Search

Abstract Background Plant microbiome confers versatile functional roles to enhance survival fitness as well as productivity. In the present study two pearl millet panicle microbiome member species Bacillus subtilis PBs 12 and Bacillus paralicheniformis PBl 36 found to have beneficial traits including plant growth promotion and broad-spectrum antifungal activity towards taxonomically diverse plant pathogens. Understanding the genomes will assist in devising a bioformulation for crop protection while exploiting their beneficial functional roles. Results Two potential firmicute species were isolated from pearl millet panicles. Morphological, biochemical, and molecular characterization revealed their identities as Bacillus subtilis PBs 12 and Bacillus paralicheniformis PBl 36. The seed priming assays revealed the ability of both species to enhance plant growth promotion and seedling vigour index. Invitro assays with PBs 12 and PBl 36 showed the antibiosis effect against taxonomically diverse plant pathogens (Magnaporthe grisea; Sclerotium rolfsii; Fusarium solani; Alternaria alternata; Ganoderma sp.) of crops and multipurpose tree species. The whole genome sequence analysis was performed to unveil the genetic potential of these bacteria for plant protection. The complete genomes of PBs 12 and PBl 36 consist of a single circular chromosome with a size of 4.02 and 4.33 Mb and 4,171 and 4,606 genes, with a G + C content of 43.68 and 45.83%, respectively. Comparative Average Nucleotide Identity (ANI) analysis revealed a close similarity of PBs 12 and PBl 36 with other beneficial strains of B. subtilis and B. paralicheniformis and found distant from B. altitudinis, B. amyloliquefaciens, and B. thuringiensis. Functional annotation revealed a majority of pathway classes of PBs 12 (30) and PBl 36 (29) involved in the biosynthesis of secondary metabolites, polyketides, and non-ribosomal peptides, followed by xenobiotic biodegradation and metabolism (21). Furthermore, 14 genomic regions of PBs 12 and 15 of PBl 36 associated with the synthesis of RiPP (Ribosomally synthesized and post-translationally modified peptides), terpenes, cyclic dipeptides (CDPs), type III polyketide synthases (T3PKSs), sactipeptides, lanthipeptides, siderophores, NRPS (Non-Ribosomal Peptide Synthetase), NRP-metallophone, etc. It was discovered that these areas contain between 25,458 and 33,000 secondary metabolite-coding MiBiG clusters which code for a wide range of products, such as antibiotics. The PCR-based screening for the presence of antimicrobial peptide (cyclic lipopeptide) genes in PBs 12 and 36 confirmed their broad-spectrum antifungal potential with the presence of spoVG, bacA, and srfAA AMP genes, which encode antimicrobial compounds such as subtilin, bacylisin, and surfactin. Conclusion The combined in vitro studies and genome analysis highlighted the antifungal potential of pearl millet panicle-associated Bacillus subtilis PBs12 and Bacillus paralicheniformis PBl36. The genetic ability to synthesize several antimicrobial compounds indicated the industrial value of PBs 12 and PBl 36, which shed light on further studies to establish their action as a biostimulant for crop protection.

The texture evolution during hot compression of extruded ZK60A-T5 magnesium alloy plate loaded along the extrusion direction (ED) and the normal direction (ND) has been examined with the help of pole figures obtained on specimens deformed in the ranges of 200 °C to 500 °C and 0.0003 s−1 to 10 s−1. The results are interpreted in terms of the operating slip systems and mechanisms identified based on processing maps developed for the above two initial specimen orientations. The processing map for the initial ED orientation exhibited three domains. In Domains 1 and 3, first-order pyramidal slip {10 1 ¯ l} 2 ¯ 0> occurs, while in Domain 2, second-order pyramidal slip {11 2 ¯ 2} 2 ¯ 3> occurs. The pole figures obtained on specimens deformed in Domains 1 and 3 are strikingly similar, indicating that the operating slip system controls the texture evolution. Compression in Domains 1 and 3 nearly randomizes the intense basal texture in the as-received specimens, while a new texture is generated in Domain 2 with basal poles at 45° to ND or transverse direction (TD). This new texture will promote basal slip when loaded in a transverse direction. When loaded in the normal direction (ND), the processing map exhibited four domains. In Domains 1 and 4, {10 1 ¯ l} 2 ¯ 3> slip occurs, while {11 2 ¯ 2} 2 ¯ 3> slip occurs in Domains 2 and 3. The pole figures obtained from specimens deformed in Domains 1 and 4 have similar features, while those deformed in Domains 2 and 3 exhibited similar features to one another, confirming that the operating slip systems control the texture development since they are the same in each pair. The compression along ND produces strong basal textures with the basal planes normal to the ED. The texture gets intensified with increased temperature of deformation and causes strong anisotropy in mechanical properties.

