Genetic enhancement of okra [Abelmoschus esculentus (L.) Moench] germplasm through wide hybridization.

Introduction: The introgression of genetic material from one species to another through wide hybridization and repeated back-crossing, plays an important role in genetic modification and enriching the cultivated gene-pool with novel genetic variations. Okra (Abelmoschus esculentus [(L.) Moench)] is a popular vegetable crop with high dietary fibre and protein, rich in essential amino acids, lysine and tryptophan. The wild Abelmoschus genepool has many desirable traits like ornamental value, short internodal length, more number of productive branches, extended bearing, perennation tendency, reduced fruit length (more consumer preferred trait), high mucilage content (medicinal value), abiotic stress tolerances such as drought, high temperature and biotic stress resistances such as okra Yellow Vein Mosaic Virus (YVMV) and Enation Leaf Curl Virus (ELCV) diseases. The repeated use of elite breeding lines led to narrowing of the genetic base of the okra crop, one of the major factors attributed to breakdown of resistance/ tolerance to biotic stresses. YVMV and ELCV are the two major diseases, causing significant yield loss in okra. Hence, wide hybridization was attempted to transfer tolerance genes from wild species to the cultivated genepool to widen the genetic base. Material and methods: The screening of germplasm of wild Abelmoschus species at hotspots led to the identification of tolerant species (Abelmoschus pungens var. mizoramensis, A. enbeepeegeearensis, A. caillei, A. tetraphyllus and A. angulosus var. grandiflorus), which were further used in a wide-hybridization programme to generate interspecific hybrids with the cultivated okra. Presence of pre- and post-zygotic barriers to interspecific geneflow, differences in ploidy levels and genotype specific variations in chromosome numbers led to varying degrees of sterility in F1 plants of interspecific crosses. This was overcome by doubling the chromosome number of interspecific hybrids by applying Colchicine at the seedling stage. The 113 cross derivatives generated comprising amphidiploids in the F1 generation (30), F3 (14), one each in F2 and F4 generations, back cross generation in BC1F2 (03), BC1F3 (25), and BC2F3 (02), crosses between amphidiploids (27), multi-cross combinations (07) and inter-specific cross (between A. sagittifolius × A. moschatus subsp. moschatus) selfed derivatives at F8 generation (03) were characterized in the present study. Besides they were advanced through selfing and backcrossing. Results and Discussion: The amphidiploids were found to possess many desirable genes with a considerable magnitude of linkage drag. Majority of the wide cross derivatives had an intermediate fruit morphology and dominance of wild characters viz., hispid fruits, stem, leaves, tough fruit fibre, vigorous perennial growth habit and prolonged flowering and fruiting. The fruit morphology of three BC progenies exhibited a high morphological resemblance to the cultivated okra, confirming successful transfer of useful genes to the cultivated okra genepool. The detailed morphological characteristics of the various combinations of Abelmoschus amphidiploids and the genetic enhancement of the genepool achieved in this process is reported here.

Macrolactin A mediated biocontrol of Fusarium oxysporum and Rhizoctonia solani infestation on Amaranthus hypochondriacus by Bacillus subtilis BS-58.

