Tropical Plant Research
An International Journal by Society for Tropical Plant Research
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Abstracted and Indexed
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2020, VOLUME 7 ISSUE 2Pages: 388-395
Detection and identification of microbial contaminants from plant tissue culture
Mohit Dangariya, Dharam Khandhar, Jagdishchandra Monpara, Kiran Chudasama* and Vrinda Thaker
*Plant Physiology and Molecular Biology Lab, UGC-CAS Department of Biosciences, Saurashtra University, Gujarat, India
Abstract:
Microbial contamination is a major problem, which affects the growth and development of the plant in micropropagation techniques. In the present study, the aim was to investigate the source of microbial contamination in the tissue culture laboratory. In the present work, microbes were isolated from the contaminated different plant tissue culture tubes. Isolated fungi were inoculating on Potato Dextrose Agar and incubated for 5 days at 37ºC in case of fungi and bacteria on Nutrient Agar medium incubated for 2 days at 37ºC. Total of 28 isolates obtained which include nineteen fungi and nine bacteria. It was sub-cultured for isolation, identified by morphological and molecular techniques. For identification of the strain, genomic DNA was isolated and amplified by universal primer and amplified DNA fragments were separated by electrophoresis. Bacteria and fungi were identified by 16S rDNA and 28S rDNA gene sequencing, respectively. Purified PCR product was preceded for cycle sequencing and sequenced on 3130 Genetic analyzer (Applied Biosystem, USA). All the obtained sequences were submitted in NCBI database. These fungal and bacteria were found to cause the death of the culture material. Probable sources of contamination in plant tissue culture laboratory were discussed.
Microbial contamination is a major problem, which affects the growth and development of the plant in micropropagation techniques. In the present study, the aim was to investigate the source of microbial contamination in the tissue culture laboratory. In the present work, microbes were isolated from the contaminated different plant tissue culture tubes. Isolated fungi were inoculating on Potato Dextrose Agar and incubated for 5 days at 37ºC in case of fungi and bacteria on Nutrient Agar medium incubated for 2 days at 37ºC. Total of 28 isolates obtained which include nineteen fungi and nine bacteria. It was sub-cultured for isolation, identified by morphological and molecular techniques. For identification of the strain, genomic DNA was isolated and amplified by universal primer and amplified DNA fragments were separated by electrophoresis. Bacteria and fungi were identified by 16S rDNA and 28S rDNA gene sequencing, respectively. Purified PCR product was preceded for cycle sequencing and sequenced on 3130 Genetic analyzer (Applied Biosystem, USA). All the obtained sequences were submitted in NCBI database. These fungal and bacteria were found to cause the death of the culture material. Probable sources of contamination in plant tissue culture laboratory were discussed.
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