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. The UWA Institute of Agriculture The UWA Institute of Agriculture
WHITE RUST DISEASE Racial status & mechanisms of host resistance
Parwinder Kaur
Supervisors: Prof. Martin Barbetti and Prof. K. Sivasithamparam School of Plant Biology, FNAS The UWA Institute of Agriculture Outline
¾ General introduction ¾ Objective of PhD study ¾ Racial status of Albugo candida in WA ¾ Relation among the different pathotypes present in WA ¾ Mechanism of host resistance studies ¾ Significance of results
The UWA Institute of Agriculture Brassica juncea extending oilseed production to lower rainfall regions
Better adapted than Canola (Brassica napus) to hotter and drier areas
The UWA Institute of Agriculture White rust caused by Albugo candida
Local infections Systemic infections The UWA Institute of 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 ¾ Proteome analysis ¾ qRT-PCR
The UWA Institute of Agriculture Racial status
For different strains of A. candida available in WA
The UWA Institute of 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. rapa 3. B. oleracea, 4. B. tournefortii 5. Raphanus raphanistrum (Wild Radish) 6. R. sativa (Radish) 7. Eruca sativa 8. Capsella bursa-pastoris (Shepherd’s purse) 9. Sisymbrium irio (London rocket)
The UWA Institute of Agriculture 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
The UWA Institute of Agriculture 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
The UWA Institute of Agriculture Host specificity and variability in virulence of Albugo candida isolates from 9 different host species in Western Australia
Brassica oleracea Raphanus sativus isolate isolate
B. juncea cv. B. juncea cv. Vulcan Vulcan
B. nigra Capsella bursa-pastoris Raphanus Brassica B. napus cv. isolate FAN 189 raphanistrum isolate juncea isolate B. juncea cv. Commercial Brown
Brassica Brassica rapa Sisymbrium irio tournefortii isolate isolate isolate Eruca vesicaria isolate
The UWA Institute of Agriculture DNA Extraction
Nucleotide Sequencing
Phylogenetic Analysis
The UWA Institute of Agriculture AY929824 Albugo amaranthi 0.05
Other Australian isolates Wild radish and Radish isolates showed least similarity
Other Continents isolates
WA isolates
Brassica tournefortii isolate formed a separate clade
The UWA Institute of 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
The UWA Institute of Agriculture Proteomics
To define overall protein expression profiles
The UWA Institute of Agriculture Screening trials
A highly susceptible and resistant variety of B. juncea was selected
The UWA Institute of Agriculture Inoculation technique
10 μL drop of zoosporangial suspension
The UWA Institute of Agriculture Sample collection
Samples were collected as follows
Sc S Rc R 2 hrs 3333 4 hrs 3333 8 hrs 3333 24 hrs 3333 72 hrs 3333
S: Susceptible cultivar inoculated with A. candida Sc: Susceptible cultivar inoculated with DI water only R: Resistant cultivar inoculated with A. candida Rc: Resistant cultivar inoculated with DI water only
The UWA Institute of Agriculture Proteomics: Method overview
The UWA Institute of Agriculture Image analysis: Comparison of overall protein expression profiles
Resistant Susceptible
The UWA Institute of Agriculture Proteomics: Method overview
The UWA Institute of Agriculture Proteins of interest
The UWA Institute of Agriculture From Proteome to Genome
For absolute quantification of proteins to support proteomics study
Real Time - Reverse Transcriptase (RT)PCRThe UWA Institute of Agriculture Zoospore
Sporangium germ‐tube Haustorium
Extrahustorial matrix GENE ACTIVATION
Intercellular Nucleus signalling
mRNA ROS PR‐5 PROTEIN SYNTHESIS Intercellular signalling Defense responses Protective measures Pathogen growth for plant metabolism ROS O2 GST H O Redox regulation 2 2 CSase PPIase RCCR catalase
RCC pFCC SODs
Pathogen The UWA Institute of Major elicitor-induced changes during the interaction of Brassica juncea and Albugo candidaAgriculture 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 Hyaloperanospora parasitica 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). The UWA Institute of Agriculture Publications arising from PhD studies
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.
The UWA Institute of Agriculture Acknowledgements
Funding
International Postgraduate Research Scholarship (IPRS) University International Stipend (UIS) UWA Grants for Research Student Training (GRST) Convocation travelling fellowship Mike Carroll travelling fellowship
Supervision
Prof. Martin Barbetti Prof. K. Sivasithamparam
Experimental guidance and technical support Dr. Ricarda Jost Dr. Hua Li Dr. Roberta Marra
Dr. Caixia Li The UWA Institute of Agriculture Many thanks to School of Plant Biology & UWA for all the wonderful opportunities & great time
The UWA Institute of Agriculture Major findings
¾ Defined the racial status of Albugo candida in WA
¾ 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 YOU all for listening !!! The UWA Institute of Agriculture