AZ31-based nanocomposites are produced by disintegrated melt deposition (DMD) processing. In this investigation, the influence of the addition of Ca to AZ31-1.5 vol.% nano-alumina composite (base) on its hot working behavior is studied to develop a processing route for manufacturing components with these composites. A processing map for the base composite in the temperature range 250–500 °C and strain rate 0.0003–10 s−1 is compared with those for composites with 1% Ca and 2% Ca. The grain size of the base composite is refined by Ca addition and the texture is strengthened. Besides nano-alumina particles, the Ca-containing composites have intermetallic particles (Mg,Al)2Ca present at grain boundaries as well as in the matrix. All the three nanocomposites exhibit three DRX domains, with one of them at high strain rate that facilitates high productivity. Addition of Ca mitigates the occurrence of wedge cracking that occurs in AZ31-1.5NAl composite. Increasing of Ca addition to 2% prevents dynamic recrystallization (DRX) at lower temperatures and strain rates and causes only dynamic recovery. At lower temperatures and higher strain rates, DRX occurs by basal + prismatic slip along with recovery via climb controlled by grain boundary self-diffusion promoted by very fine grain size in the composites.

Among newly developed TX (Mg-Sn-Ca) alloys, TX32 alloy strikes a good balance between ductility, corrosion, and creep properties. This study reports the influence of aluminum and zinc additions (0.4 wt % each) to TX32 alloy on its strength and deformation behavior. Uniaxial compression tests were performed under various strain rates and temperature conditions in the ranges of 0.0003–10 s−1 and 300–500 °C, respectively. A processing map was developed for TXAZ3200 alloy, and it exhibits three domains that enable good hot workability in the ranges (1) 300–340 °C/0.0003–0.001 s−1; (2) 400–480 °C/0.01–1 s−1; and (3) 350–500 °C/0.0003–0.01 s−1. The occurrence of dynamic recrystallization in these domains was confirmed from the microstructural observations. The estimated apparent activation energy in Domains 2 and 3 (219 and 245 kJ/mole) is higher than the value of self-diffusion in magnesium. This is due to the formation of intermetallic phases in the matrix that generates back stress. The strength of TXAZ3200 alloy improved up to 150 °C as compared to TX32 alloy, suggesting solid solution strengthening due to Al and Zn. Also, the hot deformation behavior of TXAZ3200 alloy was compared in the form of processing maps with TX32, TX32-0.4Al, TX32-0.4Zn, and TX32-1Al-1Zn alloys.

The compressive strength and creep resistance of cast Mg-6Al-3Ba-3Ca (ABaX633) alloy has been measured in the temperature range of 25 to 250 °C, and compared with that of its predecessor ABaX422. The alloy is stronger and more creep-resistant than ABaX422, and exhibits only a small decrease of yield stress with temperature. The higher strength of ABaX633 is attributed to a larger volume fraction of intermetallic particles (Al, Mg)2Ca and Mg21Al3Ba2 in its microstructure. Hot deformation mechanisms in ABaX633 have been characterized by developing a processing map in the temperature and strain rate ranges of 300 to 500 °C and 0.0003 to 10 s−1. The processing map exhibits two workability domains in the temperature and strain rate ranges of: (1) 380 to 475 °C and 0.0003 to 0.003 s−1, and (2) 480–500 °C and 0.003 to 0.5 s−1. The apparent activation energy values estimated in the above two domains (204 and 216 kJ/mol) are higher than that for lattice self-diffusion of Mg, which is attributed to the large back-stress that is caused by the intermetallic particles. Optimum condition for bulk working is 500 °C and 0.01 s−1 at which hot workability will be maximum. Flow instability is exhibited at lower temperatures and higher strain rates, as well as at higher temperatures and higher strain rates. The predictions of the processing map on the workability domains, as well as the instability regimes are fully validated by the forging of a rib-web (cup) shaped component under optimized conditions.

