Synchrotron-based phase-sensitive imaging of leaves grown from magneto-primed seeds of soybean.

Experiments were conducted to study the effects of static magnetic fields (SMFs) on the venation network of soybean leaves using the synchrotron-based X-ray micro-imaging technique. The seeds of soybean (Glycine max, variety JS-335) were pretreated with different SMFs from 50 to 300 mT in steps of 50 mT for 1 h. The phase-contrast images obtained showed that, as the strength of the SMF increased, the area, width of the midrib, area of the midrib and minor vein of the middle leaflets of third trifoliate leaves also increased up to the SMF strength of 200 mT (1 h) and decreased thereafter. Quantification of the major conducting vein also showed the differences in the major and minor vein structures of the soybean leaves as compared with control leaves. Further, the phase-retrieval technique has been applied to make the segmentation process easy and to quantify the major and minor veins in the venation network. The width and area of midrib enhancement by pre-treatment with SMF implies an enhancement in the uptake of water, which in turn causes an increased rate of photosynthesis and stomatal conductance.

Synchrotron X-ray phase contrast imaging of leaf venation in soybean (Glycine max) after exclusion of solar UV (280-400†nm) radiation.

The hydraulic efficiency of a leaf depends on its vascular structure as this is responsible for transport activities. To investigate the effect of exclusion of UVAB and UVB radiation from the solar spectrum on the micro-structure of leaves of soybean (Glycine max, variety JS-335), a field experiment was conducted using synchrotron-based phase contrast imaging (PCI). Plants were grown in specially designed UV exclusion chambers, and wrapped with filters that excluded UVB (280-315 nm) or UVAB (280-400 nm), or transmitted all the ambient solar UV (280-400 nm) radiation (filter control). Qualitative observation of high-resolution X-ray PCI images obtained at 10 keV has shown the differences in major and minor vein structures of the leaves. The mid-rib width of the middle leaflet of third trifoliate leaves, for all treatments, were obtained using quantitative image analysis. The width of the mid-rib of the middle leaflet of third trifoliate leaves of UVB excluded plants was found to be more compared to leaves of filter control plants, which are exposed to ambient UV. The mid-rib or the main conducting vein transports water and sugars to the whole plant; therefore, mid-rib enhancement by the exclusion of solar UV radiation possibly implies enhancement in the leaf area which in turn causes an increased rate of photosynthesis.

Foliar application of pyraclostrobin fungicide enhances the growth, rhizobial-nodule formation and nitrogenase activity in soybean (var. JS-335).

A field study was conducted to investigate the impact of the fungicide pyraclostrobin (F500 - Headline®; a.i. 20%) on the activity of nitrogenase in soybean (var. JS-335). Pyraclostrobin (F500) was applied on the leaves of soybean plants at 10 and 20 days after emergence (DAE) of seedlings at concentrations ranging from 0.05% to 1%. Leghemoglobin (Lb) content and nitrogenase activity in root nodules were analyzed at 45(th)day after emergence of seedlings indicated a remarkable increase in Lb content and enhanced activity of nitrogenase in the root nodules of pyraclostrobin treated plants. The fungicide also enhanced the number of nodules along with weight of nodules, root biomass and growth of shoot and leaves. Enhanced nitrogen fixation in the root nodules by pyraclostrobin improves the growth of the plant in soybean before flowering and pod formation which ultimately resulted in yield and yield attributes. These results suggest that pyraclostrobin (F500) can be successfully employed as a foliar spray under field conditions to enhance the growth, nitrogen assimilation and hence yield of soybean.

Effect of stationary magnetic field strengths of 150 and 200 mT on reactive oxygen species production in soybean.

Our previous investigation reported the beneficial effect of pre-sowing magnetic treatment for improving germination parameters and biomass accumulation in soybean. In this study, soybean seeds treated with static magnetic fields of 150 and 200 mT for 1 h were evaluated for reactive oxygen species (ROS) and activity of antioxidant enzymes. Superoxide and hydroxyl radicals were measured in embryos and hypocotyls of germinating seeds by electron paramagnetic resonance spectroscopy and kinetics of superoxide production; hydrogen peroxide and antioxidant activities were estimated spectrophotometrically. Magnetic field treatment resulted in enhanced production of ROS mediated by cell wall peroxidase while ascorbic acid content, superoxide dismutase and ascorbate peroxidase activity decreased in the hypocotyl of germinating seeds. An increase in the cytosolic peroxidase activity indicated that this antioxidant enzyme had a vital role in scavenging the increased H(2)O(2) produced in seedlings from the magnetically treated seeds. Hence, these studies contribute to our first report on the biochemical basis of enhanced germination and seedling growth in magnetically treated seeds of soybean in relation to increased production of ROS.

