EFFECTS OF THE TOXICITIES OF ZN AND FE EXPOSURE ON THE GROWTHS OF MUNG BEAN (VIGNA RADIATA): AN EXPERIMENTAL LABORATORY STUDY

Mung bean (Vigna radiata) is widely cultivated across Asia and has gained popularity in various cuisines due to its high nutritional value. Rich in protein, dietary fiber, minerals, vitamins, and bioactive compounds like polyphenols and peptides, mung beans are considered a valuable functional food. The present experiment aimed to study the effects of water stress on the growth and yield of mung beans by applying varying water treatment volumes (20, 30, 40, 50, and 60 ml). Key growth parameters measured included stomata size on the stem and leaves, the number of leaves, average plant height, and biomass, all assessed after one week of growth. Water was supplemented every two days, and progress was monitored over the course of the week. The study found that the different volumes of water had a significant impact on the growth patterns, leaf count, biomass, and stomatal opening in the stem and leaves of the mung beans. The results suggest that while an adequate amount of water is essential for maximizing mung bean yield, excessive water can hinder growth by disrupting nutrient balance and other physiological processes. Therefore, to achieve optimal growth and yield in mung beans, careful management of water supply is crucial.

Effects of UVB Radiation on Stomatal Density and Opening in Rice (Oryza sativa L.)

