Open Access Original Research Article

Effects of Exogenous Salicylic Acid on the Antioxidative System in Bean Seedling Treated with Manganese

Issam Saidi, Nasreddine Yousfi, Wahbi Djebali, Yacine Chtourou

Biotechnology Journal International, Page 93-100
DOI: 10.9734/BBJ/2015/15830

In the present study we investigated the role of salicylic acid (SA) in regulating Mn-induced oxidative stress in bean (Phaseolus vulgaris) leaves. Exposure of plants to 100 µM Mn inhibited biomass production and intensively increased Mn accumulation in leaves. Concomitantly, Mn significantly enhanced protein carbonyl, H2O2 content and lipid peroxidation as indicated by malondialdehyde (MDA) accumulation. SA (10, 50 and 100 µM) pretreatment alleviated the negative effect of Mn on plan growth and led to decrease in oxidative stress induced by Mn stress. Furthermore, SA enhanced the activities of catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APX, EC 1.11.1.11), but lowered that of superoxide dismutase (SOD, EC 1.15.1.1) and guaiacol peroxidase (POD, EC 1.11.1.7). The data suggest that the beneficial effect of SA could be related to avoidance of oxidative damage upon exposure to Mn thus reducing the negative consequences of oxidative stress caused by Mn toxicity.

Open Access Original Research Article

Characterization of New Quality Protein Maize (QPM) Varieties from a Breeding Program: Analysis of Amino Acid Profiles and Development of a Variety-Diagnostic Molecular Marker

K. K. Nkongolo, G. Daniel, K. Mbuya, P. Michael, G. Theriault

Biotechnology Journal International, Page 101-112
DOI: 10.9734/BBJ/2015/15729

Aims of the Study: This study was to determine the amino acid profile of two newly developed quality protein maize varieties and to develop variety-diagnostic molecular markers.

Methodology: Two new maize varieties, named MUDISHI 1 and MUDISHI 3 have been developed by breeders and farmers using the participatory breeding approach. Total protein and amino acid profiles of the two new lines were compared to their respective parental population and a locally released genetically improved normal maize variety. Maize accessions from the DR-Congo breeding program were analyzed using ISSR primers. Variety-diagnostic markers were identified and characterized.

Results: Protein analysis data revealed that MUDISHI 1 and MUDISHI 3 are QPM varieties that are distinct from their original population, Longe 5 QPM from NARI- Unganda and DMR-ESR-W-QPM from the International Institute for Tropical Agriculture (ITTA, Ibadan), respectively. Lysine content in MUDISHI 1, and MUDISHI 3 were 3.5 g and 3.6 g of lysine / 100 g, respectively, which represents a significant increase of 20% and 23% over the genetically improved normal maize variety (Salongo 2) that is currently released. There was a significant increase of 25% of tryptophan and 33% of methionine in MUDISHI 3 compared to its parental variety while the amount of lysine was similar for the two varieties. There were 10% and 15% decrease of lysine and tryptophan, respectively in MUDISHI 1 compared to its original parent Longe 5 QPM. Genomic DNA was extracted from different maize varieties. One ISSR diagnostic-marker of 480 bp that was identified was unique to the QPM variety MUDISHI 3. This sequence was converted to a sequence characterized amplified region (SCAR) marker using a pair of designed primers. This SCAR sequence was not specific to MUDISHI 3 as it was present in all the varieties tested.

Conclusion: The newly developed varieties are typical QPM lines. The development of an ISSR diagnostic marker indicates that it is possible to develop a molecular breeding program involving QPM and normal varieties.

Open Access Original Research Article

Establishment of Callus Derived from Jatropha curcas L. Petiole Explants and Phytochemical Screening

K. Nyembo, N. Mbaya, N. Muambi, C. Ilunga

Biotechnology Journal International, Page 113-118
DOI: 10.9734/BBJ/2015/15124

Aims: The focus of this study was to assess the production of secondary metabolites of callus derived from Jatropha curcas L. petiole explants.

Study Design: Laboratory experimental tests; Tissue culture, Lyophilisation, Phytochemical Analysis, Determination of tannins by Folin-Denis method.

Place and Duration of the Study: Department of Phytobiology, Department of Biotechnology. General Atomic Energy Commission. Regional Center of Nuclear Studies of Kinshasa P.O BOX 868 Kin.XI DR Congo during March and June 2013.

Methodology: In vitro callus cultures were initiated from petiole explants of Jatropha curcas L. on Murashige & Skoog (1962) basal medium supplemented with growth regulator formulations 4.44 μM of 6-benzylaminopurine (BAP), 4.92 μM of Indole-3-butyric acid (IBA), and 100 ml L-1 coconut milk. Callus was lyophilized. The extracts were subjected to phytochemical tests for the presence of plant secondary metabolites. The amount of tannins was estimated by Folin-Denis method.

