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Open Access
Development, characterization and application of bionanoconjugate on terminal heat stress in wheat (Triticum aestivum L.)
(CCSHAU, Hisar, 2023-04) Narender Mohan; Ajay Pal
To ensure sustainable farming practices, nanotechnology in modern agriculture must address the worldwide issues of biotic and abiotic stress (Terminal Heat Stress) brought on by climate change and the population load on farms. Numerous biopolymers, including alginate, cellulose, chitin, and chitosan, have increased use in the new nanotechnology period to create new materials useful in crop growth and protection. Chitosan is the most extensively studied biomaterial in nanotechnology because it is the second most prevalent and conveniently accessible biopolymer after cellulose. Due to its unrestricted qualities, such as biocompatibility, antibacterial, biodegradability, and non-toxicity for plants, it is employed primarily as nanochitosan instead of as bulk throughout the world. Compared to pure/bulk chitosan or direct use of ionic metals, nanochitosan biopolymer has been deemed more valuable for its involvement in chelation with metal ions, such as Zn, Cu, Fe, etc. Herein we incorporated plant hormones (Salicylic acid) and metal (Zn) ions into ongoing cross linking of chitosan and TPP to create nano-composites/conjugates, leading to more pronounced and efficient effects on seed germination and screen house. The current study shows that the genotype, species, growth stage, ambient conditions, and ingredients/nutrients affect how plants react to BNCs. It is further inferred that applying BNCs with co-encapsulated SA and Zn improved the crop's resistance to stress. In combination with the slow-release capability of BNCs, it considerably improved the germination potential, antioxidant system, crop yield, and grain quality of both WH-1124 and WH-542, according to laboratory and pot studies. Additionally, we conclude that the synthesis of these BNCs may be accelerated further to assess their efficacy on other crops, which could be advantageous for the technical translation.
Open Access
Development, characterization and efficacy of bionanocluster of indole acetic acid and cobalt chloride on seed germination and yield in Cotton (Gossypium hirsutum L.)
(CCSHAU, Hisar, 2023-09) Vikram Singh; Mandhania, Shiwani
In the present investigation, Co2+ and indole-3-acetic acid were combined to create chitosan- Co2+-IAA BNCs. The mean hydrodynamic diameter, PDI value and zeta-potential of BNCs were recorded as 481.9 nm, 0.536 and +22.00 mV, respectively. Effect of seed priming with 100 ppm BNCs and four time foliar application of 100 ppm BNCs on seed germination, seedling growth, and boll drop was studied in H 1098i genotype of G. hirsutum L. Cotton seeds subjected to BNCs priming displayed higher percent germination and seed vigor index-I (92.14 %) compared to hydroprimed seeds due to elevated activities of glyoxylate cycle enzymes i.e. isocitrate lyase and malate synthase, which increased the conversion of fats to carbohydrates. Four times foliar application of 100 ppm BNCs increased leaf area, plant height, total chlorophyll content, photosynthetic rate, ACC synthase activity, indole-3-acetic acid content, boll number, boll weight ultimately increasing seed cotton yield by 20.37 %. However, ACC oxidase, cellulase, pectinase activities and boll drop significantly decreased compared to control. It is concluded that application of BNCs comprising of chitosan, IAA and Co2+ via seed priming and foliar mode is novel, economical, and environment friendly.
Open Access
Effect of exogenous sulphur in alleviating lead toxicity in Indian mustard [Brassica juncea (L.)]
