Elaeagnus multiflora fruit extract inhibits melanin biosynthesis via regulation of tyrosinase gene on translational level

Seung Tae Kim, Yeon Jo Ha, Sam Woong Kim and Sang Wan Gal

The goal of this study obtains basal data for application as a functional cosmetic material to induce whitening by an extract of Elaeagnus multiflora fruits. Tyrosinase activity, which plays a critical role for melanin biosynthesis to regulate skin whitening, was completely inhibited by addition of 30 µg/ml the extract. Furthermore, tyrosine inhibitor(s) among the extract of E. multiflora fruits exhibited high temperature stability. The melanin biosynthesis to B16BL6 mouse melanoma cell was inhibited above 90% by addition of 3µg/ml of the extract. However, cytotoxicity was never observed to melanoma cell at high concentration of 15µg/ml of the extract. E. multiflora fruit extract was not involved in modulation of tyrosinase gene on transcriptional level but associated on translational level. All taken together, E. multiflora fruit extracts are able to modulate activity of tyrosinase via regulation on translation level. Therefore, these results suggest that E. multiflora fruit extract is a potential possibility of application as a whitening functional cosmetic material through repression of melanin biosynthesis.

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Comparative Performance between Heat-shocked Anaerobic Sludge and Clostridium butyricum TISTR 1032 Inocula in Biohydrogen Production from Food waste

Kanchanasuta Suwimon, Sirisukpoka Ubonrat and Pisutpaisal Nipon

Dark H2 fermentation of a synthetic non-sterile food waste with varying food waste content in the range of 2.5 - 10 % (w v-1) was carried out in 0.5 L batch reactors under the condition of initial pH 6 and 37oC. Comparative performance of heat-shocked anaerobic sludge and C. butyricum TISTR1032 inocula in the H2 fermentation was evaluated. No CH4 was detected in all cases. Food waste content influenced the hydrogen production potential, specific rate and yield. The H2 yield was achieved maximum at 2.5% food waste and declined at the increased food waste content. The highest H2 yields of 84.0 and 133.9 mL g-1 VS were obtained in the anaerobic sludge and C. butyricum reactors at 2.5% food waste. Better performance of H2 fermentation was observed regarding the H2 yield and specific rate by C. butyricum over the anaerobic sludge for most food waste content. Acetate and butyrate are the main end-products at 2.5-5.0% food waste while propionate and ethanol are predominant at higher food waste content for both inocula. This work demonstrated C. butyricum inoculum achieved a better performance in the H2 fermentation than the heat-shocked anaerobic sludge which presumably contained spore-forming H2 producers. Maximum energy recovery was observed in C. butyricum fermentation at 2.5% food waste that is higher than that in anaerobic sludge at 5% food waste by 9% while the energy recovery in anaerobic sludge fermentation tended to be increased at the high organic loading (5 -7.5% food waste).

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Effects of Light Quality on the Growth, Development and Metabolism of Rice Seedlings (Oryza sativa L.)

Chang-Chang Chen, Meng-Yuan Huang, Kuan-Hung Lin, Shau-Lian Wong, Wen-Dar Huang and Chi-Ming Yang

The V3 seedlings of two rice cultivars, IR1552 (purple leaf) and Taichung sen 10 (TS10, green leaf) were hydroponically cultured under 12 h photoperiod at 30/25°C (day/night), 70% relative humidity and 160 μmol m−2s−1 photon flux density under red light-emitting diodes (LEDs) (R), green LEDs (G), blue LEDs (B) and red + blue LEDs (RB) inside growth chambers for 14 days (starting 2 days after sowing). The results showed that shoot elongation was induced under the exposure of R and G. The maximum health index [(stem diameter/plant height) × biomass)] occurred under B because blue light inhibited shoot elongation. The root length under RB was the shortest. Different wavelengths mediated the chlorophyll (Chl) a/b ratio of the leaves. The content of anthocyanin (Ant) in seedling leaves was observed to be highest in RB but less in R and B, the latter pair being even lower than in G. B light LEDs enhanced effective quantum yield of PSII photochemistry (ΦPSII) and photochemical quenching (qP), but reduced non-photochemical quenching (NPQ) of seedling leaves. B LEDs also showed higher total protein content in the tested leaves compared to B plus R. In summary, precise management of irradiance and wavelength may hold promise in maximizing the economic efficiency of plant growth, development and metabolic potential of rice seedlings grown in controlled environments.

