Identification of smut-responsive genes in sugarcane using cDNA-SRAP

Abstract

ABSTRACT. Sugarcane smut, caused by the fungus Sporisorium scitamineum, is one of the main diseases that affect sugarcane worldwide. In the present study, the cDNA-SRAP technique was used to identify genes that are likely to be involved in the response of sugarcane to S. scitamineum infection. In total, 21 bands with significant differential expression during cDNA-SRAP analysis were cloned and sequenced. Real-time qPCR confirmation demonstrated that expression of 19 of these 21 differential bands was consistent with the expression observed during cDNA-SRAP analysis, with a deduced false positive rate of 9.5%. Sequence alignment indicated that 18 of 19 differentially expressed genes showed homologies from 19% to 100% to certain genes in GenBank, including the following genes: topoisomerase (EU048780), ethylene insensitive (EU048778), and tetraspanin (EU048770). A real-time qPCR assay showed that during 0-72 h after pathogen infection, expression of the topoisomerase and 6809 Smut-responsive genes in sugarcane ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (2): 6808-6818 (2015) the ethylene insensitive genes was upregulated, whereas expression of the tetraspanin gene was downregulated, identical to the expression patterns observed under salicylic acid treatment. Therefore, all three genes are thought to play a role during S. scitamineum challenge, but with different functions. To our knowledge, this is the first report on the application of cDNA-SRAP in differential gene expression analysis of sugarcane during a sugarcane-S. scitamineum interaction. The results obtained also contribute to a better understanding of the molecular mechanisms associated with sugarcane-S. scitamineum interactions.

Development of an RAPD-based SCAR marker for smut disease resistance in commercial sugarcane cultivars of Pakistan

“…In a recent study based on AFLP analysis, two DNA bands or markers were found to be specific to gamma-radiation induced smut-resistant mutants of sugarcane variety GT 54-9 (Esh and Nasr, 2014). The first demonstration of cDNA-SRAP method for the purpose of isolating 19 differentially expressed genes in smut-resistant sugarcane variety NCo376 under smut attack, has also been documented recently (Huang et al, 2015). Hence AFLP and cDNA-SRAP techniques are proven to be quite proficient in determining different aspects of smut resistance mechanism employed by sugarcane.…”

Peluang Pengembangan Pengendalian Penyakit Luka Api pada Tebu di Indonesia

“…Analisis ekspresi gen yang berkaitan dengan ketahanan ini dapat dilakukan melalui pendekatan dengan metode molekuler seperti sequence-related amplified polymorphism (cDNA-SRAP), RT-qPCR, serta RNA-seq (Huang et al, 2015;Su et al, 2015;McNeil et al 2018). Melalui metode cDNA-SRAP, Huang et al (2015)…”

Comparative analysis of protein and differential responses of defense-related gene and enzyme activity reveals the long-term molecular responses of sugarcane inoculated with Sporisorium scitamineum

“…Molecular studies recognized specific changes in protein synthesis and identified genes that are differentially expressed during the early stage of sugarcane-S. scitamineum interaction. Many of these genes and proteins are considered to play an essential role in the host plant defense responses (Orlando et al 2005;Lao et al 2008;Que et al 2009;Huang et al 2015;Barnabas et al 2016;Singh et al 2017), but their functions are not studied in genetically different sugarcane germplasm and production environment. Also, the molecular and biochemical mechanism (s) underpinning smut resistance is not well understood (Sundar et al 2012;Sundar et al 2015).…”