Genome-Wide Assessment of Polygalacturonases-Like (PGL) Genes of Medicago truncatula, Sorghum bicolor, Vitis vinifera and Oryza sativa Using Comparative Genomics Approach.

The polygalacturonases (PG) is one of the important members of pectin-degrading glycoside hydrolases of the family GH28. In plants, PG represents multigene families associated with diverse processes. In the present study, an attempt has been made to investigate the diversity of PG genes among monocots and dicots with respect to phylogeny, gene duplication and subcellular localization to get an insight into the evolutionary and functional attributes. The genome-wide assessment of Medicago truncatula, Vitis vinifera Sorghum bicolor, and Oryza sativa L. ssp. japonica genomes revealed 53, 49, 38 and 35 PG-like (PGL) genes, respectively. The predominance of glyco_hydro_28 domain, hydrophilic nature and genes with multiple introns were uniformly observed. The subcellular localization showed the presence of signal sequences targeting the secretory pathways. The phylogenetic tree constructed marked uniformity with three distinct clusters for each plant irrespective of the variability in the genome sizes. The site-specific selection pressure analysis based on K a/K s values showed predominance of purifying selection pressures among different groups identified in these plants. The functional divergence analysis revealed significant site-specific selective constraints. Results of site-specific selective pressure analysis throw light on the functional diversity of PGs in various plant processes and hence its constitutive nature. These findings are further strengthened by functional divergence analysis which reveals functionally diverse groups in all the four species representing monocots and dicots. The outcome of the present work could be utilized for deciphering the novel functions of PGs in plants.

Single temperature liquefaction process at different operating pHs to improve ethanol production from Indian rice and corn feedstock.

Conventional grain ethanol manufacturing is a high-temperature energy-intensive process comprising of multiple-unit operations when combined with lower ethanol recovery results in higher production cost. In liquefaction, jet cooking accounts for significant energy cost, while strong acid or base used for pH adjustment presents a safety hazard. A need is felt for sustainable ethanol manufacturing process that is less hazardous, consumes lower energy, and operates in a low pH range of 4.50-5.50. A single temperature liquefaction (STL) process that could efficiently operate at lower liquefaction temperature over a pH range of 4.50-5.50 was developed using rice and corn feedstock. Ethanol recovery witnessed at pH 4.5, 5.0, and 5.5 are 481.2 ± 1.5, 492.4 ± 1.5, and 493.6 ± 1.5 L MT-1 rice, respectively. Similarly, ethanol recovery witnessed at pH 4.5, 5.0, and 5.5 are 404.6 ± 1.3, 413.9 ± 0.8, and 412.4 ± 1.8 L MT-1 corn, respectively. The improvement in ethanol recovery is attributed to higher starch conversion by alpha-amylase even at pH as low as 4.50. Thus, the STL process operated at pH lower than 5.20 is poised to enhance sustainability by offering dual advantage of energy as well as chemical saving.

In vitro assessment of allergenicity features and localization of probable IgE binding regions.

Rice is cultivated as a staple grain crop in many countries, especially in Asia. In the present study, recombinant rice chitinase was expressed, purified and characterized by in silico and immunobiochemical methods. Rice chitinase was affinity purified and it resolved at 24 kDa on SDS-PAGE. Purified protein was analyzed for pepsin resistance, heat stability, and IgE binding using atopic patients' sera. Chitinase was resistant to pepsin digestion and heat treatment at 90 °C for 1 h. It showed significant IgE binding with 7 of 110 patients' sera positive to different food allergens. Homology modeled 3D structure of rice chitinase was used for B cell epitope prediction. In silico predicted B cell peptides were assessed for IgE binding by ELISA using food allergic patients' sera, epitope RC2 showed IgE binding comparable to chitinase. In conclusion, chitinase was identified as a potential allergen and may share cross reactive epitopes with food allergens.

Laccase production optimization by response surface methodology with Aspergillus fumigatus AF1 in unique inexpensive medium and decolorization of different dyes with the crude enzyme or fungal pellets.

In the present study, alkaline pretreatment was applied for the enhanced laccase production from rice straw. Various process parameters including sodium hydroxide concentration, pH and fermentation temperature were optimized via response surface methodology (RSM) with a Box-Behnken design (BBD). Through regression analysis, it was found that laccase activity was well fitted by a quadratic polynomial equation (R(2)=0.998, Adj R(2)=0.995), and the fermentation temperature was the most significant factor influencing laccase activity. The optimized process conditions found were NaOH concentration of 0.39 mol L(-1), pH 3.12 and temperature 25.43 °C, under which laccase activity reached 142,198 ± 3586 U L(-1). Further studies were carried out to probe different dyes decolorization ability of laccase produced by Aspergillus fumigatus, A. fumigatus pellets and whole fermentation broth (WFB) using sodium hydroxide pretreated rice straw as sole carbon source. Results showed that pure laccase demonstrate limited decolorization ability to all the studied dyes, while crude laccase, A. fumigatus pellets and WFB exhibit significant decolorization ability to all the studied dyes with WFB being the most excellent one. Effectiveness of degradation was confirmed by uv-vis and phytotoxicity studies, which indicated that A. fumigatus transformed the dyes into non-toxic metabolites.

