Isolates S4 Research Articles

ISOLASI DAN KARAKTERISASI SENYAWA FENOLIK DARI DAUN PUTAT (Planchonia valida Blume) (ISOLATION AND CHARACTERIZATION OF PHENOLIC COMPOUND FROM PUTAT LEAVES (Planchonia valida Blume))

Gallic acid have been isolated from the leaves of the Putat plant (Planchonia valida Blume). This study aimed to isolate and characterize isolates from Putat leaves by phytochemical and FTIR spectrophotometry. The research procedures included extraction by maceration, partitioning by fractionation, vacuum liquid chromatography (KCV), gravity column chromatography (KKG), thin layer chromatography (TLC), and FTIR analysis. A total of 1400 grams of putat leaf powder was macerated to produce 159.243 grams (11.38%) of methanol extract. A total of 101 grams of methanol extract was dechlorophyllated to obtain 21.430 grams (21.21%) of dechlorophyllated extract. The dechlorophyllated extract was partitioned gradually with n-hexane and ethyl acetate to produce n-hexane fractions of 0.341 grams (1.591%), and ethyl acetate of 10.143 grams (47.33%), and methanol of 9.170 grams (42.79%). A phytochemical test showed that in the dechlorophyllated extract, the methanol and ethyl acetate fractions contained alkaloids, phenolics, flavonoids, and terpenoids while the n-hexane fraction contained alkaloids and flavonoids. A total of 7 grams of ethyl acetate fraction was fractionated by vacuum liquid chromatography to produce fractions B1 to B8. The B4 fraction was continued to the separation step by gravity column chromatography so that the fractions S1 to S9 were obtained. The S4 fraction was continued to the separation stage using preparative thin layer chromatography (TLC) to produce isolates S4.1 to S4.5. Isolate S4.1 was tested for purity by two-dimensional TLC showed that the isolate was not pure. The results of the phytochemical test with 5% FeCl3 spray reagent indicated a positive phenolic compound which was strengthened by FTIR characterization of the presence of –OH, -CH aromatic, -CH aliphatic, C=C, C=O and C-O goups. Based on these results, it was concluded that the isolate from the dechlorophyllated extract of Putat leaves belonged to phenolic compounds.

Pathogenic and genomic characterisation of a rabbit sourced Pasteurella multocida serogroup F isolate s4

BackgroundPasteurella multocida is one of the most significant pathogens for a number of animals. In rabbits, the infection is generally associated with the P. multocida serogroups A and D, and the knowledge about the serogroup F is limited. In the present study, a P. multocida serogroup F isolate designated s4 was recovered from the lungs of rabbits died of respiratory disease in Fujian, in the southeast of China. The pathogenicity and genomic features of the s4 were then determined.ResultsThe serotype and sequence type of s4 were F:L3 and ST12, respectively. The s4 was pathogenic for rabbits, but it was a low virulent strain comparing to the previously reported highly pathogenic P. multocida serogroup F strains J-4103, C21724H3km7, P-4218 and HN07. The whole genome of the s4 was then sequenced to understand the genomic basis for pathogenicity. Particularly, a large-sized fragment of approximate 275 kb in length was truncated from the chromosome to form a plasmid. Moreover, the in-frame deletion of natC and N-terminal redundance of gatF would resulted in the production of a mutant L3 outer core structure that was distinct from those of the other P. multocida strains belonging to the lipopolysaccharide genotype L3. We deduced that these features detected in the genome of s4 might impair the pathogenicity of the bacterium.ConclusionsThis study evaluated the pathogenicity and determined the genomic features of the rabbit sourced P. multocida serogroup F isolate s4, the observations and findings would helpful for the understanding of the pathogenicity variability and genetic diversity of P. multocida.

Identification of Endoglucanase Gene Responsible for Cellulose Degradation Using Aspergillus flavus

Twentytwo of cellulose-degrading fungal isolates were isolated from five differentsamples; rotted sugar cane bagasse, rotted plant, termites, soil and animalmanure. Out of 22 isolates, 21 strains showed hydrolyzing zone on agar platescontaining carboxy methyl cellulose after iodine staining. The fungal isolateNo. S4, exhibited the highest endoglucanase (CMCase) activity with (0.165IU/ml) in cellulase production culture. The bestexoglucanase (FPase) and endoglucanase (CMCase) of fungal isolate S4 wasobtained after incubation at 30°C for 7 days. Sugar cane bagasse (SCB) inducedthe production of FPase and CMCase with maximum activity of 3.6 fold and 2.2fold, respectively more than that of the maximum yield in case ofcarboxy methyl cellulose (CMC). The fungal isolate S4 was identified as Aspergillusflavus on the basis of 18S rRNA and ITS region sequence analysis.

