Synthesis of green zinc-oxide nanoparticles and its dose-dependent beneficial effect on spermatozoa during preservation: sperm functional integrity, fertility and antimicrobial activity.

Introduction: The development of an effective extender is important for semen preservation and the artificial insemination (AI) industry. This study demonstrates the beneficial effect of zinc oxide nanoparticles (ZnO-NPs) as an additive to semen extenders to improve semen quality, fertility, and antibacterial activity during liquid preservation in a boar model. Methods: Initially, to find out the safe concentration of ZnO-NPs in sperm cells, a wide range of ZnO-NP concentrations (0, 5, 10, 50, 100, 500, and 1,000 µM) were co-incubated with sperm at 37°C for a cytotoxic study. These NP concentrations were compared to their salt control zinc acetate (ZA) at the same concentrations and to a control group. The effect of the different concentrations of ZnO-NPs on sperm motility, membrane integrity, mitochondrial membrane potential (MMP), and apoptosis was assessed. Accordingly, the non-toxic dose was selected and supplemented in MODENA extender to determine its beneficial effect on the boar semen parameters mentioned and the lipid peroxidation (LPO) levels during liquid preservation at 16°C for 6 days. The non-cytotoxic dosage was subsequently chosen for AI, fertility investigations, and the evaluation of the antibacterial efficacy of ZnO-NPs during preservation hours. An antibacterial study of ZnO-NPs and its salt control at doses of 10 µM and 50 µM was carried out by the colony forming unit (CFU) method. Results and discussion: The cytotoxic study revealed that 5, 10, and 50 µM of ZnO-NPs are safe. Consequently, semen preserved in the MODENA extender, incorporating the non-toxic dose, exhibited 10 and 50 µM ZnO-NPs as the optimal concentrations for beneficial outcomes during liquid preservation at 16°C. ZnO-NPs of 10 µM concentration resulted in a significantly (p < 0.05) improved conception rate of 86.95% compared to the control of 73.13%. ZnO-NPs of 10 and 50 µM concentrations exhibit potent antimicrobial action by reducing the number of colonies formed with days of preservation in comparison to the negative control. The investigation concluded that the incorporation of 10 µM ZnO-NPs led to enhancements in sperm motility, membrane integrity, and MMP, attributed to a reduction in the malondialdehyde (MDA) levels. This improvement was accompanied by a concurrent increase in fertility rates, including farrowing rate and litter size, during the liquid preservation process. Furthermore, ZnO-NPs exhibited an antimicrobial effect, resulting in decreased bacterial growth while preserving boar semen at 16°C for 6 days. These findings suggest that ZnO-NPs could serve as a viable alternative to antibiotics, potentially mitigating antibiotic resistance concerns within the food chain.

Comparison of dehulling efficiency and grain nutritional parameters of two cultivated barnyard millet species (Echinochloa spp.).

Due to increased awareness regarding the health-promoting profile of millets, inclination towards their consumption has increased considerably. In the Himalayan region of India, cultivars of the two species of barnyard millet, namely Indian (Echinochloa frumentacea) and Japanese barnyard millet (E. esculenta), are grown. To compare the dehulled grain recovery, grain physical parameters, nutritional profile and antioxidant activity, an experiment was carried out at ICAR-VPKAS, Almora, Uttarakhand hills for two years using released and popular cultivars of Indian barnyard millet (VL 207 and VL 172) and Japanese barnyard millet (PRJ-1). The results indicated that the whole grain yield of Japanese barnyard millet cultivar PRJ-1 was significantly higher than Indian Barnyard millet cultivars VL 172 and VL 207; however, the dehulled grain recovery was considerably higher in VL 172 and VL 207 than PRJ-1. Similarly, the physical grain parameters were significantly higher in PRJ-1, but most dehulled grain parameters were at par in cultivars of both species. The nutritional estimation of dehulled grains of both species did not show remarkable differences for most traits. Still, crude fibre, Mn, and Zn were high in PRJ-1, while total digestible nutrients and phosphorous were high in VL 172 and VL 207. Dehulled grains exhibited much more crude protein, ash, minerals, and total digestible nutrients, but the husk accumulated significantly higher crude fibre and total polyphenols.

Identification and expression pattern of aluminium-responsive genes in roots of rice genotype with reference to Al-sensitivity.

