De novo transcriptome assembly and analysis of gene expression in different tissues of moth bean (Vigna aconitifolia) (Jacq.) Marechal.

BACKGROUND: The underutilized species Vigna aconitifolia (Moth Bean) is an important legume crop cultivated in semi-arid conditions and is valued for its seeds for their high protein content. It is also a popular green manure cover crop that offers many agronomic benefits including nitrogen fixation and soil nutrients. Despite its economic potential, genomic resources for this crop are scarce and there is limited knowledge on the developmental process of this plant at a molecular level. In the present communication, we have studied the molecular mechanisms that regulate plant development in V. aconitifolia, with a special focus on flower and seed development. We believe that this study will greatly enrich the genomic resources for this plant in form of differentially expressed genes, transcription factors, and genic molecular markers. RESULTS: We have performed the de novo transcriptome assembly using six types of tissues from various developmental stages of Vigna aconitifolia (var. RMO-435), namely, leaves, roots, flowers, pods, and seed tissue in the early and late stages of development, using the Illumina NextSeq platform. We assembled the transcriptome to get 150938 unigenes with an average length of 937.78 bp. About 79.9% of these unigenes were annotated in public databases and 12839 of those unigenes showed a significant match in the KEGG database. Most of the unigenes displayed significant differential expression in the late stages of seed development as compared with leaves. We annotated 74082 unigenes as transcription factors and identified 12096 simple sequence repeats (SSRs) in the genic regions of V.aconitifolia. Digital expression analysis revealed specific gene activities in different tissues which were validated using Real-time PCR analysis. CONCLUSIONS: The Vigna aconitifolia transcriptomic resources generated in this study provide foundational resources for gene discovery with respect to various developmental stages. This study provides the first comprehensive analysis revealing the genes involved in molecular as well as metabolic pathways that regulate seed development and may be responsible for the unique nutritive values of moth bean seeds. Hence, this study would serve as a foundation for characterization of candidate genes which would not only provide novel insights into understanding seed development but also provide resources for improved moth bean and related species genetic enhancement.

Construction of genetic linkage map and genome dissection of domestication-related traits of moth bean (Vigna aconitifolia), a legume crop of arid areas.

The moth bean (Vigna aconitifolia), possibly the most primitive crop of the genus Vigna, is a highly drought- and heat-resistant legume grown in arid areas. Moth bean domestication involved phenotypic changes, including reduction of seed dormancy and pod shattering, increased organ size, and earlier flowering and maturity. However, the genetics of the domestication process in moth bean is not known. In this study, we constructed a genetic linkage map for moth bean and used the map to identify quantitative trait loci (QTL) for domestication-related traits of an F2 population of 188 individuals produced from a cross of wild moth bean (TN67) and cultivated moth bean (ICPMO056). The genetic linkage map comprised 11 linkage groups (LG) of 172 simple sequence repeat markers and spanned a total length of 1016.8 centiMorgan (cM), with an average marker distance of 7.34 cM. A comparative genome analysis showed high genome synteny between moth bean and mungbean (Vigna radiata), adzuki bean (Vigna angularis), rice bean (Vigna umbellata), and yardlong bean (Vigna unguiculata). In total, 50 QTLs and 3 genes associated with 20 domestication-related traits were identified. Most of the QTLs belonged to five LGs (1, 2, 4, 7, and 10). Key traits related to domestication such as seed dormancy and pod shattering were controlled by large-effect QTLs (PVE > 20%) with one or two minor QTLs, whereas all other traits were controlled by one-seven minor QTLs, apart from seed weight, which was controlled by one major and seven minor QTLs. These results suggest that a small number of mutations with large phenotypic effects have contributed to the domestication of the moth bean. Comparative analysis of QTLs with related Vigna crops revealed that there are several domestication-related large-effect QTLs that had not been used in moth bean domestication. This study provides a basic genetic map and identified genome regions associated with domestication-related traits, which will be useful for the genetic improvement of the moth bean and related Vigna species.