Plant diseases are one of the main hurdles for successful crop production and sustainable agriculture development world-wide. Though several chemical measures are available to manage crop diseases, many of them have serious side effects on humans, animals and the environment. Therefore, the use of such chemicals must be limited by using effective and eco-friendly alternatives. In view of the same, we found a Bacillus subtilis BS-58 as a good antagonist towards the two most devastating phytopathogens, i.e., Fusarium oxysporum and Rhizoctonia solani. Both the pathogens attack several agricultural crops (including amaranth) and induce a variety of infections in them. The findings of scanning electron microscopy (SEM) in this study suggested that B. subtilis BS-58 could inhibit the growth of both the pathogenic fungi by various means such as perforation, cell wall lysis, and cytoplasmic disintegration in the fungal hyphae. Thin-layer chromatography, LC-MS and FT-IR data revealed the antifungal metabolite to be macrolactin A with a molecular weight of 402 Da. Presence of the mln gene in the bacterial genome further endorsed that the antifungal metabolite produced by BS-58 was macrolactin A. Pot trial conducted in the present study showed that seed treatment by BS-58 effectively reduced seedling mortality (54.00 and 43.76%) in amaranth, when grown in pathogen infested soil (F. oxysporum and R. solani, respectively), when compared to their respective negative controls. Data also revealed that the disease suppression ability of BS-58 was almost equivalent to the recommended fungicide, carbendazim. SEM analysis of roots of the seedlings recovered from pathogenic attack substantiated the hyphal disintegration by BS-58 and prevention of amaranth crop. The findings of this study conclude that macrolactin A produced by B. subtilis BS-58 is responsible for the inhibition of both the phytopathogens and the suppression of the diseases caused by them. Being native and target specific, such strains under suitable conditions, may result in ample production of antibiotic and better suppression of the disease.

Effect of plant growth promoting Bacillus spp. on nutritional properties of Amaranthus hypochondriacus grains.

Amaranth (Amaranthus hypochondriacus Linn.) is an important pseudocereal crop having important nutrients along with the indispensable amino-acids. The present study was aimed to study the effect of plant growth promoting bacilli on proximate constituents of amaranth grains, including three of the essential amino acids (methionine, lysine and, tryptophan). The combination of Bacillus pumilus and Bacillus subtilis showed a significant increase in different proximate constituents, including crude protein (22.13%), dry matter (32.25%), fat (30.77%), and carbohydrate (49.08%) in amaranth grains. Similarly, a significant increase in essential amino-acids (methionine 47.68%, lysine 59.41% and, tryptophan 38.05%) was recorded. This study suggests that the combination of Bacillus pumilus BS-27 and Bacillus subtilis BS-58 provides the natural, persistent and durable potential to enhance the nutritive value of the crop. Therefore, present study was designed to explore the enhancement of most desirable amino acid synthesis in amaranth due to application of plant growth promoting Bacillus spp.

Isolation of antibacterial protein from Lactobacillus spp. and preparation of probiotic curd.

The study was aimed to isolate antagonistic lactobacilli and the molecules responsible for their antagonistic ability from curd. Preparation of probiotic curd and the ability of the selected lactobacilli to suppress the pathogen therein was also assessed. All the 116 isolates were identified as Lactobacillus spp. based on morphological, biochemical and curdling assays. Five of these lactobacilli (Lb-17, Lb-33, Lb-108, Lb-112, and Lb-N3) were found most promising to inhibit all test pathogens (Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Salmonella typhi and Shigella sonnei). The cell-free culture supernatants of these five lactobacilli were recorded as thermo-tolerant when subjected to heat treatment at 100 °C for 20 min. The loss in the activity after protease treatment indicated the proteinaceous nature of the antimicrobial molecule present in the culture supernatants. Active protein (19 kDa) produced by lactobacilli was confirmed by SDS-PAGE followed by agar-overlay method. Antibiotic sensitivity assay revealed that the selected Lactobacillus spp. isolates were resistant to methicillin and vancomycin. Probiotic curd prepared by using Lb-108 and Lb-N3 was found to be superior to rest of the three isolates based on organoleptic tests and shelf-life. Complete inhibition of all the test pathogens in curd was shown by Lb-108 and Lb-N3. Inhibition spectrum, production of thermostable protein and preparation of quality curd suggest Lb-108 and Lb-N3 as promising candidates to prepare probiotic curd.

Reconceptualizing INRM in understanding environmental risks from food production systems.