Green gram (Vigna radiata) is considered the chief legume in Pakistan. Thus, current study was conducted to examine the ameliorating effect of phytohormones pre-treatments under salt stress on proteome profile of green gram by sodium-dodecyl-sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The soluble green gram seedlings proteins were resolved on 4% stacking and 12% resolving gels. The SDS-PAGE resolved 24 polypeptide bands ranging from 200 to 17kDa. Among these, 12 out of 24 bands of proteins were essentials house-keeping or growth proteins of green grams. While, 120, 114.6, 51.8, 29.1, and 22.8 kDa bands were over-expressed under 50 to 350mM salt with phytohormones treatments. The others 104.5 kDa, 99.8 kDa, 95.3 kDa, 91.0 kDa, 55 kDa, 46 kDa, and 17kDa bands were related to the GAᴣ, IAA, and SA induced tolerance. Overall 120 kDa, 114.6 kDa, 104.5 kDa, 99.8, 95.3 kDa, 51.8 kDa, 29.1 kDa and 22.8kDa bands were first time identified in the current study. The information retrieved from NCBI protein database, the resolved peptides were principally belonging to 7S and 8S vicilin, 2S, 8S, 11S, and 16.5S globulins. It is determined that seed priming with SA enhanced tolerance in green gram by rapidly synthesizing stress alleviating peptides.Key word: Cluster analysis, dendrogram, mungbean, salt stress, SDS-PAGEINTRODUCTIONVarious world-wide health concerning organization recommended the use of high graded plant protein such as legumes to prevent the risk of metabolic disorder (Hou et al., 2019). Legumes are most important protein crop on the earth. Among the legumes, the green gram is the major pulses. Its seeds are rich in superior quality storage protein, which account 85% of the total protein while, another 15% have not been broadly studied (Yi-Shen et al., 2018). The soluble storage protein comprises of 60% globulins, 25% albumin and 15% prolamins. Globulins are further divided into 3.4% basic-type (7S), 7.6% legumin-type (11S), and 89% vicilin-type (8S) (Mendoza et al., 2001; Itoh et al., 2006). Other than proteins, the green gram seeds also contain starch, fiber, phenolic compound, saponins, vitamins, calcium zinc, potassium, folate, magnesium, manganese and very low in fat that made it meager man’s meat (Hou et al., 2019). It is also a good source of green manure and fodder (El-Kafafi et al., 2015). Its root has ability to fix 30 to 50 Kg/ha atmospheric nitrogen in the soil which is essential for maintaining soil fertility (Chadha, 2010). The green gram is the valuable and the major Rabi pulse crop of Pakistan. Its cultivation area in 2016-2017 was about 179,000 hectares with seed yield of 130,000 tones. In comparison during 2017-2018, it was cultivated on 161,800 hectares land with 118,800 tones seed yield (GOP, 2018). One of the reasons of this 9% decrease in both land and productivity is the shortage of irrigated land due to soil salinity. The salinity induce oxidative bust in the mungbean cells, caused by responsive oxygen species (ROS) such as hydrogen peroxide, singlet oxygen, hydroxyl radical and superoxide radical. The ROS create hindrance in various metabolic processes of plant via interacting with macromolecules like proteins (Alharby et al., 2016). However, phytohormones like gibberellic acid (GAᴣ), indole acetic acid (IAA), and salicylic acid (SA) take part in the biosynthesis of salt tolerance proteins under salinity. These salt tolerance proteins acclimate plants under salinity stress. Application of biotechnology plays a significant role in agriculture (Khan et al., 2017). Therefore, production of particular proteins under salt stress is a specific response of cell which can be analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). SDS-PAGE is the simple, valid, and cost-effective biochemical marker (Mushtaq et al., 2018). This marker has been widely used to determine the extent of evolutionary variations in crops (El-Kafafi et al., 2015).OBJECTIVES The present study was directed first time with the aim to investigate the toxic effect of sodium chloride (0-350 mM) and stress acclimation by pre-treatment of GAᴣ, IAA, and SA on the proteome profile of NM-92 cultivar of a Pakistani green gram.MATERIALS AND METHODSThe present study was replicated thrice in the plant laboratory of Department of Genetics, Faculty of Science, and University of Karachi. The seeds of mung bean cultivar NM-92 were acquired from National Agricultural Research Centre (NARC), Islamabad. These freshly collected 15 seedsˉ1 treatment / replication were divided into two sets. The first was named as sodium chloride (SC) stress treatments were imbibed in sterile distilled water (DW) whereas, second set soaked in gibberellic acid (GAᴣ) (BDH Chemicals, England), indole acetic acid (IAA) (Fluka, Switzerland), and salicylic acid (SA) (J.T. Baker, Holland) in the separate beaker for 24 hours under dark condition. After 24 hours, given ample time to both the sets at room temperature. After recovery, all 20 treatments were sown in the 150 X 30 mm sized petri-dishes containing 0, 50, 150, 250, and 350 millimolar (mM) sodium chloride solution (Fisher Scientific, UK) for 72 hours.Protein extraction: Protein extraction was done by taking 0.3g of seedlings in an ice chilled mortar and crushed by adding 600µL 0.2 M Tris-HCl buffer having pH 7.5 contained 5% SDS (w/v) and 5% 2-mercaptoethanol (v/v). The homogenate was incubated at 0oC for 30 min., boiled in the water bath for 3 min. at 100oC. Samples were centrifuged in Heraeus Biofuge D-37520, Germany for 30 min. at 8000 rpm. The protein supernatant was saved at below 0°C for quantitative and qualitative determination with minor modifications. The total soluble protein content of the samples was estimated via “Bovine Serum Albumin (BSA) standard curve” and explicit in µg protein milligramˉ1 fresh weight of mung seedlings.Bovine serum albumin standard curve (2000 μg/mL): Total protein standard curve was made by dissolving 0.05g of Bovine Serum Albumin (BSA) in 25mL of distilled water. Ten serial dilutions were made from 0.1 mL to 1mL by BSA solution then performed Lowry. A standard curve of total proteins was plotted by taking BSA absorbance at Y-axis and 2000 μg BSA / mL at X-axisSample preparation for SDS-PAGE: For qualitative assessment of total proteins; the 35μL of saved protein supernatant was combined with 15μL of sample diluting buffer (SDB). The SDB was made up of 0.0625 M Tris-HCl pH 6.8 with 2% of SDS, 10% of glycerol, 0.003% of bromophenol blue dye and 5% of 2-mercaptoethanol. Boil the 50μL protein SDB supernatant at 100oC in water bath for 3 min., centrifuged at 6000 rpm for 4 min. The supernatant was loaded on SDS-PAGE gel with the given formulae. The SDS- PAGE: Total proteins were fractionated via SDS-PAGE with 4% stacking and 12% resolving gel. The resolving gel of 12% was made by taking 6mL solution A, 1.8 mL 3 M Tris 1 M HCl buffer pH 8.8, 144μL 10% SDS, 5.74 mL sterile distilled water, 720μL 1.5% ammonium persulphate (APS) in deionized water and 10μL TEMED. While, stacking was composed of 1.25mL of solution A, 2.5mL of 0.5M Tris 1M HCl buffer pH 6.8, 100μL 10% SDS, 1.8 mL of distilled water, 500μL 1.5% APS and 12μL TEMED. Solution A was prepared by conjoining 30% acrylamide and 0.8% N, N’-methylene-bisacrylamide in deionized water. To avoid polymerization in the beaker; the prepared solution was quickly poured into the 3 mm thick gel plates after adding TEMED. The stacking was lined over resolving gel, then combs were inserted between the gel plates of SCIE-PLAS TV-100 separation system, UK, and allowed to polymerize for ½ an hour. After polymerization gel was placed in the tank which were filled with Tris-Glycine buffer (electrode buffer) pH 8.4 then combs were removed. The electrode buffer contained 0.3% Tris, 1.41% Glycine and 0.1% SDS in 2000mL d/w. The gel was pre-run for 15 min. at 60 volts and 120 mA currents. The prepared SDS-PAGE samples were loaded in wells with BlueStepTM Broad Range Protein Marker, AMRESCO, USA as standard and run at 60 volts & 120 mA for about 45 min. When samples entered in resolving gel, and then gave 100 volts and 200 mA currents for around 2.5 hours. Furthermore, electrophoresis was carried out at a constant watt.The Gel was washed with 30% ethanol on Uni Thermo Shaker NTS-1300 EYELA, Japan at the constant shaking for 30 min. Then gels were placed in 10% glacial acetic acid in 50% methanol solution (Fixative) for 24 hours. SDS Gel was stained until protein bands were visible thereat placed as 5% of Methanol in 7.5% acetic acid glacial solution to destain the bands background. SDS-PAGE stain composed of 0.125% coomassie brilliant blue R-250 dissolved in 40% of Methanol and 7% acetic acid glacial solution. The stain was stirred on Magnetic stirrer & hot plate M6/1, Germany for 6-10 hours before used. Photographs were taken by Sanyo digital camera VPC-T1284BL and bands were scored through numbering pattern. Gels preserved in 10% acetic acid solution at 4°C.Interpretation of bands and data analysis: The total soluble protein bands relative mobility calculated by below formulae and Dendrogram was constructed via SPSS v. 20Where,F=(Migrated distance of protein band)/(Migrated distance of dye front)Slop=(Log MW of protein marker lower limit band–log〖MW of protein marker upper limit band )/(RF protein marker lower limit band –RF of protein marker upper limit band)RESULTS:The total soluble proteins extracted from green gram were perceived by SDS-PAGE Blue StepTm broad range biochemical markers. The protein-based marker was used to evaluate the toxic effect of sodium chloride along with pre-treatments of GAᴣ, IAA, and SA on proteome assay. In the current work, seedlings total soluble proteome resolved 24 polypeptide bands ranging from 200 to 17.1 kDa were recognized by using SDS-PAGE. The figure 1 showed Dendrogram assay, which classified the 20 treatments of SC, GAᴣ, IAA and SA into two major cl