Aluminium (Al) is the third most abundant element in the Earth's crust. Globally, acidic soil occupies 30-40% of ice-free land areas; Al toxicity is a major threat to crops. The first symptom of Al toxicity is the inhibition of root growth followed by poor root hair development, swollen root apices, necrosis of leaves and reduced yield. Although Rice (Oryza sativa) is an Al toxicity tolerant crop, it shows considerable variations among rice genotypes to Al exposure. Therefore, it is pertinent to understand Al toxicity and underlying mechanisms for Al tolerance in Rice. In the present study, 63 rice genotypes screened under Al stress showed significant variations of root growth. Expression stability of endogenous control genes (ECGs) revealed sulphite reductase (SR) as the most stable ECG that can be used as a reference gene for quantitative real-time PCR (qRT-PCR). Expression patterns of Al-responsive genes suggest genes associated with cytoskeletal dynamics, metabolism, and ion transporter could play significant roles in Al adaptation and tolerance in rice. The results showed Motodhan, Vietnam-1, Yimyu and N-861 as Al-toxicity tolerant, while Lespah, RCPL-13, VL-31329, and UPR2919-141-1 as most Al-sensitive genotypes among the studied rice lines cultivated in North-East India.

Optimizing raffinose family oligosaccharides content in plants: A tightrope walk.

Plants synthesize various compounds for their growth, metabolism, and stress mitigation, and one such group of compounds is the raffinose family of oligosaccharides (RFOs). RFOs are non-reducing oligosaccharides having galactose residues attached to a sucrose moiety. They act as carbohydrate reserves in plants, assisting in seed germination, desiccation tolerance, and biotic/abiotic stress tolerance. Although legumes are among the richest sources of dietary proteins, the direct consumption of legumes is hindered by an excess of RFOs in the edible parts of the plant, which causes flatulence in humans and monogastric animals. These opposing characteristics make RFOs manipulation a complicated tradeoff. An in-depth knowledge of the chemical composition, distribution pattern, tissue mobilization, and metabolism is required to optimize the levels of RFOs. The most recent developments in our understanding of RFOs distribution, physiological function, genetic regulation of their biosynthesis, transport, and degradation in food crops have been covered in this review. Additionally, we have suggested a few strategies that can sustainably reduce RFOs in order to solve the flatulence issue in animals. The comprehensive information in this review can be a tool for researchers to precisely control the level of RFOs in crops and create low antinutrient, nutritious food with wider consumer acceptability.

Spatio-temporal expression pattern of Raffinose Synthase genes determine the levels of Raffinose Family Oligosaccharides in peanut (Arachis hypogaea L.) seed.

Raffinose family oligosaccharides (RFOs) are known to have important physiological functions in plants. However, the presence of RFOs in legumes causes flatulence, hence are considered antinutrients. To reduce the RFOs content to a desirable limit without compromising normal plant development and functioning, the identification of important regulatory genes associated with the biosynthetic pathway is a prerequisite. In the present study, through comparative RNA sequencing in contrasting genotypes for seed RFOs content at different seed maturity stages, differentially expressed genes (DEGs) associated with the pathway were identified. The DEGs exhibited spatio-temporal expression patterns with high RFOs variety showing early induction of RFOs biosynthetic genes and low RFOs variety showing a late expression at seed maturity. Selective and seed-specific differential expression of raffinose synthase genes (AhRS14 and AhRS6) suggested their regulatory role in RFOs accumulation in peanut seeds, thereby serving as promising targets in low RFOs peanut breeding programs. Despite stachyose being the major seed RFOs fraction, differential expression of raffinose synthase genes indicated the complex metabolic regulation of this pathway. The transcriptomic resource and the genes identified in this study could be studied further to develop low RFOs varieties, thus improving the overall nutritional quality of peanuts.

Alleviating aluminum toxicity in plants: Implications of reactive oxygen species signaling and crosstalk with other signaling pathways.

Aluminum (Al) toxicity is a major limiting factor for plant growth and productivity in acidic soil. At pH lower than 5.0 (pH < 5.0), the soluble and toxic form of Al (Al3+ ions) enters root cells and inhibits root growth and uptake of water and nutrients. The organic acids malate, citrate, and oxalate are secreted by the roots and chelate Al3+ to form a non-toxic Al-OA complex, which decreases the entry of Al3+ into the root cells. When Al3+ enters, it leads to the production of reactive oxygen species (ROS) in cells, which are toxic and cause damage to biomolecules like lipids, carbohydrates, proteins, and nucleic acids. When ROS levels rise beyond the threshold, plants activate an antioxidant defense system that comprises of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione S-transferase (GST), ascorbic acid (ASA), phenolics and alkaloids etc., which protect plant cells from oxidative damage by scavenging and neutralizing ROS. Besides, ROS also play an important role in signal transduction and influence many molecular and cellular process like hormone signaling, gene expression, cell wall modification, cell cycle, programed cell death (PCD), and development. In the present review, the mechanisms of Al-induced ROS generation, ROS signaling, and crosstalk with other signaling pathways helping to combat Al toxicity have been summarized, which will help researchers to understand the intricacies of Al-induced plant response at cellular level and plan research for developing Al-toxicity tolerant crops for sustainable agriculture in acid soil-affected regions of the world.

