Open Access Original Research Article

Characterization of Brassica napus (Canola) Germplasm Based on Microsatellite Markers

Muhammad Asif, Zinnia Mansoor, Syed Bilal Hussain, Muhammad Arshad, Muhammad Babar, Tahir Naqqash

Biotechnology Journal International, Page 1-7
DOI: 10.9734/bji/2021/v25i630154

Brassica napus L. is a major oilseed crop all over the world. The aim of this study was to investigate the genetic diversity of B. napus germplasm by using simple sequence repeats (SSR) markers. In the current study, ten SSR markers were used for studying genetic diversity of ten Brassica cultivars. Out of 110 total bands, 68 bands were polymorphic with 52.11% average polymorphism. Mean value of Nei’s genetic diversity and Polymorphism Information Content was 1.7, and 0.2630, respectively. These mean values show that there are moderate allelic differences between Brassica cultivars. The Nei’s genetic distance among various cultivars was 0.3281 and 0.125 which showed that germplasm of Brassica cultivars are different from each other, which is probably due to anthropogenic interventions and environmental factors. Thus, genetically different lines identified in this study could be employed in breeding programmes to develop higher-quality canola inbred varieties in future.

Open Access Original Research Article

Induction of Embryogenic Callus in Cashew (Anacardium occidentale L.) by Cotyledon, Nucellus and Testa with Antioxidants Effect

Bénédicte S. A. Akakpo, Jerome Anani Houngue, Bienvenu T. Badou, Serge S. Houédjissin, Gilles Habib Todjro Cacaï, Joseph Dossou, Corneille Ahanhanzo

Biotechnology Journal International, Page 8-21
DOI: 10.9734/bji/2021/v25i630155

Aims: This work aims to evaluate the influence of different growth regulators combined with antioxidants on the induction of callus with different explants in cashew.

Materials and Methods: Nucellar tissues, cotyledons and testa from elite tree nuts are cultured on media that differ in their concentration of 6-Benzylaminopurine (BAP: 0 mg/l; 0.25 mg/l; 0.5 mg/l) and Acid 2,4 dichlorophenoxyacetic (2,4-D: 0 mg/l; 0.8 mg/l). Therefore, for the control of the browning of the explants, the effect of activated charcoal and polyvinylpyrrolidone was tested. The amount of callus formed is evaluated after 28 days and after 90 days by simple observation according to a given scale. 

Results: Analysis of variance of callus formation 28 days after the culture of explants shows that the interaction between growth regulators and antioxidants significantly influences (p < 0.05) the induction of callus. The combination BAP 0.25mg/l and 2,4-D 0.8mg/l produces on average more callus (0.47 ± 0.00). There is a very significant difference (P < 0.05 to p<0.001) in the effect of growth regulators and antioxidants on obtaining callus after 90 days of cultivation. The 0.25 mg/l combination of BAP and 0.8 mg/l of 2,4-D still appears to be the best combination of growth regulators for callus with 58% of callus formed. Cotyledons in the presence of PVP respond better than activated charcoal. The nucellus are the explants that respond better in the presence of activated charcoal. The test for Least Significant Difference reveals that PVP significantly promotes (p <0.05) the production of a large number of calli (77%) unlike activated charcoal (9%) after 90 days of culture.

Conclusion: Summary, for obtaining viable and embryogenic callus in cashew, the combination of BAP at 0.25 mg/L and 0.8 mg/L of 2,4-D is the best and this in the presence of PVP with cotyledons.

Open Access Original Research Article

Assessment of Some Ripening Parameters (Antioxidant and Enzymes) during Storage of Banana (Musa acuminata) after Treatment with Calcium Chloride

Eugene Tafre-Phounzong, Jean Aghofack-Nguemezi

Biotechnology Journal International, Page 22-35
DOI: 10.9734/bji/2021/v25i630156

The valorisation of the large scale production of bananas in the producing countries depends on the safeguarding of the quality during exports throughout the world. The objective of the present study was to extend the shelf life of bananas by elucidating some physical and physiological phenomena that accompany the ripening process. To achieve this, bananas were treated by soaking in solutions of tween 20 for 10 minutes and calcium chloride at 2, 4, 6 and 8% for 30 minutes. The following parameters were evaluated generally in the peel and pulp as a function of time during ripening: green life, firmness of bananas, water content, Brix index, pigments, antioxidant compounds such as ascorbic acid and flavonoids, activity of two enzymes, chlorophyllase and pectin-methylesterase. The green life was 25 days for bananas treated with tween 20 at 4% calcium chloride and almost 15 days for controls. Firmness was lower in bananas treated with tween 20 at 6 and 8% calcium chloride as well as in control bananas. While the Brix index was higher in the latter, photosynthetic pigment contents such as chlorophylls a and b were lower over time. Lycopene, β-carotene and ascorbic acid contents increased significantly during ripening. Flavonoid content in the skin varied less during the sampling period. However, a regression of the latter was noted in the pulp. The enzymatic activity of both chlorophyllase and pectin-methylesterase was increasing during the whole experiment. The effect of the treatments on the majority of the evaluated parameters was elucidated. Although it was not always statistically significant. The treatments with tween 20 at 2 and 4% calcium chloride were the best, certainly because of an adequate integration of calcium chloride in the tissues. On the other hand, the 6 and 8% treatments showed an unexpected result, similar to the control. The integration of a high calcium content would lead to a tearing of the tissues and consequently to a disorganisation of the membrane, resulting in a faster ripening.