Biopolymer generation from sweet sorghum juice: screening, isolation, identification, and fermentative polyhydroxyalkanoate production by Bacillus aryabhattai

This study aims to use sweet sorghum juice (SSJ) to produce an environmentally friendly biopolymer of polyhydroxyalkanoates (PHAs) by bacteria isolated from soil. Among 18 isolates, only 3 isolates, coded as S4, S6, and S11, were chosen to produce PHA via batch fermentation compared to 3 strains of Alcaligenes eutrophus, A. latus, and Hydrogenophaga sp., respectively, under controlled conditions at pH 7, 35 °C, and 20 g/L of initial total sugar in the SSJ. The results clearly showed that the isolate S4 can produce maximum PHAs of 1.74 g/L with 57.62% dry cell weight and yield maximum hourly productivity at 0.097 g/L. S4 was further identified by full-length 16S rDNA gene sequence homology, and the findings showed that it was closely related to Bacillus aryabhattai (99.70% similarity). Interestingly, there has been as of yet no report on the use of this strain to produce an added-value biopolymer of PHA.

Molecular Cloning, Sequencing and Phylogenetic Analysis of Coat Protein Gene of a Biologically Distinct Citrus Tristeza Virus Isolate Occurring in Central India

Citrus tristeza virus (CTV) culture, collected from a fifteen-year-old wilted and declined mosambi sweet orange [Citrus sinensis (L) Osb] plant, was maintained under green house into acid lime [C. aurantifolia Swing] and mosambi seedlings. It gave positive reaction in ELISA, both with CTV specific polyclonal antibodies (G604) and monoclonal antibody MCA13, which specifically detects severe CTV strains. Coat protein gene (CPG) of the virus was amplified by RT PCR using CPG specific primers yielding an amplicon of 672 bp. Sequence analysis of the CPG amplicon showed 97% nucleotide sequence homology with Chinese isolate CTV-0002 (Acc. No. AJ518841) and isolate S4 (Acc. No. EF063109). In phylogenetic analysis, the present CTV isolate was displayed in different clade than other reported Indian CTV isolates, but it shared a separate clade with isolates from China, Israel, Jordan and New Zealand.

MOLECULAR ANALYSIS OF A CHINESE CITRUS TRISTEZA VIRUS ISOLATE SHOWING ANOMALOUS SEROLOGICAL REACTIONS

A Chinese isolate of Citrus tristeza virus (CTV), denoted HB1, was not recognized by a polyclonal antiserum (PAb-I) raised to the Italian CTV isolate 0002 and ten monoclonal antibodies to the Chinese isolate S4 (MAbs-S4). However, it reacted with a polyclonal antiserum (PAb-L5) raised to the Chinese isolate L5 in both ELISA and Western blots. To understand this unusual serological behaviour, the CP gene sequence of HB1 and other CTV isolates was determined. The results showed that HB1 was closely related to L5 at the nucleotide and amino acid levels. However, three amino acid residues in the HB1 CP differed from those of L5, which were conserved in all other isolates sequenced, including CTV-0002 and S4. This suggested that the failure of PAb-I and MAbs-S4 to recognise HB1 could depend on these molecular changes. Prediction of Antigenic Indexes and Surface Probability in four isolates (HB1, S4, L5 and 0002) with the Peptidestructure and Plotstructure softwares, showed that the HB1 tracing contained two small peaks at aa 84 and 190 that did not appear in any of the other profiles, but no apparent difference in aa 98. However, HB1 and L5 showed a similar small peak at the 5' end which did not occur in the S4 and CTV-0002.

