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Identification of I-7 expands the repertoire of genes for resistance to Fusarium wilt in tomato to three resistance gene classes

Gonzalez-Cendales, Yvonne ; Catanzariti, Ann-Maree ; Baker, Barbara ; Mcgrath, Des J. ; Jones, David A.

Molecular plant pathology, 2016-04, Vol.17 (3), p.448-463 [Periódico revisado por pares]

England: Blackwell Publishing Ltd

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  • Título:
    Identification of I-7 expands the repertoire of genes for resistance to Fusarium wilt in tomato to three resistance gene classes
  • Autor: Gonzalez-Cendales, Yvonne ; Catanzariti, Ann-Maree ; Baker, Barbara ; Mcgrath, Des J. ; Jones, David A.
  • Assuntos: Alleles ; Amino Acid Sequence ; Chromosomes, Plant - genetics ; Conserved Sequence - genetics ; Disease Resistance - genetics ; Fusarium - physiology ; Fusarium oxysporum ; Fusarium oxysporum f. sp. lycopersici ; Fusarium wilt ; Genes ; Genes, Plant ; leucine-rich repeat ; Lycopersicon esculentum ; Lycopersicon esculentum - genetics ; Lycopersicon esculentum - microbiology ; Multigene Family ; Original ; plant disease resistance gene ; Plant Diseases - genetics ; Plant Diseases - microbiology ; Plant Proteins - chemistry ; Plant Proteins - genetics ; Plant Proteins - metabolism ; Polymorphism, Single Nucleotide - genetics ; receptor-like protein ; Sequence Analysis, RNA ; Solanum ; Solanum lycopersicum ; Solanum pennellii
  • É parte de: Molecular plant pathology, 2016-04, Vol.17 (3), p.448-463
  • Notas: Australian Research Council Linkage Project grant - No. LP100100172
    Australian Research Council postdoctoral fellowship - No. DP1095157
    ark:/67375/WNG-KSC1Q96W-6
    Australian Postgraduate Award (Industry) scholarship
    Fig. S1 Marker analysis on Tristar, M82 and 15 homozygous-resistant and 16 homozygous-susceptible Tristar × M82 F3 or F4 lines using CAPS7774 primers (Table S2) targeting the Solyc08g077740 gene. Polymerase chain reaction (PCR) amplification generates an 808-bp product with all genomic DNA templates (not shown) and, after digestion with AgeI, Tristar shows digested products of 612 and 196 bp, whereas M82 products remain undigested. Resistant lines all show the Tristar banding pattern, whereas susceptible lines all show the M82 banding pattern, indicating complete linkage between I-7 and the CAPS7774 marker. Fig. S2 Polymerase chain reaction (PCR) screening of MM-7774Tristar T2 plants from two lines for the presence of the Solyc08g077740 transgene from Tristar using 7774F4 (Table S4) and M13 reverse primers to detect the transgene (top band) and CAPS7774 primers (Table S2) to detect the transgene and the endogenous Solyc08g077740 gene in transgenic plants, or the endogenous Solyc08g077740 gene alone in non-transgenic sibs (bottom band). Asterisks indicate plants carrying the transgene. Controls included a genomic DNA sample from Moneymaker (MM), a no tempate control (Water) and plasmid DNA containing the transgene (pL2-Tristar). Fig. S3 (A) Disease assays on T2 plants from two additional lines segregating for the Solyc08g077740 gene from Tristar in a Moneymaker (MM) background. T2 plants, together with Tristar (resistant) and Moneymaker (susceptible) control plants, were inoculated with Fol race 3. Photographs were taken at 21 days post-inoculation (dpi). (B) Disease scores for the plants shown in (A). Fig. S4 (A) Disease assays on MM-7774Tristar #1 T2 plants carrying the Solyc08g077740 gene from Tristar in a Moneymaker (MM) background. T2 plants, together with Tristar (resistant) and Moneymaker (susceptible) control plants, were inoculated with Fol race 1. Photographs were taken at 20 days post-inoculation (dpi). (B) Disease scores for the T2 plants shown in (A). Fig. S5 (A) Disease assays on MM-7774Tristar #1 T2 plants carrying the Solyc08g077740 gene from Tristar in a Moneymaker (MM) background. T2 plants, together with Tristar (resistant) and Moneymaker (susceptible) control plants, were inoculated with Fol race 2. Photographs were taken at 20 days post-inoculation (dpi). (B) Disease scores for the T2 plants shown in (A). Fig. S6 (A) Polymerase chain reaction (PCR) screening for the presence of EDS1 and the eds1 mutation in Tristar × sun1-1 F2 plants using the primers as described in the main text. Plants containing the EDS1 gene produce a PCR product of 644 bp, whereas plants carrying the eds1 mutation produce a PCR product