Mungbean [Vigna radiata (L.) R. Wilczek], is an Indian-origin legume crop and is emerging as an excellent food for human health and nutrition globally. However, the narrow genetic base in its released varieties is a bottleneck in mungbean yield enhancement. Therefore, 3903 accessions of mungbean were characterized to identify trait-specific donors, understand potential traits for breeding purposes, and select a core collection for enhancing germplasm utilization in the mungbean breeding program. A common set of 28 phenotypic traits was used for evaluation at two locations for two years (for 2019 and 2020) in an augmented block design. A substantial amount of phenotypic variation was observed in mungbean collections as revealed by diversity indices, frequency distribution, hierarchical clustering, and principal component analysis. Traits such as plant biomass, days to 50% flowering, days to maturity, plant height, and number of seeds/pod showed higher levels of heritability and genetic gains. The core development strategy involved the development of five independent core sets of 400 accessions based on different statistical methods. Among the five sets, a core collection i.e., EN100 (average entry-to-nearest-entry distance with 100% weightage), was identified as the best among all five core sets based on quality evaluation indices. The chosen core collection was further compared with the entire collection using various statistical parameters. The core collections showed optimum results based on the various quality assessment parameters such as mean difference (MD, 66.67%), variance difference (VD, 100%), coincidence rate of range (CR, 97.4%), variable rate of range (VR, 119.6%), class coverage (100%) and a higher level of coefficient of variation (CV) for all the quantitative traits. These findings provide insights into the mungbean germplasm diversity, which will play a significant role in the mungbean improvement programs. Additionally, the diversity-rich core collection will be a ready resource for breeders and researchers to address various research problems.

The present study provides the first integrative genomic comparison of five Vigna species Vigna angularis, V. umbellata, V. mungo, V. radiata, and V. unguiculata to characterize their genomic architecture and diversity. It gives a valuable understanding to the evolutionary history and genetic diversity of these legumes that may have wide inference for crop improvement under breeding programs and ultimately contribute to the food and nutrition security. Protein and genomic fasta sequences were retrieved from the NCBI GenBank. A total of 8194 single-copy and 23,560 unique genes were identified. Vigna umbellata, V. angularis, V. radiata, and V. mungo exhibited closest relationship, while V. unguiculata showed a higher degree of genomic divergence. The study reported that 496 gene clusters were linked to response to salt stress, 406 for response to water-deficit stress, and 319 for response to oxidative stress, while 112 genes were identified for response to cold exhibiting the robustness of these legume species against abiotic stresses. The gene family expansion of the Vigna species indicated a significant increase in the gene families compared to their common ancestor. The finding unravels the evolutionary relationships and genome evolution of the five Vigna species. The study revealed the divergence of V. angularis, V. umbellata, V. mungo, and V. radiata from the common ancestor that have experienced evolutionary pressures favoring diversification and increased adaptability. Therefore, these Vigna species may be used to develop improved varieties of other Vigna species being cultivated for food purposes.

Moth bean is a small-seeded, protein-rich, and abiotic stress-resilient legume grown in arid and semi-arid areas of South Asia. Cultivation of this underutilized legume can reduce the dependency on commercialized Vigna species for food and nutrient security in underdeveloped countries. However, low yield remains one of the major limitations in the wider adoption of the crop among farmers in South Asian countries. To address this challenge, the present study employs genome-wide association studies to dissect the genetic basis of seed weight, a very important trait directly related to seed size and yield. The study used a diverse panel of 327 genotypes, which were phenotyped for 100-seed weight trait across three environments. By employing four multi-locus GWAS methods and 8518 SNP markers, we identified 17 significant SNPs across the genome associated with seed weight. The important seed size/seed development-associated candidate genes anchoring these SNPs were S-protein-like protein, purple acid phosphatase, kinetochore protein spc25 isoform X2 and CBL-interacting protein kinase 8-like protein. Four trait-associated SNPs/genomic regions identified in this study exhibited significant effects on the 100-seed weight trait. Genes associated with two of these SNPs (SNP2374: “g20155.t2” encoding purple acid phosphatase, and SNP5696: “g29506.t1” encoding kinetochore protein spc25 isoform X2) showed prominent upregulation in flower and early developing seed tissues/pod development stages suggesting their role in regulating seed size in moth bean. Trait-based markers developed from these candidate genes could potentially accelerate the marker-assisted breeding program to enhance seed weight and overall productivity of moth bean.