The inhibitory effect of metals and other ions on acid phosphatase activity from Vigna aconitifolia seeds.

Sensitivity of acid phosphatase from Vigna aconitifolia seeds to metal ions, fluoride, and phosphate was examined. All the effectors had different degree of inhibitory effect on the enzyme. Among metal ions, molybdate and ferric ion were observed to be most potent inhibitors and both exhibited mixed type of inhibition. Acid phosphatase activity was inhibited by Cu2+ in a noncompetitive manner. Zn and Mn showed mild inhibition on the enzyme activity. Inhibition kinetics analysis explored molybdate as a potent inhibitor for acid phosphatase in comparison with other effectors used in this study. Fluoride was the next most strong inhibitor for the enzyme activity, and caused a mixed type of inhibition. Phosphate inhibited the enzyme competitively, which demonstrates that inhibition due to phosphate is one of the regulatory factors for enzyme activity.

Release of an encrypted, highly potent ACE-inhibitory peptide by enzymatic hydrolysis of moth bean (Vigna aconitifolia) protein.

Aim: Dietary approaches for the regulation of blood pressure are the need of the hour. Hence, identifying the foods possessing such activity is gaining importance. With this aim, moth bean (Vigna aconitifolia), an underutilized pulse, was explored for the presence of antihypertensive activity in terms of angiotensin converting enzyme (ACE)-inhibition bioactivity. Methods: Defatted moth bean protein concentrate was hydrolyzed by using different proteases including Alcalase, papain, and trypsin, to identify the enzyme producing highly potent ACE inhibitory peptides. The hydrolysate showing the highest ACE inhibitory activity was further fractionated using an ultrafiltration membrane (10, 3 and 1 kDa) based on ACE inhibitory activity. The active fraction was further subjected to the ion-exchange chromatography followed by RP-HPLC and LC-MS/MS analysis for the enrichment and identification of ACE inhibitory peptides. Finally, based on the bioinformatic analysis, few peptides were synthesized and evaluated for ACE inhibitory activity, followed by docking study and molecular dynamic simulation of a peptide with the highest ACE inhibitory activity. Results and discussion: Out of the three proteases, Alcalase-derived hydrolysate showed the highest (~59%) ACE inhibition activity. Molecular weight-based fractionation revealed that <1 kDa fraction possessed the highest ACE inhibitory activity. Activity guided separation of 1 kDa fraction using ion-exchange chromatography, RP-HPLC and LC-MS/MS showed the presence of about 45 peptides. Based on the bioinformatic analysis, 15 peptides were synthesized and evaluated for ACE inhibitory activity. Among these, a novel octapeptide FPPPKVIQ showed the highest ACE inhibitory activity (93.4%) with an IC50 of 0.24 µM. This peptide retained about 59% activity post gastrointestinal digestion simulation. A Dixon plot as well as docking studies revealed the uncompetitive inhibitory nature of this peptide with a Ki value of 0.81 µM. Molecular dynamic simulation studies till 100 ns ensured the stability of the ACE-peptide complex. Conclusion: Thus, present study identified a novel potent ACE inhibitory peptide from moth bean that can be incorporated in a functional dietary formulation for regulation of hypertension.

Natural Retinol Analogs Potentiate the Effects of Retinal on Aged and Photodamaged Skin: Results from In Vitro to Clinical Studies.

