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Dive into the research topics where Carolyn H. Keiffer is active.

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Featured researches published by Carolyn H. Keiffer.


Plant Molecular Biology Reporter | 2001

DNA extraction from plants: The use of pectinase

Steven H. Rogstad; Brian Keane; Carolyn H. Keiffer; Fred Hebard; Paul Sisco

Several earlier protocols for extracting plant DNA or RNA do not work well for a variety of plants because contaminating substances coprecipitate with the nucleic acids and, thus, are present even at the last DNA-hydration step. While DNA extraction protocols have been published in which pectinase is employed to break down these contaminating substances, here we present an alternative modified pectinase protocol that potentially uses fewer steps and avoids the use of ethylene glycol monoethyl ether and phenol. DNA analyses results are described for 6 plant species demonstrating that the method works across distantly related plant taxa.


Journal of Applied Ecology | 2013

Soil preparation methods promoting ectomycorrhizal colonization and American chestnut Castanea dentata establishment in coal mine restoration

Jenise M. Bauman; Carolyn H. Keiffer; Shiv Hiremath; Brian C. McCarthy

Summary 1. The objective of this research was to evaluate soil subsurface methods that may aid in seedling establishment and encourage root colonization from a diverse group of ectomycorrhizal (ECM) fungi during restoration projects. 2. American chestnut Castanea dentata Marsh. Borkh. and backcrossed chestnuts seedlings were planted on a reclaimed coal mine site in central Ohio, USA. Roots from chestnut seedlings planted in the plots that were cross-ripped, plowed and disked, or a combination of treatments were sampled for ECM fungi and compared with control plots. The presence and identification of native ECM were determined by fungal DNA sequencing of the internaltranscribed (ITS) region. 3. After two growing seasons, mechanical soil treatments resulted in seedlings with significantly more ECM species when compared to seedlings grown in the control plots (P <0 0001). A nonmetric multidimensional scaling ordination followed by a permutational MANOVA confirmed significant dissimilarities in community composition between the control and mechanically treated plots (F =0 24, P =0 015). 4. Ectomycorrhizal root colonization was significantly higher on the chestnut seedlings sampled from the mechanically treated plots when compared to the control plots (F =1 063, P <0 0001). Differences did not exist among the three mechanical treatments. There was also a significant increase in above-ground seedling growth in the plots that were treated with a surface soil method (F =1 572, P <0 0001). It is not clear whether ECM activity was the driver of plant growth; regardless, both are strong indicators of healthy tree establishment. 5. Synthesis and applications. This study illustrates that the use of soil subsurface methods increased ectomycorrhizal (ECM) activity and seedling growth. Employing methods that encourage the root colonization by beneficial ECM and promote healthy seedling establishment may aid the long-term survival of chestnuts in restoration projects. This can be applied to other hardwood seedlings used in reforestation in soils compacted after anthropogenic disturbances.


Journal of The Torrey Botanical Society | 2012

Seed production of mature forest-grown American chestnut (Castanea dentata (Marsh.) Borkh)1

Keith E. Gilland; Carolyn H. Keiffer; Brian C. McCarthy

Abstract Gilland, K. E. (Department of Environmental and Plant Biology, Ohio University, Athens OH 45701), C. H. Keiffer (Department of Botany, Miami University, Oxford OH 45042), and B. C. McCarthy (Department of Environmental and Plant Biology, Ohio University, Athens OH 45701). Seed production of mature forest-grown American chestnut (Castanea dentata (Marsh.) Borkh). J. Torrey Bot. Soc. 139: 283–289. 2012.—Few records concerning pre-blight seed production of forest grown American chestnut (Castanea dentata) exist. As blight resistant American chestnut becomes available to use in restoration efforts, many conservation groups are interested in reintroducing the species as a wildlife food resource. As part of a long-term ecological study involving a disjunct population of American chestnut in Wisconsin, we examined seed production of mature forest grown trees in fall 2005 and 2006. Eleven large diameter (> 23 cm DBH), overstory position, fruiting American chestnuts were selected and measured for height, crown spread, crown volume, and dbh. Because seed predation is intense on this species, we indirectly measured seed production by counting the large opened and spiny cupules (i.e., burs) beneath the tree canopy, along with any cupules remaining on the tree. Castanea dentata typically produces three seeds per cupule, allowing for a reasonably accurate estimate of seed production, and cupules are rarely moved by seed predators. One half of the total projected canopy area was sampled plus an additional meter beyond the canopy edge. This method requires no estimation for losses due to predation or trap damage as found in traditional seed trap experiments. Mean seed production per m2 of basal area was 69,489 in 2005 and 74,825 in 2006. A significant relationship between total number of seeds produced and DBH was found in 2005 but not in 2006. Using basal area measurements from the stand, we would expect 1.04×106 seeds produced per hectare with 2758.5 kg ha−1 of mast production. Based on a sample of seeds we were able to collect from the forest floor, mean seed weight was 2.65 g (± 0.039 SE). Overall, the results indicate that the decline of American chestnut after infection by the chestnut blight likely had a very significant impact on the availability of important wildlife food resources in stands where the species was dominant.


