Bhendi Yellow Vein Mosaic Virus and Bhendi Yellow Vein Mosaic Betasatellite Cause Enation Leaf Curl Disease and Alter Host Phytochemical Contents in Okra.

Whitefly (Bemisia tabaci)-transmitted begomoviruses cause severe diseases in numerous economically important dicotyledonous plants. Okra enation leaf curl disease (OELCuD) has emerged as a serious threat to okra (Abelmoschus esculentus L. Moench) cultivation in the Indian subcontinent. This study reports the association of a monopartite begomovirus (bhendi yellow vein mosaic virus; BYVMV) and betasatellite (bhendi yellow vein mosaic betasatellite; BYVB) with OELCuD in the Mau region of Uttar Pradesh, India. The BYVMV alone inoculated Nicotiana benthamiana and A. esculentus cv. Pusa Sawani plants developed mild symptoms. Co-inoculation of BYVMV and BYVB resulted in a reduced incubation period, an increased symptom severity, and an enhanced BYVMV accumulation by Southern hybridization and quantitative real-time PCR. This is the first study that satisfies Koch's postulates for OELCuD in its natural host. Activities of various antioxidative enzymes were significantly increased in the virus-inoculated okra plants. Differential responses in various biochemical components (such as photosynthetic pigments, phenol, proline, and sugar) in diseased okra plants were observed. This change in phytochemical responses is significant in understanding its impact on virus pathogenesis and disease development.

Genotyping-by-sequencing based genetic mapping reveals large number of epistatic interactions for stem rot resistance in groundnut.

KEY MESSAGE: Genetic mapping identified large number of epistatic interactions indicating the complex genetic architecture for stem rot disease resistance. Groundnut (Arachis hypogaea) is an important global crop commodity and serves as a major source of cooking oil, diverse confectionery preparations and livestock feed. Stem rot disease caused by Sclerotium rolfsii is the most devastating disease of groundnut and can cause up to 100% yield loss. Genomic-assisted breeding (GAB) has potential for accelerated development of stem rot resistance varieties in short period with more precision. In this context, linkage analysis and quantitative trait locus (QTL) mapping for resistance to stem rot disease was performed in a bi-parental recombinant inbred line population developed from TG37A (susceptible) × NRCG-CS85 (resistant) comprising of 270 individuals. Genotyping-by-sequencing approach was deployed to generate single nucleotide polymorphism (SNP) genotyping data leading to development of a genetic map with 585 SNP loci spanning map distance of 2430 cM. QTL analysis using multi-season phenotyping and genotyping data could not detect any major main-effect QTL but identified 44 major epistatic QTLs with phenotypic variation explained ranging from 14.32 to 67.95%. Large number interactions indicate the complexity of genetic architecture of resistance to stem rot disease. A QTL of physical map length 5.2 Mb identified on B04 comprising 170 different genes especially leucine reach repeats, zinc finger motifs and ethyleneresponsive factors, etc., was identified. The identified genomic regions and candidate genes will further validate and facilitate marker development to deploy GAB for developing stem rot disease resistance groundnut varieties.

Advances in the development and use of DREB for improved abiotic stress tolerance in transgenic crop plants.

Abiotic stresses negatively influence the survival, biomass production, and yield of crops. Tolerance to diverse abiotic stresses in plants is regulated by multiple genes responding differently to various stress conditions. Genetic engineering approaches have helped develop transgenic crops with improved abiotic stress tolerance including yields. The dehydration-responsive element binding protein (DREB) is a stress-responsive transcription factor that modulates the expression of downstream stress-inducible genes, which confer simultaneous tolerance to multiple stresses. This review focuses on advances in the development of DREB transgenic crops and their characterization under various abiotic stress conditions. It further discusses the mechanistic aspects of abiotic stress tolerance, yield gain, the fate of transgenic plants under controlled and field conditions and future research directions toward commercialization of DREB transgenic crops.

Morphological and toxigenic variability in the Aspergillus flavus isolates from peanut (Arachis hypogaea L.) production system in Gujarat (India).

