Parwinder Kaur
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The UWA Institute of Agriculture Parwinder Kaur School of Plant Biology Born and educated in India, Parwinder wanted to do something for the farming industry she completed her Bachelor of Agricultural Science with Honors in Plant protection, at Punjab Agricultural University in India followed by a Master’s in Entomology. After completing her Master's degree she looked for hands-on experience in the plant pathology area, as plants have an equal threat from pathogens as they have from insect-pests. Parwinder won an international postgraduate research scholarship to undertake her PhD at UWA, looking into mechanisms of host resistance in Brassica juncea to an oomycete pathogen Albugo candida by understanding host-pathogen interaction using proteome and transcriptome analysis.
Transcript of Parwinder Kaur
The UWA Institute of
Agriculture
Parwinder KaurSchool of Plant Biology
Born and educated in India, Parwinder wanted to do
something for the farming industry she completed her
Bachelor of Agricultural Science with Honors in Plant
protection, at Punjab Agricultural University in India followed
by a Master ’s in Entomology. After completing her Master's
degree she looked for hands-on experience in the plant
pathology area, as plants have an equal threat from
pathogens as they have from insect-pests. Parwinder won
an international postgraduate research scholarship to
undertake her PhD at UWA, looking into mechanisms of host
resistance in Brassica juncea to an oomycete pathogen
Albugo candida by understanding host-pathogen interaction
using proteome and transcriptome analysis.
The UWA Institute of
Agriculture
The UWA Institute of
Agriculture
WHITE RUST DISEASE
Racial status & mechanisms of host resistance
Parwinder KaurParwinder Kaur
Supervisors:Prof. Martin Barbetti and Prof. K. Sivasithamparam Prof. Martin Barbetti and Prof. K. Sivasithamparam School of Plant Biology,School of Plant Biology,FNASFNAS
The UWA Institute of
Agriculture
Outline
General introduction
Objective of PhD study
Racial status of Albugo candida in WA
Relation among the different pathotypes present in WA
Mechanism of host resistance studies
Significance of results
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Agriculture
Brassica juncea extending oilseed production to lower rainfall regions
Better adapted than Canola (Brassica napus) to hotter and drier areas
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Agriculture
Local infectionsLocal infections Systemic infectionsSystemic infections
White rust caused by Albugo candida
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Agriculture
Objectives
Two main objectives of the PhD studies were to define:
1. Racial status of Albugo candida in WA
2. Mechanism of host-resistance
Proteome analysis
qRT-PCR
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Agriculture
Racial status Racial status
For different strains of A. candida available in WA
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Agriculture
Sample collection
Nine isolates of A. candida from different hosts collected during field surveys in WA from 2007-2009 were used in this study:
1. Brassica juncea (Mustard)2. B. rapa3. B. oleracea,4. B. tournefortii5. Raphanus raphanistrum (Wild Radish)6. R. sativa (Radish)7. Eruca sativa8. Capsella bursa-pastoris (Shepherd’s purse)9. Sisymbrium irio (London rocket)
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Morphological trials
A total of 16 cruciferous differentials were used to characterize the pathogenic behaviour of A. candida
On the basis of the occurrence of white rust
disease in WA
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Reactions of different cruciferous host differentials to Western Australian isolates of Albugo candida obtained presented with the mean disease severity in parenthesis.
Race 2V in
Australia
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Agriculture
Brassica oleracea isolate
Raphanus sativus isolate
Raphanus raphanistrum isolate
Host specificity and variability in virulence of Albugo candida isolates from 9 different host species in Western Australia
Brassica juncea isolate
B. juncea cv. Vulcan
B. napus cv. FAN 189
B. nigra
B. juncea cv. Commercial Brown
Capsella bursa-pastoris isolate
B. juncea cv. Vulcan
Sisymbrium irio isolate
Brassica rapa isolate
Eruca vesicaria isolate
Brassica tournefortii isolate
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DNA Extraction
Nucleotide Sequencing
Phylogenetic Analysis
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AY929824 Albugo amaranthi0.05
WA isolates
Other Australian
isolates
Other Continents
isolates
Brassica tournefortii isolate formed a separate clade
Wild radish and Radish isolates showed least similarity
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Agriculture
Racial status of A. candida in WA
Race 1 Radish
Race 2V Brassica juncea (Mustard)
Race 4 Capsella-bursa-pastoris (Shepherd’s purse)
Race 5 ? Sisymbrium officinale / Sisymbrium irio
Race 7A or 7V ? Brassica rapa
Race 9 Brassica oleracea
Race ?? Brassica tournefortii
Race ?? Eruca sativa
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ProteomicsProteomics
To define overall protein expression profiles