Genetic diversity analysis of specialty glutinous and low-amylose rice (Oryza sativa L.) landraces of Assam based on Wx locus and microsatellite diversity.

The sticky rice of Assam is traditionally classified as bora (glutinous) and chokuwa (semi-glutinous) based on their stickiness after cooking. The Waxy (Wx) gene encodes for granule-bound starch synthase (GBSS) that controls the synthesis of amylose, which is a key determinant of rice end-use quality attributes. In this report, we analysed the level of variation in grain quality traits in a collection of bora and chokuwa cultivars, and examined the nucleotide diversity at the Wx locus of selected rice accessions to identify the possible cause of low-amylose in these rice cultivar groups. The Wx gene sequencing from 24 bora and chokuwa cultivars revealed several nucleotide variations that can explain the variation in the amylose phenotypes. The nucleotide polymorphisms in the downstream intron regions were similar to those reported in Bangladeshi Beruin cultivars. Among the Wx polymorphisms, the CTn microsatellite in exon 1 and G/T SNP in intron 1 (G/T-Int1) should be considered for marker assisted breeding involving bora cultivars. The Wx gene tree, classified the bora accessions possessing the G/T-Int1 SNP as japonicas. However, cluster analysis using microsatellite markers classified the bora and chokuwa cultivars as indica, and intermediate of indica-aus. The findings of this study supplemented our understanding on the evolution of the Wx gene under human selection. The results will assist plant breeders to effectively improve the bora and chokuwa landraces.

The potential of arbuscular mycorrhizal fungi in C cycling: a review.

Arbuscular mycorrhizal fungi (AMF) contribute predominantly to soil organic matter by creating a sink demand for plant C and distributing to below-ground hyphal biomass. The extra-radical hyphae along with glomalin-related soil protein significantly influence the soil carbon dynamics through their larger extent and turnover period need to discuss. The role of AMF is largely overlooked in terrestrial C cycling and climate change models despite their greater involvement in net primary productivity augmentation and further accumulation of this additional photosynthetic fixed C in the soil. However, this buffering mechanism against elevated CO2 condition to sequester extra C by AMF can be described only after considering their potential interaction with other microbes and associated mineral nutrients such as nitrogen cycling. In this article, we try to review the potential of AMF in C sequestration paving the way towards a better understanding of possible AMF mechanism by which C balance between biosphere and atmosphere can be moved forward in more positive direction.

Revisiting the plant growth-promoting rhizobacteria: lessons from the past and objectives for the future.

Plant beneficial rhizobacteria (PBR) is a group of naturally occurring rhizospheric microbes that enhance nutrient availability and induce biotic and abiotic stress tolerance through a wide array of mechanisms to enhance agricultural sustainability. Application of PBR has the potential to reduce worldwide requirement of agricultural chemicals and improve agro-ecological sustainability. The PBR exert their beneficial effects in three major ways; (1) fix atmospheric nitrogen and synthesize specific compounds to promote plant growth, (2) solubilize essential mineral nutrients in soils for plant uptake, and (3) produce antimicrobial substances and induce systemic resistance in host plants to protect them from biotic and abiotic stresses. Application of PBR as suitable inoculants appears to be a viable alternative technology to synthetic fertilizers and pesticides. Furthermore, PBR enhance nutrient and water use efficiency, influence dynamics of mineral recycling, and tolerance of plants to other environmental stresses by improving health of soils. This report provides comprehensive reviews and discusses beneficial effects of PBR on plant and soil health. Considering their multitude of functions to improve plant and soil health, we propose to call the plant growth-promoting bacteria (PGPR) as PBR.

Rice bean: a lesser known pulse with well-recognized potential.