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Isolation and identification of drought stress-related proteins expressed in rye leaves using proteome analysis

Ki-Won Lee, Dong-Gi Lee, Jin-Yong Mun, Gi Jun Choi, Ki-Yong Kim, Hee Chung Ji and Sang-Hoon Lee

The present study investigated rye leaf proteomes in response to drought stress. Rye seedlings were exposed to drought conditions via water withholding and samples were collected seven days after treatment. Two-dimensional gel electrophoresis coupled with mass spectrometry (MS) was used to identify drought-responsive proteins in rye leaves. Twenty-five spots changed their expression levels under drought conditions and 16 proteins were identified by peptide mass fingerprinting using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). These proteins were classified into several categories including defense, metabolism, signal transduction, energy and protein synthesis. The investigation of rye leaf proteome expression and its identification against drought stress could be useful in better understanding molecular drought stress responses in plants.

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Purification and Characterization of Recombinant Methyl Parathion Hydrolase from Pseudomonas aeruginosa HS-D38

Yongliang Zheng, Lifang Long , Jianping Fang and Shuzhen Wang

Organophosphate hydrolase plays an important role in the bioremediation of organophosphate chemicals contaminating the environment. In this study, some characteristics of recombinant methyl parathion hydrolase (MPH) cloned from Pseudomonas aeruginosa HS-D38 were investigated. The optimum conditions of the MPH reaction were determined i.e. 30 °C, pH 10.0. Some metal ions such as Zn2+, Fe3+, Cr2+ and Co2+ activated the activity whereas others, including Cu2+, K+ and Mg2+, inhibited it. SDS-PAGE analysis indicated a molecular weight of 35 kDa for the recombinant MPH protein and the enzyme activity was two-fold higher compared to that in the wild-type strain.

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Development and Quality Evaluation of New Canned Date-Chickpea Product

Al-Jasass Fahad Mohammed, Siddiq Muhammad and Greiby Ibrahim

Our objectives were to develop a new canned date-chickpea product and evaluate its antioxidant, physical and sensory quality. Chickpeas were canned using a standard thermal process with 0 (control), 10, 20 and 30% added dates. Phenolic profile indicated that chickpea with 30% added dates had the highest total phenolics (93 mg GAE/100g). Antioxidant activity in chickpea with 0, 10, 20 and 30% of dates as measured by ATBS was 24.3, 117.8, 190.0 and 247.6 µM TE/g respectively. There were highly positive linear correlations between total phenolics and the levels of added dates and also for the antioxidant activities measured by ATBS, DPPH, FRAP and ORAC assays. Significant differences in Hunter color L, a and b values were observed between control and dates added samples. The sensory attribute scores canned date-chickpeas did not differ from control indicating that adding dates to canned chickpeas did not affect the quality negatively.

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Modified CTAB protocol for genomic DNA extraction from mature leaves of Ziziphus nummularia

Yadav R., Verma O. P. and Padaria Chatrath J.

Ziziphus nummularia, a tree species of India desert has attracted attention of molecular biologists due to its tolerance to various abiotic stresses. High quality and quantity of DNA from field grown leaves is an essential requirement to carry out any studies on genomics of Ziziphus nummularia but the thick cell wall and high content of secondary metabolite such as polysaccharide, phenolic compounds, alkaloids, flavonoids, terpenoids and saponin etc. present in mature leaves of Ziziphus hamper the isolation of high quality and quantity of DNA. In the present study a simple and efficient modified cetyl trimethylammonium bromide (CTAB) protocol for isolation of good quality DNA from mature leaves of Ziziphus nummularia has been developed. Different extraction buffers with varying concentration of CTAB or SDS, PVP and β mercaptoethanol were used and buffer with 2.5% CTAB, 3% PVP and 0.5% β mercaptoethanol was observed to give the best yield of 116.75µg of DNA per gram of tissue. The DNA thus isolated was of good quality and suitable for downstream molecular biology processes like Polymerase Chain Reaction, restriction digestion and cloning.

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Modeling of biomass ramets growth patterns using fractal topography analysis of Scirpus grossus L. on peat and paddy soils

Ali Majrashi, Baki Bin Bakar, Boyce Amru Nasrulhaq and Khandaker Mohammad Moneruzzaman