Morphological and toxigenic variability in 187 Aspergillus flavus isolates, collected from a major Indian peanut production system, from 10 districts of Gujarat was studied. On the basis of colony characteristics, the isolates were grouped as group A (83%), B (11%) and G (6%). Of all the isolates, 21%, 47% and 32% were found to be fast-growing, moderately-fast and slow-growing respectively, and nosclerotia and sclerotia production was recorded in 32.1% and 67% isolates respectively. Large, medium and small number of sclerotia production was observed in 55, 38 and 34 isolates respectively. Toxigenic potential based on ammonia vapour test was not found reliable, while ELISA test identified 68.5%, 18.7% and 12.8% isolates as atoxigenic, moderately-toxigenic and highly-toxigenic, respectively. On clustering, the isolates were grouped into 15 distinct clusters, 'A' group of isolates was grouped distinctly in different clusters, while 'B' and 'G' groups of isolates were clustered together. No association was observed between morphological-diversity and toxigenic potential of the isolates. From the present investigation, most virulent isolates were pooled to form a consortium for sick-plot screening of germplasm, against Aspergillus flavus. In future, atoxigenic isolates may be evaluated for their potential to be used as bio-control agent against toxigenicisolates.

Antiangiogenic effect of docetaxel and everolimus as individual and dual-drug-loaded micellar nanocarriers.

PURPOSE: The in vitro inhibitory effect of Docetaxel (DTX) and Everolimus (EVR) alone and together in poly(ethylene glycol)-block-poly(D,L-lactic acid) (PEG-b-PLA) nanocarriers on angiogenic processes and acute toxicity in mice was evaluated. METHODS: PEG-b-PLA DTX and/or EVR nanocarriers were characterized for size, drug loading, stability, and drug release. Cell proliferation, tubule formation, and migration studies were performed in Human Umbilical Vein Endothelial Cells (HUVEC) and Maximum Tolerated Doses (MTD) studies were in mice. RESULTS: DTX and EVR loading was 1.93 and 2.00 mg/mL respectively with similar solubilities for dual-drug micelles. All micelles were below 30 nm with diffusion controlled drug release. The IC50 for DTX, EVR micelles were, 6.80 ± 0.67, 18.57 ± 2.86 and 0.65 ± 0.11 nM respectively with a synergistic inhibitory effect for dual-drug nanocarriers. Significant inhibition of tube formation occurred upon treatment with dual-drug nanocarriers as compared to individual micelles. EVR presence in dual-drug nanocarriers was able to significantly increase the inhibition of the migration of HUVEC by DTX. The MTDs for EVR, DTX and dual-drug micelles were 50, 30 and 20 mg/kg for each respectively. CONCLUSIONS: DTX-EVR dual-drug nanocarriers have antiangiogenic effects in vitro mediated through cellular angiogenic process and possess clinically relevant MTD.

A 5-methylcytosine DNA glycosylase/lyase demethylates the retrotransposon Tos17 and promotes its transposition in rice.

DNA 5-methylcytosine (5-meC) is an important epigenetic mark for transcriptional gene silencing in many eukaryotes. In Arabidopsis, 5-meC DNA glycosylase/lyases actively remove 5-meC to counteract transcriptional gene silencing in a locus-specific manner, and have been suggested to maintain the expression of transposons. However, it is unclear whether plant DNA demethylases can promote the transposition of transposons. Here we report the functional characterization of the DNA glycosylase/lyase DNG701 in rice. DNG701 encodes a large (1,812 amino acid residues) DNA glycosylase domain protein. Recombinant DNG701 protein showed 5-meC DNA glycosylase and lyase activities in vitro. Knockout or knockdown of DNG701 in rice plants led to DNA hypermethylation and reduced expression of the retrotransposon Tos17. Tos17 showed less transposition in calli derived from dng701 knockout mutant seeds compared with that in wild-type calli. Overexpression of DNG701 in both rice calli and transgenic plants substantially reduced DNA methylation levels of Tos17 and enhanced its expression. The overexpression also led to more frequent transposition of Tos17 in calli. Our results demonstrate that rice DNG701 is a 5-meC DNA glycosylase/lyase responsible for the demethylation of Tos17 and this DNA demethylase plays a critical role in promoting Tos17 transposition in rice calli.

