Plant basal resistance - Universiteit Utrecht

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Chapter 2 were collected at 6 h (for SA) and 4 h (for JA or MeJA) after treatment. RNA extraction, RNA blotting, and labelling of specific probes for PR-1 and PDF1.2 were performed as previously described by Ton et al. (2002). Equal loading was verified by ethidium bromide staining of the gels. Gene expression assays by reverse transcription-quantitative PCR (RT-qPCR) Basal TF gene expression profiles in accessions were based on 3 biologically replicate samples, each consisting of 3 to 5 rosettes from 5-week-old plants. TF gene expression profiling of water- and BABA-treated Col-0 plants were based on 3 similar biologically replicate samples, collected at 2 days after soil-drench treatment of 4-week-old plants with water or 80 μM BABA. Analysis of PDF1.2 and VSP2 gene induction was based on 3 biologically replicate samples, each consisting of 6 leaves of similar age from 3 different plants of 5 weeks old, which were collected at the indicated time-points after spraying 0.01 % (v/v) Silwet L-77 solution with 0, 200, or 500 μM JA, or after mechanical wounding by forceps (1 wounding site per leaf), with or without 5 μL caterpillar regurgitant pipetted onto the wounded leaf areas. Gene expression analysis of RILs was performed by cultivating 15 – 18 plants of each RIL (maximally 20 RILs per screen) along with both parental accessions. Leaves of 4-week- old plants were sprayed with water, 200 uM JA, or 0.5 mM SA, each supplemented with 0.01% (v/v) Silwet L-77. At 4 h and 7 h after treatment, 3 biologically replicate samples, each consisting of 6 leaves from 3 different plants, were collected for analysis of PDF1.2 and PR-1 gene expression, respectively. RNA extraction, cDNA synthesis and qPCR reactions were performed as described by Van der Ent et al. (2009). Primers were similar as described previously (Czechowski et al., 2004; Czechowski et al., 2005), or designed using Primer3 software (http://frodo.wi.mit.edu/primer3/input.htm) with a T m between 60.5 and 62, and a product size
39 Genetic dissection of basal defence responsiveness one day later seedlings were treated with 0.01 mM flg22 or 0.01% chitosan. Cotyledons (8 to 15 from different plants) were collected at 24 h after PAMP treatment, stained with alinine blue, and quantified for callose intensity as described by (Luna et al., 2011). Cluster analysis of TF gene expression profiles TF gene profiles were analysed using TIGR Multi-experiment Viewer (tmev) software (Saeed et al., 2003). Analyses were based on the log-transformed values of the fold inductions of each gene, relative to the mean expression value of three independent, un-induced Col-0 samples. Differences in TF gene expression between accessions were tested for statistical significance using a Student’s t-tests, or a non-parametric Wilcoxon Mann-Whitney test when values did not follow normal distributions. QTL mapping analysis QTL mapping was performed with the Bur-0 x Col-0 core population from INRA Versailles Genomic Resource Centre (Bouchabke et al., 2008). This mapping population was genotyped with 87 molecular markers at an average genetic marker distance of 4.4 cM (~1.4 Mb) and has a global allelic equilibrium of 51.3 % of Col-0 and 48.7 % Bur-0 (http://dbsgap.versailles. inra.fr/vnat/Documentation/20/DOC.html). GenStat software (12 th edition) was used for analysis of genetic linkage. Gene expression values (2 ΔCt ), callose intensities, and rosette diameters for each RIL were standardised to corresponding average values from the parental accessions in each screen and up-loaded as phenotypic data. After calculation of the genetic predictors, an initial genome scan produced candidate QTL positions by simple interval mapping, which were used as cofactors in a subsequent genome scan by composite interval mapping. A logaritham of odds (LOD) score of 2.94 was used as threshold of significance, corresponding to a genome-wide significance of P = 0.05 for normally distributed data. Comparison of Col-0 and Bur-0 genomic sequences Genomic polymorphisms between Col-0 and Bur-0 were based on the fully sequenced genome of accession Bur-0 (Ossowski et al., 2008) and visualized using polymorph (http:// polymorph.weigelworld.org/). RESULTS Natural variation in JA-induced PDF1.2 expression is associated with basal resistance against the nectrophic fungus Plectosphaerella cucumerina and the generalist herbivore Spodoptera littoralis Six Arabidopsis accessions were selected on the basis of previously reported natural variation in basal resistance (Supplementary information Table S1). To test the response of
Page 1 and 2: Plant basal resistance: genetics, b
Page 3 and 4: Most of the research described in t
Page 5: Promotor: Prof. dr. Ir. C.M.J. Piet
Page 10 and 11: 10 CHAPTER 1 General Introduction A
Page 12 and 13: Chapter 1 (PRRs; Gomez-Gomez and Bo
Page 14 and 15: Chapter 1 by avirulent pathogens (R
Page 16 and 17: Chapter 1 Selective non-pathogenic
Page 18 and 19: Chapter 1 an endogenous plant signa
Page 20 and 21: Chapter 1 between defence signallin
Page 22 and 23: Chapter 1 rarely provides complete
Page 24 and 25: Chapter 1 genetic resources for Ara
Page 26 and 27: Chapter 1 metabolites are biosynthe
Page 28 and 29: Chapter 1 constitutively produced i
Page 30: Chapter 1 latest insights. This Cha
Page 33 and 34: ABSTRACT 33 Genetic dissection of b
Page 35 and 36: 35 Genetic dissection of basal defe
Page 37: 37 Genetic dissection of basal defe
Page 45 and 46: Natural variation in responsiveness
Page 57: 57 Genetic dissection of basal defe
Page 61 and 62: ABSTRACT 61 Role of benzoxazinoids
Page 63 and 64: 63 Role of benzoxazinoids in maize
Page 67 and 68: Chemical treatments and callose qua
Page 69 and 70: Aphid infestation stimulates apopla
Page 81 and 82: Table S1: Primers used for RT-qPCR
Page 86 and 87: 86 CHAPTER 4 Benzoxazinoids in root
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Chapter 4 INTRODUCTION Plants have
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Chapter 4 Our study presents eviden
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Chapter 4 Maize - P. putida FBC004
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Chapter 4 P. putida is tolerant to
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Chapter 4 Impact of DIMBOA on the P
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Chapter 4 PP5286 dut deoxyuridine 5
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Chapter 4 DIMBOA-producing wild-typ
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Chapter 4 DISCUSSION The rhizospher
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Chapter 4 patterns, we considered t
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108 CHAPTER 5 General Discussion
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Chapter 5 QTL was caused by a polym
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Chapter 5 Figure 1: CYP81D11 regula
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Chapter 5 et al., 2009), and allele
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Chapter 5 defence against different
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Chapter 5 occurred at time-points l
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Chapter 5 rhizospheric signals that
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Chapter 5 glucosinolate profiles. 1
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References References Abramovitch R
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References QTL mapping and characte
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References Gianoli E, Niemeyer HM (
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References Kliebenstein DJ, Kroyman
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References Studies in Natural Produ
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References Geniostoma antherotrichu
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References Todesco M, Balasubramani
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References Walters DR, Paterson L,
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Summary Summary Basal resistance in
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Samenvatting Samenvatting Basale re
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Samenvatting inzichten opgeleverd i
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Acknowledgments Acknowledgements In
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Curriculum vitae Shakoor was born o
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