ISSN 2756-3707
Research Article
International Journal of Histology and Cytology ISSN 2756-3707 Vol. 6 (6), pp. 001-008, June, 2019. © International Scholars Journals
Full Length Research Paper
Rapid and efficient production of transgenic East African Highland Banana (Musa spp.) using intercalary meristematic tissues
Leena Tripathi1*, Jaindra Nath Tripathi1 and Wilberforce Kateera Tushemereirwe2
1International Institute of Tropical Agriculture, PO Box 7878, Kampala, Uganda.
2Kawanda Agriculture Research Institute, PO Box 7065, Kampala, Uganda.
Accepted 16 April, 2019
Abstract
East Africa is the largest banana producing and consuming region in Africa. In particular, the East African Highland Banana serves as the major staple crop of countries like Uganda, but production is constrained by a number of serious pests and diseases. Banana breeding is a very difficult and slow process, so genetic engineering offers an alternative approach to improvement. A transformation system using intercalary meristematic tissues was developed using Agrobacterium strain EHA105 harboring the binary vector pCAMBIA2301 containing the gusA reporter gene and nptII as selectable marker. In this paper, a new transformation protocol is described that yields kanamycin-resistant, GUS-expressing banana plants from roughly 10% of the initial explants. The resulting fully-rooted transgenic plants do not appear to be chimeras since they can be stably propagated, GUS activity is observed uniformly throughout the plants including the germline cells of the meristem, and PCR and Southern blots indicate stable integration of the genes into the genome.
Key words: Agrobacterium, intercalary meristem, genetic transformation, banana.
Leena Tripathi*, Jaindra Nath Tripathi and Wilberforce Kateera Tushemereirwe
Research Article
International Journal of Histology and Cytology ISSN 2756-3707 Vol. 6 (5), pp. 001-006, May, 2019. © International Scholars Journals
Full Length Research Paper
Use of an adsorbent and antioxidants to reduce the effects of leached phenolics in IN VITRO plantlet regeneration of faba bean
Rabha Abdelwahd1, Najat Hakam1, Mustapha Labhilili1 and Sripada M. Udupa2*
1Biotechnology Unit, Institut National de la Recherche Agronomique (INRA), Avenue de la Victoire, B.P. 415, Rabat, Morocco.
2ICARDA-INRA Cooperative Research Project, International Center for Agricultural Research in the Dry Areas (ICARDA), B.P. 6299, Rabat, Morocco.
Accepted 21 February, 2019
Abstract
Development of a reliable IN VITRO regeneration protocol is necessary to facilitate genetic transformation of faba bean. However, leaching of phenolics from the explants of most genotypes of faba bean to the culture medium causes browning, and eventually kills the explants, hindering IN VITRO regeneration. This study is aimed to minimize the effect of phenolics and to identify the most suitable types of explants for IN VITRO regeneration. We pre-treated faba bean seeds in polyvinylpyrrolidone (PVP), then cultured different types of explants on tissue culture media supplemented with an adsorbent (activated charcoal) and antioxidants (ascorbic acid, cysteine and silver nitrate). Our results showed that treating the over night soaked seed (after removing the seed coat) with PVP solution (1000 mg/l) for 1 h, followed by culturing in Murashige and Skoog medium (MS medium) with 3% (w/v) sucrose, 0.8% (w/v) agar, 2 mg/l 6-benzylaminopurine and 2 mg/l thidiazuron, supplemented with ascorbic acid (1 mg/l) or activated charcoal (10 g/l), greatly reduced lethal browning in explants and improved shoot regeneration. The shoots rooted on half-strength MS medium supplemented with 0.5 mg/l α-naphthaleneacetic acid. The cotyledonary node is the most suitable type of explant for regeneration. Regenerated plantlets were successfully established in pots and set seeds in the green house.
Key words: Adsorbent, antioxidants, faba bean, in vitro regeneration, phenolics.
Mustapha Labhilili and Sripada M. Udupa*, Rabha Abdelwahd, Najat Hakam
Research Article
A simple and efficient method for extraction of genomic DNA from tropical tuber crops
International Journal of Histology and Cytology ISSN 2756-3707 Vol. 6 (5), pp. 001-005, May, 2019. © International Scholars Journals
Full Length Research Paper
A simple and efficient method for extraction of genomic DNA from tropical tuber crops
Kamal Sharma, Ajay Kumar Mishra and Raj Shekhar Misra*
Central Tuber Crops Research Institute, Thiruvananthapuram, Kerala, India-695017.
Accepted 18 February, 2019
Abstract
DNA extraction in many plants is difficult because of metabolites that interfere with DNA isolation procedures and subsequent applications, such as DNA restriction, amplification and cloning. We have developed a reliable and efficient method for isolating genomic DNA free from polysaccharide, polyphenols and protein contaminants from tropical tuber crops (Elephant foot yam, Cassava, Sweet potato Taro and Tannia). The method involves inactivation of contaminant proteins by using CTAB/Proteinase K and precipitation of polysaccharides in the presence of high concentration of salt. The purity of genomic DNA was confirmed by A260/280 and A260/230 ratios calculated from the spectrophotometric readings and further by restriction analysis of the isolated DNA using restriction enzymes ECORI and HIND III. The described protocol also resulted in the isolation of sufficiently higher yield of DNA from leaf sample of tropical tuber crops. The new protocol can be successfully used for both small and large scale preparation of genomic DNA from leaf tissues of tuber crops which is highly suitable for further down stream processes like PCR amplification and restriction digestion analysis.
Key words: DNA isolation, RAPD, restriction enzyme digestion, tuber crops.