Enhancement of germination, growth, and photosynthesis in soybean by pre-treatment of seeds with magnetic field.

Experiments were conducted to study the effect of static magnetic fields on the seeds of soybean (Glycine max (L.) Merr. var: JS-335) by exposing the seeds to different magnetic field strengths from 0 to 300 mT in steps of 50 mT for 30, 60, and 90 min. Treatment with magnetic fields improved germination-related parameters like water uptake, speed of germination, seedling length, fresh weight, dry weight and vigor indices of soybean seeds under laboratory conditions. Improvement over untreated control was 5-42% for speed of germination, 4-73% for seedling length, 9-53% for fresh weight, 5-16% for dry weight, and 3-88% and 4-27% for vigor indices I and II, respectively. Treatment of 200 mT (60 min) and 150 mT (60 min), which were more effective than others in increasing most of the seedling parameters, were further explored for their effect on plant growth, leaf photosynthetic efficiency, and leaf protein content under field conditions. Among different growth parameters, leaf area, and leaf fresh weight showed maximum enhancement (more than twofold) in 1-month-old plants. Polyphasic chlorophyll a fluorescence (OJIP) transients from magnetically treated plants gave a higher fluorescence yield at the J-I-P phase. The total soluble protein map (SDS-polyacrylamide gel) of leaves showed increased intensities of the bands corresponding to a larger subunit (53 KDa) and smaller subunit (14 KDa) of Rubisco in the treated plants. We report here the beneficial effect of pre-sowing magnetic treatment for improving germination parameters and biomass accumulation in soybean.

Assessment of impact of solar UV components on growth and antioxidant enzyme activity in cotton plant.

A field experiment was conducted to study the impact of solar UV-B (280-315 nm) and UV-A (315-400 nm) components on the growth and antioxidant enzyme activity in cotton plant (Gossypium hirsutum var. Vikram). Solar UV components were excluded by filtering the sunlight through selective UV-B (<315 nm) and UV-B/A (<400 nm) absorbing polyester films. Plants grown under filters that transmitted solar UV served as controls. Exclusion of UV-B and UV-B/A enhanced plant height, leaf area and total biomass of plants. The activity of antioxidant enzymes superoxide dismutase (SOD), ascorbic acid peroxidase (APX), glutathione reductase (GR) and guaiacol peroxidase (GPx) assayed in the leaves were lesser in the UV excluded plants. The level of ascorbic acid and UV absorbing substances were also decreased by the exclusion of UV. Solar UV components exerted a limitation on the potential growth of cotton plants. Reduction in the antioxidant enzyme activity and ascorbic acid after UV exclusion indicated that ambient UV components exert a significant stress on cotton plants. Reduction in the production of UAS indicated a changed pattern of metabolism leading to improved primary metabolism. Exclusion of UV components is advantageous from the agricultural point to enhance the growth of cotton plants.

Foliar application of fungicide-opera alleviates negative impact of water stress in soybean plants.

The modulatory effect of opera was investigated on the physiological and morphological aspects in soybean thriving in water stress environment. The data procured from current investigation indicated that water stress significantly declined the plant growth, leaf area in addition to photosynthetic efficiency, nitrate reductase activity and crop yield at various stages of growth such as vegetative (VS), flowering (FS) and pod filling stage (PFS). However, foliar application of opera (0.15%) was effective to enhance the the leaf area (42%), rate of photosynthesis (194%), and nitrate reductase activity (68%) at FS stage while the maximum enhancement in biomass accumulation (92%) and yield (119%) was observed at PFS stage as compared to their control plants. The opera is applied as foliar spray in field experiments to augment the assimilation of nitrogen and carbon in soybean which contributes to increased crop development and productivity under water stress conditions.

Similarities in the biochemical changes between solar UV exclusion and GA application in Amaranthus caudatus.

Seedlings of Amaranthus caudatus grown under UV exclusion filters (-UV -B <320nm; -UV -B/A <400nm) in the ambient solar radiation showed an enhancement in growth. This was accompanied by reduction in the synthesis of betacyanin but enhancement in the level of amino acids and proteins. The growth promotion and the accompanying biochemical changes were similar to the external application of GA3 to the seedlings under ambient solar radiation. The results are discussed in the light of similarities between UV exclusion and GA3 application in terms of diversion of amino acid precursors towards enhanced primary metabolism.

