Analysis of expressed sequence tags derived from pea leaves infected by Peronospora viciae f. sp. pisi

• Published in: Annals of applied biology Vol. 161; no. 3; pp. 214 - 222
• Main Authors: Feng, J, Chang, K.F, Hwang, S.F, Strelkov, S.E, Conner, R.L, Gossen, B.D, McLaren, D.L, Chen, Y.Y
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01-11-2012; Blackwell; Wiley Subscription Services, Inc
• Subjects: Airborne microorganisms; Arabidopsis; Biological and medical sciences; cDNA libraries; cDNA library; clones; complementary DNA; Crop diseases; Downy mildew; EST; expressed sequence tags; Fundamental and applied biological sciences. Psychology; Fungal plant pathogens; gene expression regulation; Genes; Infection; Leaves; pathogenesis; Pathogens; peas; Peronospora viciae; Phytopathology. Animal pests. Plant and forest protection; Polymerase chain reaction; SSH; suppression subtractive hybridisation; suppression subtractive hybridization; Stramenopila; Plant production; Molecular hybridization; Mycosis; Plant pathogen; Peronospora viciae; EST; SSH; Plant leaf; Molecular marker; Gene library; Oomycota; Pea; cDNA library; Suppression subtractive hybridization; Infection; suppression subtractive hybridisation; Applied biology; Analysis; Vegetative apparatus; Downy mildew; Expressed sequence tag
• ISSN: 0003-4746; 1744-7348
• DOI: 10.1111/j.1744-7348.2012.00566.x

A cDNA library was constructed from field pea leaves infected by the downy mildew pathogen, Peronospora viciae f. sp. pisi, using a suppression subtractive hybridisation approach. The library consists of 399 expressed sequence tags, from which 207 unisequences were obtained after sequence assembly. Of the unisequences, six were shown to be of Peronospora viciae f. sp. pisi origin. The remaining unisequences were subjected to gene ontology analysis and their functions were predicted in silico. Eleven of these unisequences (representing 24 clones) shared significant sequence similarities with Arabidopsis genes known to be involved in downy mildew resistance, including the well‐characterised genes RPP5, RPP6 and RPP27. Expression analysis of five selected unisequences by real‐time PCR indicated that all five were up‐regulated during downy mildew pathogenesis, suggesting a significant role for these genes in the host response to downy mildew infection.