Ajay Kumar Mishra and Raj Shekhar Misra*, Kamal Sharma
Research Article
International Journal of Histology and Cytology ISSN 2756-3707 Vol. 6 (4), pp. 001-009, April, 2019. © International Scholars Journals
Full Length Research Paper
Tissue culture in PINUS CARIBAEA Mor. var. HONDURENSIS barr. and golf. II: Effects of two auxins and two cytokinins on callus growth habits and subsequent organogenesis
Akaneme, Florence Ifeoma1* and Eneobong, Effiom Eneobong2
1Department of Botany, University of Nigeria, Nsukka, Nigeria.
2Michael Okpara University of Agriculture, Umudike, Nigeria.
Accepted 11 February, 2019
Abstract
The growth habits of calluses of PINUS CARIBAEA Mor. were investigated to determine their correlation with organogenesis for micropropagation purposes. Two forms of callus growth habits were observed; friable and compact calluses. Their development occurred over a range of auxin-cytokinin combinations. In naphthalene acetic acid (NAA) x 6-furfuryl amino purine (kinetin) cultures, 100% friable calluses were obtained while in NAA x 6-benzyl aminopurine (BAP) cultures, 100% friable callus was formed only with 1:2 NAA/BAP ratio. Many compact calluses were also formed by the various combinations of NAA and BAP. Friability of calluses was further promoted by the interaction of indole butyric acid (IBA) and BAP, but organogenesis was not achieved. However, different degrees of greening were observed in some of the cultures (both compact and friable type). Thus, greening was not associated with a particular type of callus growth habit. Anatomical studies indicated that the differences between the compact and friable calluses were in the distribution of the meristematic cells. The histological studies also revealed some important and unexpected features. These were the presence of embryo-like structures, tracheary elements, lignification and starch-grain like structures. These results have further demonstrated the potential totipotency of callus cells of P. CARIBAEA.
Key words: Auxins, cytokinins, callus, growth habits, organogenesis.
Florence Ifeoma* and Eneobong, Akaneme , Effiom Eneobong
Research Article
International Journal of Histology and Cytology ISSN 2756-3707 Vol. 6 (4), pp. 001-004, April, 2019. © International Scholars Journals
Full Length Research Paper
Effects of intraperitoneal administration of vitamins C and E or A and E combinations on the severity of Trypanosoma brucei brucei infection in rats
Umar, I. A.* Rumah, B. L., Bulus, S. L., Kamla, A. A., Jobin, A., Asueliman, B. I., Mazai, M.H., Ibrahim, M. A. and Isah, S.
Department of Biochemistry, Faculty of Science, Ahmadu Bello University, Samaru-Zaria, Nigeria.
Accepted 11 March, 2019
Abstract
The effects of intraperitoneal administration of combinations of vitamins C and E (100mg/Kg b.w each) or vitamins A and E (10,000i.u/Kg, 100mg/Kg b. w., respectively) on Trypanosoma brucei brucei infection in rats were compared. The rats were infected intraperitoneally with the same parasite load and the experiment lasted for 17 days. The two vitamin combinations significantly (P<0.05) and persistently kept the parasitaemia lower than was recorded in the untreated infected rats. Terminal parasitaemia of the infected group given the vitamins A and E combination (AE) was significantly (P<0.05) lower than that of the group given the vitamins C and E combination (CE) . Although all infected animals developed anaemia, its severity in the untreated infected animals was significantly (P<0.05) higher than observed in the two infected groups treated with the vitamin combinations; with the group given AE developing a significantly (P<0.05) less severe anaemia than those given CE. Trypanosoma brucei infection, without vitamin treatment caused general increases in serum alanine- and aspartate aminotransferases, urea and creatinine. Although CE had no significant effect on the infection levels of serum ALT and AST; AE significantly (P<0.05) prevented the disease-induced increases in these parameters. The two vitamin combinations prevented, to a significant degree, the disease-induced elevation of serum urea and creatinine; however, AE was more effective at this than CE. It was concluded that intraperitoneal administration of either vitamins C and E or vitamins A and E combinations alleviates the degenerative changes in tissues and organs associated with Trypanosoma brucei infection of rats; with the vitamins A and E combination being more effective.
Key words: Trypanosoma brucei brucei, antioxidant vitamins, anaemia, tissue damage.
M. A. and Isah, Jobin , Bulus , A. A., Asueliman , B. L., A. , S. L., Ibrahim , I. A.* Rumah, Mazai , B. I., M.H. , Kamla , Umar , S.
Research Article
Autofluorescence of routinely hematoxylin and eosin-stained sections without exogenous markers
International Journal of Histology and Cytology ISSN 2756-3707 Vol. 6 (4), pp. 001-004, April, 2019. © International Scholars Journals
Full Length Research Paper
Autofluorescence of routinely hematoxylin and eosin-stained sections without exogenous markers
Deeb S.*, Nesr K. H., Mahdy E., Badawey M. and Badei M.
Department of Pathology, Faculty of Veterinary Medicine, University of Beni Suef, Egypt.
Accepted 16 January, 2019
Abstract
Hematoxylin and eosin stained paraffin section was examined by fluorescence microscopy to study the pattern and distribution of fluorescence. Autofluorescence was sensitive and specific for detection of elastic and collagen fibers. It was concluded that analytical morphological techniques based on autofluorescence can obtain information about morphological and pathological state of tissue and cells.
Key words: Histology, pathology, fluorescence microscopy.
Nesr K. H., Badawey M. and Badei M., Deeb S.*, Mahdy E.