quantitative pcr and histopathological … pcr and histopathological assessment of cereal infection...
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![Page 1: Quantitative PCR and histopathological … PCR and histopathological assessment of cereal infection by Fusarium pseudograminearum. ... Solophenyl flavine](https://reader033.vdocuments.us/reader033/viewer/2022052711/5ad31d6f7f8b9afa798d7f66/html5/thumbnails/1.jpg)
Quantitative PCR and histopathological assessment of
cereal infection by Fusarium pseudograminearum.
Supervisors: Prof. Mark Sutherland, Dr. Anke Martin and Dr. Damian Herde
Noel Knight
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Crown Rot• Crown rot (F. pseudograminearum)
~ $100 million per annum
Aims
25um
• Examine cereal tissues affected by Fp crown rot using visual and qPCR rating methods
• Microscopically observe Fp growth through cereal tissues
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• Resistance assessed visually– Brown discolouration of internodes
• qPCR assay amplifies Fp Translation Elongation Factor α DNA
Determining Host Resistance
Internodes
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Crown Rot of Adult Plants
Genotype Species Crown Rot Resistance2-49 T.aestivum Partially Resistant
Grimmett H. vulgare Moderately SusceptiblePuseas T.aestivum SusceptibleBellaroi T.turgidum durum Very Susceptible
• 16 and 22 weeks after planting (WAP)
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Crown Rot of Adult Plants16 WAPAnthesis
2-49
Grimmett
Puseas
BellaroiFp
DN
A (n
g) /
gFp
DN
A (n
g) /
g
Pearson’s r0.86
0.5822 WAPMaturity
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Stem Tissue Types
Parenchymatous hypoderm
50µm 50µmEpidermis
Sclerenchymatous hypoderm
Pith parenchyma
Large vascular bundle
Small vascular bundle
Vascular parenchyma
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Adult Tissue Microscopy
25µm50µm
25µm25µm
Knight and Sutherland (2011) Plant Pathology
Safranin
Solophenyl flavine
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Adult Tissue Microscopy
25µm
25µm
25µm50µm
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Adult Tissue Microscopy
400µm 400µm
h
pc h
25µm
st
50µm 25µm50µm
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• Strong correlation between IN discolouration and Fp DNA at 16 WAP
• Significant differences in Fp DNA quantity between R and S cereals at 16 WAP
• Fp colonises all cell types– vascular tissues colonised
Summary
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Questions?