Mung bean (Vigna radiata L.) is a legume crop with high economic value, containing 20-25% protein and rich in vitamins and minerals. Despite its agronomic advantages, its productivity in Indonesia remains low. The quality of planting media and proper nutrient management significantly affect productivity. The use of inorganic fertilizers can negatively impact soil, necessitating alternatives such as liquid biofertilizers. One potential type of liquid biofertilizer is JAKABA, a fungal culture derived from fermented rice washing water. This study aims to determine the effect of JAKABA as a pre-planting liquid biofertilizer on the growth of mung bean plants. The experiment employed a Completely Randomized Design (CRD) with four treatments: P0 (control), P1 (JAKABA 1:5 water), P2 (JAKABA 1:10), and P3 (JAKABA 1:15 water), each replicated four times. The experimental procedure included JAKABA production, mung bean seeding, and watering for seven days. Measured parameters included plant height, stem diameter, number of leaves, leaf length, and leaf width. Data analysis used One Way ANOVA test, followed by Duncan's test at α=5% significance level if significant differences were found. Results showed that the application of JAKABA as a pre-planting liquid biofertilizer had a significant effect on mung bean growth. Application of JAKABA liquid biofertilizer in pre-planting soil significantly enhanced mung bean vegetative growth. The 1:5 water dilution optimally increased plant height (26.35±0.64 cm), stem diameter (0.22±0.05 cm), and leaf count (0.22±0.05), while the 1:15 dilution maximized leaf dimensions (length: 4.12±0.29 cm; width: 3.02±0.38 cm). To conclude, the use of JAKABA as a pre-planting liquid biofertilizer effectively enhancing the vegetative growth of mung bean plants, with varying effectiveness depending on the concentration and observed growth parameters.