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A highly susceptible and resistant variety of B. juncea was selected
Screening trials
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10 μL drop of zoosporangial
suspension
Inoculation technique
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Samples were collected as follows
Sc S Rc R
2 hrs
4 hrs
8 hrs
24 hrs
72 hrs
Sc: Susceptible cultivar inoculated with DI water only
Rc: Resistant cultivar inoculated with DI water only
S: Susceptible cultivar inoculated with A. candida
R: Resistant cultivar inoculated with A. candida
Sample collection
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Proteomics: Method overview
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Agriculture
Image analysis: Comparison of overall protein expression profiles
Resistant Susceptible
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Agriculture
Proteomics: Method overview
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Agriculture
Proteins of interest
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From Proteome to GenomeFrom Proteome to Genome
Real Time Real Time -- Reverse Transcriptase (RT)PCRReverse Transcriptase (RT)PCR
For absolute quantification of For absolute quantification of proteins to support proteomics studyproteins to support proteomics study
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Sporangium
Zoospore
germ‐tube Haustorium
Extrahustorial matrix
Nucleus
GENE ACTIVATION
PROTEIN SYNTHESIS
ROS
Protective measures for plant metabolism
Defense responses
Intercellular signalling
Intercellular signalling
O2
RCCR
RCC pFCC
H2O2
SODs
catalase
Pathogen growth
PR‐5
Redox regulationGST
CSase
Pathogen
PPIase
Major elicitor-induced changes during the interaction of Brassica juncea and Albugo candida
ROSmRNA
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Publications arising from PhD studies
Scientific papers
1. Kaur Parwinder, M. J. Barbetti and K. Sivasithamparam (2008) Pathogenic behaviour of strains of Albugo candida from Brassica juncea and Raphanus raphanistrum (Wild Radish) in Western Australia. Australasian Plant Pathology 37: 353-356.
2. Kaur Parwinder, C. X. Li and M. J. Barbetti, M. P. You, Hua Li and K. Sivasithamparam (2008) First Report of Powdery Mildew Caused by Erysiphe cruciferarum Opiz ex Junnel, on Brassica juncea (L.) Czern & Coss in Australia. Plant Disease (American Phytopathological Society) 92: 650.
3. Kaur Parwinder, Hua Li, K. Sivasithamparam and M. J. Barbetti (2010) Pre-Inoculation with Hyaloperanosporaparasitica reduces incubation time and increases severity of disease caused by Albugo candida in a Brassica juncea variety resistant to Downy mildew. Plant Disease (under review).
4. Kaur Parwinder, Ricarda Jost, M. J. Barbetti and K. Sivasithamparam (2010) Differential expression of proteins following challenge by Albugo candida in varieties of Brassica juncea resistant or susceptible to white rust. Plant Physiology (under preparation).
5. Kaur Parwinder, M. J. Barbetti and K. Sivasithamparam (2010) Racial/Pathotype status and phylogenetic relationships of Albugo candida occurring on cruciferous hosts in Western Australia. Fungal Biodiversity (under preparation).
6. Kaur Parwinder, M. J. Barbetti and K. Sivasithamparam (2010) Site of inoculation and stage of plant development determine symptom type and expression in Brassica juncea following infection with Albugo candida. Journal of Phytopathology (under preparation).
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Conference papers
1. Kaur Parwinder, M. J. Barbetti and K. Sivasithamparam (2010) Host-pathogen interactions in the Mustard-White rust pathosystem: Protein expression profiling. Proceedings: APS Annual Meeting. Abstract (accepted).
2. Kaur Parwinder, M. J. Barbetti and K. Sivasithamparam (2009) Racial status of Albugo candida in Australia and India. Proceedings: 5th International Congress on “Plant Pathology in the Globalized Era”. Abstract.
3. Kaur Parwinder, M. J. Barbetti and K. Sivasithamparam (2008) Response of Brassica juncea genotypes to races of Albugo candida in Western Australia. Proceedings: 9th International Congress of Plant Pathology (ICPP 2008). Abstract.
Publications arising from PhD studies
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Agriculture
Acknowledgements
Funding
International Postgraduate Research Scholarship (IPRS)University International Stipend (UIS)UWA Grants for Research Student Training (GRST)Convocation travelling fellowshipMike Carroll travelling fellowship
Supervision
Prof. Martin BarbettiProf. K. Sivasithamparam
Experimental guidance and technical support
Dr. Ricarda Jost Dr. Hua Li Dr. Roberta Marra Dr. Caixia Li
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Agriculture
Many thanks to School of Plant Biology & UWA for all the wonderful
opportunities & great time
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Agriculture
Major findings
Defined the racial status of Albugo candida in WA
Understanding of the host-pathogen interaction at cellular level which can be effectively used to design genetic markers to screen for the resistance in the B. juncea germplasm
Opened many novel avenues for engineering durable resistance in plants
THANK THANK YOU all for listening !!!YOU all for listening !!!