Identification of genetically diverse genotypes for photoperiod insensitivity in soybean using RAPD markers.

Most of the Indian soybean varieties were found to be highly sensitive to photoperiod, which limits their cultivation in only localized area. Identification of genetically diverse source of photoperiod insensitive would help to broaden the genetic base for this trait. Present study was undertaken with RAPD markers for genetic diversity estimation in 44 accessions of soybean differing in response to photoperiod sensitivity. The selected twenty-five RAPD primers produced a total of 199 amplicons, which generated 89.9 % polymorphism. The number of amplification products ranged from 2 to 13 for different primers. The polymorphism information content ranged from 0.0 for monomorphic loci to 0.5 with an average of 0.289. Genetic diversity between pairs of genotypes was 37.7% with a range of 3.9 to 71.6%. UPGMA cluster analysis placed all the accessions of soybean into four major clusters. No discernable geographical patterns were observed in clustering however; the smaller groups corresponded well with pedigree. Mantel's test (r = 0.915) indicates very good fit for clustering pattern. Two genotypes, MACS 330 and 111/2/1939 made a very divergent group from other accessions of soybean and highly photoperiod insensitive that may be potential source for broadening the genetic base of soybean for this trait.

UV-B induced changes in antioxidant enzymes and their isoforms in cucumber (Cucumis sativus L.) cotyledons.

To assess the role of antioxidant defense system on exposure to ultra-violet-B (UV-B) radiation, the activities of antioxidant enzymes superoxide dismutase (SOD), ascorbic acid peroxidase (APX), glutathione reductase (GR) and guaiacol peroxidase (GPX), as well as the level of antioxidants ascorbic acid (AA) and alpha-tocopherol were monitored in cucumber (Cucumis sativus L. var long green) cotyledons. UV-B enhanced the activity of antioxidant enzymes as well as AA content, but decreased the level of alpha-tocopherol. Significant increase was observed in the activities of SOD and GPX. Analysis of isoforms of antioxidant enzymes by native-PAGE and activity staining revealed three isoforms of GPX in unexposed dark-grown cotyledons (control), and their intensity was enhanced by UV-B exposure. In addition, four new isoforms of GPX were observed in cotyledons after UV-B exposure. Although no new isoforms were observed for the other antioxidant enzymes, the activities of their existing isoforms were enhanced by UV-B.

Oxyradical accumulation and rapid deterioration of soybean seeds due to field weathering.

The effect of field weathering on oxyradical accumulation and subsequent changes were studied in the seeds of soybean [Glycine max (L.) Merr.] cv. JS 71-05. Electron spin resonance (ESR) quantification of oxyradical revealed that field weathering plays an important role in acceleration of their accumulation. One week of weathering increased the accumulation of oxyradicals to almost 2-fold and triggered the deteriorative cascade, by enhancing the lipid peroxidation and membrane perturbation, leading to cell death in seed tissues and poor germinability and vigour of soybean seeds. Thus, the weather conditions at the time of physiological maturity to harvesting of crop are very crucial and the field weathering plays a critical role for the maintenance of seed quality.

Involvement of oxyradicals in promotion/inhibition of expansion growth in cucumber cotyledons.

Cytokinin-induced expansion growth of cucumber cotyledons and its interaction with UV-B (280-320 nm) was studied with reference to oxyradicals. UV-B radiation enhanced the level of oxyradicals in the cotyledons measured by EPR spectroscopy. Cytokinin promoted expansion growth was inhibited by UV-B radiation. Cytokinins reduced the level of oxyradicals in dark grown cotyledons, while promoting growth. Overproduction of oxyradicals by UV-B could not be fully accounted for the inhibition of growth of cotyledons, since quenching of radicals by cytokinins did not fully restore inhibition of growth.

Exclusion of solar UV radiation improves photosynthetic performance and yield of wheat varieties.

