Performance of garden pea varieties for their growth and yield characteristics in Vidharbha region of Maharashtra, India

An experiment was conducted in 2013 to study the performance of different varieties of garden pea under Akola condition at Department of Horticulture Dr.Punjabrao Deshmukh Krishi Vidhyapeeth, Akola, Maharashtra. Eight varieties were evaluated on black soil in replicated randomized block design and Results were found significant for all characters among these varieties. All varieties exhibited considerable variation in their performance for most of the parameters. Better growth and yield parameters in terms of plant height (cm), number of branches/plant, days to first flowering, number of green pod/plant, green pod weight, green pod length, pod yield/plant, green pod yield per plot and green pod yield per ha were noticed in all varieties. Maximum plant height was observed in Jawahar Matar-2 (72.26 cm) and minimum was in Palam Priya (28.46 cm). In case of number of pods plant-1 was maximum in PB-89 (16.43) followed by Palam Triloki (13.9) and minimum in Jawahar Matar-2 (9.83). Similarly for pod characters, average pod weight, maximum pod weight was recorded in PB-89 (6.12 g) and minimum was recorded in Arka Kartik (3.27g). Green pod yield/plant was highest in PB-89 (87.93 g), Palam Triloki (75.45 g) and Ankur (68.42 g). Whereas, maximum green pod/yield. was recorded in PB-89 (93.12q/ha) followed by Palam Triloki (76.97q/ha). Among all these varieties highest protein and Total Soluble Solid contents was recorded in Palam Triloki variety (23.06% and 17.67% respectively). PB-89, Palam Triloki and Ankur had the highest yields over the others, hence, they are recommended to farmers in semi-arid condition of Vidharba region for cultivation

Depletion of M. tuberculosis GlmU from infected murine lungs effects the clearance of the pathogen

M. tuberculosis N-acetyl-glucosamine-1-phosphate uridyltransferase (GlmUMtb) is a bi-functional enzyme engaged in the synthesis of two metabolic intermediates N-acetylglucosamine-1-phosphate (GlcNAc-1-P) and UDP-GlcNAc, catalyzed by the C- and N-terminal domains respectively. UDP-GlcNAc is a key metabolite essential for the synthesis of peptidoglycan, disaccharide linker, arabinogalactan and mycothiols. While GlmUMtb was predicted to be an essential gene, till date the role of GlmUMtb in modulating the in vitro growth of Mtb or its role in survival of pathogen ex vivo / in vivo have not been deciphered. Here we present the results of a comprehensive study dissecting the role of GlmUMtb in arbitrating the survival of the pathogen both in vitro and in vivo. We find that absence of GlmUMtb leads to extensive perturbation of bacterial morphology and substantial reduction in cell wall thickness under normoxic as well as hypoxic conditions. Complementation studies show that the acetyl- and uridyl- transferase activities of GlmUMtb are independently essential for bacterial survival in vitro and GlmUMtb is also found to be essential for mycobacterial survival in THP-1 cells as well as in guinea pigs. Depletion of GlmUMtb from infected murine lungs, four weeks post infection, led to significant reduction in the bacillary load. The administration of Oxa33, a novel oxazolidine derivative that specifically inhibits GlmUMtb, to infected mice resulted in significant decrease in the bacillary load. Thus our study establishes GlmUMtb as a strong candidate for intervention measures against established tuberculosis infections

Short Communication: Isolation and biochemical characterization of Rhizobium strains from nodules of lentil and pea in Tarai agro-ecosystem, Pantnagar, India

Abstract. Upadhayay SP, Pareek N, Mishra G. 2015. Isolation and biochemical characterization of Rhizobium strains from nodules of lentil and pea in Tarai agro-ecosystem, Pantnagar, India. Nusantara Bioscience 7: 73-76. Root nodules were collected from young and healthy seedling of Pisum sativum L and Lens culinaris L from the field at different locations of Norman E. Borloug Crop Research Center, G.B.P.U.A. &amp; T., Pantnagar, Uttarakhand state, India. Fifteen Rhizobium strains were isolated from the root nodule of P. sativum and L. culinaris and characterized by standard tests. All strains were gram-negative and did not absorb red color when cultured in YEMA containing congo red. In the ketolactose test yellowish zone of Cu2O not found. Also, isolates showed either poor or no growth on the glucose peptone medium after one day which is indicating character of rhizobia Thirteen isolates were fast grower and only two were slow growers which is confirmed by bromothymol blue test. Results confirmed that isolated strains were Rhizobium.</jats:p

Evaluation of bread wheat (Triticum aestivum L.) for terminal heat tolerance

Terminal heat tolerance of 34 wheat genotypes were analyzed for two years. Among 14 traits, canopy temperature, plot yield and days to heading were major components in clustering of genotypes. Three genotypes viz., DBW39, DBW16 and DBW14 had lowest HSI (0.34-0.36) for plot yield and were considered as heat tolerant genotypes by, both, HCA (Hierarchical Cluster Analysis) as well as DA (Discriminant Analysis). These genotypes may serve as potential donors in wheat breeding to improve the terminal heat tolerance.</jats:p

Depletion of GlmU_Mtb from infected lungs leads to clearance of pathogen.

