Theses and Dissertations

Author

Jinyan She

Issuing Body

Mississippi State University

Advisor

Mlsna, Todd E.

Committee Member

Baird, Richard E.

Committee Member

Emerson, Joseph P.

Committee Member

Gwaltney, Steven

Committee Member

Wipf, David O.

Date of Degree

1-1-2017

Document Type

Dissertation - Open Access

Degree Name

Doctor of Philosophy

Abstract

Our study focuses on the application of volatile metabolomics and chemometrics in plant pathology. Specifically, volatile metabolites or volatile organic compounds (VOCs) from the American chestnut tree (Castanea dentata) and its pathogenic fungus Cryphonectria parasitica have been investigated. The American chestnut was once a dominant tree species in the eastern forests of the United States. However, it was nearly devastated by the fungal pathogen C. parasitica. The loss of this tree species has significantly impacted the ecosystem. Therefore, preservation and restoration of American chestnut are crucial. Chapter one provides an overview of mass spectrometry based volatile metabolomics and their implementation in the investigation of plant pathology. The study of volatile metabolites profiles from virulent and hypovirulent strains of C. parasitica are presented in chapter two. The microbial volatile organic compounds (MVOCs) profiles were analyzed via nondestructive sampling method, headspace solid phase microextraction (HS-SPME), combined with gas chromatography (GC)-mass spectrometry (MS). The results indicate that the MVOCs profiles emitted from these two strains are significantly different. In general, compared with its hypovirulent strains, high emissions of sesquiterpenes were observed in the virulent strains. Furthermore, the study explored MVOCs differences associated with hypovirulence processes. The study found that both hypovirulence and aging can alter the virulent strains' MVOCs, and the process can be observed via their volatile metabolites. Chapter three describes the effects of aging, cultivation medium, and pH on fungal volatile metabolite profiles, all of which can change the strength of MVOCs emission and their composition. An acidic environment favors fungal bioactivity and therefore enhanced MVOCs emission. However, due to the inherently low MVOCs production from hypovirulent strains, the pH effect was less apparent in the hypovirulent isolates. The strength of MVOCs emission was highly correlated to the fungal expansion in virulent strains for the first 14 days. The overall emission from hypovirulent strains was relatively steady during the 28-day observation. Finally, the cultivation media are critical to the fungal MVOCs production. Among the tested media, cornmeal was least favorable for MVOCs production for both strains. Finally, Chapter Four presents a study of the total constitutive phenolic content estimation and volatile organic compounds identification from four species of chestnut tree leaf tissues. Folin Ciocalteu reagent assay with UV/Vis spectrophotometry was applied to estimate the total phenolic content in leaf tissues of American chestnut (Castanea dentata), Chinese chestnut (Castanea mollissima), and their backcross breeding generations (B3F2 and B3F3). The results from leaf tissue extraction in methanol/water (95:5 v/v), pH 2, and analyzed under the UV/Vis at 765 nm show that the variations among these tree species are significant (ANOVA, p < 0.05). The kinetics of phenolic compound solid-liquid extraction was elaborated using Peleg, second order, and power law models. Moreover, the analysis of VOCs collected from these species indicated that the distinction of American and Chinese chestnut could be archived via their VOCs, while the hybrids’ leaf VOCs are different from their parents’.

URI

https://hdl.handle.net/11668/18616

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