A hydroponics based high throughput screening system for Phytophthora root rot resistance in chickpea (Cicer arietinum L.)

Abstract Background Phytophthora root rot (PRR) caused by P. medicaginis is a major soil borne disease in chickpea growing regions of Australia. Sources of resistance have been identified in both cultivated and wild Cicer species. However, the molecular basis underlying PRR resistance is not known....

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Main Authors: Amritha Amalraj, Julian Taylor, Tim Sutton
Format: Article
Language:English
Published: BMC 2019-07-01
Series:Plant Methods
Subjects:
Online Access:http://link.springer.com/article/10.1186/s13007-019-0463-3
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author Amritha Amalraj
Julian Taylor
Tim Sutton
author_facet Amritha Amalraj
Julian Taylor
Tim Sutton
author_sort Amritha Amalraj
collection DOAJ
description Abstract Background Phytophthora root rot (PRR) caused by P. medicaginis is a major soil borne disease in chickpea growing regions of Australia. Sources of resistance have been identified in both cultivated and wild Cicer species. However, the molecular basis underlying PRR resistance is not known. Current phenotyping methods rely on mycelium slurry or oospore inoculum. Sensitive and reliable methods are desirable to study variation for PRR resistance in chickpea and allow for a controlled inoculation process to better capture early defence responses following PRR infection. Results In this study, a procedure for P. medicaginis zoospore production was standardized and used as the inoculum to develop a hydroponics based in planta infection method to screen chickpea genotypes with established levels of PRR resistance. The efficiency of the system was both qualitatively validated based on observation of characteristic PRR symptom development, and quantitatively validated based on the amount of pathogen DNA in roots. This system was scaled up to screen two biparental mapping populations previously developed for PRR studies. For each of the screenings, plant survival time was measured after inoculation and used to derive Kaplan–Meier estimates of plant survival (KME-survival). KME-survival and canker length were then selected as phenotypic traits associated with PRR resistance. Genetic analysis of these traits was conducted which identified quantitative trait loci (QTL). Additionally, these hydroponic traits and a set of previously published plant survival traits obtained from multiple PRR field experiments were combined in a model-based correlation analysis. The results suggest that the underlying genetic basis for plant survival during PRR infection within hydroponics and field disease environments is linked. The QTL QRBprrkms03 and QRBprrck03 on chromosome 4 identified for the traits KME-survival and canker length, respectively, correspond to the same region reported for PRR resistance in a field disease experiment. Conclusion A hydroponics based screening system will facilitate reliable and rapid screening in both small- and large-scale experiments to study PRR disease in chickpea. It can be applied in chickpea breeding programs to screen for PRR resistance and classify the virulence of new and existing P. medicaginis isolates.
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spelling doaj.art-3b8140bc47e64a7c8ceca782c4c650b72022-12-22T00:20:19ZengBMCPlant Methods1746-48112019-07-0115111510.1186/s13007-019-0463-3A hydroponics based high throughput screening system for Phytophthora root rot resistance in chickpea (Cicer arietinum L.)Amritha Amalraj0Julian Taylor1Tim Sutton2School of Agriculture Food and Wine, University of AdelaideSchool of Agriculture Food and Wine, University of AdelaideSchool of Agriculture Food and Wine, University of AdelaideAbstract Background Phytophthora root rot (PRR) caused by P. medicaginis is a major soil borne disease in chickpea growing regions of Australia. Sources of resistance have been identified in both cultivated and wild Cicer species. However, the molecular basis underlying PRR resistance is not known. Current phenotyping methods rely on mycelium slurry or oospore inoculum. Sensitive and reliable methods are desirable to study variation for PRR resistance in chickpea and allow for a controlled inoculation process to better capture early defence responses following PRR infection. Results In this study, a procedure for P. medicaginis zoospore production was standardized and used as the inoculum to develop a hydroponics based in planta infection method to screen chickpea genotypes with established levels of PRR resistance. The efficiency of the system was both qualitatively validated based on observation of characteristic PRR symptom development, and quantitatively validated based on the amount of pathogen DNA in roots. This system was scaled up to screen two biparental mapping populations previously developed for PRR studies. For each of the screenings, plant survival time was measured after inoculation and used to derive Kaplan–Meier estimates of plant survival (KME-survival). KME-survival and canker length were then selected as phenotypic traits associated with PRR resistance. Genetic analysis of these traits was conducted which identified quantitative trait loci (QTL). Additionally, these hydroponic traits and a set of previously published plant survival traits obtained from multiple PRR field experiments were combined in a model-based correlation analysis. The results suggest that the underlying genetic basis for plant survival during PRR infection within hydroponics and field disease environments is linked. The QTL QRBprrkms03 and QRBprrck03 on chromosome 4 identified for the traits KME-survival and canker length, respectively, correspond to the same region reported for PRR resistance in a field disease experiment. Conclusion A hydroponics based screening system will facilitate reliable and rapid screening in both small- and large-scale experiments to study PRR disease in chickpea. It can be applied in chickpea breeding programs to screen for PRR resistance and classify the virulence of new and existing P. medicaginis isolates.http://link.springer.com/article/10.1186/s13007-019-0463-3PRR resistance in chickpeaP. medicaginis zoospores productionPRR phenotyping methodHydroponics screening systemPlant survival traitsKaplan–Meier (KM) estimates of survival probability
spellingShingle Amritha Amalraj
Julian Taylor
Tim Sutton
A hydroponics based high throughput screening system for Phytophthora root rot resistance in chickpea (Cicer arietinum L.)
Plant Methods
PRR resistance in chickpea
P. medicaginis zoospores production
PRR phenotyping method
Hydroponics screening system
Plant survival traits
Kaplan–Meier (KM) estimates of survival probability
title A hydroponics based high throughput screening system for Phytophthora root rot resistance in chickpea (Cicer arietinum L.)
title_full A hydroponics based high throughput screening system for Phytophthora root rot resistance in chickpea (Cicer arietinum L.)
title_fullStr A hydroponics based high throughput screening system for Phytophthora root rot resistance in chickpea (Cicer arietinum L.)
title_full_unstemmed A hydroponics based high throughput screening system for Phytophthora root rot resistance in chickpea (Cicer arietinum L.)
title_short A hydroponics based high throughput screening system for Phytophthora root rot resistance in chickpea (Cicer arietinum L.)
title_sort hydroponics based high throughput screening system for phytophthora root rot resistance in chickpea cicer arietinum l
topic PRR resistance in chickpea
P. medicaginis zoospores production
PRR phenotyping method
Hydroponics screening system
Plant survival traits
Kaplan–Meier (KM) estimates of survival probability
url http://link.springer.com/article/10.1186/s13007-019-0463-3
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