(CCSHAU, Hisar, 2023) Manne Hemanth Kumar; Nisha Kumari
The Indian mustard [Brassica juncea (L.)] is one of the well-known plant for phytoextraction of heavy metals. The present study was designed to elucidate whether exogenously applied sulphur (S1; 100 ppm, S2; 200 ppm sulphur) alleviate lead (Pb)-induced (Pb1; 100 ppm, Pb2; 200 ppm and Pb3; 300 ppm Pb) stress in the leaves of Brassica juncea. Lead was applied in the form of lead nitrate five days prior to sampling and sulphur applied in the form of zinc sulphate at three days before sampling and samples were collected on 30, 60 and 90 DAS. Lead stress of 100, 200 and 300 ppm has considerably reduced plant growth as evidenced by increased H2O2 contents of 303.64, 317.39 and 238.89 % at 300 ppm of 30, 60 and 90 DAS, respectively, and they further caused the peroxidation of lipid membranes. This disruption increased more leakage of electrolytes, and created ionic imbalance. Further, alteration of chlorophyll pigment by replacing its central magnesium atom has reduced photosynthetic rate by 27.24 % at Pb3 stress, in respect to control. The redox status of the cell got altered by lead toxicity, depleting the AsA and GSH pool metabolites. Besides, GR, APX and POX antioxidative enzyme activities have shown highest increase under Pb3 stress by 51.97, 57.84 and 121.45 %, at 30, 60 and 90 DAS, respectively. In contrast, S application showed an increased AsA-GSH contents by 15.30, 9.40 and 7.89 % at S1 spray and 38.12, 23.44 and 33.40 %, respectively, at 30, 60 and 90 DAS at S2 spray. Sulphur-metabolites such as ATPS, OASTL and GST activities enhanced upon stress and resulted in further augmentation upon sulphur supplementation. Sulphur application to the Pb-stressed Brassica plants has reduced the oxidative stress by decreasing EL content by 11.69 % and MDA contents by 13.79 % at 90 DAS, respectively, in respect to Pb3-stressed plants. This decrease in oxidative stress biomarkers was due to an increase in AsA-GSH thiol pool and antioxidative enzyme enhancement under 100 and 200 ppm sulphur application to the Pb-stressed plants. Besides, S2 application to the Pb3+S2 stressed plants has increased phytochelatins by 522.47, 352.60 and 443.48 %, at 30, 60 and 90 DAS, respectively, in respect to Pb3 stress and called for PC-based detoxification of Pb. Furthermore, lead stress shown a decreased relative gene expression of BjSULT1.a gene and increased BjPb2.1, BjATPS1, BjOASTL2, BjABCC2 and BjGR1 gene expression. Moreover, S2 spray increased gene expression levels by 1.32, 1.49, 2.42 and 2.24 folds of BjSULT1.a, ABCC2, BjATPS1 and BjOASTL2, respectively, under Pb3 stress. Sulphur spray of S1 and S2 to the Pb-stressed Brassica juncea plants increased morpho-physiological parameters and yield and yield parameters. In the view of present findings, greater decrease of oxidative stress (H2O2, MDA and EL), increased enzymatic, non-enzymatic antioxidants, phytochelatins, enhanced relative gene expression levels and maximum seed yield was found under exogenous supplementation of 200 ppm sulphur and has ameliorated lead stress.
Open Access
Biochemical and protein profiling in response to application of herbal nano-formulation(s) on shelf life of guava (Psidium guajava L.)
