.5 Kgs /m(2), h/o ATT, smoking, and/or consuming alcohol had a higher risk of bacteriologically positive PTB. Weight loss was relatively more important symptom associated with tuberculosis among this tribal populations followed by night sweats, blood in sputum, and fever. CONCLUSION AND RELEVANCE: The overall prevalence of PTB among tribal groups is higher than the general populations with a wide variation of prevalence of PTB among the tribal groups at zone and state levels. These findings call for strengthening of the TB control efforts in tribal areas to reduce TB prevalence through tribal community/site-specific intervention programs

Polyphenol composition and antioxidant capacity from different extracts of Aster scaber

Phenolic contents and antioxidant capacities from different solvent extracts (petroleum ether, ethyl acetate, methanol, butanol and water) of Aster scaber leaf were investigated. Antioxidant activity was evaluated by three different methods, namely DPPH radical scavenging activity, reducing power assay and phosphomolybdenum activity. A total of twenty-three polyphenolic compounds were identified and quantified from A. scaber leaf extracts, including hydroxybenzoic acids, hydroxycinnamic acids, flavonols and other groups of phenolic compounds. Ultra high performance liquid chromatography (UHPLC) analysis of the leaf extract revealed that myricetin (4850.45 μg/g) was the most dominant flavonols, compared to quercetin and kaempferol. Caffeic acid was the dominant phenolic compound in A. scaber leaf extracts, it constituted about 104.20 μg/g, followed by gentisic acid (84.50 μg/g), gallic acid (61.05 μg/g) and homogentisic acid (55.65 μg/g). The total phenolic and flavonoid content was the highest in ethyl acetate extract (322.43 and 6.51 mg/g). The decreasing order of antioxidant activity among the A. scaber leaf extracts assayed through all the three methods was found to be ethyl acetate > butanol > methanol > petroleum ether > water extract

Biotechnological Approaches for Production of Artemisinin, an Anti-Malarial Drug from Artemisia annua L.

Artemisinin is an anti-malarial sesquiterpene lactone derived from Artemisia annua L. (Asteraceae family). One of the most widely used modes of treatment for malaria is an artemisinin-based combination therapy. Artemisinin and its associated compounds have a variety of pharmacological qualities that have helped achieve economic prominence in recent years. So far, research on the biosynthesis of this bioactive metabolite has revealed that it is produced in glandular trichomes and that the genes responsible for its production must be overexpressed in order to meet demand. Using biotechnological applications such as tissue culture, genetic engineering, and bioreactor-based approaches would aid in the upregulation of artemisinin yield, which is needed for the future. The current review focuses on the tissue culture aspects of propagation of A. annua and production of artemisinin from A. annua L. cell and organ cultures. The review also focuses on elicitation strategies in cell and organ cultures, as well as artemisinin biosynthesis and metabolic engineering of biosynthetic genes in Artemisia and plant model systems

Understanding health care-seeking behaviour of the tribal population in India among those with presumptive TB symptoms.

Background and objectivesUnderstanding the drivers for care-seeking among those who present with symptoms of TB is crucial for early diagnosis of TB and prompt treatment, which will in turn halt further TB transmission. While TB is a challenge among the tribal population, little is known about the care-seeking behaviour and the factors influencing care-seeking behaviour among the tribal population across India.MethodologyThis community-based descriptive study was carried out in 17 states of India across 6 zones, covering 88 villages from tribal districts with over 70% tribal population. The sample population included individuals ≥15 years old who were screened through an interview for symptoms suggestive of pulmonary TB (PTB), currently and/or previously on anti-TB treatment. Those with symptoms were then assessed on their health-seeking behavior using a semi-structured interview schedule.ResultsAmong 74532 eligible participants screened for symptoms suggestive of TB, 2675 (3.6%) were found to be presumptive TB cases. Of them, 659 (24.6%) sought care for their symptoms. While 48.2% sought care after a week, 19.3% sought care after one month or more, with no significant difference in the first point of care; 46.9% approaching a private and 46.7% a public facility. The significant factors influencing care-seeking behaviour were knowledge on TB (OR: 4.64 (3.70-5.83), p ConclusionDelayed healthcare-seeking behaviour among those with symptoms presumptive of TB in the tribal population is a major concern. Findings point to differences across gender about symptoms that prompt care-seeking in this population. Gender-sensitive interventions with health system strengthening are urgently needed to facilitate early diagnosis and treatment among this population

Development of Decadal (1985–1995–2005) Land Use and Land Cover Database for India

India has experienced significant Land-Use and Land-Cover Change (LULCC) over the past few decades. In this context, careful observation and mapping of LULCC using satellite data of high to medium spatial resolution is crucial for understanding the long-term usage patterns of natural resources and facilitating sustainable management to plan, monitor and evaluate development. The present study utilizes the satellite images to generate national level LULC maps at decadal intervals for 1985, 1995 and 2005 using onscreen visual interpretation techniques with minimum mapping unit of 2.5 hectares. These maps follow the classification scheme of the International Geosphere Biosphere Programme (IGBP) to ensure compatibility with other global/regional LULC datasets for comparison and integration. Our LULC maps with more than 90% overall accuracy highlight the changes prominent at regional level, i.e., loss of forest cover in central and northeast India, increase of cropland area in Western India, growth of peri-urban area, and relative increase in plantations. We also found spatial correlation between the cropping area and precipitation, which in turn confirms the monsoon dependent agriculture system in the country. On comparison with the existing global LULC products (GlobCover and MODIS), it can be concluded that our dataset has captured the maximum cumulative patch diversity frequency indicating the detailed representation that can be attributed to the on-screen visual interpretation technique. Comparisons with global LULC products (GlobCover and MODIS) show that our dataset captures maximum landscape diversity, which is partly attributable to the on-screen visual interpretation techniques. We advocate the utility of this database for national and regional studies on land dynamics and climate change research. The database would be updated to 2015 as a continuing effort of this study