Hydroponic culturing upregulates sucrose and glutamine metabolism by enhancing their utilization via intermediates of aerobic pathway in wheat

Two wheat genotypes were grown in hydroponic culture, containing 4 mM KNO3, NH4Cl and NH4NO3. Activities of N metabolizing enzymes, aminotransferases, carbohydrate and TCA cycle enzymes were analyzed along with protein, amino acid, N, sugar content and growth parameters in shoot and root. After 12 days, the size of shoot and root system decreased significantly when plants were supplied with NH4Cl as exclusive N source. Under NH4NO3 growth parameters, N and carbon metabolism were elevated as compared to NH4Cl but less than KNO3 source indicating inhibition of NH4+ toxicity by NO3− uptake. Our results suggested that GDH, aminotransferases and PEPC play an important role in ammonium detoxification by its incorporation into amino acids. Thus, the morphologic differences among plants growing in NH4+ or NO3− nutrition confirm the hypothesis that N source determines the growth habit of plant in wheat by modulating the endogenous levels of protein and sugar content

Salt tolerant wheat landraces and Gly II transformed lines show distinct biochemical mechanisms of stress tolerance

The present investigation was carried out to study the distinct salt tolerance mechanism in two sets of material, Gly II transgenics and Kharchia landraces. The Gly II transgenics were developed for glyoxalase II (osglyII) gene (GenBank accession no. AY054407) from Oryza sativa through Agrobacterium mediated method in the background of wheat cultivar PBW 621. Kharchia 65 is a salt tolerant landrace derivative developed from Kharchia local which is native to saline soils of Rajasthan. The six wheat genotypes, viz. Kharchia local, Kharchia 65, PBW 621, G-2-2, G-3-4 and G-1-13 were evaluated for growth parameters, antioxidant enzymes and contents of glutathione, ascorbic acid, malondialdehyde (MDA), H2O2, sugars, chlorophyll, carotenoid, electrolyte leakage (EL) and Na+, K+ under control and two salt treatments (150 mM and 250 mM NaCl). The activities of antioxidant enzymes, glutathione, sugar content increased in both GlyII and Kharchia genotypes as compared to PBW 621. The GlyII activity increased (77–84%) in GlyII genotypes alongwith content of reduced glutathione (GSH) to maintain redox homeostasis. Apparently, GlyII and Kharchia genotypes exhibited minimum oxidative stress due to low content of MDA, H2O2, diminished EL and thereby causing less growth reduction and maintaining high chlorophyll and carotenoid level as compared to PBW 621. In addition, Gly II transgenic material and Kharchia lines showed less Na+ accumulation, greater seedling biomass and sugar content due to its salt tolerance mechanism. We infer that GlyII activity enhances GSH which play significant role in detoxifying ROS to establish stress homeostasis. The route for generation of GSH is via ascorbate-glutathione pathway mediated by glutathione reductase. Hence, GlyII transgenics and Kharchia genotypes can diminish salt stress following above mechanism

Supplementation of nitrogen and its influence on free sugars, amino acid and protein metabolism in roots and internodes of wheat

Effect of different doses of nitrogen (N) (90, 120, 150 and 180 kg Nha–1) on the activities of aminotransferases and alkaline inorganic pyrophosphatase (AIP) in relation to the accumulation of proteins, amino acids and sugars in roots and internodes at 15 and 40 days post anthesis (DPA) stages was studied in six wheat genotypes namely HD 2967, GLU 1101, PBW 343, BW 9022, PH-132-4840 and PBW 550. Supra-optimal N doses (150 kg Nha–1 and 180 kg Nha–1) accentuated glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and alkaline inorganic pyrophosphatase activities in correspondence with an increase in amino acid, protein and sugar content in both roots and internodes in all the six genotypes. Activities of analyzed enzymes were significantly high at 15 days post anthesis (DPA) stage and thereafter declined at maturity (40 DPA) in parallel with decrease in amino acid contents. Maximum activity of GOT, GPT and AIP was observed in HD 2967 and GLU 1101 genotypes along with higher build up of proteins and amino acids which resulted in higher grain yield. Activity of GPT was comparatively high over GOT, indicating its major role towards protein synthesis. Grain filling processes in terms of proteins and amino acids were positively correlated with GOT and GPT activities while sugars were correlated to AIP. Thus, nitrogen acquisition and assimilation resulted in favoured utilization of N in form of amino acid and proteins accumulation while sugar content was also stimulated. Due to immense activities of aminotransferases and higher contents of amino acids and proteins in GLU 1101 and HD 2967 genotypes at optimal dose and higher dose of N, these genotypes hold future potential for developing new cultivars with better grain quality characteristics

Modulation of Polyamines during Grain Development under Different Concentrations of Nitrogen in Wheat