<p><b>(A)</b><i>Rv</i> and <i>Rv</i>∆<i>glmU</i> cultures were inoculated at an initial <i>A</i>₆₀₀ of 0.1 and the growth was monitored every day for eight days. ATc was added to the <i>Rv</i> culture on day 2 and <i>Rv</i>∆<i>glmU</i> cultures were either grown in the absence of ATc or was supplemented with ATc in the growth media on 2^nd, 4^th or 6^th day. Experiment was performed in triplicates and the error bars represent s.e.m. <b>(B)</b> Differentiated THP-1 cells were infected <i>Rv</i> or <i>Rv</i>∆<i>glmU</i> cultures and the infection was allowed to be established for 24 h. 24 h post infection ATc was supplemented in the media for <i>Rv</i> and <i>Rv</i>∆<i>glmU</i> infected cultures. As a control THP1 cells infected with <i>Rv</i>∆<i>glmU</i> were grown without ATc. CFUs were enumerated at 0, 24 and 96 h post infection. **<i>p</i><0.005, two tailed non parametric <i>t</i>-test, Experiment was performed in triplicates and the error bars represent s.e.m. <b>(C)</b> BALB/c mice (6 to 9 mice / group) were infected with <i>Rv</i> and <i>Rv</i>∆<i>glmU</i> strains and the infection was established for next 28 days. Subsequently, Dox was provided (with 5% dextrose on every alternate day) in the drinking water for <i>Rv</i> infected mice. One group of <i>Rv</i>∆<i>glmU</i> infected mice were administered with Dox and other with vehicle control for the next 56 days. CFUs were enumerated on day 1, day 28 and day 84 post infection. At 28^th days post infection mean CFUs for <i>Rv</i> and <i>Rv</i>∆<i>glmU</i> infected mice were 4.43 and 4.47 on log₁₀ scale and 84^th days post infection mean CFUs for <i>Rv</i> +Dox or <i>Rv</i>∆<i>glmU</i> +Dox and <i>Rv</i>∆<i>glmU</i>–Dox were 5.1, 0.42 and 5.2 on log₁₀ scale. ***<i>p</i><0.0005, two tailed non parametric <i>t</i>-test, mean, error bars represent s.e.m.. <b>(D)</b> Overall pathology and histopathology of infected BALB/c mice lungs. Infected lungs dissected on 84^th day after infection from <i>Rv</i> (+Dox) and <i>Rv</i>∆<i>glmU</i> (-Dox) infected mice shows clear lesions (upper panel) and granuloma with lymphocytes and foamy histiocytes (lower panel, HE stain, 100x) while <i>Rv</i>∆<i>glmU</i> (+Dox) was showing normal lung parenchyma and no granuloma. G = Granuloma, BL = Bronchial Lumen, AS = Alveolar Space. <b>(E)</b> Granuloma scores from 4 and 12 weeks of histopathology results.</p

Presence of GlmU_Mtb is obligatory for the survival of <i>Mtb</i> in the host.

<p><b>(A)</b> Confocal microscopy images of THP1 cells infected with <i>Rv</i> and <i>Rv</i>∆<i>glmU</i> in the presence or absence of ATc as indicated were taken 48 h post infection. Bacteria were stained with FITC (green) and lysosomes were stained with Lyso Tracker red DND 99 (red). Scale bars, 10 μm. <b>(B)</b> Quantification of percentage co-localization of bacterium and lysosomes in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005235#ppat.1005235.g003" target="_blank">Fig 3A</a>. n = 10 to 15. <b>(C)</b> PMA differentiated THP1 cells were infected with <i>Rv</i> and <i>Rv</i>∆<i>glmU</i> and either ATc (400 ng/ml) or the vehicle were added at 0 h. CFUs were enumerated at different time points post infection. The experiment was performed in triplicates and error bars represent s.e.m. <b>(D)</b> Guinea pigs were infected with 150–200 bacilli CFUs of <i>Rv</i> or <i>Rv</i>∆<i>glmU</i> and Dox was provided in the water as indicated. Guinea pigs (n = 2) were sacrificed on day 1 and homogenates from the lungs were plated in triplicates to determine the implantation. Guinea pigs (6 guinea pigs /group) were sacrificed four weeks post infection and CFUs were determined in the lung homogenates and results were plotted with log₁₀ /lung on the Y-axis and samples on the X-axis. At four weeks post infection mean CFUs for <i>Rv</i> +Dox or <i>Rv</i>∆<i>glmU</i> +Dox and <i>Rv</i>∆<i>glmU</i>–Dox were 5.2, 0 and 5.11 on log₁₀ scale.***<i>p</i><0.0001, two tailed non parametric <i>t</i>-test, error bars represent s.e.m. <b>(E)</b> Overall pathology (upper panel) and histopathology (lower panel, HE, 40X) of the infected guinea pig lungs four weeks post infection. Both <i>Rv</i> +Dox and <i>Rv</i>∆<i>glmU</i> -Dox were having prominent granuloma and necrotic lesions in the central and epithelioid cells and lymphocytes around it. Guinea pigs infected with <i>Rv</i>∆<i>glmU</i> +Dox were showing normal lung parenchyma with clear alveolar spaces. G = Granuloma, AS = Alveolar Space. <b>(F)</b> Granuloma scores from histopathology results.</p

