Open Access Original Research Article

Genetic Variations in Asparagus racemosus, an Endangered Medicinal Herb Endemic to India Using RAPD Markers

Mahesh Kumar, Pradeep Kumar Naik, Sarla ., Vinod Chhokar

Biotechnology Journal International, Page 1-11
DOI: 10.9734/BBJ/2016/19173

Aim: To study the genetic diversity in Asparagus racemosus germplasm using RAPD molecular markers for its better conservation and utilization.

Study Design: RAPD markers used to check genetic diversity in Asparagus racemosus using different softwares.

Place and Duration of Study: Department of Bio & Nano Technology, Guru Jambheshwar University of Science & Technology, Hisar-125001 between May 2013 to June 2014.

Methodology: A total of 60 RAPD markers used to check polymorphism at genetic level among    60 asparagus genotypes. PCR amplified bands were scored as 0 and 1 for absence and presence. The binary data so obtained used to reveal genetic polymorphism via NTSYS, POPGENE and AMOVA analysis.

Results: A significant level of genetic diversity (81.48%) among all genotypes was assessed by using RAPD molecular markers. Out of 60, 49 RAPD primers produced 425 polymorphic loci. The value of Jaccard’s coefficient varied from 0.48 to 0.97 for RAPD. OPB-15 primer proved to be the most polymorphic marker among all used. The POPGENE analysis revealed 44.44, 79.01 and 64.20% polymorphism for RAPD analysis in groups with low, intermediate and high saponin content. The overall value of Shannon’s index and Nei’s genetic diversity was 0.3402 & 0.2169 for RAPD marker system.

Conclusion: These results showed RAPD marker system useful in detecting significant genetic polymorphism among genotypes which can be used for production and conservation of improved genotypes.

Open Access Original Research Article

Investigation the Consumption of Aromatic Hydrocarbons by Anoxybacillus rupeinsis Ir3 (JQ912241) using FTIR and HPLC Analyses

Majid H. Al-Jailawi, Mayada S. Mahdi, Ayad M. A. Fadhil

Biotechnology Journal International, Page 1-12
DOI: 10.9734/BBJ/2016/18203

Aims: To confirm the ability of Anoxybacillus rupiensis strain Ir3 (JQ912241) to utilize the aromatic compounds using FTIR and HPLC analyses. 

Study Design: Experimental study.

Place and Duration of Study: Department of Biotechnology, College of Science, Al-Nahrain University. Baghdad, Iraq, between December 2012 and April 2013.

Methodology: Anoxybacillus rupiensis strain Ir3 (JQ912241), a newly thermophilic bacterium, isolated from hydrocarbon contaminated soil in Iraq, was used. Analytical experiments include HPLC (High performance liquid chromatography) and FTIR (Fourier transform infrared) were used to determine the ability of this strain to utilize the aromatic compounds.

Results: The quantitative analysis (HPLC) indicated that this bacterium showed as much as 99.62% consumption of carbazole, 99.4% of ρ-nitrophenole, 97.73% of nitrobenzene and 98.89% of naphthalene. Qualitative analysis of FTIR spectra showed that A. rupiensis strain Ir3 (JQ912241) has the ability to convert carbazole to anthranilic acid, indicating the presence of the meta cleavage enzyme, this also confirmed by using 2, 3-dihydroxybiphenyl through converting the colony color on Luria-Bertani (LB) and minimal agar plates to brown.

Conclusion: The good ability of A. rupiensis strain Ir3 to utilize the studied aromatic hydrocarbons make it a good candidate as biocatalist. Its ability to convert carbazole to anthranilic acid and to oxidize catechol to 2-hydroxymuconic semialdehyde is through the meta cleavage enzyme

Open Access Original Research Article

Exploration of Phytochemical and Antibacterial Potentiality of Anagallis arvensis L. Extract against Methicillin-Resistant Staphylococcus aureus (MRSA)

Javad Sharifi-Rad, Seyedeh Mahsan Hoseini-Alfatemi, Abdolhossein Miri, Majid Sharifi-Rad, Marzieh Sharifi-Rad, Masoumeh Hoseini, Mehdi Sharifi-Rad

Biotechnology Journal International, Page 1-8
DOI: 10.9734/BBJ/2016/20505

Context: Anagallis arvensis L. (Scarlet pimpernel) was used to treatment of several ailments in several countries.

Objective: The aim of this study was to evaluate the in vitro antimicrobial activity of leaf methanolic extract of A. arvensis against clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA).

Methods: In this study A. arvensis leaf was shade dried, powdered and extract was made by using methanol. The antimicrobial activity of methanolic extract was investigated against clinical isolates of MRSA by both the disc diffusion method and the microbroth dilution method.