Characterization of putative calcium-dependent protein kinase-1 (TaCPK-1) gene: hubs in signalling and tolerance network of wheat under terminal heat.

Calcium-dependent protein kinase (CDPK) is member of one of the most important signalling cascades operating inside the plant system due to its peculiar role as thermo-sensor. Here, we identified 28 full length putative CDPKs from wheat designated as TaCDPK (1-28). Based on digital gene expression, we cloned full length TaCPK-1 gene of 1691 nucleotides with open reading frame (ORF) of 548 amino acids (accession number OP125853). The expression of TaCPK-1 was observed maximum (3.1-fold) in leaf of wheat cv. HD2985 (thermotolerant) under T2 (38 ± 3 °C, 2 h), as compared to control. A positive correlation was observed between the expression of TaCPK-1 and other stress-associated genes (MAPK6, CDPK4, HSFA6e, HSF3, HSP17, HSP70, SOD and CAT) involved in thermotolerance. Global protein kinase assay showed maximum activity in leaves, as compared to root, stem and spike under heat stress. Immunoblot analysis showed abundance of CDPK protein in wheat cv. HD2985 (thermotolerant) in response to T2 (38 ± 3 °C, 2 h), as compared to HD2329 (thermosusceptible). Calcium ion (Ca2+), being inducer of CDPK, showed strong Ca-signature in the leaf tissue (Ca-622 ppm) of thermotolerant wheat cv. under heat stress, whereas it was minimum (Ca-201 ppm) in spike tissue. We observed significant variations in the ionome of wheat under HS. To conclude, TaCPK-1 plays important role in triggering signaling network and in modulation of HS-tolerance in wheat. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-024-03989-6.

Inhibitory effect of paclitaxel and rapamycin individual and dual drug-loaded polymeric micelles in the angiogenic cascade.

Angiogenesis is an essential process for disease progression in many solid tumors. There are several major cascade events in the angiogenic process that can be targeted to inhibit new blood vessel formation in the tumor tissue. The purpose of this work is to evaluate the inhibitory effect of paclitaxel (PTX) and rapamycin (RAP) as individual and in dual drug-loaded poly(ethylene glycol)-block-poly(d,l-lactic acid) (PEG-b-PLA) micelles on the angiogenic cascade processes of proliferation, migration, and tube formation. PEG-b-PLA PTX and/or RAP micelles were formed and characterized for size and drug loading. Sizes of individual and dual drug micelles were below 40 nm. PEG-b-PLA micelles significantly enhanced the aqueous solubility of PTX 1.80 mg/mL and RAP 1.60 mg/mL. The PTX-RAP dual drug PEG-b-PLA micelles were able to load PTX and RAP at 1.60 mg/mL for both drugs. Cell proliferation, apoptosis, tubule formation, and migration studies were performed in human umbilical vein endothelial cells (HUVEC). PTX and RAP in DMSO inhibited HUVEC proliferation with IC50 values of 0.82 ± 0.02 and 13 829 ± 681 nM, respectively, while the combination of both drugs in DMSO produced synergistic inhibition. PTX and RAP individual micelles had IC50 values of 6.3 ± 1.1 and 14 051 ± 821 nM, respectively. PTX and dual drug micelles had a synergistic inhibition effect on HUVEC proliferation through the induction of apoptosis via caspase 3/7 activity. In vitro tube formation assay demonstrated significant inhibition of tube formation upon treatment with dual drug micelles as compared to individual PTX or RAP micelles. Migration studies in HUVEC have shown that individual PTX micelles inhibited cell migration at 1 nM, while RAP micelles did not show any inhibitory effect on cell migration. Interestingly, the presence of RAP in the dual drug micelles was able to initiate the inhibition of the migration of HUVEC at 0.1 nM concentration of PTX. These results indicate that PTX-RAP dual drug micelles have antiangiogenic effects in vitro mediated through three major events in the angiogenic process and have strong potential for further development as antiangiogenic chemotherapy.

Rust (Uromyces viciae-fabae Pers. de-Bary) of Pea (Pisum sativum L.): Present Status and Future Resistance Breeding Opportunities.

