Research Journal of Biotechnology

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Research Journal of Biotechnology

Effects of Agave Plant Extracts on C. gloeosporioides and Characterization of Inhibitory Compounds

Chang Jin-mei, Zhang Lu-bin and Zhan Ru-lin

Anthracnose, a fungal infection of plants caused by Colletotrichum gloeosporioides, affects mango, causing low yield and poor fruit quality. Five species of Agave plants, each of which contained substances that are inhibitory to radial colony growth of C. gloeosporioides, were tested in this study; the species containing the most-inhibitory extract was Mexican maguey (Agave americana L.). Inhibitory substances in dried A. americana were readily extracted with ethanol but not with petroleum ether, ethyl acetate, or chloroform. The ethanol (75%) extract was very effective for controlling anthracnose caused by C. gloeosporioides in mango. Water soluble content of the extract did not diffuse through the dialysis membrane, had a minimum molecular weight of 100 and inhibited the growth of C. gloeosporioides up to 82.41% at 1,000-fold dilution. And the inhibitory substances with molecular weight more than 8,000, restrained the growth of C. gloeosporioides by 62.15% at 1,000-fold dilution. So the estimated molecular weight of the inhibitory substances lies between 100 and 500 and more than 8,000.

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Anti-bacterial activity of tomato glycosides

Li Jian-guang and Zhang Lu

Glycosides were extracted from green tomatoes with ethanol as the solvent. The extracted glycosides were purified by an HPD macroporous resin and a silica gel column, crystallised and then re-crystallized to yield white powder. The minimal inhibitory concentration (MIC) of the extracted tomato glycosides against Escherichia coli was determined to be 3.54 mg/mL through agar diffusion test. The effects of pH, temperature and reaction time on the anti-bacterial activity of the tomato glycosides were investigated. The bacteriostatic effect of the tomato glycosides was stronger than that of 1.5% sodium benzoate. The tomato glycosides exhibited the strongest anti-bacterial effect at pH 7.

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Genetic diversity and Genetic Relationships of Lilium revealed by Random Amplified Microsatellite Polymorphism (RAMP) in China

Lihua Zhao

In this study, random amplified microsatellite polymorphism (RAMP) markers were used to analyze the genetic diversity and genetic relationships of the 20 Lilium species in China. Six pairs of highly polymorphic primers were selected from 100 pairs of RAMP primers to amplify the 20 Lilium species. The results showed that a total of 147 DNA bands were amplified, 145 of which were polymorphic, accounting for 98.81%. The number of effective alleles (Ne) was 1.2684. The difference value between number of alleles (Na) and effective alleles (Ne) was 0.7180. The Nei’s genetic diversity index (H) was 0.2009 and the Shannon’s information index (I) was 0.3446. These figures indicated that there was high genetic diversity among Lilium species in China. Taking the average genetic similarity coefficient (Gs) 0.688 as the threshold, the 20 tested Lilium species were clustered into 5 categories which were not entirely consistent with the outcome of traditional clustering by morphological traits. The genetic diversity and genetic relationships in Lilium species revealed by this study can provide reference for effective germplasm resources conservation, germplasm innovation and selection of parents for breeding.

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Enhancement of the Antioxidant and Whitening Activities of Scutellaria baicalensis extract by Nano-encapsulation

Myong Ki Kim, Nam Young Kim and Hyeon Yong Lee

The aim of this study was to improve the whitening activity which was associated with increased antioxidant activities, of a 70% ethanol extract of Scutellaria baicalensis by encapsulateding it in lecithin. The average size of the nanoparticles was 100 nm, They were produced with approximately 65% encapsulation efficiency and they contained a maximum of 3.368 mg g-1 of baicalein. The level of tyrosinase inhibitory activity of the nanoparticles was estimated to be 46.2% at the maximum concentration of 1.0 mg mL-1 which was followed by 37.5% inhibition by the 70% ethanol extract (EE) obtained by extraction at 80 °C for 24 hours and by 29.8% inhibition by the water extract (WE) obtained by extraction at 100°C for 24 hours at the same concentration. Regarding the melanin-synthesis inhibitory activity, the nanoparticles exerted up to 43.2% inhibition whereas only 37.1% inhibition was displayed by the EE extract. A strong antioxidant activity of 59.4% DPPH radical scavenging activity was obtained using 0.5 mg mL-1 of the nanoparticles, which appeared to be closely correlated with their whitening activity. This study was the first to find that the whitening activities of the nanoparticles were closely related to the suppression of the expression of the TYRP-1, TYRP-2 and MITF genes that are involved in melanogenesis and were higher than that of the EE or the WE.