Bedding materials play significant role to provide comfortable microenvironment to animals under intensive production system, which can be assessed by behaviour of the animal. The present study observed the effect of bedding material in different behaviour parameters of Osmanabadi goat kids. A total number of 18 Osmanabadi goat kids between 2-3 months of age were distributed into three groups (6 kids per group): control (T0) and treatments (T1, T2). The kids in T0 were reared in concrete floor without any bedding material, whereas treatment groups T1 and T2 were composed of wood shavings and straw bedding material, respectively. At the end of experiment lying time per day of T2 group was significantly higher as compared to T0 and T1 groups. Lying time per bout was significantly higher in T2 group than T0 group, but corresponding values were similar to that of T1 group. Standing time per day was higher in concrete floor (T0) as compared to wood shavings and straw bedding group. Fighting behaviour was significantly higher in kids kept in concrete floor and wood shavings group than those kept in straw bedding group. Grooming behaviour was higher in T2 group as compared to T0, but similar to T1 group and kids had more negative behaviour in concrete floor. Overall, it can be concluded that straw bedding has a positive influence on the welfare of growing kids and therefore can be used as an alternative bedding material in growing goat kids.

Horsegram has wide genetic variability including variability in the seed colour. Significant variability for various morphological traits, namely, plant height, number of branches, number of nodes, number of pods, days to flowering, day to maturity, pod length, pod yield and seed yield, has been recorded. The genetic variability in the crop has been assessed using different methods. A detailed account on genetic improvement and genetic variability in horsegram has been discussed in this chapter.

Mung bean [Vigna radiata (L.) Wilczek] is one of the upsurging, highly economical, nutritive Asiatic leguminous crops. The crop is getting higher attention in terms of the consumption and production worldwide being an important source of amino acids, proteins, dietary fibre and unsaturated fatty acids. It possesses folate and iron in significant amount along with several phytochemicals. The short life cycle and nitrogen-fixing ability make it more suitable for sowing along with other crops. In spite of several advantages, it has got less attention in terms of development of morphophysiological and molecularly diverse varieties. Mung bean has a small genome, and fortunately it has been sequenced; therefore, it may be utilized as an exemplary plant to understand other legumes. Development of wild mung bean pool from diverse origins and environmental conditions would help to conserve the genetic wealth of the crop. Higher yields, shorter maturity period, higher harvest index, photoperiod insensitivity, resistance to major insect pests/diseases, compact canopy and synchronous maturity are some of the important objectives for crop improvement in mung bean. This chapter reviews the morphophysiological and molecular diversity of mung bean and also gives an insight about mutagenesis, plant protection and abiotic stresses associated with the crop.

Moth bean is susceptible to several diseases, but yellow mosaic virus, bacterial blight, etc. are some of the serious diseases that appear almost every year leading to reduced production and productivity of moth bean. Cloudy climates, lesser sunshine and lesser variation between the day and night temperatures are the most optimum climates favouring the diseases. Development of varieties having disease resistance potential and development of robust strategies of insect pest management are the most holistic and realistic approaches to deal with the issues related to plant protection in moth bean. Several germplasms and improved lines have been screened for disease resistance potentials in the crop. A detailed account of fungal, bacterial, viral diseases and major insect pests of the crop has been discussed in this chapter.

Horsegram can adapt harsh environmental conditions and has fairly good tolerant capacity towards various physiological and abiotic stresses. However, better seed production can be realized by addressing various issues related to physiological and abiotic stresses. Physiological aspects associated with seed colour, germination and seedling growth, seed yield, storage along with effect of heavy metal, water stress, drought condition and salt tolerance have been discussed in this chapter.