MAIN CONCLUSION: Required genetic resources for the improvement of agronomic, nutritional and economic value of rice bean are available in the world collection. International cooperative effort is required to utilize and conserve them. Rice bean [Vigna umbellata (Thunb.) Ohwi and Ohashi], a lesser known pulse among the Asiatic Vigna, has long been considered as a food security crop of small and marginal farmers of Southeast Asia. Considered as a nutritionally rich food and fodder, it is also a source of genes for biotic and abiotic stress tolerance including drought, soil acidity and storage pest. Although it spread from its centre of domestication in the Indo-China region to other parts around the world, it never became an important crop anywhere probably because of agronomic disadvantages. Crop improvement for determinate nature, good yield, less variable seed colour, pleasant organoleptic properties and lower antinutrients is required. Scanning of scientific literature indicates that genetic resources with desirable agronomic and nutritional traits exist within the current collection but are spread across countries. Genomic studies in the species indicate that except for insect resistance and aluminium toxicity tolerance, not much attention has been paid to decipher and utilize other stress tolerance and nutritional quality traits. Collaborative efforts towards improving farming, food, trade value and off-farm conservation of rice bean would not only help marginal farmers but will also help to preserve the yet to be explored genomic resources available in this sturdy pulse.

Ancient orphan legume horse gram: a potential food and forage crop of future.

MAIN CONCLUSION: Tailoring crops to withstand rising temperature and declining precipitation may be unrewarding, therefore the potential of alternative undervalued crops such as horse gram can be explored for safeguarding food and nutritional security with health benefits in the era of climate change. Horse gram [Macrotyloma uniflorum (Lam.) Verdc] under the family Fabaceae, has been cultivated for food, folklore medicine and fodder mainly by rural and tribal communities since prehistoric times in Asian and African countries. This valuable ancient legume not only offers diversification and resilience to agro-ecosystems but it also possesses high adaptation in risk-prone traditional farming systems in marginal environments of semi-arid and arid regions. Being a nutrient dense legume with remedial health-promoting effects due to the presence of various bioactive compounds, it is suitable for the development of functional food as well as for addressing micronutrient deficiencies among poor rural communities. Despite its enormous potential and a growing awareness about the utility of this underutilized crop for future climate adaptation and human well-being, this legume continues to be seriously neglected and labelled as "food of the poors". India is the major producer of horse gram and presently, cultivation of horse gram remains confined to small-scale farming systems as an inter- or mixed crop. This crop is alienated from mainstream agriculture and relegated to a status of "underutilized" due to its limited competitiveness as compared to other commercial crops. Besides a scanty basic research on this crop, no attention has been paid to the factors like improvement of plant type, yield improvement, processing, value addition to suit consumer needs and reduction of anti-nutritional factors, which restricted the diffusion of this crop outside its niche area. The present review therefore is an attempt to compile the meagre information available on crop history, evolution, genetic enhancement, nutritional and health benefits to make the crop competitive and revitalize horse gram cultivation.

Marker-trait association for low-light intensity tolerance in rice genotypes from Eastern India.

Light intensity is a crucial environmental factor that affects photosynthesis and ultimately, grain yield in rice. However, no gene or marker directly associated with improved performance under low-light intensity under field conditions has been identified till date. With an aim of identifying genes and markers associated with improved performance (measured in terms of better yields) under low-light intensity, an integrated field screening, in silico and wet lab validation analysis was performed. Field-based screening of a diverse set of 110 genotypes led to the identification of two physiological and three morphological parameters critical for low-light tolerance in rice. In silico analysis using information available in public databases led to the identification of a set of 90 potential candidate genes which were narrowed to thirteen genic targets for possible marker-trait association. Marker-trait association on the panel of 48 diverse rice genotypes varying in their response to low-light intensity led to the identification of six markers [HvSSR02-44 (biological yield), HvSSR02-52 (spikelet fertility), HvSSR02-54 (grain yield), HvSSR06-56 (spikelet fertility), HvSSR06-69 (spikelet fertility; biological yield), HvSSR09-45 (spikelet fertility)] lying on chromosomes 2, 6 and 9 showing significant association (R2 > 0.1) for traits like grain yield/plant, biological yield and spikelet fertility under low light. Eight rice genes [including member of BBX (B-box) family] lying within 10 kb distance of these identified markers already reported for their role in response to stress or change in plant architecture in rice were also identified. The eight rice genotypes, five traits, eight genes and six markers identified in the current study will help in devising strategies to increase yield under low light intensity and pave way for future application in marker-assisted breeding.

Insights into maize genome editing via CRISPR/Cas9.

Maize is an important crop for billions of people as food, feed, and industrial raw material. It is a prime driver of the global agricultural economy as well as the livelihoods of millions of farmers. Genetic interventions, such as breeding, hybridization and transgenesis have led to increased productivity of this crop in the last 100 years. The technique of genome editing is the latest advancement in genetics. Genome editing can be used for targeted deletions, additions, and corrections in the genome, all aimed at genetic enhancement of crops. The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR associated protein 9 (CRISPR/Cas9) system is a recent genome editing technique that is considered simple, precise, robust and the most revolutionary. This review summarizes the current state of the art and predicts future directions in the use of the CRISPR/Cas9 tool in maize crop improvement.

