S. B. Agrawal

Chapter 3 Ultraviolet-B Induced Changes in Gene Expression and Antioxidants in Plants

Abstract The depletion of the stratospheric ozone layer leads to an increase in the level of ultraviolet-B radiations reaching the Earths surface. UV-B radiations are known to have damaging effects on all forms of life. In plants, the UV-B exposure leads to the generation of reactive oxygen species (ROS), eventually resulting in oxidative stress. ROS induce lipid peroxidation of biological membranes, destroy the natural lipid-soluble antioxidants, and alter the expression of several genes through nonspecific signaling pathways. The integration of the thylakoid membrane appears to be much more sensitive than the activities of the photosynthetic components bound within. However, the decrease of mRNA transcripts in the photosynthetic complexes and other chloroplast proteins are among the early events of UV-B damage. Other genes, encoding defense-related proteins are rapidly upregulated under UV-B irradiation. UV-B radiation induced production of ROS, increased the antioxidant capacity and thus, minimized the magnitude of negative impact of UV-B on plants. Specific signaling pathway includes the UVR8 component that regulates the expression of a set of genes essential for the protection of plant against UV-B. This chapter comprises information regarding the UV-B perception, signal transduction, regulation of gene expression, ROS formation, and its metabolism from various studies performed under growth chamber, green house, and field conditions.

Ultraviolet-B induced changes in morphological, physiological and biochemical parameters of two cultivars of pea (Pisum sativum L.)

Increase in perception of solar ultraviolet-B (UV-B) radiation on Earths surface due to anthropogenic activities has potential in causing detrimental effects on plants. The present study was performed to evaluate the effect of elevated UV-B on Pisum sativum L., a leguminous plant with emphasis on nitrogen metabolism, flavonoids and hormonal changes. Elevated UV-B (ambient+7.2 kJ m(-2) day(-1)) negatively affected the growth, biomass, yield and its quality by generating oxidative stress directly or due to elevation of salicylic acid in two cultivars with higher magnitude being observed in HUP-2 as compared to HUDP-15. The increased accumulation of flavonoids (quercetin and kaempferol) under elevated UV-B neither provided sufficient protection to the photosynthetic machinery nor helped in elevation of biological nitrogen fixation. Nitrogen fixation and its assimilation were negatively affected under elevated UV-B as observed by the decline in nitrogenase, nitrate reductase, nitrite reductase activities and leghaemoglobin contents. Higher accumulation of salicylic acid in HUP-2 might be associated with its higher degree of sensitivity against UV-B, while higher induction of jasmonic acid and antioxidative enzymes (superoxide dismutase, catalase and ascorbate peroxidase activities) provided resistance to HUDP-15 against applied stress vis-a-vis exhibited less reduction in biomass, yield and quality of produce.

Combined effects of Psoralens and ultraviolet-B on growth, pigmentation and biochemical parameters of Abelmoschus esculentus L.

The effects of pre-treatment of Psoralens (furocoumarin compounds) and supplemental ultraviolet-B (sUV-B) were studied on plant growth, photosynthetic and non-photosynthetic pigments, protein, phenylalanine ammonia lyase (PAL) activity and antioxidative defense potential as well as their ultimate effects on biomass production in Abelmoschus esculentus L. (Okra) plants. Psoralens are capable of absorbing radiant energy and stimulating the pigmentation of human skin when photo-activated in presence of UV-A or UV-B making them beneficial in the treatment of vitilago. Pre-treatment of Psoralens against sUV-B (pUV-B), stimulates higher production of UV-B protective pigments (flavonoids and carotenoids) and helps in maintaining its biomass against UV-B stress. Antioxidative defense system in the test plant was activated by combined treatment of Psoralens and sUV-B as evidenced by the enhanced activity of enzymatic (ascorbate peroxidase-APX, superoxide dismutase-SOD, POX) and non-enzymatic (ascorbic acid and phenol) antioxidants. Individual treatments of Psoralens and sUV-B showed inhibitory effect on various morphological traits i.e. reduction in plant height, leaf area and ultimately on biomass production. Our results clearly indicated that adverse effect of sUV-B on biomass production was ameliorated by pre- treatment with Psoralens.

