Open Access Short Research Article

Relationship between Neutral Invertase Activity and Sugar Contents in Tomato Fruit and Its Functional Prediction Analysis

Wang Hao-Ran, Yao Jia-Yu, Zhang Tong-Tong, Cui Na, He Sheng, Zhang Rui, Xu Si-Qiong

Biotechnology Journal International, Page 1-6
DOI: 10.9734/BJI/2017/37195

Aims: Neutral invertase (NI) probably plays an important role in sucrose metabolism of tomato, but main function and mechanism are unclear. In this study, contents of soluble sugar and NI activity were measured at different stages in tomatoes of Solanum chmielewskii and Solanum lycopersicumc, meanwhile analyzed the correlation between contents of soluble sugar and NI activity, also cloned NI gene and predicted by bioinformatics for further making clear the function of NI in tomato sugar metabolism.

Study Design: In order to analyze the function and action mechanism of NI, we used the tomato as materials, measured the contents of soluble sugars, activity of NI, and cloned the gene of NI. We also analyzed the NI with bioinformatics, and predicted the molecular function of NI for further making clear the function of NI in tomato sugar metabolism.

Place and Duration of Study: College of Biological Science and Technology, between February 2016 and March 2017.

Methodology: The contents of soluble sugar in tomato fruits were measured with HPLC (High performance liquid chromatography). The function structure and interacting proteins with NI family were predicted by bioinformatics.

Results: The contents of glucose and fructose were significantly higher in Solanum lycopersicum than those in Solanum chmielewskii, and greatly increased along with the process of fruit mature period. Nevertheless, the content of sucrose was raised obviously in the mature period of wild type fruit. In normal cultivated tomato, NI activity was positively correlated with fructose and glucose contents, and negatively correlated with sucrose content. In order to further study the function of neutral invertase, the NI gene was cloned and analyzed by bioinformatics. The analysis results showed that the NI was glycosyl hydrolases with Glyco_hydro_100 conserved domain.

Conclusion: The contents of glucose and fructose were significantly higher in Solanum lycopersicum than those in Solanum chmielewskii, and greatly increased along with the process of fruit mature period. The content of sucrose was raised obviously in the mature period of wild type fruit. NI activity in normal cultivated tomato was positively correlated with fructose and glucose contents, and negatively correlated with sucrose content. The NI was glycosyl hydrolases with Glyco_hydro_100 conserved domain, and might play an important role in the regulation of soluble sugar contents in tomato fruit.

Open Access Original Research Article

In vitro Pepper (Capsicum annuum L.) Anther Culture: Can be Affected Via Vitamins B?

Tugce Ozsan, Ahmet Naci Onus

Biotechnology Journal International, Page 1-13
DOI: 10.9734/BJI/2017/37102

Pepper (Capsicum annuum L.) is a vegetable in family Solanaceae and has a very important place and development potential in the field of plant breeding and biotechnology. It is at the forefront of haploid plant acquisitions in plant biotechnology, especially in the sense that it helps traditional wisdom. In vitro haploid studies have shown that the genetic structure is 'recalcitrant' and the rate of embryo formation is low during the vegetative stages of embryo formation and applications are being made to increase it. In this study, four different pepper cultivars and vitamin B compounds, which are thought to play an important role as antioxidants recently, have been used to see effect of vitamin B on pepper androgenesis. When the results of the study were evaluated, it was observed that not only differences were found in cultivars but also different vitamin B compounds added to the media were positively affecting androgenesis.

Open Access Original Research Article

Molecular Differentiation of Five Quinoa (Chenopodium quinoa Willd.) Genotypes Using Inter-simple Sequence Repeat (ISSR) Markers