Green bean plants (Vigna radiata L.) are a staple food crop in Indonesia and are commonly consumed as legume food crops. Based on the Directorate General of Food Crops findings, there has been a decrease in the yield of green bean crops in Riau Province; therefore, measures must be taken to enhance productivity—green bean plant cultivation in the Riau Province. One potential method for improving the productivity of green bean plants involves the application of fertilizers. This study aimed to evaluate the impact of the interaction between liquid organic fertilizer rabbit urine and NPK fertilizer, as well as the individual effects of liquid organic fertilizer rabbit urine and NPK fertilizer, on the growth and yield of green bean plants (Vigna radiata L.) to determine the optimal treatment combination. The findings indicated no statistically significant impact on the development and production of green bean plants due to the combined application of rabbit urine LOF and NPK. The Rabbit urine Point of Concern (LOF) factor substantially impacts plant growth, including height, the abundance of productive branches, the number of full pods, and the weight of dry seeds per plot. The NPK fertilizer factor significantly influences various aspects of plant growth and productivity, including plant height, leaf count, root nodules effectiveness, branch productivity, pod yield, and seed weight per plot. The optimum treatment combination of 200 ml.l-1 rabbit urine POC and 300 kg.ha-1 NPK fertilizer has been found to enhance green bean plants' growth and yield significantly.

In Zea mays L., Bryophylum laxiflorum Bak., Gossypium hirsutum L., Helianthus annuus L., Oryza sativa L. and Vigna radiata L., a pre‐illumination in nitrogen causes transient stomatal opening upon returning the plant to darkness and normal air. In Zea mays L. hybrid INRA 508, K+ and Cl− fluctuations in the stomatal complex during this stomatal opening in darkness were similar to those observed during a light‐induced opening in normal air. These results are consistent with a two‐phase mechanism for stomatal opening: a light phase that may occur in the absence of oxygen and carbon dioxide, and a dark phase during which oxygen is necessary for ion accumulation and other mechanisms of osmotic adjustment.

Development of non-dairy probiotic drink utilizing sprouted cereals, legume and soymilk

Stomata, the microscopic pores surrounded by a pair of guard cells on the surfaces of leaves and stems, play an essential role in regulating the gas exchange between a plant and the surrounding atmosphere. Stomatal development and opening are significantly influenced by environmental conditions, both in the short and long term. The rapid rate of current climate change has been affecting stomatal responses, as a new balance between photosynthesis and water-use efficiency has to be found. Understanding the mechanisms involved in stomatal regulation and adjustment provides us with new insights into the ability of stomata to process information and evolve over time. In this review, we summarize the recent advances in research on the underlying mechanisms of the interaction between environmental factors and stomatal development and opening. Specific emphasis is placed on the environmental factors including light, CO2 concentration, ambient temperature, and relative humidity, as these factors play a significant role in understanding the impact of global climate change on plant development.

Phytotoxicity of selected herbicides to mung bean ( Phaseolus aureus Roxb.)