International Journal of Ecology | 2012

Facilitation of American Chestnut (Castanea dentata) Seedling Establishment by Pinus virginiana in Mine Restoration

Jenise M. Bauman; Carolyn H. Keiffer; Shiv Hiremath

This study evaluated the influence of planting sites on the establishment and ectomycorrhizal (ECM) colonization of American chestnut (Castanea denetata (Marsh.) Borkh.) on an abandoned coal mine in an Appalachian region of the United States. Root morphotyping and sequencing of the fungal internal transcribed spacer (ITS) region were used to identify the ECM species associated with the chestnut seedlings. Germination, survival, ECM root colonization, and growth were assessed in three habitats: forest edge, center (plots without vegetation), and pine plots (a 10-year-old planting of Pinus virginiana). Seedlings in pine plots had higher survival (38%) than the other plot types (center 9% and forest edge 5%; P = 0.007). Chestnuts found colonized by ECM within the pine plots were larger (P = 0.02), contributed by a larger root system (P = 0.03). Forest edge and pine plots had more ECM roots than seedlings in center plots (P = 0.04). ITS fungal sequences and morphotypes found among chestnut and pine matched Scleroderma, Thelephora, and Pisolithus suggesting these two plant species shared ECM symbionts. Results indicated that the presence of P. virginiana had a greater facilitative effect on growth and survival of chestnut seedlings.


New Forests | 2014

Growth performance and chestnut blight incidence (Cryphonectria parasitica) of backcrossed chestnut seedlings in surface mine restoration

Jenise M. Bauman; Carolyn H. Keiffer; Brian C. McCarthy

Anthropogenic disturbances such as mining for coal have caused significant disturbance to the Appalachian forests of North America. Recovery of these disturbances is highly dependent on restoration methods that encourage natural succession. Unfortunately, current reclamation protocols have resulted in soil compaction and aggressive herbaceous groundcovers that impede the recovery of native trees. To overcome this, methods such as deep ripping and plow and disking were applied to a reclaimed mine land in Ohio, USA Plantings of pure American chestnut (Castanea dentata) and two seedling types (BC1F3 and BC2F3) bred for resistance to chestnut blight fungus (Cryphonectria parasitica) were assessed among different soil preparation treatments over five field seasons. Seedling mortality due to natural infection by chestnut blight was recorded and related to the disease resistance potential of the BC1F3 and BC2F3 seedlings. The growth and survival of chestnuts in plots that employed either ripping, plow disking, or the combination of the two methods were significantly greater than the control plots. After five seasons, differences existed among the soil treatments; plots that applied deep ripping had the highest survival and growth. When chestnut types were compared, pure American chestnut was the tallest. However, BC2F3 chestnut seedlings had the highest survival and lower disease incidence. Results suggest that employing deep ripping with backcrossed chestnut seedlings provides a method for establishing hardwood seedlings in soils impacted by surface mining. Planting methods that promote vigorous growth can be applied more broadly to other regions where anthropogenic disturbances create soil conditions that hinder seedling establishment.