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Fermentation and evaluation of Serratia marcescens H30 and S. marcescens MG1 for 2,3-butanediol production

Liaoyuan Zhang

Two Serratia marcescens strains, S. marcescens H30 and MG1 were selected and evaluated for 2,3-butanediol production by batch fermentation. The sequential products including acetate, lactate, succinate, ethanol, acetoin and 2,3-butanediol showed that S. marcescens H30 produced higher ethanol, succinate and acetoin and lower 2,3-butanediol when compared with S. marcescens MG1. The underlying mechanism of the above observations has been investigated via the related enzymes and the internal redox state analysis. The results showed that the activity of 2,3-butanediol dehydrogenase in S. marcescens H30 was over 4-fold higher than that of S. marcescens MG1. NADH availability in S. marcescens H30 appeared to be superior to that of S. marcescens MG1. These results suggested that the amount of 2,3-butanediol dehydrogenase in S. marcescens H30 limited the conversion of acetoin into 2,3-butanediol, resulting in acetoin accumulation and NADH availability increase which activated the pyruvate-deriving pathway and promoted pyruvate into various end products such as ethanol, lactate and succinate.

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Genetic diversity of foxtail millet (Setaria italica L.) from main Asian habitats based on the rRNA 5S region

Cheng-Wu Jin, Shi-Lin Zheng, Yan-Lin Sun, Ju-Kyong Lee and Soon-Kwan Hong

Foxtail millet [Setaria italica (L.) P. Beauv.] is a crop of historical importance in some Asian and European countries. In this study, we selected the ribosomal RNA (rRNA) 5S region as the DNA marker to analyze genetic diversity and relationships of 20 foxtail millet strains collected from three representative Asian countries including China, Korea and Pakistan. According to the result of sequence alignment, strains were grouped clearly with the relevant of collected geographical region. Based on the sequence similarity and nucleotide variation, strains collected from China were grouped into two, Main China Group 1 (MCG1) and MCG2, strains collected from Korea were grouped into two, Main Korea Group 1 (MKG1) and MKG2 and strains from Pakistan were found to be close to MCG, considered to be originally transmitted from China and spread to Pakistan. Among strains from Korea, K1, K2, K5 and K10 forming MKG1 showed nearer phylogenetic relationship to MCG, considered as Chinese populations, but because of long-time environmental evolution and adaptation, MKG1 showed obvious divergence with MCG. In addition, K3 showing high similarity with Chinese strains and grouped into MCG2 was also considered as Chinese population. All strains from China showed relatively near phylogenetic relationship with each other, supporting the statement that China is one of origin areas.

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Molecular characterization and expression pattern of tobacco (Nicotiana tabacum) SD1 gene

Su Yuan, Xu Shengguang and Liu Yonggang

The complete coding sequence of one tobacco (nicotiana tabacum) gene- sterol 22-desaturase (SD1), was amplified based on some tobacco ESTs. The complete coding sequence of tobacco SD1 gene was 1,521bp which encodes a protein of 506 amino acids. Sequence analysis revealed that the SD1 of tobacco shares high homology with the SD1 of lycopersicon esculentum (86%). The prediction of transmembrane helices showed that tobacco SD1 might be a transmembrane protein. The expression profile was studied and the results indicated that tobacco SD1 gene was differentially expressed in detected tobacco tissues including leaf, stem, root and flower.

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Variable effects of Transcranial Magnetic Stimulation on protein PARP and cell death in ischemia induced rats

Kim Whi-Young

The purpose of this study was to investigate the effects of Poly (ADP-ribose) polymerase (PARP) on necrosis in cells that have undergone Transcranial Magnetic Stimulation (TMS) prior to induction of ischemia. Transcranial magnetic stimulation devices have been used mainly for diagnostic purposes by measuring the functions of the nervous system rather than for treatment purposes and has a problem of considerable energy fluctuations per repeated pulse. Control is difficult and the size is large due to the difficulty of digitalizing the energy stored in a capacitor and there are many heavy devices. In addition, there are many constraints when it is used for a range of purpose such as head and neck diagnosis, treatment and rehabilitation of nerve palsy, muscle strengthening, treatment of urinary incontinence etc. Output stabilization and minimization of the energy variation rate are required as the level of the transcranial magnetic stimulation device is dramatically improved and the demand for therapeutic purposes increases. This study developed a compact, low cost transcranial magnetic stimulation device with minimal energy variation of a high repeated pulse and output stabilization using a real time capacitor charge discharge voltage. After induction of brain ischemia, TMS was applied to acupuncture at ST 36, at 12, 24 and 48 hours. PARP expression after 24 hours significantly decreased (p<0.05) in the TMS group compared to the GI group. Based on the results of this study, TMS can be an effective method of treating dysfunction and improving function of brain cells in brain damage caused by ischemia.