(CCSHAU Hisar, 2022-09) Anju Rani; Tokas, Jayanti
The guava (Psidium guajava L) is a resilient, easily adaptable and evergreen tree of the Myrtaceae family. Guavas are nutrient rich fruit having high content of vitamin A, vitamin C, antioxidants and minerals but the post-harvest losses stand at 20-40% of the produce. Herbal NFs were used to prevent post-harvest losses. Initially, different herbal extracts of giloy (leaves and stem) and ajwain seeds were prepared using different solvents and extraction procedure. Ajwain methanol (E2), ajwain methanol-acetone (7:3) (E3), giloy methanol (E16) and giloy acetone (E17) leaves extracts were selected for NFs synthesis. Three NFs viz. sodium alginate: CaCl2 (T1), sodium alginate: CaCl2: ajwain extract (T2), and sodium alginate: CaCl2: giloy extract (T3) were selected for further treatment. Physiological, quality, minerals, cell wall and ripening related parameters were analyzed every alternate day upto 10 DAT during storage at 25±2 °C and 32±2 °C. The results revealed that physiological loss in weight increased and fruit firmness decreased during storage. The quality parameters viz., total sugars, reducing sugars total soluble solids and ascorbate increased initially and then decreased at both the storage temperatures. The percent increase in total soluble sugars, reducing sugars and TSS on 10th day was maximum in control (34.8 and 36.6%; 70.7 and 76.3%; 31.8 and 34.8%) followed by T1 (29.3 and 30.0%; 64.6 and 66.2%; 15.7 and 25.1%), T3 (17.3 and 23.1%;42.5 and 49.4%; 5.1 and 9.5%), and T2 (14.9 and 20.7%; 36.4 and 44.6%; 3.7 and 8.9%) NFs at 25±2 °C and 32±2 °C. Whereas, Titratable acidity, crude proteins and antioxidant activity reduced during storage. Minerals content decreased during ripening in all the treatments and minimum decrease was recorded in T2 NFs treated fruits. Activity of cell wall degrading enzymes cellulase and polygalaturonase increased as the storage period increased whereas pectin methyl esterase activity increased initially and then decreased. The maximum percent increase of cellulase and PG was observed in control (272.6 and 365.3%; 285.3 and 325.3%) fruits followed by T1 (225.6 and 343.6%; 271.0 and 306.5%), T3 (211.2 and 318.7%; 224.1 and 272.2%) and T2 (201.3 and 292.9%; 215.2 and 266.5%) NFs treated fruits at 25±2 °C and 32±2 °C. Ethylene peak was observed on 4th and 6th day of treatment at 25±2 °C and 32±2 °C and similar trend was observed for ACC oxidase activity. The results revealed that Sodium alginate: CaCl2: Ajwain (T2) NF was most effective in controlling post-harvest losses and increased shelf life by upto 10-12 days. Lesser quality degradation took place at 25±2 °C as compared to 32±2 °C. The, control and T2 NF treated fruits were further analyzed for proteomics study using 2D-Electrophoresis and LC-MS/MS. Maximum proteins concentrated at higher pH and more in the range of 20-120 kD in 2D gel. Analysis of LC-MS/MS results using MASCOT software revealed more than 100 differentially expressed proteins (DEPs), out of which 35 proteins were up-regulated and 20 proteins were down-regulated having fold change ≥1.5 and ≤0.12, respectively, which were further analysis using GO software. Proteins were classified on the basis of molecular, biological and cellular functions. Protein profiling elucidated the expression of some novel proteins which might play a role in delayed ripening.
Open Access
Development of herbal nano-formulations for enhancing shelf life of guava (Psidium guajava L.) fruit
(Chaudhary Charan Singh Haryana Agricultural University hisar, 2022-09) Gupta, Pernika; Tokas, Jayanti
Guava (Psidium guajava L.), member of the family Myrtaceae is an important fruit of tropical and subtropical area of world. It is also known as ―apple of the tropics‖. Nutritionally, guava fruit is a rich source of ascorbic acid but the post-harvest losses stand at 20-40% of the produce. Herbal extract based edible nano-formualtions were used to prevent post-harvest losses. Initially, different herbal extracts of ashwagandha and menthe leaves were prepared using different solvents. Three nanoformulations viz. Alg-Lys (T1), Alg-Lys:menthe extract (methanol) (T2 ), and Alg- Lys:ashwagandha extract (methanol) (T3) were selected for further treatments. Physiological, quality, cell wall components & degrading enzymes, antioxidative and ripening related parameters were analyzed every 3 day upto 12 DAT during storage at turning and mature green stage of Hisar Safeda and Hisar Surkha. The results revealed that physiological loss in weight increased and fruit firmness decreased during storage. The quality parameters viz., total soluble sugars and total soluble solids; antioxidative