Results: Excellent growth of callus was obtained. Callus was soft, friable, lush green in color and grew fast from 8 to 30 days of culture then stabilized at low growth rate. The results obtained by phyto-chemical tests revealed the presence of saponins, tannins, alkaloids, steroids and flavonoids. The tannin contents in vitro proliferated callus, leaves, stem barks, root barks, roots and latex of J. curcas were found to be 161, 173, 220, 214, 43 and 245 μg tannic acid equivalent/g of dry weight respectively.

Conclusion: The results of this study have revealed that in vitro induced callus from J. curcas petiole explants was able to produce secondary metabolites. The pharmacological activity of J. curcas reported by various researchers can be attributed to the presence of these phytochemicals.

Open Access Original Research Article

Histopathological Alterations Induced by Naja naja Crude Venom on Renal, Pulmonary and Intestinal Tissues of Mice Model

M. A. Al-Mamun, M. A. Hakim, M. K. Zaman, K. M. F. Hoque, Z. Ferdousi, M. Abu Reza

Biotechnology Journal International, Page 119-125
DOI: 10.9734/BBJ/2015/16188

Aim: Snake bite causes a significant number of mortality and morbidity throughout the world. So, the current study was carried out to estimate the extinct of damage caused by intraperitoneal introduction of cobra venom on kidney, lung and intestinal tissues of mice model using histological technique.

Place and Duration of Study: The entire study including the treatment along with preparing histological slide was conducted in protein science laboratory, Department of Genetic Engineering and Biotechnology, University of Rajshahi, Bangladesh between December 2013 to July 2014.

Methods: Twenty five mature female albino mice were divided mainly into two groups as control and envenomated group. Lyophilized Naja naja venom was dissolved in 0.9% NaCl solution and injected intraperitoneally into the mice of the envenomated group at dosages equivalent to LD50 (0.25 mg/kg). Whereas the animals from control group were not received any venomous component. Both groups of animal were sacrificed for histological study and visualized under light microscope.

Results: Injection of cobra venom induced a range of histological changes in all envenomated mice comparing with their control. Results from the histopathological examination showed mainly inflammatory cellular infiltration, vacuolation in renal tubules, shrinking of glomeruli, raising space between the walls of Bowman’s capsule in renal tissue and alveolar haemorrhage, inflammatory cellular infiltration and edema in pulmonary tissue. No significant histopathological alterations in intestinal tissue were observed without infiltration and mild hemorrhage.

Conclusion: The findings from the current study revealed that, cobra venom at lethal dose causes multiple organ failure in experimental animal which could be considered among the factors that lead to death. By observing the site and the mode of action on tissue level, these findings may help to allay the severity of damage by discovering novel anti venom drug.

Open Access Original Research Article

Production of New Almond-Peach Hybrid Rootstocks Resistance to Root-Knot Nematode

A. S. Shaltout, H. El-Wakeel, A. A. Nahla, S. Ghada

Biotechnology Journal International, Page 126-135
DOI: 10.9734/BBJ/2015/14614

Aims: This research was conducted to hybridize almond (Om El-fahm and M. Dalet) female parent with Okinawa peach rootstock as a male parent to introduce new almond rootstock resistance to root-knot nematode.

Methodology: Okinawa peach rootstock used in crosses with both of Om El-fahm and M. Dalet almond cvs. and the resulting hybrids identified by SSR and STS markers for the presence and/or absence of the expected resistance marker. Inoculation by M. incognita with suspension of 2500 eggs per plants through holes in the soil to evaluate hybrids resistance.

Results: Fruit set percentage was higher with Om El-fahm than M. Dalet while, the opposite was true for fruit drop. No significant differences were observed for seed germination with the used females. Hybrid no.3 resulting from M. Dalet x Okinawa crosses showed the highest significant value of vegetative growth parameters. The STS marker; the OPA11 primer pair with Okinawa x Om El-fahm showed the resistant marker at the expected size (166 bp) for all of the tested hybrids except for hybrids no. 7 and 8. Meanwhile, only one hybrid (no. 8) of Okinawa x M. Dalet failed to amplify the resistant marker. The Okinawa parent was clustered with hybrids no. 1, 2, 3, 4 and 14 for Om El-fahm and hybrids no. 1, 2, 3, 4, 5 and 6 for M. Dalet indicating a common genetic resistance for root-knot nematode.

Conclusion: Hybrid rootstocks slightly varied in their resistance to the root-knot nematode Meloidogyne incognita. However, line no. 6 (Okinawa × Om El-fahm) could be considered as highly resistant (HR).