Sustainable agri-development is composed of soil fertility maintenance, nutrient use efficiency and resource use efficiency. In this regard, the present study aims to address the complex dynamics of farm-soil and the water nexus for integrated natural resource management by understanding the fertilizer use decision of farmers. In addition to the monitoring of soil quality parameters, that is, the total nitrogen and nitrate concentrations, factor analysis was used to explore the drivers of high fertilizer use in the most agriculturally intensive region of India. The results classified economic benefits, intrinsic and extrinsic motivations, resource-related aspects, institutional set-up and passivity towards environmental quality as major entry points for integrated natural resource management. Although the economic benefit in the form of the affordability of nitrogenous fertilizers was found to be the most critical rationale for fertilizer use, passivity towards environmental quality emerged as a factor of interest. Because short-term economic benefits outweigh concerns for environmental quality, our research suggests that outreach educational programs on sustainable farm practices can be instrumental in resource management. Support of the farming community by governmental/non-governmental agricultural agencies to formulate site-specific recommendations can significantly mitigate fertilizer-induced non-point pollution.

Corneal swelling caused by conventional and new-design low-Dk soft contact lenses following a 10-day daily wear trial regime.

PURPOSE: To investigate the efficacy and safety of a fenestrated and channelled soft contact lens (F-SCL) compared to a standard and non-fenestrated soft contact lens (S-SCL) in experienced soft contact lens (SCL) wearers. METHODS: This was a randomised, crossover, single-blinded (subject), and multicentre clinical trial. Sixteen experienced SCL wearers were randomly divided into two groups (FS and SF). The FS group first wore F-SCLs followed by S-SCLs, each for 10 days, separated by a 1-week washout period, whereas the SF group wore the S-SCLs first and crossed over to F-SCLs in the same manner. The F-SCLs were designed with three equally spaced, symmetrical fenestrations and a partial-thickness, connecting, circumferential channel on the back surface of the mid-periphery of the lens. Measurement of central corneal thickness using ultrasonic pachymetry was performed on the day of screening, after the 1-week washout period, and after 10 days of wearing each kind of lens, based on which central corneal swelling was calculated and compared. One eye in each subject was chosen at random for analysis. RESULTS: Central corneal swelling was 1.92±1.73% vs. 5.26±2.14% in F-SCLs vs. S-SCLs wearers, which was statistically significant (P<0.001). There was no significant difference between the groups in terms of SCL-corrected visual acuity or SCL-related adverse events. CONCLUSION: The use of F-SCLs led to reduced corneal swelling compared to S-SCLs. The newly incorporated features appear to improve tear mixing and thereby the oxygen supply to the cornea, which results in reduced corneal oedema.

Changes in anterior and posterior corneal parameters in patients with keratoconus after intrastromal corneal-ring segment implantation.

PURPOSE: To compare changes in the anterior and the posterior corneal parameters and the anterior chamber depth (ACD) measured by a rotating Scheimpflug camera and scanning-slit topography before and after intrastromal corneal ring segment (ICRS) implantation. METHODS: Twenty-five eyes of 21 keratoconus patients who had Intacs (Addition Technology, Sunnyvale, CA) ICRS implantation were included. The anterior segment parameters of the participants were measured before and 3 months after Intacs ICRS implantation with a rotating Scheimpflug camera and scanning-slit topography. Outcome measures included uncorrected (UDVA) and corrected (CDVA) distance visual acuities, refraction and corneal topography. RESULTS: The UDVA improved in 21 eyes (84%) and CDVA in 18 eyes (72%). The anterior average keratometry decreased from 51.80 D preoperatively to 49.59 D postoperatively (p < 0.001) using the rotating Scheimpflug camera and Sim K decreased from 51.21 to 47.44 D using the scanning-slit topography. The posterior average keratometry decreased from -8.03 D preoperatively to -8.42 D postoperatively (p < 0.001) using the rotating Scheimpflug camera. ACD decreased from 3.44 to 3.32 mm (p < 0.001) and from 3.28 to 3.19 mm (p < 0.001) using the rotating Scheimpflug camera and scanning-slit topography, respectively. CONCLUSIONS: The implantation of Intacs segments induced corneal flattening of the anterior surface and corneal steepening of the posterior surface. Both anterior corneal flattening and posterior corneal steepening contributed to the decrease in the keratometric power.