Open Access Original Research Article

Establishment of Regeneration and Transformation System of Lycopersicon esculentum MicroTom

Zhengjing Wu, Shuwei Sun, Fenghua Wang, Dalong Guo

Biotechnology Journal International, Page 53-60
DOI: 10.9734/BBJ/2011/356

In this article in order to build up an efficient regeneration system for cotyledons or hypocotyls of tomato MicroTom, two kinds of seed disinfectant (3% NaClO and 0.1% HgCl2 used for 5, 10, 15, 20 min), two types of basal medium (MS: Murashige and Skoog,1964, and B5: Gamborg et al.,1968), different ratio of Indole Butyric Acid (IBA: 0.05, 0.1, 0.2 mgL-1) and 6-benzylaminopurine (6-BA: 1.0, 1.5, 2.0, 3.0 mgL-1) were tested. As for gene transform system, kanamycin (Kan: 0, 25, 50, 75, 100, 150mgL-1), carbenicillin, cephalosporin, cefoperazone sodium and sulbactam sodium for injection (Carb, CS, CSSS respectively) at the rate 100, 200, 300, or 500 mgL-1, respectively were added in medium B5 in order to find the suitable concentration for bacteriostasis and shoot regeneration. Moreover, cell concentration of Agrobacterium EHA105 (OD600: 0.3, 0.5, or 0.8)and infection time (5, 10, 15 min) were screened and optimized in this article.
The results showed that 3% NaClO for 20 min is optimal as for the disinfect efficiency on the seed surface. The most suitable medium to induce adventitious bud is basal medium B5+0.05 mgL-1 IBA+1.5 mgL-1 6-BA. The cotyledons and hypocotyls were cultivated in culture medium B5+IBA @ 0.05 mgL-1 for 2 d, then infected by Agrobacterium EHA105 (OD600=0.5) for 10 min, then plantlets were transferred to a fresh regeneration medium which contained 50 mgL-1 Kan and 300 mgL-1 CSSS, which was proved to be the most suitable transformation system for MicroTom.

Open Access Original Research Article

Studies on Cyanobacterial Biodiversity in Paper Mill and Pharmaceutical Industrial Effluents

Sanjay Kumar Dubey, J. Dubey, A. J. Viswas, P. Tiwari

Biotechnology Journal International, Page 61-67
DOI: 10.9734/BBJ/2011/395

A thorough knowledge of the physical, chemical and biological characteristics of an industrial waste is a crucial requirement for any attempt at chemical and/or biological treatment of the waste. Hence the present study was aimed to assess the physicochemical characteristics and cyanobacterial study on different industrial effluents. In the present study, effluents from two different places, paper mill and pharmaceutical industries, were selected to determine the cyanobacterial biodiversity. It was revealed that the physicochemical characteristics of both effluents studied were more or less similar. Total 25 species of cyanobacteria were found to be distributed in two different effluents in which twenty two were found in paper mill and fourteen were in pharmaceutical industries. Some of the species of cyanobacteria like Microcystis aeruginosa, Oscillatoria curviceps, O. princeps, Phormidium ambiguum, P. corium and few more were recorded in both the effluent analyzed. The dominant genus was recorded to be Oscillatoria and among themselves its six species were recorded. The abundance of cyanobacteria in these effluents was due to favorable contents of organic matter, rich calcium and nutrients such as nitrates and phosphates with less dissolved oxygen. Therefore, it can clearly stated that physicochemical characters together with biological monitoring of industrial effluents provided converging lines of evidences for evaluation of polluted habitats in this case as in some other studies reported by many researcher. This type of study would be valuable for future pollution abatement programmes.