of about 850 bp. The presence of the EDS1 gene is represented by a capital E and the presence of the eds1 allele by a lower case e. Plants heterozygous for eds1 (with an Ee genotype shown in white) and homozygous for eds1 (with an ee genotype shown in red) were inoculated with Fol race 3 and screened for the presence of I-7 as shown in (B). (B) PCR screening for the presence of I-7 among the Ee and ee plant DNA samples shown in (A) using the CAPS7774 primers (Table S2). I-7 generates 612- and 196-bp products after digestion with AgeI, whereas the i-7 allele from sun1-1 generates an 808-bp product that remains undigested. Plants carrying I-7 are indicated. In both (A) and (B), the PCR products amplified from genomic DNA of the parent lines (Tristar and sun1-1) are shown (boxed). Fig. S7 (A) Authentication of IL8-1, IL8-2 and IL8-3 lines using chromosome 8 cleaved amplified polymorphic sequence (CAPS) and sequence characterised amplified region (SCAR) markers diagnostic for each line (Table S5, see Supporting Information). (B) Analysis of the Tristar, M82, IL8-1, IL8-2 and IL8-3 lines using the CAPS7774 marker (Table S2), showing that IL8-2 carries the LA716 allele (i-7LA716) of the I-7 gene. (C) Disease assays on IL8-1, IL8-2 and IL8-3 plants inoculated with Fol race 3 or mock inoculated. Photographs were taken at 21 days post-inoculation (dpi). Fig. S8 Ligation-independent cloning (LIC) using the in-house binary vector pL2 derived from pGreenII showing: (A) steps in the LIC process; (B) a map of pL2.Table S1 RNA-seq output, mapping and single nucleotide polymorphism (SNP) analysis data Table S2 Cleaved amplified polymorphic sequence (CAPS) markers designed for genes within the cluster of Tristar single nucleotide polymorphisms (SNPs) on chromosome 8 Table S3 Genes present in the introgressed and neighbouring regions of Tristar chromosome 8. The introgressed region encompasses genes Solyc08g077520 to Solyc08g077800, genes which show a high frequency of single nucleotide polymorphisms (SNPs) unique to Tristar. Table S4 Primers used for amplification, cloning and sequencing of Solyc08g077740 genes from Tristar and M82. Table S5 Cleaved amplified polymorphic sequence (CAPS) and sequence characterised amplified region (SCAR) markers used to authenticate Solanum pennellii LA716 chromosome 8 introgression lines IL8-1, IL8-2 and IL8-3.
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  • Descrição: Summary The tomato I‐3 and I‐7 genes confer resistance to Fusarium oxysporum f. sp. lycopersici (Fol) race 3 and were introgressed into the cultivated tomato, Solanum lycopersicum, from the wild relative Solanum pennellii. I‐3 has been identified previously on chromosome 7 and encodes an S‐receptor‐like kinase, but little is known about I‐7. Molecular markers have been developed for the marker‐assisted breeding of I‐3, but none are available for I‐7. We used an RNA‐seq and single nucleotide polymorphism (SNP) analysis approach to map I‐7 to a small introgression of S. pennellii DNA (c. 210 kb) on chromosome 8, and identified I‐7 as a gene encoding a leucine‐rich repeat receptor‐like protein (LRR‐RLP), thereby expanding the repertoire of resistance protein classes conferring resistance to Fol. Using an eds1 mutant of tomato, we showed that I‐7, like many other LRR‐RLPs conferring pathogen resistance in tomato, is EDS1 (Enhanced Disease Susceptibility 1) dependent. Using transgenic tomato plants carrying only the I‐7 gene for Fol resistance, we found that I‐7 also confers resistance to Fol races 1 and 2. Given that Fol race 1 carries Avr1, resistance to Fol race 1 indicates that I‐7‐mediated resistance, unlike I‐2‐ or I‐3‐mediated resistance, is not suppressed by Avr1. This suggests that Avr1 is not a general suppressor of Fol resistance in tomato, leading us to hypothesize that Avr1 may be acting against an EDS1‐independent pathway for resistance activation. The identification of I‐7 has allowed us to develop molecular markers for marker‐assisted breeding of both genes currently known to confer Fol race 3 resistance (I‐3 and I‐7). Given that I‐7‐mediated resistance is not suppressed by Avr1, I‐7 may be a useful addition to I‐3 in the tomato breeder's toolbox.
  • Editor: England: Blackwell Publishing Ltd
  • Idioma: Inglês
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  • Realização: ABCD-USP USP   Logos de Redes Sociais: Freepik

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