Biodegradation of s-triazine herbicides at low concentrations in surface waters

Fourteen bacterial strains were isolated from industrial waste (SL) agricultural soil (S), surface water (B) or water treatment filter material (WT) following enrichment in a minimal salts medium (MSM) containing 5–10 mg 1 −1 of both atrazine and simazine. Biodegradation by the isolates of 1 μg 1 −1 of atrazine and 1 μg 1 −1 simazine in 100 ml MSM or reservoir surface water was assessed by comparison with non-inoculated controls. Extraction with dichloromethane recovered 100 ± 5% of both s-triazines in non-inoculated MSM and reservoir surface water. Recovery was not reduced by adsorption of the s-triazines to bacteria, and no abiotic degradation occurred over the incubation periods. Isolate S4 degraded 20 ± 7% of the atrazine and isolates S7 and S8 degraded 40 ± 12% of the simazine in MSM after 7d at 30°C. Decreasing the NH 4NO 3-N in the MSM from 35 mg 1 −1 to 1 mg 1 −1 stimulated biodegradation of atrazine by a fourth isolate (SL3). At 10°C isolates BI, SL2, S4 and S6 each biodegraded 20 ± 1% of the atrazine and isolate SL2 biodegraded 30 ± 7% of the simazine in surface water after 15 d. No biodegradation occurred in MSM at 10°C. Granular activated carbon (GAC, 1 g 1 −1) was added to surface water (10 μg 1 −1 atrazine and 10 μg 1 −1 simazine) which adsorbed 9.7 μgg −1 s-triazine and provided sites for bacterial attachment. Recovery of adsorbed s-triazines (1, 10 and 40 μg 1 −1) by Soxhlet extraction of non-inoculated GAC was 97 ± 1.5%. Inoculation of s-triazine-degraders into surface water containing GAC reduced the solution concentration (0.3 μg 1 −1 atrazine and 0.3 μg 1 −1 simazine) by up to 86 ± 18% (S8) after incubation for 15 d at 10°C. Biodegradation of 25 μg 1 −1 atrazine and 28 μg 1 −1 simazine in surface water amended with 0.5 g 1 −1 GAC and inoculated with isolate WT1 was 50 ± 4% (atrazine) and 38 ± 4% (simazine) after 21 d at 10°C. Biodegradation by WT1 attached to 0.5 g GAC in surface water (0.5 μg 1 −1 atrazine and 6 μg 1 −1 simazine) was increased from zero atrazine and 23 ± 7.6% simazine to 40 ± 5.3% atrazine and 71 ± 5.6% simazine following transfer of bioaugmented GAC into fresh surface water. Inoculation of GAC filters with selected strains has potential as a biotreatment for surface water containing s-triazine herbicides.

An eclipse of pea seed‐borne mosaic virus in vegetative tissue of pea following repeated transmission through the seed

SummaryFour isolates of pea seed‐borne mosaic virus (PSbMV) representing pathotypes P1 (isolates US and Q) and P4 (isolates S4 and S6), and groups III (US and Q) and V (S4 and S6) have been used in a study of the survival and partitioning of PSbMV under conditions of continuous seed transmission in the commercial pea cultivar Dundale.Assays suitable for detecting virus in small tissue samples were developed, and included dot‐immunobinding assay with antisera to both PSbMV and cytoplasmic inclusion body (CIB) protein, and dot hybridisation assay (DHA) with cDNA transcribed from virus RNA. Under the conditions of our experiments, seed transmission occurred at rates exceeding 90% for all virus isolates.Virus was detectable by serology and symptoms in inoculated plants, and in all vegetative tissue of second generation plants raised from seed of the inoculated plants. However, in the third, fourth and fifth sequential generations raised from seed, all plants were symptomless. Neither virus nor CIB were detectable in leaf, stem or roots by serology, but both were readily detectable in some floral parts, and in immature and mature seed. Mature seed contained virus and CIB antigen in the testa, cotyledon and embryo. Inoculum prepared from whole seeds was infectious. The testa was shown not to be involved in transmission between generations, thus implicating the embryo alone in vertical transmission. Virus antigen could not be detected in the emerging cotyledons of germinating seed and all true leaves by serology, but the leaves contained PSbMV RNA detectable by DHA.These results show that PSbMV infection can be transferred through the vegetative phase at a subliminal level, and reaches relatively high concentrations in floral parts and seeds. Thus PSbMV may be maintained at a high level of infection in seed in the absence of any apparent symptoms in the plant, and without a requirement for horizontal transmission between plants by vectors. Such a mechanism may explain the high levels of infection commonly reported in pea breeding lines.