Field studies were conducted to determine the potential for alterations in photosynthetic performance and grain yield of four wheat (Triticum aestivum) varieties of India- Vidisha, Purna, Swarna and Naveen Chandausi by ambient ultraviolet radiation (UV). The plants were grown in specially designed UV exclusion chambers, wrapped with filters that excluded UV-B (<315 nm), UV-A/B (<400 nm) or transmitted ambient UV or lacked filters. The results indicated that solar UV exclusion increased the leaf mass per area ratio, leaf weight ratio and chlorophylls per unit area of flag leaves in all the four varieties of wheat. Polyphasic chlorophyll a fluorescence transients from the flag leaves of UV excluded wheat plants gave a higher fluorescence yield. Exclusion of solar UV significantly enhanced photosynthetic performance as a consequence of increased efficiency of PS II, performance index (PIABS) and rate of photosynthesis in the flag leaves of wheat varieties along with a remarkable increase in carbonic anhydrase, Rubisco and nitrate reductase activities. This additional fixation of carbon and nitrogen by exclusion of UV was channelized towards the improvement in grain yield of wheat varieties as there was a decrease in the UV-B absorbing substances and an increase in soluble protein content in flag leaves of all the four varieties of wheat. The magnitude of response for UV exclusion for all the measured parameters was higher in two varieties of wheat Vidisha and Purna as compared to Swarna and Naveen Chandausi. Cumulative stress response index (CSRI) for each variety was developed from the cumulative sum of physiological and yield parameters such as leaf mass area ratio of flag leaf, total chlorophyll content, performance index at absorption basis, rate of photosynthesis and grain yield. All the varieties had a negative CSRI, demonstrating a negative impact of ambient UV radiation. Naveen Chandausi and Swarna are less sensitive to ambient UV radiation; Vidisha is more sensitive to both UV-A and UV-B and Purna is more sensitive to ambient UV-B radiation.

Oxyradicals under UV-b stress and their quenching by antioxidants.

Formation of oxyradicals under UV-B stress was investigated using cucumber cotyledons. UV-B radiation induced production of free radicals which were analyzed by ESR spectroscopy. Evidence was obtained for the formation of superoxide and hydroxyl radicals in the tissues by comparing PBN-adducts formed with radicals obtained by chemical autooxidation of KO2 and Fenton's reaction. Addition of superoxide dismutase (SOD) to the reaction mixture partially reduced the intensity of signals confirming the production of superoxide radical as well as hydroxyl radicals. These radicals were quenched in vitro by the natural antioxidants alpha-tocopherol, ascorbic acid and benzoquinone. Changes in the level of antioxidants were also monitored under UV-B stress. The endogenous level of ascorbic acid was enhanced and alpha-tocopherol level was reduced in the tissue after exposure to UV-B radiation. The present report happens to be the first direct evidence obtained for the formation of superoxide and hydroxyl radicals in plant tissues exposed to UV-B radiation.

Enhancement of growth, photosynthetic performance and yield by exclusion of ambient UV components in C3 and C4 plants.

A field experiment was conducted under tropical climate for assessing the effect of ambient UV-B and UV-A by exclusion of UV components on the growth, photosynthetic performance and yield of C3 (cotton, wheat) and C4 (amaranthus, sorghum) plants. The plants were grown in specially designed UV exclusion chambers, wrapped with filters that excluded UV-B (<315nm), UV-A+B (<400nm), transmitted all the UV (280-400nm) or without filters. All the four plant species responded to UV exclusion by a significant increase in plant height, leaf area, leaf biomass, total biomass accumulation and yield. Measurements of the chlorophyll, chlorophyll fluorescence parameters, gas exchange parameters and the activity of Ribulose-1,5-bisphosphate carboxylase (Rubisco) by fixation of (14)CO2 indicated a direct relationship between enhanced rate of photosynthesis and yield of the plants. Quantum yield of electron transport was enhanced by the exclusion of UV indicating better utilization of PAR assimilation and enhancement in reducing power in all the four plant species. Exclusion of UV-B in particular significantly enhanced the net photosynthetic rate, stomatal conductance and activity of Rubisco. Additional fixation of carbon due to exclusion of ambient UV-B was channeled towards yield as there was a decrease in the level of UV-B absorbing substances and an increase in soluble proteins in all the four plant species. The magnitude of the promotion in all the parameters studied was higher in dicots (cotton, amaranthus) compared to monocots (wheat, sorghum) after UV exclusion. The results indicated a suppressive action of ambient UV-B on growth and photosynthesis; dicots were more sensitive than monocots in this suppression while no great difference in sensitivity was found between C3 and C4 plants. Experiments indicated the suppressive action of ambient UV on carbon fixation and yield of C3 and C4 plants. Exclusion of solar UV-B will have agricultural benefits in both C3 and C4 plants under tropical climate.

Enhancement of root growth and nitrogen fixation in Trigonella by UV-exclusion from solar radiation.