Evaluating the response of two high yielding Indian rice cultivars against ambient and elevated levels of ozone by using open top chambers

A continuous increase in the background level of tropospheric ozone (O(3)) has become a major challenge for present and future agricultural productivity at worldwide. Present study was designed to assess the impact of ambient (present) and elevated (future) concentrations of O(3) on two cultivars of Indian rice (Oryza sativa L. cvs Malviya dhan 36 and Shivani). Shoot and root lengths, number of leaves and total leaf area were severely affected by both ambient and elevated concentrations of O(3). Photosynthetic rate, stomatal conductance and photosynthetic efficiency (F(v)/F(m)) were also reduced by O(3) with more drastic effects under elevated levels of O(3). Leaf proteome showed reduction of some major proteins due to O(3). Pollen viability, viable florets plant(-1) and economic yield also showed significant negative impact under O(3)-exposure in both the test cultivars. The experimental findings depict that both the cultivars of rice demonstrate differential response against O(3), and it may help the plant breeders in selection of resistant cultivars for the area having higher concentrations of O(3).

Supplemental UV‐B induced changes in leaf morphology, physiology and secondary metabolites of an Indian aromatic plant Cymbopogon citratus (D.C.) Staph under natural field conditions

Cymbopogon citratus (D.C.) Staph, an aromatic perennial herb, is known to have great potential as a medicinal plant. In our study, we investigated in realistic field conditions the effect of supplemental ultraviolet‐B (sUV‐B) at two levels (+1.8 and +3.6 kJ m−2 d−1 above ambient) on various growth, physiological and biochemical characteristics of C. citratus. A higher dose of sUV‐B produced a 17.6% reduction in biomass while at a lower dose the characteristics remained unaffected. The sUV‐B imparted its negative effects to chlorophyll contents while an increment was noticed for carotenoids and phenolic compounds. No significant change in the rate of photosynthesis was noticed. The SEM observation revealed dense waxy deposition on the adaxial surface of exposed leaves at a lower dose of sUV‐B to increase epidermal transmittance. This study clearly indicated the beneficial effect of sUV‐B on volatile oils production in C. citratus plants grown under a low dose of sUV‐B.

Effects of ambient O3 on wheat during reproductive development: Gas exchange, photosynthetic pigments, chlorophyll fluorescence, and carbohydrates

The current concentrations of O3 have been shown to cause significant negative effects on crop yield. The present levels of ozone may not induce visible symptoms in most of plants, but can result in substantial losses in reproductive output. This paper considers the impact of ambient O3 on gas exchange, photosynthetic pigments, chlorophyll (Chl) fluorescence and carbohydrate levels in the flag leaf of wheat plants during various stages of reproductive development using open-top chambers. Mean O3 concentration was 45.7 ppb during wheat growth and 50.2 ppb after flag leaf development. Reproductive stage showed higher exceedence of O3 above 40 ppb compared to the vegetative stage. Diurnal variations in net photosynthetic rate (PN) and stomatal conductance (gs), intercellular CO2 concentration (Ci), Fv/Fm ratio, photosynthetic pigments, soluble sugars, and starch were measured at 10, 30, and 50 days after flag leaf expansion (DAFE). The results showed reductions in PN, gs, Fv/Fm ratio, photosynthetic pigments and starch, and increases in Ci, F0, and soluble sugars in nonfiltered chambers (NFCs) compared to filtered chambers (FCs). Maximum changes in measured parameters were observed at 50 DAFE (i.e. grain filling and setting phase). Diurnal variation in PN showed double peaked curve in both FCs and NFCs, but delayed peak and early depression in NFCs. Stomatal conductance was significantly lower in NFCs. The study suggests that higher prevalence of ambient O3 during reproductive development led to significant alteration in physiological vitality of wheat having potential negative influence on yield.