Open Access Original Research Article

Screening of Yeasts Other than Saccharomyces for Amino acid Decarboxylation

L. C. Nnodim, N. N. Odu, D. N. Ogbonna, D. B. Kiin-Kabari

Biotechnology Journal International, Page 36-47
DOI: 10.9734/bji/2021/v25i630157

This study is aimed at screening non-Saccharomyces for amino acids decarboxylation potentials. The yeasts were isolated from banana fruit and honey purchased from markets in Rivers State. The isolation and molecular identification of yeast isolates were according to standard microbiological procedures. A plate assay method for amino acid decarboxylation (biogenic amine production) screening was used. Wild Non-Saccharomyces yeast (NSY) were identified as Candida tropicalis Pe 1 (B7), Candida tropicalis WC65-1 (B10), Candida tropicalis WC57 (H4), Clavispora lusitaniae WM03 (H7), and a Commercial Wine yeast (CY) identified as Candida tropicalis zhuan4 (CY). The NSYs and CY were biogenic amine producers, from L-histidine and glutamic acid; strain variability from glycine, proline, glutamine, and asparagine decarboxylation; while L-arginine, lysine, tyrosine, cysteine, leucine, and phenylalanine were not decarboxylated at a concentration of 0.1 %. The increase in amino acid concentration influenced the number of amino acids decarboxylated - phenylalanine and leucine; L-histidine, glycine, asparagine and glutamic acid were decarboxylated by wild NSY and CY, while the strain variability of phenylalanine, proline, leucine and glutamic acid decarboxylation. The amino acids L-arginine, lysine, tyrosine, and cysteine were not decarboxylated. In terms of the concentration of amino acids, L-histidine and glutamic acid were decarboxylated and arginine, lysine, tyrosine, and cysteine were not decarboxylated by wild NSY isolates and CY. The Chi-square Test and Kendall’s Test of concordance suggest that there is no association between the amino acid concentrations (0.1 and 1 %) and biogenic amine production (P-value > 0.05). The wild NSY and CY are biogenic amine-producers, and the increase in amino acid concentration influences biogenic amine production concerning some amino acids.

Open Access Original Research Article

Molecular Characterization and Activity Analysis of Promoters from Two Cucumber Translationally Controlled Tumor Protein Genes (CsTCTPs)

Yongbo Yu, Xiangnan Meng, Shumin Jia, Mengqi Qu, Zhangtong Ma, Haiyan Fan

Biotechnology Journal International, Page 48-58
DOI: 10.9734/bji/2021/v25i630158

Aims: The aim of the paper was to isolate and characterize the promoters of two cucumber TCTP genes (CsTCTP1 and CsTCTP2) and evaluate their active domains.

Study Design: CsTCTP1 and CsTCTP2 promoter activity were analyzed under treatments with different exogenous hormones.

Place and Duration of Study: In 2017, these experiments were conducted in College of Bioscience and Biotechnology of Shenyang Agricultural University (Lab 240).

Methodology: CsTCTPpro::GUS constructs were generated by using double digests method. Transient expression was mediated by Agrobacterium tumefaciens GV3101. Histochemical and Fluorometric GUS Assays were follow by the biochemical method.

Results: Bioinformatics analysis revealed some hormone- and defense-related response elements. Histochemical and fluorometric GUS assays demonstrated that the 0.7-kb CsTCTP1 promoter (proT1-0.7kb) and 0.7-kb and 1.3-kb CsTCTP2 promoters (proT2-0.7kb and proT2-1.3kb) had strong transcriptional activity. In addition, we used exogenous hormones (abscisic acid [ABA], salicylic acid [SA], and ethylene [ETH]) for treatment. The results showed that proT1-0.7kb and proT2-1.3kb activity were upregulated in the ABA treatment group, suggesting that these promoter sequences may contain ABA-related response elements. However, in the SA and ETH treatment groups, the activity of all the promoter fragments of CsTCTP1 and CsTCTP2 declined to different degrees, suggesting that SA and ETH may have negative regulatory effects on CsTCTP1 and CsTCTP2 promoters.

Conclusion: Taken together, these results suggest that the proT1-0.7kb promoter of CsTCTP1 and proT2-1.3kb promoter of CsTCTP2 may contain ABA-related response elements, and SA and ETH may have negative regulatory effects on the CsTCTP1 and CsTCTP2 promoters. This study will help to further understand the expression patterns and the regulatory mechanism of gene transcriptional regulation.