Cast Mg–1Zn–1Ca alloy (ZX11) has been tested to evaluate its compressive strength between 25 °C and 250 °C, and workability in the range of 260–500 °C. The ultimate compressive strength of this alloy is about 30% higher than that of creep-resistant alloy Mg–3Sn–2Ca (TX32) between 25 °C and 200 °C, and exhibits a plateau between 100 °C and 175 °C, similar to TX32. This is attributed to Mg2Ca particles present at grain boundaries that reduce their sliding. The processing map, developed between 260 and 420 °C in the strain rate limits of 0.0003 s−1 to 1 s−1, exhibited two domains in the ranges: (1) 280–330 °C and 0.0003–0.01 s−1 and (2) 330–400 °C and 0.0003–0.1 s−1. In these domains, dynamic recrystallization occurs, with basal slip dominating in the first domain and prismatic slip in the second, while the recovery mechanism being climb of edge dislocations in both. The activation energy estimated using standard kinetic rate equation is 191 kJ/mol, which is higher than the value for lattice self-diffusion in magnesium indicating that a large back stress is created by the presence of Ca2Mg6Zn3 intermetallic particles in the matrix. It is recommended that the alloy be best processed at 380 °C and 0.1 s−1 at which prismatic slip is favored due to Zn addition. At higher strain rates, the alloy exhibits flow instability and adiabatic shear band formation at 400 °C.

Pigeon pea ( Cajanus cajan L. Millsp. ) is a legume crop resilient to climate change due to its tolerance to drought. It is grown by millions of resource-poor farmers in semiarid and tropical subregions of Asia and Africa and is a major contributor to their nutritional food security. Pigeon pea is the sixth most important legume in the world, with India contributing more than 70% of the total production and harbouring a wide variety of cultivars. Nevertheless, the low yield of pigeon pea grown under dry land conditions and its yield instability need to be improved. This may be done by enhancing crop nodulation and, hence, biological nitrogen fixation (BNF) by supplying effective symbiotic rhizobia through the application of "elite" inoculants. Therefore, the main aim in this study was the isolation and genomic analysis of effective rhizobial strains potentially adapted to drought conditions. Accordingly, pigeon pea endosymbionts were isolated from different soil types in Southern, Central, and Northern India. After functional characterisation of the isolated strains in terms of their ability to nodulate and promote the growth of pigeon pea, 19 were selected for full genome sequencing, along with eight commercial inoculant strains obtained from the ICRISAT culture collection. The phylogenomic analysis [Average nucleotide identity MUMmer (ANIm)] revealed that the pigeon pea endosymbionts were members of the genera Bradyrhizobium and Ensifer . Based on nodC phylogeny and nod cluster synteny, Bradyrhizobium yuanmingense was revealed as the most common endosymbiont, harbouring nod genes similar to those of Bradyrhizobium cajani and Bradyrhizobium zhanjiangense . This symbiont type (e.g., strain BRP05 from Madhya Pradesh) also outperformed all other strains tested on pigeon pea, with the notable exception of an Ensifer alkalisoli strain from North India (NBAIM29). The results provide the basis for the development of pigeon pea inoculants to increase the yield of this legume through the use of effective nitrogen-fixing rhizobia, tailored for the different agroclimatic regions of India., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Jorrin, Maluk, Atoliya, Kumar, Chalasani, Tkacz, Singh, Basu, Pullabhotla, Kumar, Mohanty, East, Ramachandran, James, Podile, Saxena, Rao and Poole.)

