Differential Gene Expression in Response to Papaya ringspot virus Infection in Cucumis metuliferus Using cDNA- Amplified Fragment Length Polymorphism Analysis

Abstract

A better understanding of virus resistance mechanisms can offer more effective strategies to control virus diseases. Papaya ringspot virus (PRSV), Potyviridae, causes severe economical losses in papaya and cucurbit production worldwide. However, no resistance gene against PRSV has been identified to date. This study aimed to identify candidate PRSV resistance genes using cDNA-AFLP analysis and offered an open architecture and transcriptomic method to study those transcripts differentially expressed after virus inoculation. The whole genome expression profile of Cucumis metuliferus inoculated with PRSV was generated using cDNA-amplified fragment length polymorphism (cDNA-AFLP) method. Transcript derived fragments (TDFs) identified from the resistant line PI 292190 may represent genes involved in the mechanism of PRSV resistance. C. metuliferus susceptible Acc. 2459 and resistant PI 292190 lines were inoculated with PRSV and subsequently total RNA was isolated for cDNA-AFLP analysis. More than 400 TDFs were expressed specifically in resistant line PI 292190. A total of 116 TDFs were cloned and their expression patterns and putative functions in the PRSV-resistance mechanism were further characterized. Subsequently, 28 out of 116 candidates which showed two-fold higher expression levels in resistant PI 292190 than those in susceptible Acc. 2459 after virus inoculation were selected from the reverse northern blot and bioinformatic analysis. Furthermore, the time point expression profiles of these candidates by northern blot analysis suggested that they might play roles in resistance against PRSV and could potentially provide valuable information for controlling PRSV disease in the future.

Differentially Expressed Genes in Resistant and Susceptible Common Bean (Phaseolus vulgaris L.) Genotypes in Response to Fusarium oxysporum f. sp. phaseoli

“…Differential display is useful at generating transcript-derived fragment (TDF) libraries from specific treatments, genotypes and disease conditions during varying stages of the infection process and disease reaction in the plant. TDF libraries have been used extensively to characterize the differentially expressed genes at different levels of disease development in order to analyze their function and correlation with disease resistance having the advantage of producing cloned fragments useful for further experiments[ 18 ]. As a result, morphological, molecular and anatomical studies have then been carried out to evaluate whether differential display gene expression within specific host tissues is correlated with the onset as well as the progression of the pathogen in the plant[ 19 ].…”

Identification of Genes Differentially Expressed between Resistant and Susceptible Tomato Lines during Time-Course Interactions with Xanthomonas perforans Race T3

“…Moreover, compared with hybridization-based approaches including DNA chips and microarrays, cDNA-AFLP has a relatively low startup cost and enables distinguishing expression patterns of highly homologous gene family members [24] . The cDNA-AFLP technique has been successfully employed to identify differentially expressed genes in various plant-pathogen systems [22] , [23] , [25] – [27] . In this study, we isolate 71 genes differentially expressed in resistant line PI 114490 and susceptible line OH 88119 during time-course of response to bacterial spot race T3 using the cDNA-AFLP technique.…”

Differential Gene Expression Pattern of Importin β3 and NS5 in C6/36 Cells Acutely and Persistently Infected with Dengue Virus 2

“…aegypti mosquitoes successfully [ 17 , 18 , 19 , 20 ]. A variation of this technique, referred to as cDNA-AFLP, is an easy tool to evaluate genome-wide expression levels and does not require prior knowledge of gene coding sequences [ 21 , 22 , 23 , 24 ].…”