Genetic diversity studies have immense importance as it provides information for selection of suitable genotype for hybridization. Cowpea is a self-pollinated crop, and pollination in the flower usually occurs before the opening of flowers. Therefore, variation in the DNA polymorphism is not much expected. The crop has been widely exploited for genetic assessment using various molecular markers. A brief account of various marker techniques, namely, random amplified polymorphic DNA (RAPD), simple sequence repeats (SSR), inter simple sequence repeats (IRRs), DNA amplification fingerprinting (DAF), start codon targeted (SCoT), sequence-related amplified polymorphism (SRAP), restriction fragment length polymorphism (RFLP), along with biochemical markers used for different analyses in cowpea, has been described. Besides, these genetic linkage and genetic map studies carried out in cowpea have also been described in this chapter.

Background: Bone grafting is a preferred treatment option for the healing of large diaphyseal bone defects and is useful in the management of nonunion, delayed union, and tumor resection. Aims: To investigate a decellularization protocol of bovine cancellous bone for xenogenic implantation in radial bone defects in rabbits. Methods: Bovine bone scaffolds fabricated with various decellularization protocols viz phosphate buffer saline (PBS), 1% sodium dodecyl sulfate (SDS), and rapid freeze and thaw technique. The manufactured scaffolds were characterized by biomechanical testing, histological staining, and scanning electron microscopy. A 10 mm rabbit radius bone defect was repaired with autograft and SDS treated and rapid freeze and thaw in groups A, B, and C respectively. Healing was evaluated by radiography and histopathology at 0, 30, 60, and 90 days. The grafts were also checked for host tissue reaction and incorporation into the defect. Results: The freeze and thaw group showed complete elimination of all cellular nuclei, regular arrangement of collagen fiber, and no significant difference in tensile strength compared to 1% SDS treated and native groups. The in vivo radiographic and histopathological study showed that the rapid freeze and thaw group had complete bridging of the bone gap defect with new bone formation and they were immunologically less reactive compared to group B. Conclusion: The in vitro and in vivo evaluation of the grafts suggested that freeze and thaw technique was most superior to all other techniques for effective decellularization and augmentation of bone healing with better integration of the graft into the host.

Species composition and diversity dynamics of the actinomycetes was studied in five salt basins of arid and semi-arid areas of Rajasthan, India. A novel approach integrating molecular (16S rRNA gene) and diversity indices was applied to reveal species composition and diversity dynamics. Fifty-three actinomycetes isolates were isolated from five arid and semi-arid salt basins. Molecular characterization resulted in the identification of actinomycetes species belonging to three genera namely, Streptomyces, Nocardiopsis, and Actinoalloteichus. The diversity study among actinomycetes species validates their universal occurrence in arid and semi-arid regions of Rajasthan. The species N. dassonvillei subsp. albirubida was omnipresent in all the five salt basins but its relative manifestation was not static across habitats. The study revealed that three species N. chromatogenes, S. durbertensis, and S. mangrovicola are being reported for the first time from India. The maximum species of actinomycetes were recorded from Pachpadra (14) and the minimum from Didwana area (6). This study not only documents the hitherto wealth of actinomycetes species in arid and semi-arid salt basins of Rajasthan but also reveals the composition and diversity dynamics of actinomycetes.

Twenty-six morphotypes of actinomycetes bacteria were isolated from the soils of arid zone of Indian Thar desert, Rajasthan. A significant and positive correlation was found between density of actinomycetes isolates and availability of nitrogen in sandy soil of arid zone suggesting the influence of soil nitrogen on occurrence and propagation of actinomycetes in this region. Molecular identification based on 16S rRNA gene sequencing revealed that the bacterial isolates belong to four actinomycetes genera, viz. Streptomyces (22 species), Nocardiopsis (two species), Saccharomonospora (one species) and Actinoalloteichus (one species). The preliminary screening of 26 isolates against five human pathogenic bacteria, viz. Escherichia coli, Vibrio cholera, Salmonella enterica typhimurium, Staphylococcus aureus and Enterococcus faecalis, showed that only four isolates, viz. Streptomyces sp. (ITD-27), S. enissocaesilis (ITD-29), S. Malachitospinus (ITD-35) and Streptomyces sp. (ITD-47), had antibacterial activity. The secondary screening of these four isolates revealed that the isolate S. malachitospinus (ITD-35) showed the maximum growth inhibition zone and inhibited the growth of all tested gram-positive and gram-negative pathogenic bacteria. Gas chromatography–mass spectrometry analysis of S. malachitospinus (ITD-35) cultural filtrate in n-butanol solvent identified three antibacterial compounds of medicinal significance, viz. 3-octanone, neopentyl isothiocyanate and 2-methyl butyl isothiocyanate.

