Open Access Original Research Article

Refined Factors in Multi-needle-assisted Transformation of Soybean

Biao Zhang, Ren-Gao Xue

Asian Journal of Biotechnology and Genetic Engineering, Page 1-7

Previous studies have improved Agrobacterium-mediated soybean [Glycine max (L.) Merrill] cotyledonary node method by the development of a simple multi-needle-assisted wounding method using cotyledonary node cells of 1-day-old half seeds as target tissue. The goal of this study was to investigate the factors affecting the efficiency of the multi-needle-assisted transformation of soybean cotyledonary node cells (MNAT). The factors were studied by the GUS activity using a binary vector pCAMBIA1301 containing both a gus-intron gene and a hpt (hygromycin phosphotransferase) selectable marker. All of the factors affecting the transformation efficiency were determined after the 1-day-old half seeds punctured 2 times with the multi-needle. The transformation efficiency based on transient expression of the gus gene was significantly affected by the concentration of antioxidants, density of Agrobacterium suspension, infection time and the concentration of acetosyringone (AS). The frequency of the transformed cotyledonary node cells was also affected by soybean genotypes.

Open Access Original Research Article

Performance of Tomato (Solanum lycopersicum L.) Genotypes Based on Agro-morphogenic Traits under Drought Condition

Rozina Akter, Md. Ehsanul Haq, Bilkish Begum, Naheed Zeba

Asian Journal of Biotechnology and Genetic Engineering, Page 1-10

A pot experiment was conducted in the net house of the Department of Genetics and Plant Breeding, Sher-e-Bangla Agricultural University, Dhaka-1207, during November 2013 to March 2014 to observe the performances of fifteen tomato genotypes under three different drought treatments. Two factorial experiments included fifteen tomato genotypes viz. G1 (BD-7759), G2 (BD-7292), G3 (BD-7760), G4 (BD-7258), G5 (BD-7762), G6 (BD-7761), G7 (BD-7289), G8 (BD-7291), G9 (BD-7301), G10 (BARI Tomato-11), G11 (BARI Tomato-9), G12 (BARI Tomato-8), G13 (BARI Tomato-7), G14 (BARI Tomato-3) and G15 (BARI Tomato-2) and three drought treatments, T1 (Control), T2 (30 days withholding of water) and T3 (45 days withholding of water) were outlined in completely randomized design (CRD) with three replications. The results showed that both, the different tomato genotypes and drought treatments had significant influence independently and also in interaction on agro-morphogenic traits of the tomato plant. Almost all traits responded negatively as the drought level increased except days to first flowering, maturity. Considering the yield and yield contributing characters, genotype G4, G5 and G6 showed tolerance at moderate drought stress and G6, G7 and G13 showed tolerance at prolonged and severe drought stress. These genotypes could be recommended to the farmers for cultivation in the drought-prone areas of Bangladesh and also could be used in future hybridization or other gene transfer programs.

Open Access Original Research Article

Antibacterial Effect of Honey and Lime Extract on Selected Pathogens

Nnachi, Raphael Chukwuka, Oluwatosin Oluwatoyin Olaosebikan, Nwagu Kingsley Ekene

Asian Journal of Biotechnology and Genetic Engineering, Page 1-11

This study was carried out to determine the in vitro antibacterial activities of honey and lime extract on pathogens isolated from stool samples using ciprofloxacin as control. Identification of the isolates was carried out using molecular characterisation, the sequence analysis of the 16S rRNA region of the isolates using GenBank Basic Local Alignment Search Tool showed that the isolates are Bacillus cereus (ACCESSION MK011879.1), Lysinibacillus xylanilyticus (ACCESSION MK011878.1) and Bacillus anthracis (ACCESSION MK011880.1). The antibacterial susceptibility test was determined using the agar well diffusion method while the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined using the broth dilution and drop plate methods, respectively. The maximum mean zone diameter of inhibition was observed using lime extracts on B. anthracis with a mean zone diameter of inhibition (16 mm), followed by B. cereus (10 mm) and L. xylanilyticus (6 mm). Honey had antibacterial effect on B. cereus only with a mean zone diameter of inhibition of 4 mm. Lime extract had higher inhibitory and bactericidal effects than honey on the isolates with an MIC and MBC values of 20% v/v and 50% v/v, respectively which was observed on B. anthracis. Honey showed inhibitory and bactericidal effects on B. cereus only with an MIC and MBC values of 70%v/v and 99%v/v respectively. The findings showed that there was significant variation (P < 0.05) in the combination and single use of lime extract and honey. This suggests that crude extracts of lime and Honey can be used as alternative antibacterial therapy for the treatment of infections caused by the isolates.

