Volume 3, Issue 2

Advancements in Life Sciences, volume 3, issue 2

Published online: 25-February-2016
ISSN 2310-5380 


Full Length Research Articles:

Characterization of mercury resistant and growth promoting Enterobacter sp. from rhizosphere to use as a biofertilizer
Nageena Mobeen, Zakia Latif, pages 36-41
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 Background: Mercury occurs naturally in environment, it is heavy metal that exists in three chemical forms like elemental mercury, inorganic mercury and organic mercury. All forms of mercury are problematic for living organism. Currently, the contamination of agricultural land and water systems with mercury has become one of the major environmental issues. The cheapest mode to remove mercury metal and its other forms from the ecosystem is the use of microorganisms. Methods: In this study, initially bacterial species were isolated and purified from nodule like structures on roots and stems of plants on MacConkey agar medium. Further screening for resistance to mercury was done on N- agar medium supplemented with different concentration of HgCl2 (20, 30, 40 and 50 µg/mL). Well plate method was used for the determination of bacterial strains having maximum ability to detoxify mercury. Selected bacterial strains were subjected to different biochemical tests for characterization and other metabolic tests were also performed to characterize their capabilities. Results: All strains were highly resistant to HgCl2 at the concentration of 20 µg/mL and moderately resistant at 30 µg/mL. Bacterial strain S-2 was moderately resistant and S-3 was least resistant at 40 µg/ml whereas S-2 was least resistant at 50 µg/ml. Selected bacterial strains were positive for nitrogen fixation and protease production, negative for phosphate solubilization but only S-1 was positive for hydrogen cyanide (HCN). Bacterial species were molecular characterized by 16S rDNA sequencing as Enterobacter cloacae (KJ857483, KJ857484 and KJ857485: NCBI GeneBank). Conclusion: Selected Enterobacter sp. exhibiting multiple characteristics can be used as biofertilizer in mercury polluted land for sustainable agriculture. 

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Insight of Tp53 Mutations and their effect on Protein in Different Feline and Canine Neoplasms
Rashid Saif, Ezza Khan, Arooj Azhar, Shahnaz Choudhary, Tanveer Hussain, Masroor Ellahi Babar, Ali Raza Awan, Muhammad Tayyab, Saeeda Zia, Muhammad Wasim, pages 42-50
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 Background: Mutations in the Tp53 gene, a tumor suppressor gene, may cause dysfunction in growing cells and hinder the phenomenon of apoptosis, an alleged cause of tumorigenesis. It is involved in conservation of the genome and DNA repair, mutations of this gene may cause the damaged cells to grow continuously. Methods: The type of molecular changes in Tp53 gene and their effects on physiochemical and structural properties of this protein in various Canine and Feline cancers were observed in this study by using online bioinformatics tools. Results: Our results indicated that lymphomas and perianal adenocarcinomas (PAC) have the same mutation at c. 104, while mammary tumors and canine transmissible venereal tumor (CTVT) contain different mutations. Referring to changes in protein, synonymous mutations in granulomas were observed while certain mutations in squamous cell carcinoma (SCC) and head & neck tumors were detected in Canis familiaris. In Felis catus, the mutant protein was similar to wild type protein with exception of mutant 5 of mammary tumor, which had a deletion at the 287 amino acid position. Conclusion:  The insight gathered on the p53 mutant proteins in both species aided our understanding of the in-vivo fate of the p53 protein and its isoforms and the effects that morphological changes can have on the fate of cells. Furthermore, isolation of this protein may augment our understanding about the structural biology of these proteins. 

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Screening for drought tolerance: comparison of maize hybrids under water deficit condition
Qurban Ali, Muhammad Ahsan, Saif-ul-Malook, Naila Kanwal, Fawad Ali, Arfan Ali, Wazir Ahmed,  Muhammad Ishfaq, Muhammad Saleem, pages 51-58

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 Background: Maize is an important cereal crop, grown throughout the globe for human food and livestock feed, but biotic and abiotic factors had shown adverse effects on biomass and grain yield. Changing climatic conditions have imposed drought (water scarcity) as a major problem to combat with yield losses and biomass in maize and other cereals. Methods: The prescribed study was conducted to evaluate F1 hybrids at seedling subject to 45% moisture level. The data was recorded and subjected to multivariate analysis to find the significant variation attributed by various traits under stress conditions for efficient root/shoot development. Results: Significant differences were found in F1 hybrids for all studied traits. Higher heritability was found for root length, shoot length and fresh shoot weight, while fresh shoot weight, dry shoot weight and dry root weight showed higher genetic advance. Significant correlation was found for dry root weight to fresh shoot length and fresh root length, fresh root weight to fresh root/shoot weight ratio and fresh shoot weight. The hybrids Sh-139×B-316, Raka-poshi×B-316, B-327×B-316, Sh-139×EV-340, EV-1097Q×EV-347, EV-1097Q×EV-340, EV-1097Q×Pop/209 and B-327×EV-340 showed higher and positive heterosis and heterobeltiosis for most of the studied traits. To assess the overall variation on dependent structure, we used multivariate analysis, an important tool in breeding program, for efficient selection. Conclusions: EV-1097Q×Pop/209 and Sh-139×EV-340 showed significant results for root and shoot development under various water stress regimes at seedling stage, thus further studies should be carried out to find out the known and un-known loci regarding root and shoot development traits in high yielding maize cultivars under arid/semi-arid regions. 