The genus Nocardiopsis is well known to produce secondary metabolites especially antibacterial bioactive compound. Isolation and characterization of bioactive compounds producing novel isolates from unusual habitats are crucial. The present study was aimed to explore Didwana dry salt lake of Rajasthan state in India for the isolation and characterization of actinomycetes. The isolated actinomycetes isolates were characterized based on culture characteristics, biochemical tests and 16S rRNA gene sequencing. The 16S rRNA gene sequence analysis revealed that all the five isolates inhabiting soil of the said dry salt lake of Didwana, Rajasthan belonged to four species of Nocardiopsis viz., N. synnemataformans, N. potens, N. prasina and N. dassonvillei subsp. albirubida. The molecular identification based on 16S rRNA gene sequences was found accurate and robust. The phylogram generated through multiple sequence alignment of all the test isolates of Nocardiopsis revealed that the isolates aroused from a single branch and validated monophyletic association. The present study is the first report of exploring Nocardiopsis isolates from the dry salt lake. These characterized Nocardiopsis isolates isolated from Didwana dry salt lake habitat are novel stains and can be of significance in the detection and utilization of novel bioactive compounds.

Most soil fungi are omnipresent and produce metabolites for their survival. These metabolites are of considerable significance to the service of humankind. Nevertheless, some of them are toxic and harmful. The significance of these secondary metabolites in pharmaceuticals, food, agriculture, and day-to-day life is well documented. Laboratory-synthesized metabolites are of limited access and have several limitations. However, fungi can play a greater role in the supply of metabolites. Fungal-derived products have been found to possess several useful pharmacological and medicinal properties. Some are antibiotic, hypocholesterolemic, anti-cancerous, and immunosuppressive, and are used in products such as pigments, sweeteners, fungal enzymes, and lectins. Presently, many fungi are a major component of a large number of industries and have enormous capacity for improving microbial products. Discovery of novel metabolites will help to secure our health against resistant microbial species responsible for causing deadly diseases. This chapter reviews current applications of metabolites of soil-borne fungi and their prospects.

Aim : The aim of the present study was to isolate antifungal protein from seeds of Acacia senegal in order to develop a new, effective and environmental friendly biofungicide. Methodology : Antifungal protein from A. senegal seeds was extracted and purified through ammonium sulphate precipitation, dialysis, ion exchange and gel filtration chromatography. The novel antifungal was characterized employing SDS-PAGE, chitinase activity and antifungal efficacy. The purified protein was also characterized through MALDI-TOF MS/MS. Results : The yield of purified antifungal protein was estimated to be 0.96 mg 25 g(-1) seeds and its molecular mass determined by SDS PAGE was 52.9 kDa. The purified protein exhibited antifungal activity against phytopathogenic fungi viz., Macrophomina phaseolina and Fusarium oxysporum also possessed chitinase activity. The purified protein was characterized through MALDI-TOF MS/MS and its spectra revealed 14 peptides with their specific amino acid sequences. Interpretation : The antifungal protein isolated from A. senegal seeds has broad-spectrum antifungal activity with chitinase activity against pathogenic fungi that can be exploited for management of fungal disease as biopesticide to promote sustainable agriculture.