Development and characterization of a new set of genomic microsatellite markers in rice bean (Vigna umbellata (Thunb.) Ohwi and Ohashi) and their utilization in genetic diversity analysis of collections from North East India.

Rice bean [Vigna umbellate (Thumb.) Ohwi and Ohashi] is an underutilized crop believed to be domesticated in the Myanmar-Thailand region of Asia. In India, rice bean is mainly cultivated in the North-Eastern Hills, which is a hotspot for biological diversity. A 5' anchored PCR was used to develop microsatellite markers in rice bean. Twenty-eight specific primer pairs were designed and employed to characterize sixty five ricebean accessions collected from North East India. A total of 179 alleles were amplified with an average of 6.393 alleles per locus. The gene diversity was high (mean 0.534) in the accessions collected from Darjeeling, Nagaland and Manipur, which are bordering areas with East Nepal and Myanmar, respectively. Exceptionally high outcrossing rate was observed in the entire population. Population structure analysis identified three distinct clusters in which accessions collected from areas bordering Myanmar and East Nepal grouped separately. Using a combination of STRUCTURE and Principal Coordinate Analysis, relative affinity of the intermediate accessions could be established. However, differences in allelic counts among populations were non-significant. The results showed that there is a high level of genetic diversity within the accessions, with high outcrossing rate.

Advances in Maize Transformation Technologies and Development of Transgenic Maize.

Maize is the principal grain crop of the world. It is also the crop where genetic engineering has been employed to a great extent to improve its various traits. The ability to transform maize is a crucial step for application of gene technology in maize improvement. There have been constant improvements in the maize transformation technologies over past several years. The choice of genotype and the explant material to initiate transformation and the different types of media to be used in various stages of tissue culture can have significant impact on the outcomes of the transformation efforts. Various methods of gene transfer, like the particle bombardment, protoplast transformation, Agrobacterium-mediated, in planta transformation, etc., have been tried and improved over years. Similarly, various selection systems for retrieval of the transformants have been attempted. The commercial success of maize transformation and transgenic development is unmatched by any other crop so far. Maize transformation with newer gene editing technologies is opening up a fresh dimension in transformation protocols and work-flows. This review captures the various past and recent facets in improvement in maize transformation technologies and attempts to present a comprehensive updated picture of the current state of the art in this area.

Stability Performance of Inductively Coupled Plasma Mass Spectrometry-Phenotyped Kernel Minerals Concentration and Grain Yield in Maize in Different Agro-Climatic Zones.

Deficiency of iron and zinc causes micronutrient malnutrition or hidden hunger, which severely affects ~25% of global population. Genetic biofortification of maize has emerged as cost effective and sustainable approach in addressing malnourishment of iron and zinc deficiency. Therefore, understanding the genetic variation and stability of kernel micronutrients and grain yield of the maize inbreds is a prerequisite in breeding micronutrient-rich high yielding hybrids to alleviate micronutrient malnutrition. We report here, the genetic variability and stability of the kernel micronutrients concentration and grain yield in a set of 50 maize inbred panel selected from the national and the international centres that were raised at six different maize growing regions of India. Phenotyping of kernels using inductively coupled plasma mass spectrometry (ICP-MS) revealed considerable variability for kernel minerals concentration (iron: 18.88 to 47.65 mg kg(-1); zinc: 5.41 to 30.85 mg kg(-1); manganese: 3.30 to 17.73 mg kg(-1); copper: 0.53 to 5.48 mg kg(-1)) and grain yield (826.6 to 5413 kg ha(-1)). Significant positive correlation was observed between kernel iron and zinc within (r = 0.37 to r = 0.52, p < 0.05) and across locations (r = 0.44, p < 0.01). Variance components of the additive main effects and multiplicative interactions (AMMI) model showed significant genotype and genotype × environment interaction for kernel minerals concentration and grain yield. Most of the variation was contributed by genotype main effect for kernel iron (39.6%), manganese (41.34%) and copper (41.12%), and environment main effects for both kernel zinc (40.5%) and grain yield (37.0%). Genotype main effect plus genotype-by-environment interaction (GGE) biplot identified several mega environments for kernel minerals and grain yield. Comparison of stability parameters revealed AMMI stability value (ASV) as the better representative of the AMMI stability parameters. Dynamic stability parameter GGE distance (GGED) showed strong and positive correlation with both mean kernel concentrations and grain yield. Inbreds (CM-501, SKV-775, HUZM-185) identified from the present investigation will be useful in developing micronutrient-rich as well as stable maize hybrids without compromising grain yield.