A. M. M. Al-Naggar, R. M. Abd El-Salam, A. E. E. Badran, Mai M. A. El-Moghazi

Biotechnology Journal International, Page 1-12
DOI: 10.9734/BJI/2017/37053

Knowledge of genetic diversity is one of the important tools used for genetic management of quinoa accessions for plant breeding. This research aimed to molecularly characterize five quinoa genotypes using ISSR markers to reveal genetic polymorphism and identify unique markers for each genotype. Analysis of inter-simple sequence repeats (ISSR) revealed that 10 ISSR primers produced 53 amplicons, out of them 33 were polymorphic and the average percentage of polymorphism was 61.83%. The number of amplicons per primer ranged from 3 (HB-13, HB-10, HB-8 and 17898A) to 10 (HB-15) with an average of 5.3 fragments/primer across the different quinoa genotypes. Data showed a total number of unique ISSR markers of 24; eleven of them were positive and 13 were negative. Using ISSR analysis, we were able to identify some unique bands associated with quinoa genotypes. The genetic similarity ranged from 49% (between Ollague and each of QL-3 and Chipaya) to 76% (between CICA-17 and CO-407). The results indicated that all the five quinoa genotypes differ from each other at the DNA level where the average of genetic similarity (GS) between them was about 59%. The dendrogram separated the quinoa genotypes into two clusters; the first cluster included two genotypes (QL-3 and Chipaya). The second cluster was divided into two groups; the first group included two genotypes (CICA-17 and CO-407) and the second group included only one genotype (Ollague).

Open Access Original Research Article

In vitro Salinity Evaluation Studies in Golden Berry (Physalis peruviana L.)

Fatma Burcu Celikli, Pınar Akkelle, Ahmet Naci Onus

Biotechnology Journal International, Page 1-8
DOI: 10.9734/BJI/2017/37097

Aim: Present study, therefore, was conducted in in vitro conditions to study effect of NaCl at varied levels on growth and chlorophyll content of golden berry shoot apices grown in in vitro conditions.

Place and Duration: The study was carried out in the Department of Horticulture, Faculty of Agriculture, Akdeniz University, Antalya, Turkey, between March-June, 2017.

Methodology: In this study in vitro salinity  of golden berry shoot apex culture were studied within the Murashige and Skoog nutrient medium with 1 mg L-1 indole acetic acid (IAA)+ 3% sucrose and 0.7% agar supplemented with NaCl (0; 25; 50; 75  and 100 mM). The explants were incubated at 25±2°C for 4 weeks and related parameters, such as shoot, leaf and root formation, were measured.

Results: Experimental results revealed that different level of salinity treatments in in vitro culture had notable effect on above stated growth parameters. These parameters decreased significantly by increasing salinity level for excluding shoot diameters.

Open Access Original Research Article

Production and Characterization of Halo Tolerant Alkaline Protease of Enterobacter aerogenes Strain ZB and Its Application

Mohammed A. Almalki, D. S. Deepa Dhas, Rakesh Varghese, Ponnuswamy Vijayaraghavan

Biotechnology Journal International, Page 1-8
DOI: 10.9734/BJI/2017/37466

Aims: The present investigation was carried out to isolate a halo tolerant Enterobacter aerogenes strain ZB for the production of alkaline protease to degrade protein wastes from the tannery wastewater.

Study Design: Halo tolerant protease producing bacteria was isolated from the salt pan. It was used for the production of halo tolerant protease in submerged fermentation. The crude protease thus obtained was characterized and its protein removal efficacy was tested in tannery wastewater.

Place and Duration of Study: Smykon Biotech Private LtD, Nagercoil, Kanyakumari District, Tamilnadu, India between January 2017 and March 2017. 

Methodology: The halo tolerant bacterial isolates were screened for the production of alkaline protease using skimmed milk agar plates. Then a potent bacterial isolate was selected for the production of halo tolerant protease in submerged fermentation. Crude protease was used treat tannery wastewater and the protein removal efficacy of the enzyme was evaluated.     

Results: A halo tolerant strain able to grow at wide sodium chloride concentration was isolated from a salt pan and identified as E. aerogenes strain ZB. The isolate was the potential to produce protease which showed catalytic activity under high saline conditions. The E. aerogenes strain ZB protease activity was found to be maximum at pH 8.0 and enzyme activity was reduced at pH 10.0. The protease from E. aerogenes strain ZB was highly stable upto pH 8.0 and declined at higher pH. Enzyme activity was found to be maximum at 50°C and was stable upto 40°C. Enzyme was highly active in the presence of ions like Ca2+, Na+ and Mg2+ ions (0.005 M) and the enzyme activity was 101.3%, 109% and 102% respectively. This halo tolerant enzyme showed the protein degrading capacity in tannery wastewater at 1% and 5% enzyme concentrations.

Conclusion: This halo tolerant enzyme effectively removed proteinaceous waste from the tannery wastewater. A maximum protein removal was achieved at 5% enzyme concentration. The proteases produced by E. aerogenes strain ZB could have potential application in tannery wastewater treatment.