Open Access Original Research Article

The Adsorption Performance of Urtica dioica on the Removal of Cadmium from Aqueous Solutions

Bengü Ertan, Derya Efe

Asian Journal of Biotechnology and Genetic Engineering, Page 1-7

Many contaminants (dyes, heavy metals and some inorganic substances) occur naturally through processes such as the weathering of rocks and human activities including the mining, processing and smelting of ore, and the nuclear and automotive industries. The release of contaminants, especially heavy metals to the media without any remediation process, has a very harmful effect on the environment and human health. Recently, scientists have been researching various methods to purify heavy metals from wastewater. Adsorption is an efficient method and many of materials are used as an adsorbent; activated carbons, clays, biopolymers, agricultural wastes etc. Among them, biosorbents have been preferred as they are cheap, practical, easy to find and environmentally friendly. In this study, the adsorption potential of Urtica dioica (U. dioica) on the removal of cadmium, one of the most dangerous poisons for the organisms and their environment, was investigated. The fact that the plant has low specific weight, rich functional groups and also lignocellulosic, porous structure led to the idea that it may be suitable for adsorption. U. dioica was used without any chemical modification only after being dried and passed through a 0.5 mm sieve. The effect of pH (4-10), adsorbent dose (0.2-0.8 g) and metal solution concentration (50-150 mgL-1) on adsorption was investigated. The great adsorption performance of 95% was obtained at pH 10 with 150 mgL-1 metal solution concentration and 0.8 g adsorbent dose.

Open Access Original Research Article

Screening of PRSV-P Resistance and Profiling of Defensive Secondary Metabolites in Carica papaya and Interspecific Hybrid of Vasconcellea

M. R. Razean Haireen, A. Mohd Zulkhairi, M. Razali, S. Rogayah, M. A. Mohd Shukri, H. Mohd Azhar, A. Nurul Ain

Asian Journal of Biotechnology and Genetic Engineering, Page 1-11

Aims: To Screen for PRSV-P resistance in Carica papaya and interspecific hybrid of Vasconcellea (IR) and to determine the secondary metabolites difference between PRSV-P resistant and susceptible papaya using LCMS-QTOF.

Study Design: The experiment was carried out using completely randomized design (CRD).

Place and Duration of Study: MARDI Headquarters, Persiaran MARDI-UPM, 43400 Serdang, Selangor, Malaysia between January 2017 to June 2019.

Methodology: C. papaya lines L33, L90, L13 and Eksotika were germinated from seed whilst IR was imported in tissue culture from Griffith University Australia. Screening for PRSV-P resistance was carried out using completely randomized design in the glasshouse of Agrobiodiversity and Environment Research Centre, MARDI. Percentage of disease incidence and disease severity of inoculated papaya plant were evaluated weekly for eight weeks. The generated data of disease incidence and disease score were then statistically analysed using SAS 9.4 software. Secondary metabolites analysis of Carica papaya and interspecific hybrid of Vasconcellea (IR) was carried out using LCMS-QTOF in phytochemical laboratory MARDI. The accurate mass compound in the MassHunter Qualitative analysis was confirmed by hits from existing databases [Metlin - Scripps, PCDL (MassHunter Personal Compound Database and Library - Agilent Technologies)].

Results: Screening for PRSV-P resistance amongst Malaysian local lines showed a significant difference (Alpha=0.05) between line L90 [disease incidence (55%); disease score (1.2)] and the PRSV-P susceptible (Eksotika) over eight weeks observation. IR that was also significantly difference to Eksotika categorized as PRSV-P resistance. In order to profile the defense-related secondary metabolites before and after PRSV-P entry in plant, C. papaya (Eksotika) and IR were used to represent PRSV-P susceptible and PRSV-P resistant respectively. IR was used to develop partial resistance to PRSV-P in C. papaya via introgression of resistance genes in other study. Analysis of secondary metabolites using LCMS-QTOF detected anthranilic acid (AA) and para-aminobenzoic acid (pABA) in C. papaya (Eksotika) and IR at day five after inoculation.  Interestingly, ϒ-aminobutyric acid (GABA) was only found in IR at rt, 1.881 min based on their accurate mass using LCMS-QTOF prior to PRSV-P inoculation.

Conclusion: Findings of this study recommended the usage of L90 in future conventional breeding with other local PRSV-P susceptible varieties such as Eksotika that has better taste and market. ϒ-aminobutyric acid (GABA) that was only found in IR was presumed to be involved in the plant defense response to PRSV-P. It potential to be developed as a resistance chemical marker could be explored in the near future.