The polyamine (PAs), putrescine (Put), spermidine (Spd) and spermine (Spm) are small aliphatic amines that ubiquitous in all living organisms. PA metabolism in higher plants is involved in both biotic and abiotic stress responses, and also influenced by nutrient deficiency. Accumulated evidence suggests that in plants the cellular titers of PAs are affected by various nitrogenous compounds. Therefore, the present study analyzed the effects of different nitrogen levels viz. recommended doses of nitrogen (RDN, 120 kg N/ha), sub-optimal N dose (RDN −25%, 90 kg N/ha) and supra-optimal N dose (RDN +25%, 150 kg N/ha) on PA metabolism in grains of six wheat genotypes at 15 days post anthesis (DPA) and 30 DPA. The activities of polyamine synthesizing enzymes (arginine decarboxylase, ornithine decarboxylase), catabolizing (diamine oxidase, polyamine oxidase) and the PA content were increased at supra-optimal nitrogen dose as compared to RDN. Whereas at sub-optimal nitrogen dose, higher activity of polyamine catabolizing enzymes results in speeding up oxidation of various PAs to cope up with nitrogen deficiency in plant. In relation to PA content, Put was found to be higher at early stage whereas Spd and Spm were found to be higher towards mature stages of grain indicating the use of Put in grain filling process. Highly significant correlation was observed between PA metabolism, yield and nitrogen use efficiency at sub-optimal N dose as compared to supra-optimal dose

Modulation of antioxidant defense system and polyamine catabolism in rice leaves under two planting conditions

Aerobic rice offers an attractive alternative approach over transplanting system as it consumes less water with low labour expenses. Flag leaf of six rice cultivars, viz. PR 120, PR 115, PR 116, Feng Ai Zan, PAU 201 and Punjab Mehak 1 was analysed for antioxidant defence mechanism and polyamine catabolism under the aerobic and the transplanting conditions. Ascorbate peroxidase (APX), guaiacol peroxidase (GPX), catalase (CAT), superoxide dismutase (SOD), diamine oxidase (DAO) and polyamine oxidase (PAO) activities increased gradually from tillering to anthesis stage and then declined towards maturity stage under both planting conditions. Apparently, contents of ascorbic acid, α-tocopherol, proline and polyamines (PAs) also revealed similar trend. The aerobic condition elevated activities of PAO, SOD as well as contents of PAs, lipid peroxide and H2O2 whereas the transplanting condition had higher levels of APX, GPX, CAT and total antioxidant activities and contents of ascorbate, α-tocopherol and proline. Cultivars Feng Ai Zan, PR 115 and PR 120 exhibited superior tolerance over other cultivars by accumulating higher contents of PAs with increasing levels of PAO and SOD activities under the aerobic condition. However, under the transplanting condition PR 116 and PAU 201 showed higher activities of antioxidative enzymes with decreasing contents of lipid peroxide and H2O2. We infer that under the aerobic condition, enhancement of PAs and PAO activity enabled rice cultivars to tolerate oxidative stress, while under the transplanting condition, antioxidative defence system with decreasing of lipid peroxide content was closely associated with the protection of flag leaf by maintaining membrane integrity. In crux, results indicated that H2O2 metabolic machinery was strongly up-regulated especially at the anthesis stage

Water and Salt Stress Metabolomics for Wheat Genotypes of India

Shoots of ten day old seedlings of nineteen wheat genotypes were evaluated for proline metabolism, H2O2, thiobarbituric acid reactive substances (TBARS) and 2,2 diphenyl-picrylhydrazyl (DPPH) radical scavenging activity under water deficit, water withholding and salinity stress conditions. Principle component analysis demarcated four groups: i.e. drought tolerant (Excalibar, Krichauff, Babax, Drysdale, Gladius and C306), salt tolerant (Kharchia, Type11, Krl 1-4 and Krl 19), low stress tolerant (C273, C518 and C591) and susceptible (HD2967, PBW621, WH1105, HD3086, PBW660 and PBW175). Salt stress treatment affected the length, fresh weight and dry weight of seedlings of all studied genotypes in comparison to water deficit and water withholding condition which may be due to higher contents of TBARS. Shoots of salt and drought tolerant genotypes possessed higher proline content and DPPH radical scavenging activity alongwith reduced content of TBARS in parallel with decreased H2O2 content under water stress conditions. The activities of proline synthesizing enzymes i.e. pyrroline-5-carboxylate synthetase (P5CS) and pyrroline-5-carboxylate reductase (P5CR) were significantly higher than proline degrading enzyme viz. proline dehydrogenase (PDH) under water stress as compared to salinity stress conditions. Overall, results indicated that P5CS, P5CR and PDH activities led to higher build up of proline under water stress, which might play a significant role in improving membrane stability by increasing radical scavenging activity and finally imparting stress tolerance in specific wheat genotypes