Scirpus grossus L. is a principal rhizomatous weed in the rice fields and wasteland in Malaysia. This study describes to use fractal ramets to create probability distributions fractal of ramets system and growth patterns with fertilizer under different soil types. The NPK fertilizer application at 100:30:30 ha-1 resulted in more robust aerial plant growth with ca. 253.5 ramets m-2(mean dry aerial bioamass of 23.2 gplant-1) compared with 235.6 ramets m-2 (16.3 g plant-1) in unfertilized peat soils 24 weeks after planting of the mother plant. The parallel figures for plants growing on paddy soils of the Jawa series were ca. 97.08 ramets m-1 (12.19 g plant-1)(fertilized paddy soils) and 83.67 ramets m-1 (10.89 g plant-1)(unfertilized paddy soils) 24 weeks after planting of the mother plant. Mean ramets mortality was significantly higher in unfertilized paddy soils at 121.3 ramets m-2, while in the fertilized paddy soils this was only 34.7 ramets m-2 resulting respective net populations of ca. 218.8 ramets m -2 and 114.3 ramets m-2 in fertilized and unfertilized plots. In paddy soils mean ramets mortality in unfertilized paddy soils was ca. 8.58 ramets m-2 while this was only ca. 5.67 ramets m-2, leading to the respective resultant net populations of 91.41m-2 and 75.09 ramets m-2. The time- and space-mediated clonal growth of S. grossus did not register any significant preferential directionality and dispersion of aerial plants of fertilizer application or soil types, but rather displaying opportunistic resource capture by aerial module. In this paper we used ANOVA analysis and fractal dimension gray-level.

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HMMotif: predicting sequence motifs governing constitutive exon splicing

Jing Xing, Lei Huang, Jianhua Lu, Xiaobo Zhou, Hongwei Li and Jiawen Bian

A computationally effective algorithm HMMotif is proposed to detect two kinds of splicing regulatory factor, which are ESEs and ESSs. HMMotif can make full use of the contextual information from the vicinity of 5' and 3' splice sites on the DNA to predict the hidden state of ESEs and ESSs simultaneously. A constitutive exons database is used for the extracting the EST of the concern area from UCSC database. Experiments on this data verified the effectiveness by comparing HMMotif with the traditional threshold based method.

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Phytochemical composition, antidiabetic and antioxidant activities of Acalypha indica Linn (Euphorbiaceae) stem extract

Lekhya Priya Charles and Bhaskara Rao Kokati Venkata

The present investigation was aimed to evaluate antidiabetic and antioxidant activities of methanol extract of Acalypha indica stem. Antidiabetic activity was examined by α-glucosidase enzyme inhibitor assay fallowed by enzyme kinetics. Antioxidant activity was investigated by DPPH radical scavenging, β carotene bleaching, antilipid peroxidation, reducing power and total antioxidant assays. Preliminary phytochemical screening was carried out. Quantitative analysis of phenolic compounds and flavonoids was conducted to analyze polyphenols present in the extract. Extract presented high α-glucosidase enzyme inhibitor activity (IC50= 40.76 µg/ml) in comparision with Voglibose (IC50= 60.90 µg/ml) and has shown non competitive mode of inhibition of α- glucosidase enzyme. Extract was found to be highly antioxidant nature in the tests accomplished. Phytochemical screening revealed the presence of polyphenols, tannins, oils and fats. Extract possesses high amount of phenols (278.97 mg GAE/gm of extract) and flavanoids (225.94 mg QE/gm of extract). High Performance Thin Layer Liquid Chromatography (HPTLC) profile identified five polyphenols and HPLC analysis confirmed the presence of gallic acid, gallocatechin, naringenin-7-0- rutinoside, caffeic acid and cyanidine-3-o-rutinoside which might be responsible for antidiabetic and antioxidant ability of the extract. Since A. indica stem extract has shown significant antidiabetic and antioxidant activities, this herb can be considered as a potent antioxidant and as a source of antidiabetic drug discovery.

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Analysis of simple sequence repeats dynamics in the genic regions of wheat rust fungi (Puccinia sp.)

Singh Rajender, Pandey Bharati, Chatrath Ravish and Sharma Indu

Simple sequence repeats (SSRs) are one of the valuable sources for genetic markers because of their abundance and inherent potential for determining extensive allelic variation in the genomes. In this study, we analyzed and compared the abundance and organization of SSRs in the genic region of three important fungal pathogens of wheat, leaf rust (Puccinia triticina), stem rust (Puccinia graminis f. sp. Tritici) and stripe rust (Puccinia striiformis). The total number of SSR ranged from 4026 to 5844 representing 0.3% of genic region. The relative abundance and SSR density was highest in stem rust followed by stripe rust and leaf rust. The distribution pattern of different SSR motifs provides the evidence of greater accumulation of dinucleotide followed by trinucleotide in leaf and stripe rust, but the frequency of dinucleotide and trinucleotide repeats was same in stem rust. Among trinucleotide repeats, ACC repeats are more frequent in leaf rust with a frequency of 12.51 SSR/Mb. AAC and ATG repeats are more frequent in stem rust and stripe rust respectively with a frequency of 27.12 SSR/Mb and 17.99 SSR/Mb. Frequencies of different codon repeats vary considerably depending on the type of encoded amino acid. The information about the frequency, relative abundance, relative density and variation in length of different SSR motifs in Puccinia sp. will be useful for developing markers that can be used for analysis of genetic diversity, population genetics, race identification and acquisition of new virulence.

