Open Access Short communication

Transferability of Expressed Sequence Tag of Single Sequence Repeats Markers of Tomato Fruit in Different Vegetable Varieties

Na Cui, Yicong Shang, Jiayu Yao, Tongtong Zhang, Shuyu Pang, Yang Yu, Haiyan Fan

Biotechnology Journal International, Page 1-7
DOI: 10.9734/BJI/2017/30341

Aims: The expressed sequence tag of single sequence repeats (EST-SSR) primers designed from tomato fruit were studied for their polymorphism and transferability to other vegetable varieties.

Study Design: Primers were designed from ESTs in which the repeat bases of SSR were more than 18 bp. Twenty primer pairs chosen randomly were tested for their ability to amplify among 14 tomato varieties. The primer pairs were further used for 8 Chinese cabbage varieties, 8 muskmelon varieties and 8 eggplant varieties.

Place and Duration of Study: College of Biological Science and Technology, Shenyang Agricultural University, China, between February 2015 and March 2016.

Methodology: Twenty primer pairs selected randomly were used for the amplification of genomic DNA in 14 tomato varieties. Genomic DNA of tomato was isolated and amplified, and tested the transferability of primers.

Results: Eighteen of primer pairs showed the amplification, and 2 primer pairs showed the non-pre-amplification. Seventeen of 18 primer pairs revealed polymorphism. Eighteen primer pairs were further used for PCR to genomic DNA from 8 Chinese cabbage varieties, 8 muskmelon varieties and 8 eggplant varieties. Twelve of 18 primer pairs showed amplification in all of materials provided, and the individual ratios of amplification were 77.8%, 83.3% and 83.3%, the ratios of polymorphism of them were 65.0%, 46.7% and 80%, respectively.

Conclusion: Eighteen primer pairs showed the amplification. Seventeen of 18 primer pairs revealed polymorphism. Twelve of 18 primer pairs showed amplification in 8 Chinese cabbage varieties, 8 muskmelon varieties and 8 eggplant varieties. Tomato EST-SSR markers had highly transferability to other plant species.

Open Access Short communication

Genome-wide Characterization of MicroRNAs from Mungbean (Vigna radiata L.)

Sujay Paul, Amita Pal

Biotechnology Journal International, Page 1-9
DOI: 10.9734/BJI/2017/30984

Aims: MicroRNAs (miRNAs) are endogenous, short (~21-nucleotide), non-coding RNA molecules that play important roles in post-transcriptional gene silencing by directing target mRNA cleavage or translational inhibition. The main aim of this study is to identify and characterize miRNAs from economically important and high stress tolerant crop mungbean (Vigna radiata L.).

Study Design: Conserved miRNAs and their targets were characterized from mungbean using computational and RT-PCR approach.

Place and Duration of Study: Division of Plant Biology, Bose Institute, P 1/12 CIT Scheme VII M, Kolkata- 700054, India between January 2011- November 2015.

Methodology: Conserved miRNAs and their targets from mungbean were identified in this study using homology based strict filtering approach. Software tools such as mfold and psRNATarget were used during this study. Predicted mungbean miRNAs were validated by RT-PCR technique.

Results: In this study using recently published draft genome sequence of mungbean (Vigna radiata  L.) and applying  genome-wide computational-based approaches a total of 56 potentially conserved microRNAs belonging to 28 families were identified. 3 putative mungbean miRNAs (vra-miR160a, vra-miR162b and vra-miR398b) were successfully validated by RT-PCR. Using psRNATarget tool a total of 88 potential miRNA target transcripts were also recognized for the identified mungbean miRNAs which include a number of transcription factors.

Conclusion: For the first time 56 conserved microRNAs and 88 potential target sequences were identified in mungbean. Predicted target transcripts were found to be involved in development, metabolism and stress responses.

Open Access Original Research Article

Identification and Enzymatic Potential of Bacillus Species Isolated from Traditional Cassava Starters: Potential for Attiéké Production

Charlotte Ehon, Micael Bedikou, Souleymane Soumahoro, Sebastien Niamké

Biotechnology Journal International, Page 1-10
DOI: 10.9734/BJI/2017/29688

Aims: Microbial enzymatic activities are important in cassava dough fermentation for attiéké production. The objective of this study was to describe the molecular identification and the amylolytic, pectinolytic and cellulolytic enzymes potential of four (4) Bacillus species involved in cassava mash fermentation for the preparation of attiéké.

Place and Duration of Study: Laboratory of Biotechnology, UFR Biosciences, University Félix Houphouet-Boigny (Côte d’Ivoire), between February 2016 and April 2016.

