Silicon and acibenzolar-S-methyl induced defence responses in cotton (Gossypium hirsutum L.) infected with Fusarium oxysporum f. sp. vasinfectum

Jennifer Whan (2009). Silicon and acibenzolar-S-methyl induced defence responses in cotton (Gossypium hirsutum L.) infected with Fusarium oxysporum f. sp. vasinfectum PhD Thesis, School of Biological Sciences, The University of Queensland.

       
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Author Jennifer Whan
Thesis Title Silicon and acibenzolar-S-methyl induced defence responses in cotton (Gossypium hirsutum L.) infected with Fusarium oxysporum f. sp. vasinfectum
School, Centre or Institute School of Biological Sciences
Institution The University of Queensland
Publication date 2009-12
Thesis type PhD Thesis
Supervisor Dr Elizabeth Aitken
Dr Elizabeth Dann
Dr Linda Smith
Total pages 223
Total colour pages 6
Total black and white pages 217
Subjects 06 Biological Sciences
Abstract/Summary In previous studies silicon has been associated with reduced disease severity and incidence, the enhanced accumulation of phenolic compounds and lignin, and with changes in the defence-related enzyme activity and transcript abundance of defence and stress related genes. All of these aspects of plant defence were considered in this study on cotton infected with Fusarium oxysporum f. sp. vasinfectum (Fov), and the results obtained have greatly enhanced our understanding of the effects of silicon on this interaction. In all experiments conducted, defence responses were only significantly enhanced by silicon treatment following inoculation with Fov, strongly suggesting that silicon can prime defence responses in cotton infected with Fov. Sicot F-1 was the cultivar most resistant to Fov infection at the commencement of this research, whilst Sicot 189 was considered to have moderate resistance to the pathogen. Vascular discolouration was significantly reduced in the more resistant cultivar, Sicot F-1 following treatment with potassium silicate, compared to mock inoculated plants and inoculated plants treated with potassium sulphate or calcium sulphate. No significant differences between treatments were observed in the moderately resistant cultivar, Sicot 189, though further trials may need to be conducted to confirm this result. In both cultivars, silicon content was significantly greater in plants which had been treated regularly with liquid potassium silicate, rather than with calcium silicate powder. Histological investigation of cotton infected with Fov, with and without silicon treatment, was conducted to ascertain the effects of this element on the accumulation of fungitoxic phenolic compounds, cell ultrastructural changes and fungal infection structures. Fov proliferated through the cortex and stele of plants from both the resistant (Sicot F-1), and moderately resistant (Sicot 189) cultivars, regardless of silicon treatment. However, defences were more rapidly and intensely induced in endodermal and vascular regions of inoculated, potassium silicate treated Sicot F-1 plants. Significantly more phenolic compounds were present at seven days post infection (dpi) in root extracts of inoculated, potassium silicate treated Sicot F-1 plants. Phenolic compounds were not significantly increased in inoculated, potassium silicate treated root extracts of Sicot 189 plants at three or seven dpi. Lignin assays demonstrated that the dry weight percentage of lignin in root material from inoculated, potassium silicate treated Sicot F-1 plants was significantly higher than that of extracts from inoculated plants not receiving silicon treatment at three dpi. This trend was also observed at seven dpi; however lignin content was not significantly different in this case. Percentage lignin content in the roots of Sicot 189 plants was not significantly different between inoculated potassium silicate treated plants and those not treated with silicon. Histological alterations were not observed in mock inoculated water or potassium silicate treated plants, nor were any significant increases in phenolic compounds or lignin accumulation detected in control treatments not inoculated with the pathogen. The expression of several defence related genes was assessed with quantitative reverse transcriptase real-time polymerase chain reaction. The results obtained verify that potassium silicate can enhance defence responses in Sicot 189 and Sicot F-1 plants inoculated with Fov, with silicon having a more pronounced effect on the more resistant cultivar, Sicot F-1. Genes upregulated at three and four dpi in potassium silicate treated, Fov inoculated Sicot F-1 plants included peroxidase, cadinene synthase and polygalacturonase inhibiting protein (PGIP), with peroxidase associated with phenol oxidation and lignification and cadinene synthase with phytoalexin biosynthesis. Osmotin-like protein and chitinase class I were consistently upregulated in potassium silicate treated, inoculated Sicot 189 plants; both genes coding for pathogenesis related (PR) proteins, with chitinase also classified as an antifungal protein. In both cultivars, silicon treatment without Fov inoculation did not result in the significant up-regulation of any of the defence genes assessed, providing further evidence for the role of silicon in priming in this interaction. The activities of three defence related enzymes, peroxidase, chitinase and β-1, 3- glucanase was assessed in root and shoot material by colourimetric assays. Regular application of potassium silicate significantly increased the activity of peroxidase in root extracts from the highly resistant cultivar Sicot F-1, at three, four and seven dpi with Fov, and in root extracts from the moderately resistant Sicot 189 at three and four dpi. Significant increases in chitinase activity in inoculated, silicon treated Sicot 189 plants were observed in root extracts at three dpi, and in shoot extracts at four dpi. Soluble potassium silicate treatment resulted in significant increases in β-1, 3- glucanase activity in Sicot 189 root extracts at four dpi. Few significant differences between treatments in terms of chitinase and β-1, 3- glucanase activity were detected in Sicot F-1 plants, though higher levels of each of these enzymes were present in root and shoot extracts from this cultivar. In this study the effects of acibenzolar-S-methyl, applied in the form of Bion®, on defence gene expression and enzyme activity in cotton infected with Fov were more pronounced in plants cultivated from treated seed, rather than in plants treated via foliar spray; a finding which is particularly relevant to the industry presently. Significant up-regulation of chitinase class I, peroxidase, and β-1, 3-glucanase transcripts and enzyme activities occurred in the Bion® seed soak treatment with Fov inoculation compared to all other treatments. It was possible to compare the actions of silicon with those of Bion® in this study. Bion® primed defence responses in cotton infected with Fov, in a manner similar to that observed in silicon treated cotton. The use of silicon and Bion® treatments, both alone and in combination as part of integrated disease management programmes, may potentially contribute to increased protection against this pathogen in Australian cotton fields in the future.
Keyword Cotton
Silicon
Fusarium oxysporum f. sp. vasinfectum
Defence responses
Vascular discolouration
Transmission electron microscopy
Phenolics
Lignin
Quantitative reverse transcriptase real-time polymerase chain reaction (QRT-PCR)
Additional Notes Colour pages - 27, 30, 31, 62, 103, 107 Landscape pages - 71, 154-156, 159-161, 164-166

 
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Created: Wed, 26 May 2010, 08:40:31 EST by Miss Jennifer Whan on behalf of Library - Information Access Service