Background: Cationic antimicrobial peptides (AMPs) have attracted considerable attention to source antimicrobial as well as antibiofilm technology solutions. In vitro and in vivo antimicrobial efficacies of three short-chain AMPs, Indolidicin, Cecropin A (1–7) – Melittin (CAMA), lactoferricin (17–30), and their combination were explored for the first time against multi-drug resistant enteroaggregative Escherichia coli (MDR-EAEC). Methods and materials: Minimum Inhibitory Concentrations (MICs) and Minimum Bactericidal Concentrations (MBCs) of AMPs were determined by broth micro-dilution technique. The AMPs were tested for its stability at temperatures (70 °C and 90 °C), proteases (Proteinase-K and trypsin) and physiological concentration of cationic salts (NaCl and MgCl2); safety in sheep RBCs and cell lines (RAW 264.7 and HEp-2); adverse effect on beneficial flora and membrane permeability. In vitro dose- and time-dependent time kill assay was also carried out. Later, in vivo studies of AMPs (alone and in combination) employing Galleria mellonella (survival assay, bacterial burden, haemocyte density, melanisation assay, cytotoxicity assay and histopathological examination) was carried out. Results: All the three AMPs were stable (except for Indolicidin with trypsin), tested safe at lower MIC and no adverse effect against beneficial flora was observed. AMPs alone and in combination, exhibited bacterial membrane permeability. In vitro time-kill assay revealed concentration-cum-time dependent clearance of MDR-EAEC in the AMP-treated groups, while, in the in vivo G. mellonella experiment, the infected group treated with AMPs (alone and in combination) revealed an increased survival rate, immunomodulatory effect, reduced MDR-EAEC counts and were tested safe to the larval cells which concurred with histopathological observations. Conclusion: The AMPs seem to be promising candidates for further investigations using appropriate animal models for MDR-EAEC strains.

Retention of fetal membranes (RFM) in bovines not only lowers fertility and productivity but also leads to significant economic loss to the dairy farms. The present study compared peripartum progesterone, estradiol, cortisol, Prostaglandin F2 metabolite (PGFM) and relaxin concentrations in crossbred cows with normal parturition (NP) and with RFM. Blood samples were collected from 33 dairy cows at weekly interval from 30 days prepartum till 7th day before calving, followed by every alternate days till calving and 1st and 2nd day postpartum. Significantly higher plasma cortisol concentrations was observed between day 3 prepartum and day 2 postpartum in RFM cows (n=6) compared to NP cows (n=6). Plasma estradiol level was significantly lower in RFM group than in NP group on the day of calving while progesterone concentrations did not differ between the groups. In RFM cows, PGFM level was significantly lower on day 1 and day 2 postpartum compared to NP cows. On the day of calving and on day 1 postpartum, relaxin concentration was significantly (P<0.05) higher in NP cows compared to RFM cows. The ROC analysis, commonly used for development of diagnostic threshold value, revealed that cows with cortisol concentration above 7.35 ng/ml and PGFM concentration below 1,072 pg/ml on day-1 were associated with 5.99 times higher risk of RFM. It may be inferred that peripartum plasma cortisol and PGFM concentrations may be useful for identification of crossbred cows at the risk of developing RFM.

Transferability of SSR markers from related species/genera is a rapid and cost-effective method to enhance genomic database of genetically neglected crops. Monotypic genus Tecomella undulata, a pharmacologically important endangered timber tree, found in hyper arid regions is one such genetic resource limited species, wherein neither SSR identification nor cross transferability studies have been initiated. Transferability of 69 cross-genera SSR primers selected from other members of family bignoniaceae (Incarvillea sinensis, I. mairei, Jacaranda copaia, Tabebuia aurea and Arrabidaea chica) showed 40.58% (28 primers) transferability amplifying 1–10 amplicons ranging in the size from 80to 600 bp in 24 accessions of T. undulata. Per cent marker transferability varied from 66.67% (A. chica) to 38.89% (J. copaia) among the genera tested while no transferable markers could be identified from I. sinensis. Transferability of simple di-/tri-nucleotide repeat (34.2–37.5%) and complex nucleotide sequence (60.0%) based SSR motifs were higher as compared to penta-/hexa-nucleotide based repeat motifs. Within the 28 transferable markers, 26 (92.86%) were polymorphic while 2 (7.14%) were monomorphic. Average number of alleles per primer was 5.39 while average polymorphic alleles per locus were 5.11. The PIC values of SSR markers ranged from 0.23 to 0.85 with an average of 0.59. Efficiency of the identified cross species SSR markers was tested for diversity analysis using a set of 24 trees (8 each of yellow, orange and red flower colour morphotypes). The 24 samples clustered into six groups with similarity coefficient ranging from 0.29 to 0.67, while six accessions formed distinct out groups. Distribution of accessions in the SSR dendrogram showed no correlation with flower colour. More diversity (99%) was observed within populations compared to that existing between populations (1%). The present study can enrich the genomic background of T. undulata by identifying suitable polymorphic markers from confamiliar species that can be applied for genetic diversity studies, mapping of QTLs and cultivar identification as well.