Methodology: Four Bacillus strains (Bas 04, Bas 13, Bas 58 and Bas 66) used for this study were isolated from the traditional cassava starter ‘’mangnan’’. By using the PCR amplification of ribosomal 16S genes, Bacillus strains were identified. After culture in nutrient agar, strains were suspended in a tryptone salt solution. When we obtain Bacillus cells absorbance (turbidity) of 1.00 at 600 nm, 100 µL of this suspension were used to inoculate 5 mL of liquid medium containing peptone (1%), meat extract (1%), NaCl (0.3%) and 1% of starch or Pectin or Carboxylmethyl cellulose for enzymes production. The enzymatic activities were studied at different temperatures ranged from 25°C to 50°C and pHs (4.5, 5.0 and 5.5).  

Results: Bacillus thuringiensis was identified with 99% to 100% of similarity, referring to the NCBI database. The best amylase, cellulase and pectinase activities were obtained with Bas 66 (3.80 U/mg), Bas 13 (0.45 U/mg) and Bas 58 (1.09 U/mg) at different optimum temperatures (25, 40 and 50°C) and pHs (4.5, 5.0 and 5.5), respectively.

Conclusion: These enzyme activities are important for cassava dough fermentation, using these Bacillus strains, contributing to the softening of the mash thus improving texture and allowing the digestibility of attiéké.

Open Access Original Research Article

Mutagenicity of Oil Drilling Fluid (Potassium Chromate) on the Seedlings of Vigna unguiculata L. (Walp). in the Niger Delta, Nigeria

Ajah Obiageri Florence, Obute Gordian Chibuzo

Biotechnology Journal International, Page 1-12
DOI: 10.9734/BJI/2017/28755

The morphological, leaf epidermal and anatomical distortions caused by the exposure of Vigna unguiculata L. to potassium chromate was investigated using 1-4%w/w of an oil field chemical Potassium chromate and Control (0). Results showed that 2-4%w/w concentrations elicited noticeable effects on the plant. Variations occurred in plant height, leaf area, number of leaves between the treated and the control plants. V. unguiculata treated with 1%w/w concentration had malformed and chlorotic leaves. The treated plant had reduced stomatal indices in both epidermal surfaces: adaxial (18.3%) and abaxial (66.3%) as against (26.3%) and (75.3%) observed in the control. The mid rib of the treated plants had thicker cuticle than the control plant. There were 6 layers of collenchymatous cells in the mid rib of the treated plant as against 5 layers of collenchymatous cells in the control. The petiole of the treated plant also had thick sclerenchyma cells around the vascular bundle while the sclerenchymatous cells around the vascular bundles in the control plant were thin. Oxalate crystals were seen in the parenchyma cells of both the treated and the control plants but more of the crystals were observed in the treated plants. Obviously some mutagenic effects were elicited on the test organism indicating that the oil field chemical should be excluded where this plant is grown so as to avoid losses or other adverse consequences of potassium chromate on V. unguiculata.

Open Access Original Research Article

Computational Insight into PCR Amplified Pectate Lyase Genes from Different Species of Aspergillus

Manish Kumar, Jeya Nasim, Sangeeta Yadav, Aiman Tanveer, Dinesh Yadav

Biotechnology Journal International, Page 1-13
DOI: 10.9734/BJI/2017/29140

Pectate lyase (PL) is an important member of pectinase group of enzyme associated with cleaving α -1, 4 linkages in pectate polymers by a β-elimination mechanism, producing 4, 5–unsaturated oligogalacturonates. In the present study, PCR amplification of pectate lyase genes from different species of Aspergillus (namely A. oryzae, A. flavus, A. terreus, A. fumigatus and A. niger) was attempted. The expected size bands resulting from amplification using different sets of primers were gel eluted and sequenced using gene specific primers. The sequences of putative 23 pectate lyase (PL) genes subjected to BLAST analysis revealed its identity to pectate lyase sequences available in databases. Ten sequences of PCR amplified PL genes of A. oryzae namely PL-001 to PL-010 representing five identified PL genes from two different strains revealed maximum identity with the available sequences of A. oryzae RIB40 pectate lyases, A. flavus NRRL 3357 in the range of 98-100%. The phylogenetic tree constructed based on protein sequence of putative PL genes representing different strains of A. oryzae, A. terreus, A. flavus, A. fumigatus revealed several clusters and sub-clusters. A total of 12 PL gene sequences representing A. oryzae MTCC 3782,             A. oryzae MTCC 6993, A. fumigatus MTCC 3070, A. fumigatus MTCC 2584 and Aspergillus flavus MTCC 8835 were submitted to GenBank and were assigned accession numbers KP869835 to KP869846.