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Successful DNA Profiling for Identification of burnt Families from their bones using AmpFlSTR Identifiler® Plus Kit
Muhamamd Shahzad, Muhammad Shafique, Manzoor Hussain, Muhammad Adnan Shan, Rukhsana Perveen, Ziaurehman, Muhammad Idrees, pages 59-62

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 Background: DNA profiling plays a vital role in the identification of dead bodies during mass disasters. Severe fragmentation, decomposition, burning and intermixing of the remains can occur in the mass disasters. DNA analysis faces many challenges especially when the dead bodies are completely decomposed or burnt. This report presents the identification of 32 completely burnt individuals including three families from their remains in a bus using AmpFlSTR Identifiler Plus® Kit and AmpFlSTR Y-filer® Kit. Methods: DNA was extracted from provided remains of burnt bodies and reference samples by organic extraction procedure. The extracted quantity of DNA was calculated on ABI SDS7500 real time PCR with Quantifiler® Human DNA Quantification Kit (Applied Biosystems). DNA samples of 32 completely burnt individuals including three families were amplified using AmpFlSTR Identifiler Plus® Kit and AmpFlSTR Y-filer® Kit. The genotyping of these amplified samples was performed on ABI 3130xl Genetic Analyzer. Results: The resulting data obtained from Genetic Analyzer was analyzed using GeneMapper ID software version 3.2 (Applied Biosystems). Seventeen burnt individuals including 3 burnt families were identified with the help of 16 autosomal STRs and 6 were identified through Y-STR analysis by allele sharing of their provided reference samples of parents and brothers respectively. Conclusion: For the identification of unknown individuals particularly burnt deceased victims, STR analysis has become the gold standard in forensic science. Successful DNA profiling through the amplification of STR markers of AmpFlSTR Identifiler Plus® Kit proved to be very helpful in identifying the remains of burnt individuals even in the presence of inhibition observed in the Real Time PCR. 

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Gene Profiling for Invertase Activity: Assessment of Potato Varieties for Resistance towards Cold Induced Sweetening
Arfan Ali, Mazhar Iqbal, Qurban Ali, Abdul Razzaq, Idrees Ahmad Nasir, pages 63-70

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 Background: Potato is the most important staple food in the world. Cold-induced sweetening occurs when potatoes are stored at low temperature for longer period of time. Due to non- enzymatic Millard reaction it causes unwanted changes in colour, taste and in flavor when fried and roasted at high temperature. However, long-term cold storage is mandatory to keep an adequate supply of potatoes throughout the year. The cause of cold-induced sweetening is invertase enzyme. Methods: Five potato varieties (Hermes (A) Lady Rosetta (B) Oscar (C) Kuroda (D) and Multa (E))   were investigated for invertase activity during two month cold storage at 4°C. Crude protein was extracted by PD Midi Trap G25 column technique. Quantification of mRNA expression was employed through QPCR. Determination of sucrose, reducing sugars and organic acids was simply done by 80% ethanol method and concentration were find out by using HPLC with already set standards. The correlation between invertase enzyme, sugar content and mRNA expression was calculated through Statistical methods. Results: Significant activity of invertase was observed at 4ºC with up to 6.3 nmol/min/mg of protein in the type-1 & 4 (cv. Hermes and Kuroda); 2.5 times less in type 2 (Rosetta) and 3.5 times less in type 3 (Multa) when compared with same at 4ºC. In addition, malic acid concentration was found positively correlated with invertase activity at 4ºC as compared with its concentration at harvesting time. However, citric acid and oxalic acid concentrations were independent of invertase enzyme activity. The transcript level of invertase enzyme was found significantly high in potato tubers stored at 4ºC in result 1 & 4 type, less in result type 2(C) and negligible in result 3(E) potato variety when revealed through reverse transcription PCR.Conclusions: In conclusion, Oscar (C) and Multa (E) were found more resistant to CIS at 4ºC storage and may be used for future variety improvement programs for CIS resistant through breeding and molecular approaches. 

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