Uromyces viciae-fabae Pers. de-Bary is an important fungal pathogen causing rust in peas (Pisum sativum L.). It is reported in mild to severe forms from different parts of the world where the pea is grown. Host specificity has been indicated in this pathogen in the field but has not yet been established under controlled conditions. The uredinial states of U. viciae-fabae are infective under temperate and tropical conditions. Aeciospores are infective in the Indian subcontinent. The genetics of rust resistance was reported qualitatively. However, non-hypersensitive resistance responses and more recent studies emphasized the quantitative nature of pea rust resistance. Partial resistance/slow rusting had been described as a durable resistance in peas. Such resistance is of the pre-haustorial type and expressed as longer incubation and latent period, poor infection efficiency, a smaller number of aecial cups/pustules, and lower units of AUDPC (Area Under Disease Progress Curve). Screening techniques dealing with slow rusting should consider growth stages and environment, as both have a significant influence on the disease scores. Our knowledge about the genetics of rust resistance is increasing, and now molecular markers linked with gene/QTLs (Quantitative Trait Loci) of rust resistance have been identified in peas. The mapping efforts conducted in peas came out with some potent markers associated with rust resistance, but they must be validated under multi-location trails before use in the marker-assisted selection of rust resistance in pea breeding programs.

Transcriptional Regulation of Small Heat Shock Protein 17 (sHSP-17) by Triticum aestivum HSFA2h Transcription Factor Confers Tolerance in Arabidopsis under Heat Stress.

Heat shock transcription factors (HSFs) contribute significantly to thermotolerance acclimation. Here, we identified and cloned a putative HSF gene (HSFA2h) of 1218 nucleotide (acc. no. KP257297.1) from wheat cv. HD2985 using a de novo transcriptomic approach and predicted sHSP as its potential target. The expression of HSFA2h and its target gene (HSP17) was observed at the maximum level in leaf tissue under heat stress (HS), as compared to the control. The HSFA2h-pRI101 binary construct was mobilized in Arabidopsis, and further screening of T3 transgenic lines showed improved tolerance at an HS of 38 °C compared with wild type (WT). The expression of HSFA2h was observed to be 2.9- to 3.7-fold higher in different Arabidopsis transgenic lines under HS. HSFA2h and its target gene transcripts (HSP18.2 in the case of Arabidopsis) were observed to be abundant in transgenic Arabidopsis plants under HS. We observed a positive correlation between the expression of HSFA2h and HSP18.2 under HS. Evaluation of transgenic lines using different physio-biochemical traits linked with thermotolerance showed better performance of HS-treated transgenic Arabidopsis plants compared with WT. There is a need to further characterize the gene regulatory network (GRN) of HSFA2h and sHSP in order to modulate the HS tolerance of wheat and other agriculturally important crops.

Morpho-biochemical characterization of a RIL population for seed parameters and identification of candidate genes regulating seed size trait in lentil (Lens culinaris Medik.).

The seed size and shape in lentil (Lens culinaris Medik.) are important quality traits as these influences the milled grain yield, cooking time, and market class of the grains. Linkage analysis was done for seed size in a RIL (F5:6) population derived by crossing L830 (20.9 g/1000 seeds) with L4602 (42.13 g/1000 seeds) which consisted of 188 lines (15.0 to 40.5 g/1000 seeds). Parental polymorphism survey using 394 SSRs identified 31 polymorphic primers, which were used for the bulked segregant analysis (BSA). Marker PBALC449 differentiated the parents and small seed size bulk only, whereas large seeded bulk or the individual plants constituting the large-seeded bulk could not be differentiated. Single plant analysis identified only six recombinant and 13 heterozygotes, of 93 small-seeded RILs (<24.0 g/1000 seed). This clearly showed that the small seed size trait is very strongly regulated by the locus near PBLAC449; whereas, large seed size trait seems governed by more than one locus. The PCR amplified products from the PBLAC449 marker (149bp from L4602 and 131bp from L830) were cloned, sequenced and BLAST searched using the lentil reference genome and was found amplified from chromosome 03. Afterward, the nearby region on chromosome 3 was searched, and a few candidate genes like ubiquitin carboxyl-terminal hydrolase, E3 ubiquitin ligase, TIFY-like protein, and hexosyltransferase having a role in seed size determination were identified. Validation study in another RIL mapping population which is differing for seed size, showed a number of SNPs and InDels among these genes when studied using whole genome resequencing (WGRS) approach. Biochemical parameters like cellulose, lignin, and xylose content showed no significant differences between parents and the extreme RILs, at maturity. Various seed morphological traits like area, length, width, compactness, volume, perimeter, etc., when measured using VideometerLab 4.0 showed significant differences for the parents and RILs. The results have ultimately helped in better understanding the region regulating the seed size trait in genomically less explored crops like lentils.

