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Accueil du site → Master → Etats Unis → 2014 → Analysis of the microbial community in bladder cells of micropropagated Atriplex canescens (Pursh) Nutt. through microscopy and metagenomics

New Mexico State University (2014)

Analysis of the microbial community in bladder cells of micropropagated Atriplex canescens (Pursh) Nutt. through microscopy and metagenomics

Deswood, Helena

Titre : Analysis of the microbial community in bladder cells of micropropagated Atriplex canescens (Pursh) Nutt. through microscopy and metagenomics.

Auteur : Deswood, Helena

Université de soutenance : New Mexico State University

Grade : Master of Sciences (MS) Plant and Environmental Sciences 2014

Résumé
Atriplex canescens is an important forage shrub in arid rangelands and is valued for reclamation in disturbed areas. As with most plants, endophytes are important component of A. canescens because endophytes are known to improve overall plant fitness. A. canescens have a salt secreting trichome to remove the excess salt from within the plant. The epidermal trichome has two parts, the stalk cell and bladder cell ; the bladder cell is where the salt is sequestered in the vacuole. Despite the importance of bladder cells to A. canescens, few studies have focused on endophytes in the bladder cells. This study examined the diverse microbial community in micropropagated Atriplex canescens bladder cells using laser scanning confocal microscopy and DNA sequencing. Two types of dyes, Live/Dead viability stain and Calcofluor white stain, were used with confocal microcopy and revealed fungal hyphae between the bladder cells. Florescence of both dyes indicated presence of microbes in the bladder cells. Atriplex canescens bladder cells were isolated and plated on microbial and plant culture media but there were no microbial growth in the media. DNA sequencing analysis used PCR amplification of internal transcribed spacer (ITS), 16S bacterial gene and small subunit (SSU) ribosomal DNA sequences from bladder cells. Eight specific primer pairs were used to amplify DNA for fungal, bacterial, plant. Three maximum likelihood trees were obtained via PhyML with Jukes Cantor substation model. A low diversity of microbial community was found in the bladder cells and the OTUs for fungal and bacteria phyla were as the following : Ascomycota as 84%, Basidiomycota at 6%, unidentified fungi at 10% and Preoteobacteria at 100%. Atriplex canescens bladder cells were colonized by 8 different orders and the most common orders were Dothideales at 84% and Pleosprales at 16%. This study shows a high presence of Alternaria and Aureobasidium in Atriplex canescens bladder cells which have been reported to have better adaptation to the arid ecosystems.

Sujets : Fourwing saltbush — Microbiology. Trichomes — Microbiology. Endophytes — Microbiology

Présentation (NMSU Library)

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