A field experiment was conducted to study the impact of solar UV on root growth and nitrogen fixation in Trigonella foenum-graecum. Plants were grown in iron mesh cages covered with polyester filters that could specifically cut off UV-B (280-315 nm) or UV-A + B (280-400 nm) part of the solar spectrum. The control plants were grown under a polythene filter transmissible to UV. Root biomass, number of nodules and nodule fresh weight were enhanced after exclusion of solar UV. Nitrogenase activity was significantly enhanced by 120% and 80% in the UV-B and UV-A + B excluded plants respectively. Along with nitrogenase there was concomitant increase in leghemoglobin and hemechrome content in the nodules after exclusion of solar UV. These components of sunlight limits nitrogen fixation and their elimination can enhance nitrogen fixation with agricultural advantages like reduction in the use of fertilizers.

Intraspecific variations in growth, yield and photosynthesis of sorghum varieties to ambient UV (280-400 nm) radiation.

A field study was conducted to investigate the impact of ambient solar UV on the various growth, physiological and yield parameters of four sorghum (Sorghum bicolor L.) varieties-Indore-12, Indore-26, CSV-23 and Indore-27 by excluding either UV-B (<315 nm) or UV-A/B (<400 nm) components of solar spectrum. Exclusion of UV significantly enhanced plant height, area and specific leaf weight of flag leaf, biomass accumulation, yield parameters and harvest index in all the sorghum varieties. Chlorophyll b was significantly enhanced and chlorophyll a increased to a lesser extent, UV-B absorbing substances and chlorophyll a/b ratio were significantly decreased by the exclusion of solar UV. The enhancement in the vegetative growth and yield by UV exclusion might be linked to the remarkable increase in rate of photosynthesis in sorghum varieties. The magnitude of the response was high in I-26 and I-27 as compared to CSV-23 and I-12 after exclusion of solar UV. All the varieties of sorghum had a negative cumulative stress response index (CSRI), the sensitivity of the sorghum varieties was in the following sequence I-12>CSV-23>I-26>I-27. Thus I-27 was the most sensitive and I-12 the least sensitive variety to present level of solar UV radiation. The differences in UV sensitivity identified among sorghum varieties might be useful in breeding programs for increased tolerance to UV-B radiation.

Oxyradicals and PSII activity in maize leaves in the absence of UV components of solar spectrum.

The regulation of oxyradicals and PSII activity by UV-B (280-315 nm) and UV-A (315-400 nm) components were investigated in the leaves of maize [Zea mays L. var: HQPM.1]. The impact of ambient UV radiation on the production of superoxide (O(2)(·-)) and hydroxyl ((·)OH) radicals were analysed in the leaves of 20-day-old plants. The amount of O(2)(·-) and (·)OH radicals and the radical scavenging activity were significantly higher in the leaves exposed to ambient UV radiation as compared to the leaves of the plants grown under UV exclusion filters. Smaller amount of oxyradicals in the leaves of UV excluded plants was accompanied by a substantial increase in quantum yield of electron transport (φ(Eo)), rate of electron transport (ψ(o)) and performance index (PI(ABS)), as indicated by chlorophyll a fluorescence transient. Although higher amounts of oxyradicals invoked higher activity of antioxidant enzymes like superoxide dismutase and peroxidase under ambient UV, they also imposed limitation on the photosynthetic efficiency of PSII. Exclusion of UV components (UV-B 280-315 nm; UV-A 315-400 nm) translated to enhanced photosynthesis, growth and biomass. Thus, solar UV components, especially in the tropical region, could be a major limiting factor in the photosynthetic efficiency of the crop plants.

Betacyanin biosynthesis in the isolated hypocotyls bei Acetabularia mediterranea.

Isolated hypocotyls synthesize betacyanin after light exposure in Amaranthus caudatus L. Pigment synthesizing capacity is reduced in the hypocotyls with increased incubation of seedlings in dark after 24h. External feeding of precursors of betacyanin L-tyrosine and DOPA enhances pigment synthesis in the isolated hypocotyls to equal that of intact hypocotyls. Cotyledons are probably the source of precursors while both cotyledons and hypocotyls are the sites of betacyanin synthesis. Betacyanin synthesizing capacity is progressively lost from the base of the hypocotyl and precursors could not induce pigment synthesis in these regions.

Recovery of gibberellic acid inhibition of betacyanin biosynthesis by pigment precursors.

Gibberellic acid inhibition of betacyanin biosynthesis has been studied in Amaranthus caudatus L. using the pigment precursors L-tyrosine and L-dihydroxyphenylalanine. Precursors fed to gibberellic acid (GA) treated seedlings completely recovered betacyanin synthesis while the GA induced growth enhancement remained unaltered. Inhibition of betacyanin biosynthesis by GA is related to depletion of metabolites/amino acids and their diversion to support changed pattern of metabolism leading to growth.