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.

Utilization of intron-flanking EST-specific markers in the genetic characterization of Artemisia annua genotypes from the trans-Himalayan region of Ladakh, India.

Genetic variation was assessed utilizing intron-flanking EST-specific markers among genotypes of Artemesia annua collected from two sampling sites viz. Nubra (9,600 ft) and Leh (11,500 ft) valleys of the trans-Himalayan region, Ladakh, India. The available ESTs (3,60,906) sequences of A. annua were aligned with the genomic sequences of Arabidopsis to developed 'intron-flanking' EST-PCR based primers. These primers anneal with the conserved region of exon (flanking to the intron) and amplified the introns. Out of the 39 primers selected and tested on 20 genotypes of A. annua, we successfully exploited 81 codominant intron length polymorphism (ILP) markers, with an average of 2.08 markers per primer and 92.04% polymorphism detection. Clustering of genotypes revealed distribution of genotypes into 2 distinct clusters with respect to their site of collection. Significantly, this study demonstrates that Arabidopsis genome sequence can be useful in developing gene-specific PCR-based markers for other non-model plant species like A. annua in the absence of genome sequences.

Gene-Based Resistance to Erysiphe Species Causing Powdery Mildew Disease in Peas (Pisum sativum L.).

Globally powdery mildew (PM) is one of the major diseases of the pea caused by Erysiphe pisi. Besides, two other species viz. Erysiphe trifolii and Erysiphe baeumleri have also been identified to infect the pea plant. To date, three resistant genes, namely er1, er2 and Er3 located on linkage groups VI, III and IV respectively were identified. Studies have shown the er1 gene to be a Pisum sativum Mildew resistance Locus 'O' homologue and subsequent analysis has identified eleven alleles namely er1-1 to er1-11. Despite reports mentioning the breakdown of er1 gene-mediated PM resistance by E. pisi and E. trifolii, it is still the most widely deployed gene in PM resistance breeding programmes across the world. Several linked DNA markers have been reported in different mapping populations with varying linkage distances and effectiveness, which were used by breeders to develop PM-resistant pea cultivars through marker assisted selection. This review summarizes the genetics of PM resistance and its mechanism, allelic variations of the er gene, marker linkage and future strategies to exploit this information for targeted PM resistance breeding in Pisum.

Unraveling the pathways influencing the berry color and firmness of grapevine cv. Flame Seedless treated with bioregulators using biochemical and RNA-Seq analysis under semi-arid subtropics.

Plant bioregulators (PBRs) regulate developmental and physiological processes in plants. In this study, biochemical and transcriptomic analyses were conducted to evaluate the influence of PBRs [abscisic acid (ABA), benzothiadiazole (BTH), ethephon, and prohexadione-calcium (Pro-Ca)] on the grapevine cv. Flame Seedless under semi-arid subtropics. This study aims to see the effect of exogenous application of PBRs on overall berry quality, including uniformity of berry color. Uniform colored berries, the maximum total soluble solids (TSS) and total antioxidant activity (TAoA), and the highest total phenolics (TPC) and flavonoids (TFC) contents were obtained with the treatments, namely, 400 mg L-1 ethephon and 400 mg L-1 ABA. Further, RNA-Seq analysis has also identified some key DEGs like UFGT (VIT_16s0039g02230), GST (VIT_04s0079g00690), and chalcone synthase (CHS) (VIT_05s0136g00260) which were part of the anthocyanin biosynthesis pathway controlling grape berries color. Thus, ethephon (400 mg L-1) and ABA (400 mg L-1) were found promising for attaining greater uniformity in berry color development because of increased total anthocyanins content. In addition, they were also found associated with enhanced TAoA, TPC, and TFC. Hence, ethephon and ABA can be recommended for improving the berry quality.