Deficit irrigation and magnetically treated water can be sustainable agronomic techniques to improve water use efficiency, plant development, and agricultural crop yield. A pot study was conducted to investigate the impact of distinct deficit irrigation regimes using magnetically treated and untreated water on the development and grain yield of mungbean [Vigna radiata (L.) R. Wilczek.] crops in a clayey soil of the Brazilian Cerrado. Mungbean plants were irrigated with magnetized water (MW) and non-magnetized water (N-MW) using four deficit irrigation regimes [20, 40, 60, and 80% of soil field capacity (FC)], considering a 2×4 factorial scheme, with four repetitions. The plants were maintained under distinct deficit irrigation regimes with MW or N-MW throughout the crop's entire growth and development cycle (80 days). Plant growth rate, relative chlorophyll index, plant morphological traits, production components, and crop grain yield were measured. The results showed that the deficit irrigation regime of 60% and 80% of FC improves the rate of plant growth and development and grain yield of the mungbean crop. Although mungbean is considered a drought-tolerant crop, low soil water availability causes severe inhibition of plant growth and development and drastically reduces the crop's grain yield. Irrigation with magnetized water had no beneficial effect on mungbean crops' plant growth and development and grain yield. Plant growth rate, plant height, shoot dry matter, relative chlorophyll index, and number of grains per pod positively correlate with grain yield. In contrast, root length has a highly negative correlation with the grain yield of mungbean crops.

OST1 Kinase Modulates Freezing Tolerance by Enhancing ICE1 Stability in Arabidopsis

Phytoremediation of lead in Mineral, distilled and surface water using Pennisetum purpureum and Allium fistulosum

Root-infecting fungal pathogens and also parasites, which do not cause major disease symptoms cause problems of contamination in pot cultures of arbuscular mycorrhizal (AM) fungi. We investigated the effect of the AM fungus, Glomus coronatum Giovannetti on disease caused by binucleate Rhizoctonia sp. (BNR) and R. solani in mung bean in the absence (P0) and presence (P1) of added soil phosphorus (P). When G. coronatum and BNR or R. solani were inoculated at the same time, G. coronatum improved the growth of the plants and reduced colonization of roots by BNR, but not by R. solani. R. solani reduced the growth of non-mycorrhizal mung bean in P0 soil 6 weeks after inoculation, whereas BNR had no effect on growth. G. coronatum reduced the severity of disease caused by BNR or R. solani on mung bean in both soil P treatments. When G. coronatum was established in the roots 3 weeks before BNR or R. solani was added to the potting mix, there was no significant effect of BNR or R. solani on growth of mung bean. Prior colonization by G. coronatum slightly reduced indices of disease caused by BNR or R. solani. In both experiments, addition of P stimulated plant growth and reduced the colonization of roots by BNR, but had little effect on disease severity. We conclude that the reduction of the effect of BNR or R. solani on mung bean could not be explained by improved P nutrition, but could be attributed to the presence of G. coronatum within and among the roots.

Evidence is presented in support of the hypothesis that stress ethylene formation determines ozone toxicity in plants. In studies with mung beans (Vigna radiata) ozone toxicity was reduced not only when plants had been pretreated with aminoethoxyvinylglycine (AVG) but also after pretreatment of plants with CoCl2 and ascorbic acid. While AVG prevents the enzymatic conversion of S‐adenosylmethionine (SAM) to I‐aminocyclopropane‐I‐carboxylic acid (ACC), cobalt and free radical scavengers such as ascorbic acid inhibit the subsequent conversion of ACC to ethylene. Stomatal opening was not affected by pretreatment of plants with inhibitors of ethylene biosynthesis.

The allelopathic effect of Teak (Tectona grandis Linn.f.) fruit leachates on the germination and vigour characteristics of Rice (Oryza sativa Linn.) and Green Gram (Vigna radiata (Linn.) Wilczek) seeds was studied under laboratory conditions. The Rice and Green Gram seeds were soaked directly in the Teak fruit leachates of different days or in distilled water and subsequently wetted during germination with the leachate or distilled water. The germination characteristics of Rice and Green Gram seeds increased significantly due to the application of Teak fruit leachate. Largest effect was recorded generally due to first day’s leachate and the effect declined due to subsequent days’ leachate.