Abstract
The aim of this study is to comparative study between conventional and molecular assay of isolation, identification and characterization of non-leguminous endophytic bacteria and fungi in the leguminous root samples. The plant root samples, Helianthus annuus, Carica papaya and Lycoperesicum solanum (Sunflower root and stem, pawpaw root and stem, and tomato root and stem from Adekunle Ajasin University School farm, Akungba Akoko, Ondo state, Nigeria. The isolation of endophytic bacteria were performed using the conventional method of isolation (biochemical test) and characterization were done using both the conventional and molecular method of bacteria characterization. The antibiotic susceptibility test (Antibiogram) was observed using disc diffusion. The four bacteria identified were Bacillus cereus, Enterobacter sp. Actnomycoses sp. and Aeromonas sp. for conventional method and Fusarium solani, Fusarium vortecelium and Bacillus thuringiensis for molecular method as confirmatory point of view. In this study, all isolated organisms tends to be Gram positive using the gram staining technique. Antibiogram shows the zones of inhibition with diameter ranging from 0-20 mm, Enterobacter sp. were more sensitive to the various antibiotics used. Ultraviolet spectrophotometer was also used to determine the growth dynamic as well as the death rate of the isolates, the addition of antibiotics (ciprofloxacin) to the isolates at the 24th hour speed up the death rate of the isolates from non-leguminous endophytic bacteria. After the preliminary identification of the bacteria isolates and the confirmatory identification of both bacteria and fungi isolates of the non-leguminous endophytic microorganism, it was noted that the preliminary identification was only able to achieve the genus level of taxonomic characterization, While the molecular method confirm the molecular sub level identification of isolates depletes the absolute taxonomic identification and characterization to the sub-species level. The results of this study validates the use of molecular sequencing for the assay identification and characterization of non-leguminous endophytic bacteria and fungi as the easy and best mode of identification of both bacteria and fungi isolates as a veritable tools for research purposes.
Highlights
Endophytic microbes brings nutrients to plants include those that fix atmospheric nitrogen in plant tissues
The type of non-leguminous plant root and stem sample collected was Helianthus annuus, Lycopersicum solanum, and Carica papaya, the number of sample collected was 6 and the samples were collected at the School farm, Adekunle Ajasin University, Akungba Akoko, at 2:30-3:00pm in the afternoon
It was observed that 6 plates were isolated and each plate contains inoculums of serial dilution, the highest dilution factors are the 10-10 and the lowest dilution factor are the 10-7
Summary
Endophytic microbes brings nutrients to plants include those that fix atmospheric nitrogen in plant tissues. Another type of nutritional endophytic symbiosis involves microbes that inhabit both endophytic tissues and extend out into soil. Dark septate endophytes and mycorrhizal fungi establish this kind of symbiosis with many families of plants Hyphae of these fungi grow endophytically in roots, and the mycelia extending into soil acquire nutrients and mobilize it back to plants. Another nutrient acquisition mechanism involves the liberation of nutrients from insects by microbes that extend between, or cycle between, plants and decaying insects. In this process insects consume plants, accumulating nitrogen and other nutrients in their bodies; degradation of insects by microbes that are symbiotic with plants results in transfer of nutrients to plants [2]
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