enzymes viz., superoxide dismutase, catalase, ascorbate peroxidase activity and ascorbate content increased initially and then decreased in both Hisar Safeda and Hisar Surkha. Overall decrease in total soluble solids was delayed in T3 treated guava fruits which was significantly higher than in control fruits in Hisar Safeda and Hisar Surkha. Whereas, titratable acidity, crude protein content, glutathione reductase and peroxidase activity reduced during storage. Activity of cell wall degrading enzymes cellulase and polygalaturonase increased as the storage period increased whereas pectin methyl esterase activity increased initially and then decreased. The amount of ethylene evolution and activity of ACC Oxidase in two cultivars of guava fruits increased initially and during further ripening process, the activity reduced. The results revealed that Alg-Lys:ashwagandha (T3) nanoformulation was most effective in controlling post-harvest losses and increased shelf life of Hisar Safeda upto 5 days and Hisar Surkha upto 3 days. The, control and T3 nanoformulation treated fruits were further analyzed 2D-Electrophoresis and LC-MS/MS and it was observed that maximum proteins had concentrated at higher pH and more in the range of 20-120 kD. Analysis of LC-MS/MS results using MASCOT software revealed more than 60 differentially expressed proteins (DEPs), which were involved in energy and carbohydrate metabolism, stress response, defense, biosynthesis, hydrolase, secondary metabolism, amino acid and nucleic acid, protein synthesis and biogenesis of cellular components having fold change ≥1.5 which were considered for further analysis. Using GO classification, major pathways involved in molecular, biological and cellular functions in Hisar Safeda fruit ripening were classified. Proteins were classified on the basis of molecular, biological and cellular functions. Protein profiling elucidated the expression of six novel proteins which might play a role in delayed ripening.
Open Access
Sucrose mobilization and nitrogen metabolism in aerobic and lowland indica rice (Oryza sativa L.)
(CCSHAU, Hisar, 2020) Jhandai, Sonia; Ajay Pal
Water dearth is the most critical issues in modern agriculture. Therefore, aerobic cultivation condition is sustainable concept to grow rice (Oryza sativaL.) which favors huge water impetus and labor. A better understanding of sucrose mobilization in shoots at vegetative and reproductive stages and developing grains at 7th–14th DAA may help in selection of rice genotypes to grow under aerobic condition. Herein, we have studied the effect of aerobic and conventional conditions on the activities of sucrose mobilization enzymes in four rice varieties viz. PAU201, HBC19 (lowland) and MAS26, MAS25 (aerobic) under pot condition. Aerobic condition significantly increased activities of sucrose synthesizing enzyme viz. sucrose phosphate synthase (SPS), sucrose phosphate phosphatase (SPP) and sucrose synthase (synthesis) (SSs). Activities of starch synthesizing enzymes viz.sucrose synthase (cleavage) (SSc), ADP-glucose pyrophosphorylase (AGPase) and soluble starch synthase (SSS) were reduced in concord with starch content under aerobic condition. Sucrose, soluble sugars, reducing and non-reducing sugars increased under aerobic condition in contrast with starch, UDP-glucose and ADP-glucose. UDP glucose pyrophosphorylase (UGPase) activity increased in shoot under aerobic condition to contribute sucrose synthesis while decreased in developing grains resulting in decreased starch content. Nitrogen, an essential component of all proteins, is one of the essential macronutrients required for better yield and therefore, nitrogen metabolism pathway was also studied under aerobic condition with respect to conventional condition and observed decreased activities under aerobic condition in both shoots and roots. Similar to enzymes, diminished metabolites viz. nitrite and alanine were observed under aerobic condition Crop sustainability under aerobic condition in all varieties could be attributed to osmoprotective role of sucrose and total soluble sugars. Our findings by semi-quantitative RT-PCR analysis also revealed up-regulation of SPS with down-regulation of AGPase and SUS transcript under aerobic condition is responsible for significant decrease in starch with increase in sucrose. Our results reveal decreased yield attributes under aerobic condition. Biochemical analysis, gene expression and yield attributes highlight the role of various enzymes prioritized for assimilate translocation under aerobic condition, and provide evidence of differentiation of metabolic pathway under two growing condition as well as aerobic and lowland rice varieties.