Expression of a bacterial chitosanase in rice plants improves disease resistance to the rice blast fungus Magnaporthe oryzae.

Plant fungal pathogens change their cell wall components during the infection process to avoid degradation by host lytic enzymes, and conversion of the cell wall chitin to chitosan is likely to be one infection strategy of pathogens. Thus, introduction of chitosan-degradation activity into plants is expected to improve fungal disease resistance. Chitosanase has been found in bacteria and fungi, but not in higher plants. Here, we demonstrate that chitosanase, Cho1, from Bacillus circulans MH-K1 has antifungal activity against the rice blast fungus Magnaporthe oryzae. Introduction of the cho1 gene conferred chitosanase activity to rice cells. Transgenic rice plants expressing Cho1 designed to be localized in the apoplast showed increased resistance to M. oryzae accompanied by increased generation of hydrogen peroxide in the infected epidermal cells. These results strongly suggest that chitosan exists in the enzyme-accessible surface of M. oryzae during the infection process and that the enhancement of disease resistance is attributable to the antifungal activity of the secreted Cho1 and to increased elicitation of the host defense response.

Nitrogen-efficient rice cultivars can reduce nitrate pollution.

INTRODUCTION: Environmental pollution by un-utilized nitrogenous fertilizer at the agricultural field is one of the key issues of the day. Rice-based cropping system, the mainstay of Indian agriculture, is one of the main sources of unused N-fertilizer since rice utilizes only 30-40% of total applied N, and the rest goes to waste and creates environmental as well as economic loss. METHODS: Identification of rice genotypes that can grow and yield well at low nitrogen levels is highly desirable for enhancement of nitrogen use efficiency (NUE). In the present study, we have identified large variability in the NUE of rice cultivars on the basis of plant with low, medium, and high levels of N in nutrient solution. To establish the basis of this wide variability in NUE, nitrate uptake kinetics and enzymes of nitrate assimilation were studied. RESULTS AND DISCUSSION: The data of nitrate uptake kinetics revealed that the nitrate uptake is mediated by low-affinity transporter system (LATS) in N-inefficient rice cultivars and by both LATS and high-affinity transporter systems (HATS) in N-efficient genotypes. Activities of nitrate reductase, nitrite reductase, glutamine synthetase, glutamate synthase, and the soluble protein content were found to be increased in moderately N-efficient and low N-efficient cultivars with increase in external supply of nitrogen. However, a non-significant decrease in these enzymes was recorded in high N-efficient cultivars with the increase in N supply. CONCLUSIONS: This study suggests that the HATS, high NR, and glutamine synthetase activity and the soluble protein content distribution have a key role in N efficiency of rice genotypes. These parameters may be considered in breeding and genetic engineering programs for improving the NUE of rice, which might be helpful in reducing the fertilizer loss, hence decreasing environmental degradation and improving crop productivity through improvement of nitrogen utilization efficiency in rice.

Enhanced conversion of plant biomass into glucose using transgenic rice-produced endoglucanase for cellulosic ethanol.

The catalytic domain of Acidothermus cellulolyticus thermostable endoglucanase gene (encoding for endo-1,4-beta-glucanase enzyme or E1) was constitutively expressed in rice. Molecular analyses of T1 plants confirmed presence and expression of the transgene. The amount of E1 enzyme accounted for up to 4.9% of the plant total soluble proteins, and its accumulation had no apparent deleterious effects on plant growth and development. Approximately 22 and 30% of the cellulose of the Ammonia Fiber Explosion (AFEX)-pretreated rice and maize biomass respectively was converted into glucose using rice E1 heterologous enzyme. As rice is the major food crop of the world with minimal use for its straw, our results suggest a successful strategy for producing biologically active hydrolysis enzymes in rice to help generate alcohol fuel, by substituting the wasteful and polluting practice of rice straw burning with an environmentally friendly technology.

Assessment of acetolactate synthase (ALS) tolerance to imazethapyr in red rice ecotypes (Oryza spp) and imidazolinone tolerant/ resistant rice (Oryza sativa) varieties.

Three red rice ecotypes (Oryza spp), including LA 5, MS 5 and TX 4, were evaluated for acetolactate synthase resistance/tolerance to imazethapyr. The red rice ecotypes were compared with a tolerant line (CL-121), a resistant line (CL-161) and a conventional rice variety (Cypress). Based on enzymatic activity, the mean I(50) values were 1.5, 1.1, 1.5, 1.6, 20.8 and 590.6 microM imazethapyr, respectively, for LA 5, MS 5, TX 4, Cypress, CL-121 and CL-161. CL-161 was 32 times more resistant than CL-121 and at least 420 times more resistant than the average of the red rice ecotypes and Cypress. Results from the acetolactate synthase (ALS) assay showed that red rice ecotypes and Cypress had high susceptibility to imazethapyr when compared with the tolerant CL-121 and the resistant CL-161. Measurable enzymatic tolerance to ALS-inhibiting herbicides has not yet developed in these red rice ecotypes.