Open Access Original Research Article

Genotype X Environment Interaction and Yield Performance of 43 Improved Cassava (Manihot esculenta Crantz) Genotypes at Three Agro-climatic Zones in Nigeria

M. G. Akinwale, B. O. Akinyele, A. C. Odiyi, A. G. O. Dixon

Biotechnology Journal International, Page 68-84
DOI: 10.9734/BBJ/2011/475

Aim: To determine the possible effects of environment and genotypic differences on root yield and other related traits.
Methodology: 43 improved cassava genotypes were evaluated for root yield, root number, root dry matter, cassava mosaic disease and Cassava bacterial disease. The experiments were conducted using a randomized complete-block design with four replications for two years in three representative agro-ecological zones (Mokwa, Ibadan and Onne) in Nigeria. The data collected were subjected to combined analyses of variance using the GLM procedure of Statistical Analysis System (SAS 9.2) to determine the magnitude of the main effects and interactions. GGEbiplot software (GGEbiplot, 2007) was applied for visual examination of the GEI pattern of the data.
Results: Genotype, Location and genotype by environment (GXE) interaction were highly significant for all the traits studied (P< 0.001), indicating genetic variability between genotypes by changing environments. The partitioning of GGE through GGE biplot analysis showed that PC1 and PC2 accounted for 61.3% and 28.8% of GGE sum of squares respectively for root yield, explaining a total of 90.1% variation.

Conclusion: Genotypes G4 and G15 were the highest yielding and stable genotypes. G2 and G7 were equally stable but with poor roots yield. G43, which had a mean yield similar to the grand mean, may be regarded as a desirable genotype. Mokwa and Ibadan were found to be the most discriminative and the least representative environments for root yields while Onne environment was found to be the most representative and the least discriminative.

Open Access Original Research Article

Assessment of Biological Activities: A Comparison of Pergularia daemia and Jatropha curcas Leaf Extracts

Omale James, Ebiloma Godwin Unekwojo, Agbaji Ann Ojochenemi

Biotechnology Journal International, Page 85-100
DOI: 10.9734/BBJ/2011/411

The present study was aimed to compare the wound healing, free radical scavenging and cytotoxicity potentials of Jatropha curcas and Pergularia daemia leaf extracts. Quantitative analysis for some phytochemicals; flavonoids, phenols, glycosides, tannins, saponins and alkaloids were carried out using standard methods. Herbal ointments containing 50% (w/w) methanol leaf extracts of Jatropha curcas and Pergularia daemia were formulated. Excision wound measuring 7x7 mm2 was created and the ointment applied topically on the wounded area which was measured at intervals of 4 days. Blank ointment (paraffin base) served as the negative control while Povidone iodine ointment served as the standard treatment. On the 16th day, rats treated with the standard drug (Povidone iodine) showed 82.1% wound closure; J. curcas -treated rats showed 91.3% wound healing while P. daemia treated rats exhibited 97.2% wound closure, indicating an efficacy of the formulations. The ointment formulated with P. daemia leaf extract had the best wound healing potential with very minimal scar formulation. The phytochemical screening revealed that the leaves of J. curcas and P. daemia contain tannins, alkaloids, phenols, flavonoids and glycosides. J. curcas had appreciable amount of saponins, however, saponins were not detected in P. daemia. The LC50 values for J. curcas and P. daemia were 586.79 µg/ml and 344.26 µg/ml, respectively while that of the standard (Potassium dichromate) was 62.52 µg/ml. The crude methanol extracts of J. curcas and P. daemia possessed free radical scavenging activities with IC50 of 90.83 and 214.16 µg/ml, respectively while that of the standard quercetin was 50.71 µg/ml. The results obtained in this study strongly support the verbal claims on the use of these plants for wound healing. It also indicates that J. curcas and P. daemia are potential sources of natural antioxidants and are relatively safe for the purposes utilized.

Open Access Original Research Article

Switching between Heat Shock Proteins and Cold Inducible Proteins under Temperature Fluctuation in Solanum tuberosum L. Cultivars in In Vivo Condition