INTRODUCTION: Plants are a source of natural ingredients with retinol-like properties that can deliver anti-aging benefits without the side effects typically associated with retinoid use. We hypothesized that by combining two such analogs, bakuchiol (BAK) and Vigna aconitifolia extract (VAE), with the potent retinoid retinal (RAL), the anti-photoaging potential of RAL could be enhanced without compromising its skin irritation profile. The purpose of this study was to demonstrate that BAK and VAE potentiate the anti-photoaging activity of RAL. METHODS: Gene expression profiling of full-thickness reconstructed skin was first used to examine the impact of BAK or VAE in combination with RAL on skin biology. Next, the irritative potential of this combination, and its capacity to reverse key signs of photoaging in an ex vivo model was assessed. Finally, a proof-of-concept open label clinical study was performed to evaluate the anti-photoaging capacity and skin compatibility of a cosmetic formulation (tri-retinoid complex; 3RC) containing this complex in combination with other well characterized anti-photoaging ingredients. RESULTS: In vitro profiling suggested that combining 0.1% RAL with BAK or VAE potentiates the effect of RAL on keratinocyte differentiation and skin barrier function without affecting its skin irritation profile. When formulated with other anti-photoaging ingredients, such as niacinamide and melatonin, 3RC reversed ultraviolet radiation-induced deficits in structural components of the dermal extracellular matrix, including hyaluronic acid and collagen. In vivo, it led to a reversal of clinical signs of age and photodamage, with statistically significant improvement to skin firmness (+5.6%), skin elasticity (+13.9%), wrinkle count (-43.2%), and skin tone homogeneity (+7.0%), observed within 28 days of once nightly use. Notably, the number of crow's feet wrinkles was reduced in 100% of subjects. Furthermore, 3RC was very well tolerated. CONCLUSION: These data suggest that 3RC is a highly effective and well-tolerated treatment for photoaging.

Purification, molecular docking and in vivo analyses of novel angiotensin-converting enzyme inhibitory peptides from protein hydrolysate of moth bean (Vigna aconitifolia (Jacq.) Màrechal) seeds.

The moth bean is a high-protein food legume. Enzymatic hydrolysates of food proteins demostrate health benefits. Search for diet related food protein hydrolysates is therefore within the scope of functional foods. Present study asertains to produce, screen and identify natural ACE-I inhibitory peptides derived from moth bean seed protein hydrolysates. The extracted protein was hydrolysed using alcalase, chymotrypsin, flavourzyme, papain, pepsin and trypsin respectively. Alcalase achieved the greatest degree of hydrolysis and ACE inhibition. The highest ACE-I inhibitory activity was exhibited by the peptide with the lowest molecular weight i.e. <3 kDa (IC50 11.19 ± 0.15 µg/mL). This was further separated by FPLC, followed by mass spectrometry. Molecular docking analysis showed the peptides IAWDFR and ADLPGLK bind to active sites whereas DKPWWPK and AVIPNAPNLR to non-active sites of the ACE molecule. In vivo administration of MBP hydrolysate to dexamethasone-induced hypertensive rats reduced their systolic blood pressure (125 ± 0.76 mmHg) compared with positive control (155 ± 3.13 mmHg). Moth bean protein peptides exhibit functional nutraceutical properties with adequate antihypertensive activity.

Potential Use of Ascophyllum nodosum as a Biostimulant for Improving the Growth Performance of Vigna aconitifolia (Jacq.) Marechal.

The fertilizers that are derived from seaweed are known as Seaweed Liquid Fertilizers (SLF). SLF is a modern, cheap, non-toxic, and natural bioactive fertilizer. Among different studied seaweeds, Ascophyllum nodosum is significant as having bioactive ingredients that potentially regulate the molecular, physiological, and biochemical processes of crop plants. In the present study, the effects of the application of different concentrations (0.00%, 0.01%, 0.02%, 0.05%, 0.10%, 0.50%, and 1.00%) of A. nodosum Extract (ANE) to the Vigna aconitifolia through roots (Pot Root Application, PRA) and on the leaves (Pot Foliar Application, PFA) were monitored via the plant growth. The lower concentrations of ANE in both the PRA and PFA experiments showed positive growth on V. aconitifolia. The 0.10% ANE stimulated the maximum shoot growth when applied through the roots, while 0.05% ANE in both PFA and PRA experiments led to an increase in the number of pods, nodules, organic content, and moisture percentage. The 0.10% ANE also increased the leaf numbers, leaf area, and photosynthetic pigments. Hence, the application of 0.05% and 0.10% of A. nodosum extract in two ways (i.e., Pot Foliar Application, PFA, and Pot Root Application, PRA) ameliorated the growth capabilities of V. aconitifolia.