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Computational analysis of cis-acting regulatory elements in 5’ regulatory regions of sucrose transporter gene families in wheat and Arabidopsis

Sharma Himani, Sheoran Sonia, Narwal Sneh, Malik Rekha, Sharma Indu and Chatrath Ravish

Cis-Acting regulatory elements are important molecular switches involved in the transcriptional regulation of a dynamic network of gene activities controlling various biological processes including abiotic stress responses, hormone responses and developmental processes. Bioinformatics tools were used to identify putative cis-acting regulatory elements that may be involved in the regulation of Triticum aestivum and Arabidopsis thaliana sucrose transporters. The possible cis-acting regulatory elements were predicted by scanning genomic sequences of the sucrose transporter genes translational start sites, using Plant CARE, PLACE and Genomatix Matinspector professional data bases. Several cis-acting regulatory elements that are associated with plant development, plant hormonal regulation and stress response were identified and were present in varying frequencies. Putative cis-acting regulatory elements that possibly are involved in the expression and regulation of sucrose transporter gene families in wheat and Arabidopsis thaliana during cellular development or environmental stress conditions were identified as: A-box, RY, CAT, Pyrimidine-box, Sucrose-box, ABRE, ARF, ERE, GARE, Me-JA, ARE, DRE, GA-motif, GATA, GT-1, MYC, MYB, W-box and I-box. This result explains the probable cis-acting regulatory elements that possibly are involved in the expression and regulation of sucrose transporter gene families in wheat and Arabidopsis thaliana biological processes including abiotic stress responses, hormone responses and developmental processes.

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Simultaneous production of hypocrellins and elsinochromes by Shiraia sp. SUPER-H168 with different carbon sources

Kuimin Chang, Qiansong Han, Yanrui Ding, Xiangru Liao and Yujie Cai

Shiraia bambusicolas has been used as a medicinal fungus in traditional Chinese medicine. Several secondary metabolites with biological activity have been isolated from the stroma of S. bambusicola including hypocrellin A(HA) and hypocrellin (HB). Hypocrellins, the perylenequinones from S. bambusicola, has excellent photosensitive properties and promising application in photodynamic therapy (PDT) for anticancer treatment and antivirus function. Elsinochromes (another kind of perylenequinones) were only produced by Elsinoёfawcettii according to the other works. The influence of different carbon sources on the production of perylenequinones as the second metabolite was studied. Using high-performance liquid chromatography (HPLC) and electrospray ionization mass spectrometry (ESI-MS), the ability of Shiraia sp. SUPER-H168 to produce both hypocrellins and elsinochromes was demonstrated for the first time. Eight perylenequinonoid pigments (PQPs), including hypocrellin A, hypocrellin B, elsinochrome A, elsinochrome B, elsinochrome C, were identified in the fermentation broth. The formation of HA and HB was strongly stimulated when maltose was the sole carbon source. Moreover, galactose significantly stimulated the accumulation of EC.

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Understanding new paradigm of plant metabolomes through secondary metabolite enrichment

Wany Aakanksha, Nigam V. K. and Pandey D. M.

Recent achievements in the metabolic engineering of plant secondary metabolism have helped to modulate various pathways using genes encoding biosynthetic enzymes or genes encoding regulatory proteins. To increase secondary metabolite production, different strategies such as the metabolic reactions in a pathway should overcome rate limiting steps, reduce catabolism and over-expression of regulatory genes can be pursued. New functional compounds can be discovered by using high throughput systems. The metabolic pathways and its genes involved can be determined by studying functional genomics and computational approaches. In silico analysis and docking studies of inhibitors that regulate enzymes and their role in secondary metabolite enrichment have been already elucidated. Functional genomics approaches are powerful tools to accelerate comprehensive investigations of cellular metabolism in specialized tissues or whole organisms. The technology of large-scale plant cell culture is feasible for the industrial production of plant-derived fine chemicals but economically there is low productivity of desired compounds. In recent years, metabolic engineering has opened new promising perspectives for improved production of secondary metabolites in a plant or plant cell culture and find applications in pharmaceuticals, insecticides, dyes and perfumery.

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