Open Access
Biochemical studies on antioxidative defense system in advance lines of wheat (Triticum aestivum L. em Thell.) grown under salt stress
(CCSHAU, Hisar, 2021-08) Bhadu, Surina; Tokas, Jayanti
Wheat (Triticum aestivum) is an important cereal crop for the world population. Based on productivity, wheat is the third most commonly grown cereal after maize and rice. It is essential for providing the mandatory amount of calories and protein content in the affluent human diet. Among abiotic stresses, salt stress is the chief reason which affects approximately 50% of crop productivity along with soil fertility. Under saline conditions, there is an imbalance in the uptake of the ions which disturbs the homeostasis between Na+/ K+ ratio. Due to the ions imbalance in plants, there is a generation of ionic and osmotic stress at the cellular level. The excess amount of ROS produced is the consequence of this stress which damages the membrane integrity that causes several changes at physiological, morphological, biochemical, and molecular levels. The present study was carried out on F5 and F6 generation of a conventional wheat cross between Kharchia 65 (salt-tolerant) × WH1105 (saltsensitive). The F5 generation was sown at 8 ECe while F6 was sown at 10 ECe and analyses were carried out at vegetative and reproductive stages. Under salt stress, WH1105 had higher oxidative stress components viz. H2O2, MDA which increased in response to the ROS and antioxidative defense system got activated to scavenges the excess of ROS. Kharchia 65 (salt-tolerant) showed higher antioxidative enzyme activities than WH1105 (salt-sensitive) under salt stress at both vegetative and reproductive stages. Introgress of Nax1 and Nax2 genes was much affective in increasing the antioxidative enzyme activities viz. at vegetative stage, SOD (1.33%, 4.50%), CAT(14.6%, 4.13%), APX(12.57%, 20.64%), GR (14.51%, 20.34%), GPX(13.91%, 4.34%), MDHAR (55.8%, 44.4%), DHAR (42.9%, 38.3%) and POX (10.44%, 4.29%) and at reproductive stage, SOD (3.26%, 3.25%), CAT(4.67%, 7.18%), APX(11.87%, 9.13%), GR (2.65%, 19.8%), GPX(19.16%, 8.48%), MDHAR (45.7%, 48.6%), DHAR (35.7%, 34.2%) and POX (7.58%, 7.12%) in F5 and F6 generations respectively in plants expressing Nax1 and Nax2 genes as compared to Kharchia 65 under salt stressed condition. The physiological parameter like RWC and osmotic potential declined with an increase in salinity and the Na+/K+ ratio was higher in F6 as compared to F5. The LOX activity was also higher in WH1105 indicating higher lipid peroxidation as compared to Kharchia 65. The proteomics data indicate that there is up-regulation of many proteins in the plants that expressing either Nax1 or Nax2 gene as compared to salt stressed Kharchia 65. From the present study, it is concluded that introgress of Nax1 and Nax2 genes had activated the antioxidative defense mechanism with an increase in the crop yield under salt stress.
Open Access
Biochemical and expression profiling of sorghum [Sorghum bicolor (L.) Moench] under salt stress
(CCSHAU, Hisar, 2021-04) Himani; Tokas, Jayanti
Salinity induces complex metabolic processes that involve ion toxicity, osmotic stress, biochemical and physiological perturbations. Sorghum is a gluten-free cereal crop, well adapted to semi-arid tropics, highly biomass productive, and water-efficient. Therefore, the present study aimed to assess sorghum genotypes' tolerance behavior under different salinity levels (6, 8, 10, and 12 dSm-1) concerning the performance of physiological, biochemical, and molecular responses. Amongst 23 screened sorghum genotypes, SSG59-3 was identified as salt-tolerant and PC-5 as salt susceptible genotype based on germination studies. The results revealed that at 35 and 95 DAS, morpho-physiological parameters (RWC, OP, CF, CSI, total chlorophyll content) and growth traits (plant height, fresh and dry weight, root and shoot length) decreased with increasing salt concentration. The maximum decrease was observed in PC-5 as compared to SSG 59-3 at 10 dSm-1. PC-5 accumulated higher Na+and K+ content in roots while the exclusion potential of SSG 59-3 was more in roots, so it had minimum Na+ and K+ content. The quality traits