Morphological, Molecular, and Biochemical Characterization of a Unique Lentil (Lens culinaris Medik.) Genotype Showing Seed-Coat Color Anomalies Due to Altered Anthocyanin Pathway.

This study reports the identification of a unique lentil (Lens culinaris Medik.) genotype L4717-NM, a natural mutant (NM) derived from a variety L4717, producing brown, black, and spotted seed-coat colored seeds in a single plant, generation after generation, in different frequencies. The genetic similarity of L4717 with that of L4717-NM expressing anomalous seed-coat color was established using 54 SSR markers. In addition, various biochemical parameters such as TPC (total phenolic content), TFC (total flavonoid content), DPPH (2,2-diphenyl-1-picrylhydrazyl), FRAP (ferric reducing antioxidant power), H2O2 (peroxide quantification), TCC (total carotenoids content), TAC (total anthocyanin content), and TAA (total ascorbic acid) were also studied in the seeds, sprouts, and seedlings of L4717, brown, black, and spotted seed-coat colored seeds. Stage-specific variations for the key biochemical parameters were recorded, and seedling stage was found the best for many parameters. Moreover, seeds with black seed coat showed better nutraceutical values for most of the studied traits. A highly significant (p ≤ 0.01) and positive correlation was observed between DPPH and TPC, TAA, TFC, etc., whereas, protein content showed a negative correlation with the other studied parameters. The seed coat is maternal tissue and we expect expression of seed-coat color as per the maternal genotype. However, such an anomalous seed-coat expression, which seems to probably be governed by some transposable element in the identified genotype, warrants more detailed studies involving exploitation of the anthocyanin pathway.

Heat stress tolerance in peas (Pisum sativum L.): Current status and way forward.

In the era of climate change, the overall productivity of pea (Pisum sativum L.) is being threatened by several abiotic stresses including heat stress (HS). HS causes severe yield losses by adversely affecting several traits in peas. A reduction in pod yield has been reported from 11.1% to 17.5% when mean daily temperature increase from 1.4 to 2.2°C. High-temperature stress (30.5-33°C) especially during reproductive phase is known to drastically reduce both seed yield and germination. HS during germination and early vegetative stage resulted in poor emergence and stunted plant growth along with detrimental effects on physiological functions of the pea plant. To combat HS and continue its life cycle, plants use various defense strategies including heat escape, avoidance or tolerance mechanisms. Ironically, the threshold temperatures for pea plant and its responses are inconsistent and not yet clearly identified. Trait discovery through traditional breeding such as semi leaflessness (afila), upright growing habit, lodging tolerance, lower canopy temperature and small seeded nature has highlighted their utility for greater adaptation under HS in pea. Screening of crop gene pool and landraces for HS tolerance in a targeted environment is a simple approach to identify HS tolerant genotypes. Thus, precise phenotyping using modern phenomics tools could lead to increased breeding efficiency. The NGS (next generation sequencing) data can be associated to find the candidate genes responsible for the HS tolerance in pea. In addition, genomic selection, genome wide association studies (GWAS) and marker assisted selection (MAS) can be used for the development of HS tolerant pea genotypes. Additionally, development of transgenics could be an alternative strategy for the development of HS tolerant pea genotypes. This review comprehensively covers the various aspects of HS tolerance mechanisms in the pea plant, screening protocols, omic advances, and future challenges for the development of HS tolerant genotypes.

Comparative transcriptome analysis, unfolding the pathways regulating the seed-size trait in cultivated lentil (Lens culinaris Medik.).

