Suppressive Subtractive Hybridization Approach Revealed Differential Expression of Hypersensitive Response and Reactive Oxygen Species Production Genes in Tea (Camellia sinensis (L.) O. Kuntze) Leaves during Pestalotiopsis thea Infection

• Published in: Applied biochemistry and biotechnology Vol. 168; no. 7; pp. 1917 - 1927
• Main Authors: Senthilkumar, Palanisamy, Thirugnanasambantham, Krishnaraj, Mandal, Abul Kalam Azad
• Format: Journal Article
• Language: English
• Published: New York Springer-Verlag 01-12-2012; Springer; Springer Nature B.V
• Subjects: Agricultural biotechnology; Ascomycota - physiology; Biochemistry; Biological and medical sciences; Biotechnology; Camellia sinensis; Camellia sinensis - genetics; Camellia sinensis - immunology; Camellia sinensis - metabolism; Camellia sinensis - microbiology; Chemistry; Chemistry and Materials Science; Cloning, Molecular; Cultivars; Disease resistance; Disease tolerance; Economic importance; Expressed Sequence Tags - metabolism; Fundamental and applied biological sciences. Psychology; Gene expression; Genes, Plant - genetics; Hybridization; Leaves; Pestalotiopsis; Pestalotiopsis theae; Plant Diseases - immunology; Plant Diseases - microbiology; Plant Leaves - genetics; Plant Leaves - immunology; Plant Leaves - metabolism; Plant Leaves - microbiology; Plants; Polymerase Chain Reaction; Reactive Oxygen Species - metabolism; Tea; Transcriptome; cDNA; Suppressive subtractive hybridization; EST; Oxygen; Plant leaf; Hybridization; Pestalotiopsis; Pestalotiopsis thea; Fungi; Infection; Camellia sinensis; Complementary DNA; Gene; Dicotyledones; Angiospermae; Production; Spermatophyta; Fungi Imperfecti; Theaceae
• ISSN: 0273-2289; 1559-0291
• DOI: 10.1007/s12010-012-9907-1

Tea (
Camellia sinensis
(L.) O. Kuntze) is an economically important plant cultivated for its leaves. Infection of
Pestalotiopsis theae
in leaves causes gray blight disease and enormous loss to the tea industry. We used suppressive subtractive hybridization (SSH) technique to unravel the differential gene expression pattern during gray blight disease development in tea. Complementary DNA from
P
.
theae
-infected and uninfected leaves of disease tolerant cultivar UPASI-10 was used as tester and driver populations respectively. Subtraction efficiency was confirmed by comparing abundance of
β
-actin gene. A total of 377 and 720 clones with insert size >250 bp from forward and reverse library respectively were sequenced and analyzed. Basic Local Alignment Search Tool analysis revealed 17 sequences in forward SSH library have high degree of similarity with disease and hypersensitive response related genes and 20 sequences with hypothetical proteins while in reverse SSH library, 23 sequences have high degree of similarity with disease and stress response-related genes and 15 sequences with hypothetical proteins. Functional analysis indicated unknown (61 and 59 %) or hypothetical functions (23 and 18 %) for most of the differentially regulated genes in forward and reverse SSH library, respectively, while others have important role in different cellular activities. Majority of the upregulated genes are related to hypersensitive response and reactive oxygen species production. Based on these expressed sequence tag data, putative role of differentially expressed genes were discussed in relation to disease. We also demonstrated the efficiency of SSH as a tool in enriching gray blight disease related up- and downregulated genes in tea. The present study revealed that many genes related to disease resistance were suppressed during
P
.
theae
infection and enhancing these genes by the application of inducers may impart better disease tolerance to the plants.