Biomedical subjects

Miranda M Hart

Publications and source records attributed to Miranda M Hart.

4 recordsLinked to original sources

PubMed2005

Quantitation of transgenic plant DNA in leachate water: real-time polymerase chain reaction analysis.

Roundup Ready (RR) genetically modified (GM) corn and soybean comprise a large portion of the annual planted acreage of GM crops. Plant growth and subsequent plant decomposition introduce the recombinant DNA (rDNA) into the soil environment, where its fate has not been completely researched. Little is known of the temporal and spatial distribution of plant-derived rDNA in the soil environment and in situ transport of plant DNA by leachate water has not been studied before. The objectives of this study were to determine whether sufficient quantities of plant rDNA were released by roots during growth and early decomposition to be detected in water collected after percolating through a soil profile and to determine the influence of temperature on DNA persistence in the leachate water. Individual plants of RR corn and RR soybean were grown in modified cylinders in a growth room, and the cylinders were flushed with rain water weekly. Immediately after collection, the leachate was subjected to DNA purification followed by rDNA quantification using real-time Polymerase Chain Reaction (PCR) analysis. To test the effects of temperature on plant DNA persistence in leachate water, water samples were spiked with known quantities of RR soybean or RR corn genomic DNA and DNA persistence was examined at 5, 15, and 25 degrees C. Differences in the amounts and temporal distributions of root-derived rDNA were observed between corn and soybean plants. The results suggest that rainfall events may distribute plant DNA throughout the soil and into leachate water. Half-lives of plant DNA in leachate water ranged from 1.2 to 26.7 h, and persistence was greater at colder temperatures (5 and 15 degrees C).

DNA, Plant↗

PubMed2005

Microbial gene expression in soil: methods, applications and challenges.

About 99% of soil microorganisms are unculturable. However, advances in molecular biology techniques allow for the analysis of living microorganisms. With the advent of new technologies and the optimization of previous methods, various approaches to studying gene expression are expanding the field of microbiology and molecular biology. Methods used for RNA extraction, DNA microarrays, real-time PCR, competitive RT-PCR, stable isotope probing and the use of reporter genes provide methods for detecting and quantifying gene expression. Through the use of these methods, researchers can study the influence of soil environmental factors such as nutrients, oxygen status, pH, pollutants, agro-chemicals, moisture and temperature on gene expression and some of the mechanisms involved in the responses of cells to their environment. This review will also address information gaps in bacterial gene expression in soil and possible future research to develop an understanding of microbial activities in soil environments.

Bacteria↗

PubMed2002

Does percent root length colonization and soil hyphal length reflect the extent of colonization for all AMF?

Percent root length colonization may not be an appropriate measure of root colonization by arbuscular mycorrhizal fungi (AMF) in all cases. We suggest that AMF will differ in how well percent root length colonization measures the amount of AMF colonization in the root due to differences among AMF in hyphal structure and hyphal aggregation. Although soil hyphal length accounts for hyphal density, we suggest that it does not consider differences in hyphal structure in measurements of external colonization and thus might also misrepresent the true amount of AMF in the soil. To test these suggestions, we measured and compared percent root length colonization and soil hyphal length with root ergosterol and soil ergosterol, respectively, for 21 different species of AMF from three families in a greenhouse experiment. Percent root length colonization predicted intra-radical colonization best for Glomaceae and Acaulosporaceae isolates, while soil hyphal length best represented soil ergosterol for Gigasporaceae isolates. The results show that conventional methods for estimating AMF colonization are not universal for all AMF. Caution is advised when drawing inferences for different groups of AMF.

Biomass↗

PubMed2002

Colonization of roots by arbuscular mycorrhizal fungi using different sources of inoculum.

Arbuscular mycorrhizal fungi (AMF) form a number of different infective propagules that are used to form new mycorrhizal associations. These are spores, extraradical hyphae and infected roots. However, not all fungi are equally capable of colonizing roots with all of the above-mentioned propagules and there is conflicting evidence of major differences in colonization strategy between members of the Glomineae and Gigasporineae. In this study, we tested the abilities of eight fungal species from four different genera to colonize roots using three different types of inoculum. Glomus and Acaulospora isolates colonized from all inoculum types, whereas Gigaspora and Scutellospora isolates colonized mainly from spores and to a limited degree from root fragments. Extraradical hyphae were not suitable propagules for the species of Gigaspora and Scutellospora tested. This indicates that AMF have different colonization strategies and that this is largely differentiated at the suborder level. It is unclear why there is such a difference among the fungi in inoculum types. Future research should examine differences in the anatomy and physiology to discern a mechanism for such differences in life-history strategies.

Ascomycota↗