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Inheritance of Yellow Mosaic Virus (YMV) in Mungbean [Vigna radiata (L.) Wilczek]

Anusha N., Anuradha Ch., Srinivas A. M. N. and Reddy S. Sokka

An experiment was conducted to study the inheritance of yellow mosaic virus in the cross LGG 460, PM 115 in munngbean. An infector row technique was used for evaluating F2 plant for MYMV resistance. No insecticide was sprayed in order to maintain the natural whitefly population in experimental field. In the field condition, Whitefly started landing on the plants soon after germination and the disease made its first appearance 3rd week after planting. Infection and disease severity of MYMV progressed in the next 6 weeks. According to the mean disease score, the mungbean genotypes were categorized into six groups highly resistant (HR), resistant (R), moderately resistant (MR), moderately susceptible (MS), susceptible (S) and highly susceptible (HS). 3 (Susceptible): 1 (Resistance) was observed in all F2 population and it showed the dominance of susceptibility over the resistance.

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Study on the fermentation of fumaric acid by Rhizopus oryzae

Haitao Wang, Lijun Pan, Ya Liu and Xingjiang Li

In order to increase the yield of fumaric acid and the utilization rate of glucose, the Rhizopus oryzae was used as the strain to produce fumaric acid with glucose and the optimal medium compositions and fermentation conditions in Erlenmeyer flask were investigated in this paper by using single factor experiment and response surface methodology (Box-Behnken). The results showed that the optimal medium compositions were as follows: glucose, (NH4)2SO4, KH2PO4, MgSO4, FeSO4.7H2O, ZnSO4.7H2O and CaCO3 were 100 g/L, 4.5 g/L, 0.2 g/L, 0.3 g/L, 0.02 g/L, 0.2 g/L and 60 g/L respectively. The reasonable fermentation conditions were spore incubation time of 24 hours, inoculums ratio of 10%(v/v), liquid volume of 50mL/250mL, neutralizing agent concentration of 60 g/L, fermentation temperature of 320C, rotation speed of 200rpm and fermentation period lasted for 120 hours in the Erlenmeyer flask, leading to the fumaric acid yield of 37.43 g/L which was 31.06% higher than that of original conditions. Thus, the yield of fumaric acid and utilization rate of glucose were improved effectively by this experiment.

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Cross Compatibility, Heterosis and Genetic Study of Salt Tolerant Indigenous and Modern Rice Cultivars

Md. Zakiul Hasan, Naher U.A., Panhwar Q.A. and Mohd RaziI

Rice production in saline-affected areas is a great challenge for the breeders and salt tolerant rice variety is an urgent need for these areas. There are salt tolerant lines but yield is low whereas high yielding rice varieties cannot withstand salinity stress. Therefore the objectives of this study were to evaluate, cross compatibility, heterosis and interrelationship of grain yield and associated characters by using four modern high yielding (BRRIdhan28, BRRIdhan29, BRRIdhan45 and BRRIdhan47) and three indigenous salt tolerant (Pokkali, Nonabakra and Binnatoa) rice genotypes. The experimental design was RCBD where the treatments were 14 genotypes (Four modern; three salt tolerant and seven F1s genotypes), with three replications per treatment. Successful seed set in progenies of 7 rice genotypes ranged from 8% to 48.61%. The highest cross success, 48.61%, was achieved with BRRIdhan47 × Binnatoa and the lowest (8%) with BRRI dhan28 × Nonabakra. Only one cross (BRRIdhan47×Binnatoa) exhibited positive relative heterosis (17.26%) and heterobeltiosis (1.05%) for grain yield. The crosses of BRRIdhan28×Binnatoa, BRRI dhan29×Binnatoa and BRRIdhan45× Nonabakra showed negative heterosis in plant height, percent seed set, 1000-grain weight and harvest index. A significant relative heterosis was observed for BRRIdhan28 × Pokkali. The correlation and path coefficient studies showed that filled grain panicle-1, effective tiller plant-1 and harvest index were the most important yield contributing components and that these three parameters might be important traits in selecting the lines for a higher grain yield of rice. The crosses between BRRI dhan47 × Binnatoa can produce a salt tolerant new rice variety.