Bengyella Louis, Pranab Roy

Biotechnology Journal International, Page 101-112
DOI: 10.9734/BBJ/2011/495

Cross-talking between heat shock proteins (HSPs) and cold inducible proteins (CIPs) subsequent to combinational mild heat (35°C) and cold (8°C) stress was investigated in vivo for four cultivars of Solanum tuberosum L. viz. Kufri Pukhraj (PO), Kufri Jyoti (GS), Kufri Ashoka (KF) and Kufri Chandramukhi (CM) in the order of their decreasing thermotolerance, to understand how this economic crop adapts to extreme temperature fluctuation. We showed a time-course differential genotypic expression pattern for HSPs at 35°C for 10h and CIPs at 8°C for 12h time-lapse. Remarkably, we noted the disappearance of a housekeeping protein (HKP) of about 19.8KD at 2h, 35°C in GS absent in CM, KF and PO; but strongly expressed as CIPs at 8°C for all the cultivars. Furthermore, heat-stress led to an outstanding transient induction of HSP95.9, HSP83.6, HSP78.7, HSP70.7, HSP66.0, HSP54.1, HSP48.6, HSP43, sHSP38.3, sHSP35.3, sHSP29, sHSP22.5, sHSP17.8 and sHSP9.5 in GS at 6h, while HKP58.7, HKP55.5 and HKP43.7 were stably overexpressed in CM, KF and PO. Temperature switching from 35°C to 8°C upregulated HKP43.4, HKP54.6, CIP14.1 and HKP19.9 for all the cultivars. The recovery process 24h subsequent to this archetype switching was governed by overexpression of small(s)HSPs of about 25.4KD-14.1KD, HKP58.7 and HKP43.5 for all cultivars. Results suggest cross-talk protection for this paradigm-shift in temperature is chiefly conferred by isoforms of constitutively expressed HKPs, CIP19.9 and CIP14.1 in S. tuberosum L. Explicitly, this differential proteome change within 22h signify HKPs may not participate in thermotolerance as HSPs, but participate in cold acclimation as CIPs, recovery as sHSPs and even switch-off during heat-stress in some cultivars as depicted in GS.

Open Access Original Research Article

Nutritional Evaluation, in vitro Free Radical Scavenging and in vivo Anti-inflammatory Effects of Gisekia pharnaceoides and Identification of Kaempferol as a Nutraceutical Agent

Chinnasammi Gandhimathi, Bernard W. C. Sathiyasekaran, Paramasivan T. Perumal, Chellan Rose

Biotechnology Journal International, Page 113-135
DOI: 10.9734/BBJ/2011/597

Aims: To analyse the nutritional profile and assess the antioxidant and antiinflammatory activities of Gisekia pharnaceoides wild plant used as traditional food source for dietary supplementation and to identify the bioactive compound as the nutraceutical agent.
Study design: In vitro and In vivo studies.
Place and Duration of Study: Department of Biotechnology, CSIR-Central Leather Research Institute, Chennai-600020, India and Sri Ramachandra College of Pharmacy, Sri Ramachandra University, Chennai-600116 between October 2005 and December 2007.
Methodology: Dry powder of the whole plant of Gisekia pharnaceoides was used for determining the nutritional parameters. Vitamins were estimated as per Indian Pharmacopoeia and United States Pharmacopoeia methods. Mineral composition was determined using Atomic absorption spectroscopy. Proximate analysis of plant power was carried out according to AOAC methods. Solvent extracts of the plant were assessed for free radical scavenging activities in vitro and antiinflammatory activity in vivo. UV, IR, NMR and LC-MS spectroscopy were used for identification of the bioactive compound.
Results: A markedly increased composition of vitamins such as thiamine, riboflavin, ascorbic acid and vitamin A and a wide range of minerals of metabolic importance have been estimated with high nutritive value. Additionally, the plant contained 9% protein and about 3% fat and carbohydrate content to an extent of 69%, of which 8% was crude fibre. All three extracts exhibited a high degree of free radical scavenging ability against DPPH radical, NO, OH, ABTS•+, O2• -. The methanol fraction showed increased levels of antioxidant and anti-inflammatory activities compared to the other two extracts because of the presence of a flavonoid which was identified as kaempferol using UV, FTIR, NMR and LC-MS spectroscopy.
Conclusion: Gisekia pharnaceoides could therefore serve as a potential nutraceutical to prevent or inhibit the harmful oxidation process in human pathophysiology, and, is a high value nutritive source as a dietary supplement to prevent malnutrition especially in rural population. Therefore, we suggest the dietary intake of the plant for nutritional supplementation.