Results: The result of disc diffusion method showed that the plant extract recorded different degrees of antibacterial activity on MRSA as evidenced by the inhibited zones. The MICs of the plant extract and vancomycin were >100 and 14.5±0.1 µg/mL, respectively. This results showed that the plant extract although have slightly effect on MRAS but it was not sufficient strong.

Discussion and Conclusion: A. arvensis leaf extract has anti-MRSA properties, corroborating the traditional therapeutic uses of this plant, and can be used in the therapy of infectious diseases as well as an antimicrobial supplement in food industries.

Open Access Original Research Article

Genomic Analysis of Fungal Species No.11243 Mutant Strains Provides Insights into the Relationship between Mutations and High Productivity

Makoto Matsui, Tatsuya Yokoyama, Toshitaka Kumagai, Kaoru Nemoto, Goro Terai, Masayuki Machida, Takashi Shibata, Sachiyo Aburatani

Biotechnology Journal International, Page 1-13
DOI: 10.9734/BBJ/2016/19400

Aims: FR901469 is a novel antifungal antibiotic produced by fungal species No.11243. Although we have several FR901469 high-producing mutant strains, the mechanism of high productivity is unclear. This study aims to unravel the relationship between mutations and FR901469 productivity.

Methodology: We performed genome sequence analysis of mutant strains and detected mutated genes. Subsequently, we classified mutated genes into functional categories and searched the categories in which mutated genes were accumulated as generations progressed.

Results: We found that genome regions of two scaffolds were amplified and one of those contained putative FR901469 biosynthesis gene cluster. Moreover, we detected totally 396 mutated genes from 14 mutant strains and the genes within the “Replication, recombination and repair”, “Signal transduction mechanisms” and “Transcription” categories accumulated this mutation.

Conclusion: Our study suggests that productivity improvement occurs via the following two mechanisms: the amplification of putative FR901469 biosynthesis gene cluster and mutations of genes categorized as “Replication, recombination and repair”, “Signal transduction mechanisms” and “Transcription”.

Open Access Original Research Article

Development of an Efficient Plant Regeneration System of Field Mustard (Brassica campestris)

Kishore Kumar Sarker, Fakhrul Islam Monshi, Sayeda Sultana, Delara Akhter, Mohammed Shafi Ullah Bhuiyan

Biotechnology Journal International, Page 1-10
DOI: 10.9734/BBJ/2016/21529

Aims: The present study was conducted with a view to develop an efficient protocol for high frequency plant regeneration of Brassica campestris for further crop improvement program by biotechnological manipulation and to optimize this system for regeneration of a number of             B. campestris genotypes.

Study Design: Completely Randomized Design.

Place and Duration of Study: This experiment was carried out in the Genetic Engineering Laboratory of the Department of Genetics and Plant Breeding, Sylhet Agricultural University, Bangladesh during the period of July 2013 to June 2014.

Methodology: Cotyledon and hypocotyl explants of B. campestris cv. BARI sarisha-12 were cultured on MS medium supplemented with different concentrations of 6-Benzylaminopurine (BAP) and α-Naphthaleneacetic acid (NAA) for callus initiation and shoot regeneration. Later on subsequent subculturing is done for shoot elongation and multiplication. MS medium supplemented with various concentrations of NAA were used for root formation.

Results: From a total of 15 different combinations of BAP and NAA tested, the combination of 1.0 mg L-1 BAP and 0.5 mg L-1 NAA gave the highest frequency of callus initiation (94.44%) as well as shoot regeneration (63.89%) in case of cotyledon explants where as hypocotyl explants showed 47.62% callus initiation and 19.04% shoot regeneration frequency. Four days old cotyledon explants showed the highest shoot regeneration frequency (72.22%) and higher number of shoots per explant (3.94) than those from older seedling. The shoot regeneration frequency markedly enhanced to 83.33% by the addition of 2.0 mg L-1 AgNO3 to the MS medium supplemented with 1.0 mg L-1 BAP, 0.5 mg L-1 NAA and this combination also showed the maximum number of shoots per explant (6.86). Shoot regeneration potentiality of five B. campestris genotypes were investigated and indicated that this system would be widely applicable to all the genotypes. The regenerated shoots were easily rooted on MS medium supplemented with 0.2 mg L-1 NAA and the whole plants were transferred to pot soils and grown to maturity.

Conclusion: MS medium supplemented with 1.0 mg L-1 BAP, 0.5 mg L-1 NAA and 2.0 mg L-1 AgNO3 is more efficient for multiple shoot regeneration by using cotyledon explants and it may be utilized for In vitro improvement program of B. campestris.