Open Access Short Research Article

Prediction of Solanum lycopersicum Target of Rapamycin (SlTOR) Protein

Ding Li-Na, Wang Rui, Zhang Jun, Wang Hao-Ran, Wang Xiao-Yan, Yu Lan, Cui Na

Biotechnology Journal International, Page 1-11
DOI: 10.9734/bji/2020/v24i230097

Aims: Tomato (Solanum lycopersicum) is an important protected vegetable in China. Its yield and quality receive much concern, however, its growth is often adversely affected by environmental stress, and so improving stress-resistance of tomato has become an urgent issue to be resolved in facility cultivation. Recent studies find that the TOR(Target of Rapamycin) complex acts as a central coordinator of energy, growth, hormones and stress signals, as well as plays a critical role in regulating transcription, protein synthesis, cell size, cell division, and basal metabolism. To study the mechanism of SlTOR in tomato growth and development as well as in stress responses, we did a series of bioinformatics analysis on SlTOR.

Study Design: In order to explore the mechanism of TOR in regulating tomato resistant to adverse conditions, we systematically analyzed the SlTOR gene with bioinformatics methods, and carried out the determination of its tissue-differential expression, aiming at laying down the basis for further experiment research.

Place and Duration of Study: College of Bioscience and Biotechnology, between March 2018 and April 2019.

Methodology: Bioinformatics analysis was conducted by online programmes. The expression of SlTOR gene in different tissues of tomato was determined by qRT-PCR.

Results: Our results showed that SlTOR was an evolutionarily conserved protein kinase, of which the molecular formula was C12366H19734N3490O3584S104, the relative molecular weight was 277978.19Da and the number of amino acid residues was 2470. Besides, it was predicted to be an acidic and unstable protein. SlTOR protein did not contain the signal peptide or transmembrane region, showing that it might be an intracellular protein. And SlTOR was speculated to be targeted to the chloroplast. Moreover, SlTOR had five domains including HEAT, FAT, PIKKc_TOR, FRB and FATC. The KEGG database displayed the only one SlTOR metabolism pathway related to autophagy. The STRING database found that SlTOR probably interacted with SlSnRK1 and SlPP2C. The experimental results of the expression of SlTOR gene in different tissues suggested that in the mature tomato plant, it was expressed the most highly in the root, followed by in the fruit let and in the mature fruit. Our experimental results were roughly consistent with the predicted results.

Open Access Original Research Article

In silico Analysis of Single Nucleotide Polymorphism in INHA Gene of Sheep and Goats

Rasheed B. Fatai, Mabel O. Akinyemi, Osamede H. Osaiyuwu

Biotechnology Journal International, Page 12-21
DOI: 10.9734/bji/2020/v24i230098

Inhibin A (INHA) a member of the transforming growth factor-β (TGF-β) family has been implicated in the negative feedback control mechanism of the pituitary follicle stimulating hormone (FSH). Inhibin has been reported to be associated with litter size, milk yield, fertility and reproductive traits in ruminants.

A total of ten amino acid sequences (four sheep and six goats) were downloaded from the National Centre for Biotechnology Information database (http://www.ncbi.nlm.nih.gov/snp). The amino acid sequence alignment was carried out using ClustalW algorithm of Molecular Evolutionary Genetic Analysis software version 6.0. The functional effects of eighteen (18) amino acid substitutions of INHA gene in each of sheep and goats were predicted computationally using Polyphen-2, PROVEAN and SIFT algorithms while INHA gene functions and interactions with associated genes were investigated using GeneMANIA. Variants that were consensually predicted to be deleterious by the three algorithms utilised were referred to as ‘Cmutant’ and ‘Dmutant’ in sheep and goats, respectively. The MutPred was further used to determine the tolerance degree for each amino acid substitution of both the ‘Cmutant’ and ‘Dmutant’.

GeneMANIA revealed 20 genes that co-localised, co-expressed with, play functionally similar role or has physical or genetic interaction with INHA gene. Out of the studied eighteen (18) amino acid substitutions; there was a consensus by SIFT, PROVEAN and PolyPhen-2 algorithms in the prediction of variants C99R, C264D, L237T and F14C as being deleterious in sheep and variants D77F, L190K, R223D, L240N, C322T and P326C in goats. In goats, MutPred revealed that variants R223D and D77F were not harmful while, L190K, R223D, L240N and P326C mutations were observed to be highly harmful. In sheep, the ‘Dmutants’ (C99R, C264D, F14C and L237T) were predicted to be highly harmful. The obtained findings would be useful in planning research aiming at exploring the association between INHA gene variants and economically important traits of small ruminants.