Journal of the American Society of Mining and Reclamation | 2011

The Influence of Inoculated and Native Ectomycorrhizal Fungi on Morphology, Physiology and Survival of American Chestnut

Jenise M. Bauman; Carolyn H. Keiffer; Shiv Hiremath

The objective of this study was to evaluate the influence of five different species of ectomycorrhizal (ECM) fungi on root colonization of native fungi on putatively blight resistant chestnut hybrids (Castanea dentata x C. mollissima) in a reclaimed mine site in central Ohio. The five species were Hebeloma crustuliniforme, Laccaria bicolor, Scleroderma polyrhizum, Amanita rubescens, and Suillus luteus. We used a combination of DNA sequencing of the ITS region and phylogenetic analyses to indentify fungi found on roots after 12 and 18 months in the field. Non-metric multidimensional scaling (NMDS) ordinations were used to determine if ECM community composition was influenced by the fungal inoculum used. The results of this study demonstrated that the selected ECM species do not persist on chestnut after one year in the field. In addition, these selected ECM species did not impede natural root colonization of native fungi or influence ECM community composition after two growing seasons. Although these species did not persist in the field, the presence of ECM inoculum (with the exception of Amanita) greatly contributed to the survival of hybrid chestnut seedlings. Therefore, introduced inoculum that was present in the very early stages of outplanting had persisting effects with regard to seedling establishment in the field, even if the original inoculum did not persist. ECM fungi native to the area colonized chestnuts resulting in increased growth rates. These native assemblages may contain species better able to form functional mycorrhizas under these environmental extremes. Therefore, the conservation of these species may be necessary to facilitate long-term survival of deciduous tree species historically native to these lands. Additional Keywords: root colonization of fungi, chestnut restoration.


Journal of the American Society of Mining and Reclamation | 2012

THE EFFICIENCY OF INTRODUCED PISOLITHUS TINCTORIUS INOCULUM ON BACKCROSSED CHESTNUT GERMINATION AND SURVIVAL

Jenise M. Bauman; Carolyn H. Keiffer; Shiv Hiremath

American chestnut was eliminated as a canopy tree from the Appalachian region of North America with the introduction of chestnut blight in the early 1900s. Breeding programs initiated in the 1980s have produced seedling lines that display the pure American morphology with potential resistance to chestnut blight. More work is required to assess their field performance in field sites representative of their native range. This study used American (Castanea dentata) and backcrossed hybrid chestnuts (C. dentata × C. mollissima) on an abandoned coal mine in southeastern Ohio. Half of the seeds were planted with ectomycorrhizal fungus (ECM) Pisolithus tinctorius (Pt) as a granular inoculum. Germination, survival, percent ECM colonization, and ECM community were assessed. In addition, soil data such as pH, cation exchange capacity, nutrients, texture, temperature, moisture, and organic matter were evaluated to determine their influence on ECM. After the first and second growing season, germination and survival were not influenced by the Pt inoculum or chestnut genotypes. In addition, ECM root colonization, fungal community composition, and host response were similar between seedling types, regardless of the inoculum added. This indicated: 1) backcrossed breeding produces seedlings similar to pure American chestnuts with regard to growth, establishment, and fungal symbionts, and 2) the granular inoculum had no influence on ECM colonization in this field site. Introducing spore inoculum to a field site greatly deficient in nutrients and organic matter is not an efficient method to ensure ECM symbiosis. However, natural colonization by native ECM fungi, though limited, did result in larger chestnut seedlings. Cation exchange capacity significantly contributed to the percent of root colonization of ECM fungi on chestnut. Determination of planting protocols and factors that influence ECM root colonization will be useful for in future mine restoration projects using backcrossed American chestnut as a restoration tree. Additional


Canadian Journal of Forest Research | 2006

Dendroecology of american chestnut in a disjunct stand of oak-chestnut forest

Ryan W. McEwan; Carolyn H. Keiffer; Brian C. McCarthy


Restoration Ecology | 2006

Effects of Mulch on Seedlings and Soil on a Closed Landfill

Erin Rachel Athy; Carolyn H. Keiffer; M. Henry H. Stevens


Restoration Ecology | 2007

Limited Reintroduction Does Not Always Lead to Rapid Loss of Genetic Diversity: An Example from the American Chestnut (Castanea dentata; Fagaceae)

Sarah Ann Morgan Pierson; Carolyn H. Keiffer; Brian C. McCarthy; Steven H. Rogstad

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Shiv Hiremath

United States Forest Service

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Brian Keane

Miami University Hamilton

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