Journal of Applied Biotechnology

11-19

Authors: Supri Irianti Handayani, Kusmardi, Aryo Tedjo

The development of a tumor has a strong relationship with cellular immunity mediated by T lymphocytes. Some research showed that mangiferin can increase host immunity but its scientific base was still unclear. The purpose of this study was to explain the role of mangiferin as an immunostimulator on in vitro T lymphocytes culture of mammary tumor bearing mice. T lymphocytes were isolated from the spleen of mice after two weeks of mammary tumor transplantation. The isolation was performed by using sterile nylon net and passed through nylon wool. Isolated T lymphocytes were cultured in vitro on RPMI 1640 medium. The culture of T lymphocytes was divided into three groups containing 5, 25 and 50 mg/L of mangiferin, and a control group (RPMI medium). The proportion of T lymphocytes and IL-2Rα (CD25) expressed T cells were evaluated using immunoflouorescens techniques after one and three days of lymphocytes cultures. Our study showed that 5, 25 and 50 mg/L mangiferin was effective to enhance IL-2Rα (CD25) on T lymphocytes culture and T lymphocytes prolifferation.

41-60

Authors: Reza Mohammad Emon, Adedze Yawo Mawunyo Nevame, Perry J. Gustafson, Md. Shahidul Haque, M. Jahiruddin, Mirza Mofazzal Islam

The occurrence of boron toxicity becomes a major constraint in cereal production in the world, thus cause significant yield loss particularly in wheat, one of the most susceptible crops to boron toxicity. Development of tolerant variety is now of utmost importance since agronomic management of soil boron is becoming ineffective to mitigate the toxicity. Due to narrow genetic base of the existing wheat cultivars, genetic variation for this character was poorly reported. The present study was devoted to identify such variation by using two different genetic background of wheat including Aegilops and Triticum, which could be readily used in wheat breeding program. Morphological and genetic screening revealed that two species Ae. longissima and Ae. sharonensis expressed tolerance against boron toxicity. The molecular marker analysis such as unweighted pair group method and principal coordinate analysis confirmed these two species to be more tolerant to excessive boron with higher root length. Their derivatives genotypes TL-09 and TL-17 were found to explained considerable tolerance to boron. On other hand, among the SSR markers used, Xgwm192 was the most robust in identifying boron tolerance polymorphism. The diversity and variability observed in this work could open new avenue in developing B toxicity tolerant wheat variety.

13-24

Authors: M. M. D. Katunga, J. B. B. Muhigwa

In D. R. Congo, agriculture is still managed in extensive system with crop integration especially in South-Kivu Province. Demographic pressure on the land is highly due to unequal distribution. The objective of this study is to test adaptation of improved herbaceous forage legumes selected in Tropical America by “Centro International de Agricultura Tropical” (CIAT). Agronomic performance and farmer participatory evaluation in using various agroecological zones in South-Kivu, were used to draw a model for similar areas. Forages were installed randomly in the blocks with 3 replicates. In spite of the acidic soils, forages recommended were, S. guianensis 11995, S. guianensis Cook, C. molle and D. intortum. The second step should be to adapt these forages in the farmers cropping systems. The seasons didn’t influence in general the yield production. All the accessions of V. unguiculata and C. ternatea did not adapt anywhere and C. brasiliensis was sensitive to diseases and insects.

34-44

Authors: Dey K. Kishore, W. B. Borth, M. J. Melzer, John S. Hu

Infectious clones of viruses allow elucidation of viral gene function in plants. This study presents an adaptation and expansion of the circular polymerase extension reaction that enables both cell-based and cell-free one-step assembly of a plant viral cDNA. To demonstrate its infectious nature, the generated clone was introduced into plants by Agrobacterium infiltration. This technique eliminates the cumbersome strategy of restriction enzyme digestion and ligation and instead relies on the sequence specificity of overlapping PCR fragments to assemble a complete functional infectious clone of a virus. This technique is rapid and potentially applicable for cloning virus genes which may be difficult to clone using conventional approaches due to toxicity problems that may be encountered when cloning in E. coli.

63-74

Authors: A. I. Ogbonna, P. O. Nwadiaro, A. Chuku, C. I. C. Ogbonna, F. C. Onwuliri

This study was aimed at isolation and screening of fungal species associated with Artemisia annua Plantation soils from one of the under studied areas in Plateau state, Nigeria for cellulase activity. A total of thirteen fungal species were isolated from various locations within the A. annua Plantation and were screened for cellulase production. Agar plate assay was carried out using basal medium supplemented with 1% Carboxymethylcellulose (CMC) powder and staining with 0.1% Congo red solution after the incubation period. Among these species, Aspergillus fumigatus, A. niger and A. terreus were more predominant and were recorded as cellulase producing species. They have shown to possess cellulose degrading ability and exhibited maximum zones of hydrolysis on Carboxymethylcellulose medium and were selected for enzyme assay using submerged fermentation (SmF). Enzyme production was analyzed by Dinitrosalicylic acid (DNSA) methods and the enzymes assayed for were CMCase (β-1,4-endoglucanase), β-glucosidase and FPase (total cellulose) using Carboxymethylcellulose, cellulose acetate and Filter paper as substrates respectively. The highest cellulase activity was observed on the 3rd day in A. niger with enzyme production of 0.045IU/ml and 0.040IU/ml on CMC and filter paper media respectively. A. fumigatus had high enzyme activity of 0.037IU/ml and 0.025IU/ml on filter paper and cellulose acetate media respectively. Highest enzyme production of 0.034IU/ml was recorded for A. terreus on the 3rd day on cellulose acetate medium. These fungal species could be employed specially to perform in situ environmental applications involving cellulose biodegradation of wastes.