Open Access Original Research Article

Preparation of Synthetic Seeds of Citrus jambhiri Using in vitro Regenerated Multiple Plantlets

Priyanka Sharma, Bidhan Roy

Biotechnology Journal International, Page 22-29
DOI: 10.9734/bji/2020/v24i230099

Biotechnological tools are useful for true-to-type propagation. Shoot tips encapsulation is potential for plant development from pre-existing meristematic tissue. MS medium fortified with 1 and 2 mg/L of BAP (6-bezylaminopurine) was found to be suitable for in vitro mass-multiplication of plantlets (10.18 and 13.05 plantlets/explant, respectively) of Citrus jambhiri from nodal segments. Nodal segments were more appropriate than the shoot tips for in vitro multiplication of plantlets. Synthetic seeds were prepared using 2.5% sodium alginate dropping into 3.0% CaCl2 solution. Maximum germination was recorded when beaded shoot tips were cultured on MS medium fortified with 1 and 2 mg/L of BAP (96.67 and 100.00%, respectively). However, the germination of synthetic seeds was found to be comparatively high than the earlier findings. The results support the use of encapsulated unipolar explants for synthetic seed preparation. These type of capsules could be useful in exchange of sterile material between laboratories, germplasm conservation and direct plant propagation.

Open Access Original Research Article

Effect of Tuber Size and Intra-row Spacing on Yield and Quality of Potato (Solanum tuberosum L.)

Dawinder ., Gurbax Singh, Amanpreet Singh, Jagjot Singh

Biotechnology Journal International, Page 30-34
DOI: 10.9734/bji/2020/v24i230100

A study was conducted to determine the effect of tuber size and intra row spacing on potato (Solanum tuberosum L.) yield and subsequent growth in the 2017 production season. The experiment was laid out in a 3x4 combination arranged in a split-plot design with three replications (three levels of tuber size: 20-30 mm, 30-40 mm and 40-50 mm and four levels of intra row spacing's: 8, 12, 16 and 20 cm). For the optimum emergence and successful growth of potato tubers for processable yield, a size of 40-50 mm and spacing of 20 cm between plants, respectively were identified as the best treatments to be used in the study area.

Open Access Original Research Article

A Thermostable and Alkalitolerant Arabinofuranosidase by Streptomyces lividus

Abimbola Olajide, Felicia C. Adesina, Abiodun A. Onilude

Biotechnology Journal International, Page 35-47
DOI: 10.9734/bji/2020/v24i230101

Aim: The study aimed at producing and purifying thermostable and alkalitolerant microbial arabinofuranosidase using local Palm Kernel Cake (PKC) as substrate.

Study Design: This is an experimental design in which samples were collected thrice and  subjected to laboratory analyses from which quantitative data were obtained and analysed.

Place and Duration of Study: Ibadan, Nigeria, Five months.

Methodology: Bacterial strains were isolated from degrading PKC by serial dilution and pour plate technique on formulated Modified Basal Salt Agar Medium and incubated at 50°C for enzyme activity screening. Plates were afterwards flooded with 1% congo red solution for visualization of hydrolysis zone. Its arabinofuranosidase activity was optimized in solid state fermentation in PKC. Production temperature, pH, moisture content, inoculum size and agitation were studied for optimization test. Optimal production temperature and pH for arabinofuranosidase by isolate was 45°C and pH 9. Produced arabinofuranosidase was purified to apparent homogeneity with ammonium sulphate precipitation, dialysis and column chromatography techniques. Stability of arabinofuranofuranosidase obtained to temperature, pH, substrate concentration and some ions was determined as well as its molecular weight using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

Results: Isolate with highest arabinofuranosidase activity was selected and identified as Streptomyces lividus. Purity level attained was 16.36 fold. Enzyme had a specific activity of 25.4 U/mg, and total enzyme activity of 13.2 U.  Molecular weight of enzyme appeared as a band of 30 kDa. Purified arabinofuranosidase enzyme revealed optimum temperature and pH as 60oC and 9 respectively. Enzyme was stable over a broad pH range of 3-11, and temperature of 30-80oC. Residual activity after incubating for 1 hour at 70oC was 64%. Enzyme kinetics studies showed Km and Vmax values for P-nitrophenyl arabinofuranoside were 2.3mM and 0.7U/min respectively.

Conclusion: Apart from Solid State Fermentation (SSF) of PKC being a potential fermentation technique for production of arabinofuranosidase by Streptomyces lividus, the enzyme was highly stable.