Market class, cooking time, quality, and milled grain yield are largely influenced by the seed size and shape of the lentil (Lens culinaris Medik.); thus, they are considered to be important quality traits. To unfold the pathways regulating seed size in lentils, a transcriptomic approach was performed using large-seeded (L4602) and small-seeded (L830) genotypes. The study has generated nearly 375 million high-quality reads, of which 98.70% were properly aligned to the reference genome. Among biological replicates, very high similarity in fragments per kilobase of exon per million mapped fragments values (R > 0.9) showed the consistency of RNA-seq results. Various differentially expressed genes associated mainly with the hormone signaling and cell division pathways, transcription factors, kinases, etc. were identified as having a role in cell expansion and seed growth. A total of 106,996 unigenes were used for differential expression (DE) analysis. String analysis identified various modules having certain key proteins like Ser/Thr protein kinase, seed storage protein, DNA-binding protein, microtubule-associated protein, etc. In addition, some growth and cell division-related micro-RNAs like miR3457 (cell wall formation), miR1440 (cell proliferation and cell cycles), and miR1533 (biosynthesis of plant hormones) were identified as having a role in seed size determination. Using RNA-seq data, 5254 EST-SSR primers were generated as a source for future studies aiming for the identification of linked markers. In silico validation using Genevestigator® was done for the Ser/Thr protein kinase, ethylene response factor, and Myb transcription factor genes. It is of interest that the xyloglucan endotransglucosylase gene was found differentially regulated, suggesting their role during seed development; however, at maturity, no significant differences were recorded for various cell wall parameters including cellulose, lignin, and xylose content. This is the first report on lentils that has unfolded the key seed size regulating pathways and unveiled a theoretical way for the development of lentil genotypes having customized seed sizes.

Genetic Variation for Traits Related to Phosphorus Use Efficiency in Lens Species at the Seedling Stage.

Phosphorus (P) is an essential, non-renewable resource critical for crop productivity across the world. P is immobile in nature and, therefore, the identification of novel genotypes with efficient P uptake and utilization under a low P environment is extremely important. This study was designed to characterize eighty genotypes of different Lens species for shoot and root traits at two contrasting levels of P. A significant reduction in primary root length (PRL), total surface area (TSA), total root tips (TRT), root forks (RF), total dry weight (TDW), root dry weight (RDW) and shoot dry weight (SDW) in response to P deficiency was recorded. A principal component analysis revealed that the TDW, SDW and RDW were significantly correlated to P uptake and utilization efficiency in lentils. Based on total dry weight (TDW) under low P, L4727, EC718309, EC714238, PL-97, EC718348, DPL15, PL06 and EC718332 were found promising. The characterization of different Lens species revealed species-specific variations for the studied traits. Cultivated lentils exhibited higher P uptake and utilization efficiency as compared to the wild forms. The study, based on four different techniques, identified EC714238 as the most P use-efficient genotype. The genotypes identified in this study can be utilized for developing mapping populations and deciphering the genetics for breeding lentil varieties suited for low P environments.

Diversity in Phytochemical Composition, Antioxidant Capacities, and Nutrient Contents Among Mungbean and Lentil Microgreens When Grown at Plain-Altitude Region (Delhi) and High-Altitude Region (Leh-Ladakh), India.

Mungbeans and lentils are relatively easily grown and cheaper sources of microgreens, but their phytonutrient diversity is not yet deeply explored. In this study, 20 diverse genotypes each of mungbean and lentil were grown as microgreens under plain-altitude (Delhi) and high-altitude (Leh) conditions, which showed significant genotypic variations for ascorbic acid, tocopherol, carotenoids, flavonoid, total phenolics, DPPH (1, 1-diphenyl-2-picrylhydrazyl), FRAP (ferric-reducing antioxidant power), peroxide activity, proteins, enzymes (peroxidase and catalase), micronutrients, and macronutrients contents. The lentil and mungbean genotypes L830 and MH810, respectively, were found superior for most of the studied parameters over other studied genotypes. Interestingly, for most of the studied parameters, Leh-grown microgreens were found superior to the Delhi-grown microgreens, which could be due to unique environmental conditions of Leh, especially wide temperature amplitude, photosynthetically active radiation (PAR), and UV-B content. In mungbean microgreens, total phenolics content (TPC) was found positively correlated with FRAP and DPPH, while in lentil microgreens, total flavonoid content (TFC) was found positively correlated with DPPH. The most abundant elements recorded were in the order of K, P, and Ca in mungbean microgreens; and K, Ca, and P in the lentil microgreens. In addition, these Fabaceae microgreens may help in the nutritional security of the population residing in the high-altitude regions of Ladakh, especially during winter months when this region remains landlocked due to heavy snowfall.