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Application of a perfusion bioreactor to relieve the browning of Glycyrrhiza inflate in-vitro cells

Wang G. R. and Tang Ning

Our studies were carried out in a perfusion stir-tank bioreactor with Glycyrrhiza inflata suspension cells. It indicated that the upsurge of in-vitro plan cells browning occurred mainly in the initial stage of culture and it was characterized by the increased activities of polyphenol oxidase (PPO) and peroxidase (POD). When cultivated in batch mode, the Glycyrrhiza inflata cells died of lethal browning and the activities of POD and PPO were up to 210.8 and 22 respectively. The activities of POD and PPO were positively correlated with shear rate and showed that cell browning was aggravated with mechanical injury. Perfusion culture can relieve in-vitro plant cell browning effectively by enhancing perfusion medium feeding at the initial culture stage. The experimental results also revealed that the browning of Glycyrrhiza inflata suspension cells of was prone to occur under the condition of high dissolved oxygen and low pH value.

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Isolation and Partial Characterization of Phosphate Solubilizing Bacteria from the Rhizosphere of Tomato (Lycopersicon esculentum) Plant

Dhiman Aarti and Putatunda Chayanika

Phosphorus is one of the essential macronutrients required by the plants for their growth and development. However, the phosphate anions are extremely reactive and may be immobilized through precipitation with cations such as Ca2+, Mg2+, Fe3+ and Al3+and tend to become unavailable to plants. Many soil microorganisms are able to solubilize this unavailable P through various mechanisms. Phosphate solubilizing bacteria used as inoculants increase the P uptake by the plant and crop yield. In present study, soil samples were collected from the rhizosphere of tomato plant (Lycopersicon esculentum) from various places viz. Bilaspur (Himachal Pradesh), Chandigarh, Bernala and Jalandhar (Punjab), India. A total of 33 phosphate solubilizing bacterial isolates were obtained by plating the soil samples on Pikovskaya’s medium plates. The isolates were subjected to primary and secondary screening and two isolates (PSB-TAHP5 and PSB-TAHP7) with highest phosphate solubilizing activity were selected for optimization of conditions. The incubation period of 72hrs and pH 7 were observed to be best suited for phosphate solubilization by both the bacterial isolates. Among the various nutrients, dextrose was found to be the best carbon source and ammonium sulphate was the best nitrogen source. The isolates were then tested morphologically and biochemically and both of them showed similarity with the genus Bacillus.

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Isolation, characterization, functional potential and molecular diversity of Pseudomonas fluorescens isolated from the soils of Maharashtra

Sonawane R. B., Deokar C. D. and Chimote V. P.

In the present investigation 65 fluorescent pseudomonads were isolated from soil samples of different regions of Maharashtra, India. Based on these morphological, biochemical and physiological characteristics, 34 isolates were identified as Pseudomonas fluorescens, 31 as Pseudomonas aeruginosa from soils of Maharashtra. Seven P. fluorescence isolates namely P-31 (Sawantwadi), P-51 (Malegaon), P-56 (Ahmedpur), P-121 (Sevagram), P-134 (Ramtek), P-136 (Kuhi) and P-145 (Kuhi) showed antagonistic activity superior than commercial Pseudomonas against soil borne plant pathogens namely Macrophomina phaseolina and Fusarium oxysporum. Seven Pseudomonas fluorescens and Commercial Pseudomonas were high HCN producers which turned the orange colour of the filter paper. All isolates showed the siderophore ability, however those isolates which were biocontrol effective showed larger zone than the other isolates. In RAPD analysis derived divergence studies, P. fluorescens isolates were grouped on the basis of their colony morphology i.e. with irregular, spreading and round, non-spreading types in both dendrogram and 2D scatter plot analysis. Further five of those closely grouped round, non-spreading isolates also showing HCN production with high biocontrol activity against fungi. No such grouping was noticed for any other character studied. The seven highly bio-control efficient P. fluorescens isolates identified need to be further explored under field conditions against soil borne pathogens.

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