International Journal of Histology and Cytology

International Journal of Histology and Cytology ISSN 2756-3707 Vol. 7 (3), pp. 001-004, March, 2020. © International Scholars Journals

Full Length Research Paper

Micronuclei in red blood cells of armored catfish Hypostomus plecotomus exposed to potassium dichromate

Carlos Augusto Borba Meyer Normann¹, José Cláudio Fonseca Moreira² and Valesca Veiga Cardoso¹*

¹Centro Universitário Metodista - Porto Alegre - RS –Brazil.

²Department of Biochemistry-ICBS; UFRGS - Porto Alegre - RS - Brazil.

Accepted 22 January, 2020

Abstract

The present work aims to evaluate the impact of potassium dichromate in armored catfishes’ (Hypostomus plecotomus) erythropoiesis, using piscine micronucleus test. Armored catfishes (n = 30) were subjected to 12 mg/L of potassium dichromate, with an equal control group (n = 30). For each 2,000 red blood cells of animals subjected to treatment, it was possible to observe 8.25 ± 0.02% cells with micronuclei and 17 ± 0.005 mg/kg of total chrome in tissues, detected through atomic absorption spectrophotometry. In the control group, the average was 0.75 ± 0.03% for micronuclei and 1.08 ± 0.04 mg/kg of chrome in tissues. The results confirm the clastogenic effect of potassium dichromate. The sensibility to this ion and the economic relevance of H. plecotomus show that armored catfish can be used to monitor acute effects of metallic pollutant spills in freshwater.

Key words: Piscine micronucleus test, aquatic pollution, potassium dichromate, Hypostomus plecotomus.

International Journal of Histology and Cytology ISSN 2756-3707 Vol. 7 (3), pp. 001-007, March, 2020. © International Scholars Journals

Full Length Research Paper

Isolation and characterization of LHY homolog gene expressed in flowering tissues of Tectona grandis (teak)

B. Norlia¹*, M. Norwati¹, A. Norwati¹, H. Mohd Rosli¹ and M. S. Norihan²

¹Genetic Unit, Forest Research Institute Malaysia (FRIM), 52109 Kepong, Selangor, Malaysia, Forest Biotechnology Division, Forest Research Institute Malaysia (FRIM), 52109 Kepong, Selangor, Malaysia.

²Cell and Molecular Biology Department, Faculty of Biotechnology and Biomolecular Science, Universiti Putra Malaysia (UPM), 43400 Serdang, Selangor, Malaysia.

Accepted 13 November, 2019

Abstract

Floral initiation of teak through molecular biology approach is being studied for better understanding of teak flower development. Through PCR subtractive hybridization method, LHY homolog gene has been isolated from teak flowering tissues. The full-length cDNA of the gene was 2948 base pair (bp) and potentially encoded for 768 amino acids. It was named Tectona grandis LHY (Tg-LHY), as the gene was similar to the LHY gene of some species. Amino acid sequence alignment revealed that Tg-LHY was similar to LHY of Castanea sativa, LHY of Phaseolus vulgaris and LHY of Arabidopsis thaliana. The highly conserved region found in Tg-LHY was the MYB protein, which is the DNA-binding protein responsible in negative feedback loop reaction of central oscillator in plant circadian clock system. The level of gene expression was found to be high four hours after dawn in flowering shoots and flower. This paper reported the isolation and characterization of the gene.

Key words: LHY gene, circadian clock, floral gene, Tectona grandis.

International Journal of Histology and Cytology ISSN 2756-3707 Vol. 7 (4), pp. 001-006, February, 2020. © International Scholars Journals

Full Length Research Paper

Phenolic changes during in vitro organogenesis of cotton (Gossypium hirsutum L.) shoot tips

Ibrahim Ilker Ozyigit

Marmara University, Science and Arts Faculty, Department of Biology, 34722 Goztepe Istanbul/Turkey, E-mail: [email protected], Tel.: (+90216) 3487759/1182; Fax: (+90216) 3478783.

Accepted 10 November, 2019

Abstract

Browning and subsequent death of the cultured explants are major problems for many tissue culture non/hard-adaptive species and usually depended on the phenolic compounds and the quantity of total phenols. Many studies, using shoot tips as explant source indicated various problems such as phenolic exudation, media discoloration, rooting deficiencies and explant browning and death. The phenols are synthesized by the plants and in many tissue culture studies, excreted and then oxidized phenols effect in vitro proliferation negatively. In addition, amount of phenolics can be more or less in different stages of organogenesis due to metabolic actions. Therefore determination of phenolics and calculating the amounts of phenols may be another research area for many tissue culture studies. In this study, total phenol amounts of shoot tip cultures were evaluated during in vitro organogenesis of cotton (Gossypium hirsutum L.). Cotton var. Nazilli 84S was used as explant source and total phenols of young leaves, shoots and the MS (Murashige and Skoog) media (for excreted phenols from explants to medium) were calculated in 7-14-21 and 28 days of culturing period. Seeds were germinated in hormone free MS media in 7 days. After germination, 7 day old meristematic shoot tips were dissected out from seedlings and cultured on MS media, supplemented with 0.1 mg/L Kinetin (KIN). They were grown at 25^oC under fluorescent light (7500 lx) 16 h light and 8 h dark for 3 weeks. Singleton-Rossi method based Folin-Ciocaelteu reactive was used for determination of total phenol amount of the young plantlets and it was observed that different parts of the plantlets synthesized more or less amounts of phenolics in different stages of organogenesis.

Key words: Cotton, tissue culture, phenolics, browning, organogenesis, shoot tip.

International Journal of Histology and Cytology ISSN 2756-3707 Vol. 7 (1), pp. 001-008, January, 2020. © International Scholars Journals

Full Length Research Paper

Leaf storage conditions and genomic DNA isolation efficiency in Ocimum gratissimum L. from Kenya

Lexa G. Matasyoh¹*, Francis N. Wachira², Miriam G. Kinyua³, Anne W. Thairu Muigai¹ and Titus K. Mukiama⁴

¹Department of Botany, Jomo Kenyatta University of Agriculture and Technology, P. O. Box 62000, Nairobi, Kenya.

²Department of Biochemistry, Egerton University, P. O. Box 536, Njoro, Nakuru, Rift Valley, Kenya.

³Kenya Agricultural Research Institute, P.O. Njoro, Nakuru, Kenya.

⁴Department of Botany, Nairobi University, P. O. Box 29053, Nairobi, Kenya.

Accepted 16 October, 2019

Abstract

Storage of plant tissues for DNA is important to avoid degradation of DNA. Preliminary studies were conducted on Ocimum gratissimum L. in order to establish the storage conditions for the collected samples before DNA extraction. Secondly, the aim was to determine the best protocol for the extraction of high quality DNA, which would later be used for molecular analysis. DNA was extracted from the samples one month after field sampling. During the DNA extraction, four protocols were used; the modified hexadecyltrimethyl ammonium bromide (CTAB) mini preparation method described by Doyle and Doyle (1990), with reductants either mercaptoethanol or dithiothreitol; the modified sodium dodecyl sulphate (SDS) mini preparation method of Edwards et al. (1991) with redundant either mercaptoethanol or dithiothreitol. The DNA was purified, treated with RNase, quantified and examined for intactness using gel electrophoresis method. Good quality and high yield DNA could only be extracted with the buffer containing the detergent SDS and the reducing agent dithiothreiotol.

Key words: Ocimum gratissimum L., Sodium dodecyl sulphate (SDS), hexadecyltrimethl ammonium bromide (CTAB), dithiothreitol, mercaptoethanol.