SuperSAGE revealed different classes of early resistance response genes in Capsicum chinense plants harboring L3-resistance gene infected with Pepper mild mottle virus
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We used SuperSAGE, an improved version of serial analysis of gene expression, to explore transcriptome changes early in the L3-mediated resistance response of pepper plants against a tobamovirus. Capsicum chinense plants homozygous for the L3 resistance gene were infected with virulent and avirulent strains of Pepper mild mottle virus (PMMoV). Plants were maintained at a temperature nonpermissive for the resistance gene to allow the viruses to spread, then transferred to a permissive temperature for 3 h and subsequently analyzed. In the incompatible reaction, we selected 152 SuperSAGE tags (each 26 nucleotides long) possibly corresponding to upregulated genes, and 84 tags for downregulated genes. Approximately 70% of tags had matching ESTs in the genus Capsicum, other genera within the Solanaceae and/or other families of plants. More than 90% of tags with EST matches could be annotated with either functionally characterized or uncharacterized proteins. We compared genes annotated by SuperSAGE tags and those annotated by partial cDNA that was obtained using the SuperSAGE tag sequences as rapid amplification of the cDNA ends-PCR primers. Of genes annotated by SuperSAGE tags, c. 90% were consistent with those annotated by longer cDNA sequences. We cloned 17 full-length cDNAs from different SuperSAGE tags and confirmed that these genes were upregulated during normal infection in the incompatible interaction. We identified several early resistance response genes including a Ran/TC4 protein and a β-oxidation multifunctional protein, indicating that SuperSAGE is a powerful tool for investigating plant–pathogen interactions.
KeywordsEarly response L3 PMMoV Resistance SuperSAGE Transcriptome
We thank Dr. K. W. Kinzler for SAGE2000 software, Harumi Takahashi and Kazue Obara for technical assistance and all members of IBRC for fruitful discussion. This study was supported in part by the Iwate Prefecture Government and in part by a grant-in-aid for Scientific Research (C) (18580047) from the Japan Society for the Promotion of Science.
- Boukema IW (1982) Resistance to a new strain of TMV in Capsicum chacoense Hunz. Capsicum Newsl 1:49–51Google Scholar
- Boukema IW (1984) Resistance to TMV in Capsicum chacoense Hunz. is governed by allele of the L-locus. Capsicum Newsl 3:47–48Google Scholar
- Dellagi A, Birch PRJ, Heilbronn J, Avrova AO, Montesano M, Palva ET, Lyon GD (2000) A potato gene, erg–1, is rapidly induced by Erwinia carotovora ssp. atroseptica, Phytophthora infestans, ethylene and salicylic acid. J Plant Physiol 157:201–205Google Scholar
- Farmer EE (2000) Adding injury to insult: pathogen detection and responses. Genome Biol 1:1012.1–1012.3Google Scholar
- García-Luque I, Ferrero ML, Rodríguez JM, Alonso E, de la Cruz A, Sanz AI, Vaquero C, Serra MT, Díaz-Ruíz JR (1993) The nucleotide sequence of the coat protein genes and 3′ non-coding regions of two resistance-breaking tobamoviruses in pepper shows that they are different viruses. Arch Virol 131:75–88PubMedCrossRefGoogle Scholar
- Gilroy EM, Hein I, van der Hoorn R, Boevink PC, Venter E, McLellan H, Kaffarnik F, Hrubikova K, Shaw J, Holeva M, López EC, Borras-Hidalgo O, Pritchard L, Loake GJ, Lacomme C, Birch PRJ (2007) Involvement of cathepsin B in the plant disease resistance hypersensitive response. Plant J 52:1–13PubMedCrossRefGoogle Scholar
- Nasir KHB, Takahashi Y, Ito A, Saitoh H, Matsumura H, Kanzaki H, Shimizu T, Ito M, Fujisawa S, Sharma P, Ohme-Takagi M, Kamoun S, Terauchi R (2005) High-throughput in planta expression screening identifies a class II ethylene-responsive element binding factor-like protein that regulates plant cell death and non-host resistance. Plant J 43:491–505PubMedCrossRefGoogle Scholar
- Weststeijn EA (1981) Lesion growth and virus localization in leaves of Nicotiana tabacum cv. Xanthi nc. after inoculation with Tobacco mosaic virus and incubation alternately at 22°C and 32°C. Physiol Plant Pathol 18:357–368Google Scholar