Pigeon pea, a legume crop native to India, is the primary source of protein for more than a billion people in developing countries. The plant can form symbioses with N 2 -fixing bacteria; however, reports of poor crop nodulation in agricultural soils abound. We report here a study of the bacterial community associated with pigeon pea, with a special focus on the symbiont population in different soils and vegetative and non-vegetative plant growth. Location with respect to the plant roots was determined to be the main factor controlling the bacterial community, followed by developmental stage and soil type. Plant genotype plays only a minor role. Pigeon pea roots have a reduced microbial diversity compared to the surrounding soil and select for Proteobacteria , especially for Rhizobium spp., during vegetative growth. While Bradyrhizobium , a native symbiont of pigeon pea, can be found associating with roots, its presence is dependent on plant variety and soil conditions. A combination of 16S rRNA gene amplicon survey, strain isolation, and co-inoculation with nodule-forming Bradyrhizobium spp. and non-N 2 -fixing Rhizobium spp. demonstrated that the latter is a much more successful colonizer of pigeon pea roots. Poor nodulation of pigeon pea in Indian soils may be caused by a poor Bradyrhizobium competitiveness against non-nodulating root colonizers such as Rhizobium . Hence, inoculant strain selection of symbionts for pigeon pea should be based not only on their nitrogen fixation potential but, more importantly, on their competitiveness in agricultural soils. IMPORTANCE Plant symbiosis with N 2 -fixing bacteria is a key to sustainable, low-input agriculture. While there are ongoing projects aiming to increase the yield of cereals using plant genetics and host-microbiota interaction engineering, the biggest potential lies in legume plants. Pigeon pea is a basic food source for a billion low-income people in India. Improving its interactions with N 2 -fixing rhizobia could dramatically reduce food poverty in India. Despite the Indian origin of this plant, pigeon pea nodulates only poorly in native soils. While there have been multiple attempts to select the best N 2 -fixing symbionts, there are no reliable strains available for geographically widespread use. In this article, using 16S rRNA gene amplicon, culturomics, and plant co-inoculation assays, we show that the native pigeon pea symbionts such as Bradyrhizobium spp. are able to nodulate their host, despite being poor competitors for colonizing roots. Hence, in this system, the establishment of effective symbiosis seems decoupled from microbial competition on plant roots. Thus, the effort of finding suitable symbionts should focus not only on their N 2 -fixing potential but also on their ability to colonize. Increasing pigeon pea yield is a low-hanging fruit to reduce world hunger and degradation of the environment through the overuse of synthetic fertilizers.

Nuclear Magnetic Resonance (NMR) is an analytical technique extensively used in almost every chemical laboratory for structural identification. This technique provides statistically equivalent signals in spite of using spectrometer with different hardware features and is successfully used for the traceability and quantification of analytes in food samples. Nevertheless, to date only a few internationally agreed guidelines have been reported on the use of NMR for quantitative analysis. The main goal of the present study is to provide a methodological pipeline to assess the reproducibility of NMR data produced for a given matrix by spectrometers from different manufacturers, with different magnetic field strengths, age and hardware configurations. The results have been analyzed through a sequence of chemometric tests to generate a community-built calibration system which was used to verify the performance of the spectrometers and the reproducibility of the predicted sample concentrations., (Copyright © 2020 Elsevier B.V. All rights reserved.)

USP's peptide reference standards content is typically determined using an HPLC assay against an external standard for which the purity was determined by a mass balance approach. To explore the use of other analytical methods, the USP Biologics Department conducted a multi-laboratory collaborative study. The study determined the inter-laboratory variability for peptide quantitation using the following methods: HPLC assay, quantitative nuclear magnetic resonance (qNMR) spectroscopy, or amino acid analysis (AAA). The three methods were compared with regard to their suitability for quantitation of the nonapeptide oxytocin. In this study, the HPLC assay method using the same peptide bulk material as the standard showed the lowest inter-lab variability. The coefficient of variation (%CV) was calculated without counting the uncertainty associated with the purity assignment of the standard with mass balance. The proton qNMR method is a direct measurement of the peptide against an internal standard, which is not difficult to perform under common laboratory conditions. Because of the simpler operation and shorter analytical time, qNMR as a primary method for peptide reference standard value assignment deserves further exploration., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Protein-tyrosine kinases (PTKs) are of vital importance in a variety of cell functions. Recent studies have provided considerable insight into the binding of growth factors to tyrosine kinase receptors and the consequent induction of signal pathways that lead to a biologic response. Future studies will further delineate the signals that result in a proliferative response and those that induce a differentiation response. Current studies, reviewed here, indicate an important biologic role for PTKs in the regulation of megakaryocyte development and maturation. Whether PTKs function in megakaryocytes in signaling pathways that are similar to pathways in other cells will need to be examined in future studies.