Acetyl and uridyltransferase activities are independently essential.

<p><b>(A)</b> and <b>(B)</b> Schematic and cartoon representation of GlmU_Mtb depicting different domains, active site residues and the deletion mutants. <b>(C)</b> GlmU_Mtb and GlmU_Mtb-mutants were purified as described earlier [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005235#ppat.1005235.ref038" target="_blank">38</a>] and the 1 μg of the purified proteins were resolved on 10% SDS-PAGE and stained with coomassie. <b>(D)</b> Uridyltransferase (left panel) and acetyltransferase (right panel) activities were carried out as describe in Methods using 0.5 to 20 pmoles of wild type or mutant GlmU_Mtb proteins. Activity was defined as μM product formed / min / pmole of enzyme. Relative activities of the mutants were calculated with respect to the activity of GlmU_Mtb, which was normalized to 100%. The experiment was repeated three times and the error bars indicate s.e.m. <b>(E)</b> Wild type and mutated GlmU_Mtb genes were cloned into pNit vector without any N- or C- terminal tag. pNit-<i>glmU</i>_wt or pNit-<i>glmU</i>_mutant constructs were electroporated into <i>Rv</i>∆<i>glmU</i>, and the WCLs prepared from <i>Rv</i>∆<i>glmU and Rv</i>∆<i>glmU</i>::<i>glmU</i>_{<i>mutant</i>} cultures were resolved and probed with anti-GlmU and anti-GroEL1 antibodies. Bands corresponding to FLAG-GlmU_Mtb, complemented GlmU_{<i>wt/mutant</i>} and the deletion fragments of GlmU are indicated. <b>(F)</b><i>Rv</i>∆<i>glmU and Rv</i>∆<i>glmU</i>::<i>glmU</i>_{<i>mutant</i>} cultures were seeded at an initial A₆₀₀ of 0.1 and grown for five days in the absence or presence of ATc. The experiment was performed in triplicates and the error bars represent s.e.m.</p

Effect of GlmU_Mtb depletion of dormant bacteria.

<p><b> (A)</b> Schematic outline of the experiment. <b>(B)</b><i>Rv</i> and <i>Rv</i>∆<i>glmU</i> cultures were seeded at an initial <i>A</i>₆₀₀ of 0.1 in 1.5 HPLC tubes or in 500 ml flasks containing penetrable caps. Establishment of hypoxia was monitored with the help of methylene blue color change (blue to colorless). ATc was added to <i>Rv</i> on 20^th day and to <i>Rv</i>∆<i>glmU</i> on 20^th and 40^th day. CFUs were enumerated on day 0, day 20 and day 42. *<i>p</i><0.05 or ***<i>p</i><0.005, two tailed non parametric <i>t</i>-test, Error bars indicate s.e.m. and the experiment was performed in triplicates (n = 3). <b>(C)</b> INH (50 ng/ml) was added to the hypoxic <i>Rv</i> cultures on 20^th and 40^th day. CFUs were enumerated on day 42. *<i>p</i><0.05 or NS: non- significant, two tailed non parametric <i>t</i>-test, Error bars indicate s.e.m. and n = 3. <b>(D)</b> Large scale hypoxic cultures were pelleted down on day 42 and processed for scanning electron microscopy (upper panel) or transmission electron microscopy (lower panel) imaging. (SEM: scale bars: 1 μm; TEM scale bar: 20 nm). Cell wall thickness is indicated by red lines. <b>(E)</b> Cell wall thickness was measured in nm for 20 to 54 cells from the TEM images of different samples (representative image shown in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005235#ppat.1005235.g003" target="_blank">Fig 3C</a>). ***<i>p</i>< 0.0001 and 0.0005, two tailed non parametric <i>t</i>-test, Error bars indicate s.e.m.</p