Syringaldazine peroxidase stimulates lignification by enhancing polyamine catabolism in wheat during heat and drought stress

Six wheat cultivars, namely PBW 343, PBW 550 (stress susceptible), PBW 621, PBW 175 (drought tolerant), C 306 and HD 2967 (heat tolerant), were used in this study to evaluate the effect of heat and drought stress on the activities of peroxidases (POXs), diamine oxidase (DAO), polyamine oxidase (PAO) and arginine decarboxylase (ADC) and ornithine decarboxylase (ODC) in relation to contents of polyamines (PAs), lipid peroxide and lignin. High temperature (HT) elevated activities of syringaldazine peroxidase (SPX), guaiacol peroxidase (GPX) and coniferyl alcohol peroxidase (CPX) in heat tolerant cultivars while, drought stress accentuated ADC/ODC activities in drought tolerant cultivars. Both heat and drought stress enhanced activities of DAO and PAO alongwith contents of H2O2 in PBW 175 and C 306. Amongst studied POXs, SPX activity was relatively more and coincided well with lignin content under HT stress while, the levels of ADC/ODC paralleled with putrescine and spermidine contents under drought stress. Higher build up of thiobarbituric acid reactive substances in cultivars PBW 343 and PBW 550 indicated their membrane instability during both the stresses. Our results revealed that SPX mediated lignification leading to higher cell wall rigidity under heat stress and drought increased PAs involved in ROS scavenging due to presence of positive charges which can bind strongly to the negative charges in cellular components such as proteins and phospholipids and thereby stabilize the membranes under stress conditions

Apical stem culturing to enhance cell sap assimilates towards grain sucrose and glutamine metabolism in wheat

Apical stem culturing offers an alternative approach of manipulating cell sap for wheat grain carbon and nitrogen metabolism in a near in vivo conditions. Employing this technique, role of sucrose and glutamine in transport stream on sucrose metabolism, ammonia assimilation and aminotransferase activities were assessed towards starch and protein accumulation in two wheat genotypes PBW 343 (low yield) and PBW 621 (high yield). At mid-milky stage, detached tillers were cultured in complete liquid medium containing varied concentration of glutamine and sucrose for seven days during year 2012–13 and 2013–14. Increasing glutamine concentration from 17 to 25 mM in the culture medium having 117 mM sucrose enhanced activities of nitrate reductase, glutamate synthase, glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) at 4 days after culturing (DAC) in correspondence with an increase in soluble protein content. However, at 7 DAC content of soluble protein decreased whereas starch accumulation increased showing, thereby a compensatory effect on carbon and nitrogen metabolism. Apparently, activities of sucrose synthase, soluble acid and neutral invertase significantly decreased. Increasing sucrose concentration from 117 to 125mMled to an increase in transformation of sugars to starch in grain but protein content decreased. PBW621 showed high protein content due to higher activities of GOT, GPT at 4 DAC which subsequently increased carbon skeleton of proteins towards starch synthesis at 7 DAC. Grain filling processes in terms of soluble sugars/starch were strongly correlated to invertase activities whereas proteins to aminotransferases

Controlling water deficit by osmolytes and enzymes: Enhancement of carbohydrate mobilization to overcome osmotic stress in wheat subjected to water deficit conditions

The present study demonstrates the effect of polyethylene glycol-6000 (PEG) induced water-deficit stress on growth, physiological and biochemical responses in six wheat (Triticum aestivum L.) cultivars viz. C 306, C 273, PBW 175 (drought tolerant), PBW 534, PBW 343 and PBW 550 (drought susceptibile) at seedling stage for six days. Stress caused short stature in shoot and elongated roots in all cultivars. Significant reduction in chlorophyll content and increase in membrane injury index and lipid peroxide content were observed in all the cultivars. Water stress accentuated sucrose synthase/phosphate synthase activities while sucrolytic enzymes namely; acid, neutral and sucrose synthase (cleavage) got repressed. Higher build up of sucrose and lower activity of amylase in root and shoot further decreased reducing sugar (glucose, fructose) content in stressed seedlings. Though the sugar supply (sucrose) under stress was not a limiting factor for germination but the poorer ‘metabolic conversion efficiency’ via sucrolytic enzymes in root and shoot resulted in their favoured utilization in amino acid, proline and glycine betaine. It may therefore be concluded that certain traits namely: membrane stability parameters, sugars, enzymes and osmolytes might help the cultivars to tolerate water deficit conditions.Keywords: Carbohydrates, chlorophyll, sucrose metabolism, membrane stability parameters, proline, protein, starch, Triticum aestivum, water-deficit.African Journal of Biotechnology, Vol 13(19), 2072-208