1.Community structures of total bacterial DNA, cultivable bacteria and prototrophs in bulk soil and rhizospheres
Neelawan Pongsilp ; Pongrawee Nimnoi ; Saisamorn Lumyong
Malaysian Journal of Microbiology 2016;12(1):1-14
Aims: It has been hypothesized that root exudates can be a nutritional factor influencing the bacterial community
structure as well as the occurrence of prototrophs and auxotrophs in rhizospheres. The present study was performed to
examine the community structures of total bacterial DNA, cultivable bacteria and prototrophs in 3 soil samples with
different levels of abundance of root exudates.
Methodology and results: Denaturing gradient gel electrophoresis (DGGE) was performed to examine the community
structures of total bacterial DNA, cultivable bacteria and prototrophs in 3 soil samples including bulk soil, rhizosphere of
a single plant species and rhizosphere of multiple plant species. For clustering analysis, a dendrogram generated from
the DGGE patterns revealed the different bacterial community structures in these soil samples. Both rhizospheres
claded together, separating from bulk soil. The DGGE patterns of cultivable bacteria showed particular fingerprints
corresponding to kinds of media and soil samples. Nutrient agar (NA) medium, isolation medium for prototroph (IMP)
and IMP supplemented with soil extracts were used for bacterial cultivations. Prototrophs were isolated and examined by
random amplified polymorphic DNA (RAPD) and 16S rRNA gene sequence analysis. The genetic diversity of
prototrophs in 3 soil samples was similar (approximately 5% to 10% similarities) and most of them (13 of 28 strains)
were members of Pseudomonas with 97% to 100% identities.
Conclusion, significance, and impact of study: The present study provides a strong evidence of the influence of root
exudates and plant species on bacterial community structures.
Denaturing Gradient Gel Electrophoresis
2.A New Report on Edible Tropical Bolete, Phlebopus spongiosus in Thailand and Its Fruiting Body Formation without the Need for a Host Plant
Jaturong KUMLA ; Nakarin SUWANNARACH ; Saisamorn LUMYONG
Mycobiology 2020;48(4):263-275
Phlebopus spongiosus is a well-known edible ectomycorrhizal mushroom indigenous to southern Vietnam. The mushroom specimens collected from northern Thailand in this study were identified as P. spongiosus. This identification was based on morphological characteristics and the multi-gene phylogenetic analyses. Pure cultures were isolated and the relevant suitable mycelial growth conditions were investigated. The results indicated that the fungal mycelia grew well on L-modified Melin-Norkans, and Murashige and Skoog agar all of which were adjusted to a pH of 5.0 at 30 C. Sclerotia-like structures were observed on cultures.The ability of this mushroom to produce fruiting bodies in the absence of a host plant was determined by employing a bag cultivation method. Fungal mycelia completely covered the cultivation substrate after 90–95 days following inoculation of mushroom spawn. Under the mushroom house conditions, the highest amount of primordial formation was observed after 10–15 days at a casing with soil:vermiculite (1:1, v/v). The primordia developed into a mature stage within one week. Moreover, identification of the cultivated fruiting bodies was confirmed by both morphological and molecular methods. This is the first record of P. spongiosus found in Thailand and its ability to form fruiting bodies without a host plant.