viz. HCN, CP, IVDMD, CPY, DDM, and cell wall components (except HC, which decreased with plant maturity) also followed a similar pattern. SSG 59-3 maintained the higher GFY, DMY, and yield components at 10 dSm-1 (at physiological maturity). The imposition of salinity induces the activities of ROS scavenging enzymes viz. SOD, CAT, POX, APX, GPX, GR, MDHAR, and DHAR and the content of ROS scavenging metabolites viz. ascorbate, glutathione, and carotenoids which were maximum in SSG 59-3 while lower in PC-5 at 10 dSm-1. The oxidative stress as judged by accumulation of MDA and H2O2 content was maximum in PC-5 and minimum in SSG 59-3. A significant increase was observed in compatible osmolytes viz. proline, glycine betaine, and total carbohydrates, which was more pronounced in SSG 59-3 than PC-5 at 10 dSm-1. Analysis of the LFQ results using MASCOT software revealed more than 100 differentially expressed proteins, out of which 40 proteins were upregulated, 29 proteins were down regulated and the majority was involved in catalytic activity, binding proteins, metabolic inter-conversion enzymes and ion transporters. The relative expression of CIPK24, LEA3, BADH1 was highly upregulated at 10 dS m-1while NCED3, SNAC1, GPX, H+-PPase, and P5CS1 were up-regulated at 12 dS m-1. The fold change was maximum in SSG 59-3 while the expression level of transcripts was less in PC-5. Hence, SSG 59-3 seems physiologically and biochemically promising and may be exploited in plant breeding programs aimed at developing salt-tolerant sorghum genotypes.
Open Access
Biochemical studies on chromium toxicity and its amelioration in sorghum (Sorghum bicolor L.)
(CCSHAU, Hisar, 2019-10-31) Praveen Kumar; Singal, H.R.
The main aim of the present research work was to determine the effects of Cr (VI) toxicity (2 ppm & 4 ppm) and its amelioration by glycine betaine (GB) and AMF at two growth stages viz. vegetative stage (35 DAS) and grain filling stage (95 DAS) in three varieties of sorghum viz. HJ 541 (single-cut), HJ 513 and SSG 59-3(multi-cut). The effect of exogenous application of GB (spiked in soil & foliar spray) and AMF (10 g) either individually or in combination was investigated for their ameliorative capability. The ameliorative effects were studied in terms of biochemical parameters (viz. structural and non-structural carbohydrates, crude protein, chlorophyll, chromium, hydrocyanic acid, IVDMD, pectin, PME, PG and cellulase), morphophysiological parameters (viz. root length, shoot length, number of leaves/tillers, grain yield, plant height and plant biomass), antioxidative defense system parameters (viz. enzymes – SOD, APX, CAT, GR, POX and metabolites – proline, glutathione, ascorbate, β-carotene) and indices of oxidative stress parameters (viz. PPO, H2O2 and MDA). The results revealed that the biochemical and morphophysiological parameters (except hydrocyanic acid, chromium, non-structural carbohydrates and pectin) decreased with increasing concentration of Cr (VI) in all the varieties at both the growth stages, and maximum decrease was observed in HJ 541 variety at 4 ppm concentration of Cr (VI) as compared to HJ 513 and SSG 59-3 variety at both growth stages. The antioxidative defense system parameters (enzymes & metabolites) and indices of oxidative stress parameters increased with increasing concentration of Cr (VI) at both the growth stages, in all the varieties. Further increase in antioxidative defense system parameters on exogenous GB (spiked in soil & foliar spray) and AMF application, either individually or in combination ameliorated the toxicity and decreased indices of oxidative stress parameters and increased IVDMD, total sugars, crude protein, β-carotene and nutritionally quality of sorghum in all the varieties at both the growth stages. The variety SSG 59-3 had lower hydrocyanic acid, non-structural carbohydrates, chromium, indices of oxidative stress parameters and more IVDMD and nutritional quality parameters as compared to HJ 513 and HJ 541 variety, at both the growth stages. Thus, among the three varieties (viz. HJ 541, HJ 513 and SSG 59-3), SSG 59-3 variety was found more tolerant as compared to HJ 541 and HJ 513 variety.

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