Application of ethylene diurea (EDU) in assessing the response of a tropical soybean cultivar to ambient O3: Nitrogen metabolism, antioxidants, reproductive development and yield

The present study deals with assessment of response of a tropical soybean cultivar to O3 in relation to photosynthetic pigments, chlorophyll fluorescence kinetics, antioxidative capacity, N assimilation enzymes, metabolites, growth and yield using ethylene diurea (EDU) given as a soil drench (400) ppm at an interval of 10 days after germination up to maturity. Mean O3 concentration was 42 ppb and accumulated threshold above 40 ppb (AOT 40) was 9.07 ppm h. Lipid peroxidation and total phenolics reduced, while increases in activities of antioxidative and nitrogen assimilation enzymes, ascorbic acid, protein, photosynthetic pigments, Fv/Fm ratio, number of leaves, flowers, pods, branches and yield attributes were found in EDU treated plants. EDU alleviated the negative effects of O3 by enhancing the first line of defense against ROS and protecting N assimilation enzymes at flowering and maintaining adequate supply of photosynthates to developing pods during pod filling stage. EDU provided maximum protection between flowering to pod filling stage.

A Genome‐Wide Association Study Reveals ARL15, a Novel Non‐HLA Susceptibility Gene for Rheumatoid Arthritis in North Indians

OBJECTIVE Genome-wide association studies (GWAS) and their subsequent meta-analyses have changed the landscape of genetics in rheumatoid arthritis (RA) by uncovering several novel genes. Such studies are heavily weighted by samples from Caucasian populations, but they explain only a small proportion of total heritability. Our previous studies in genetically distinct North Indian RA cohorts have demonstrated apparent allelic/genetic heterogeneity between North Indian and Western populations, warranting GWAS in non-European populations. We undertook this study to detect additional disease-associated loci that may be collectively important in the presence or absence of genes with a major effect. METHODS High-quality genotypes for >600,000 single-nucleotide polymorphisms (SNPs) in 706 RA patients and 761 controls from North India were generated in the discovery stage. Twelve SNPs showing suggestive association (P < 5 × 10(-5)) were then tested in an independent cohort of 927 RA patients and 1,148 controls. Additional disease-associated loci were determined using support vector machine (SVM) analyses. Fine-mapping of novel loci was performed by using imputation. RESULTS In addition to the expected association of the HLA locus with RA, we identified association with a novel intronic SNP of ARL15 (rs255758) on chromosome 5 (Pcombined = 6.57 × 10(-6); odds ratio 1.42). Genotype-phenotype correlation by assaying adiponectin levels demonstrated the functional significance of this novel gene in disease pathogenesis. SVM analysis confirmed this association along with that of a few more replication stage genes. CONCLUSION In this first GWAS of RA among North Indians, ARL15 emerged as a novel genetic risk factor in addition to the classic HLA locus, which suggests that population-specific genetic loci as well as those shared between Asian and European populations contribute to RA etiology. Furthermore, our study reveals the potential of machine learning methods in unraveling gene-gene interactions using GWAS data.

Effects of ambient and elevated level of ozone on Brassica campestris L. with special reference to yield and oil quality parameters.

Tropospheric ozone (O(3)) has become a serious threat to growth and yield of important agricultural crops over Asian regions including India. Effect of elevated O(3) (ambient+10ppb) was studied on Brassica campestris L. (cv. Sanjukta and Vardan) in open top chambers under natural field conditions. Eight hourly mean ambient O(3) concentration varied from 26.3ppb to 69.5ppb during the growth period. Plants under O(3) exposure showed reductions in photosynthetic rate, reproductive parameters, yield as well as seed and oil quality. Cultivar Sanjukta showed more reduction in photosynthetic characteristics, reproductive structures and seed and oil quality. However, total yield was more affected in Vardan. Exposure of O(3) increased the degree of unsaturation and level of PUFA, ω-6fatty acid, linolenic acid and erucic acid in oil indicating the deterioration of its quality. The study further confirmed that there is a correspondence between O(3) induced change in photosynthetic processes, reproductive development and yield and did not find any compensatory response in the final yield.

Use of ethylenediurea for assessing the impact of ozone on mung bean plants at a rural site in a dry tropical region of India

A field study was conducted to evaluate the usefulness of ethylenediurea (EDU) in assessing the impact of ozone (O3) on mung bean plants (Vigna radiata L Malviya Janpriya) grown in a rural area of a dry tropical region of India. Mean monthly O3 concentrations varied between 52.9-64.5 ppb. Significant reductions in growth, yield, soluble protein, pigments, and ascorbic acid in non-EDU-treated plants were observed when compared with EDU-treated plants. This clearly showed that EDU treatment provided protection against negative effect of O3 on mung bean and therefore can be effectively used as a tool to assess phytotoxic O3 concentrations in rural areas.