Development of a rapid enzymatic cDNA amplification test for the detection of Apple Scar Skin Viroid (ASSVd) in apple trees from eastern Anatolia, Turkey

Abstract

A validated RT-PCR method was used to investigate the presence of Apple scar skin viroid (ASSVd) in the commercial apple orchards of eastern Anatolia. Among three modified and simplified silica-capture based extraction methods, one was used for mass extraction to ascertain the presence of ASSVd. The test was initially performed from an ASSVd-infected source and then applied to total RNA preparations from fresh leaf tissues of apple trees collected from eastern Anatolia. ASSVd was found to occur in apple trees. Among 263 apple samples, 121 were positive for ASSVd. The infected trees showed no apparent disease symptoms on the leaves other than scarring on fruit skin. Overall incidence of ASSVd was 46% in eastern Anatolia. The presence of ASSVd reported for the first time in Turkey. Among three total RNA preparation methods, Method II was determined to be the best procedure for large scale routine analysis. The improved test can be used in a certification or clean stock program to contribute to the prevention of the spread of ASSVd in the eastern Anatolia.

Viroids of the Mediterranean Basin

“…PLMVd was occasionally detected in the Malatya region of Turkey in persimmon, walnut, and sorghum [ 3 , 231 ]. ASSVd was detected in apple with an incidence of 46% in apple orchards in Eastern Anatolia [ 24 ].…”

“…Table 1 shows the geographical distribution and host range of these viroids. Lebanon apple [10] Israel, Spain pomegranate [11] Greece, Turkey fig [12][13][14] AHVd Italy, Spain apple, loquat [15][16][17] ASSVd Greece apple, sweet cherry, pear, wild pear, wild apple [18][19][20][21][22][23] Italy pear [19] Turkey apple [24] AGVd Greece, Italy, Tunisia, Turkey grapevine [25][26][27][28] ASBVd Greece, Israel, Spain, Turkey avocado [29][30][31][32] CChMVd France chrysanthemum [33] Table 1. Cont.…”

Small Heat Shock Protein (sHsp22.98) from Trialeurodes vaporariorum Plays Important Role in Apple Scar Skin Viroid Transmission

“…The ASSVd genome consists of circular, single-stranded RNA (ssRNA) with ~330 nucleotides, and it autonomously replicates in the host cell nucleus. ASSVd is known to cause apple scar skin and dappling diseases in apple, as well as rusty skin, fruit crinkling, and fruit dimple diseases in pear [6][7][8][9]. ASSVd spreads from infected to healthy plants naturally through grafting and mechanically through pruning tools.…”

Correction: A Universal Oligonucleotide Microarray with a Minimal Number of Probes for the Detection and Identification of Viroids at the Genus Level

“…Many diagnostic techniques have been developed for plant viroid detection. These include bioassays [1][2][3][4], return-polyacrylamide gel electrophoresis (Return-PAGE) [5], reverse transcriptionpolymerase chain reaction (RT-PCR) [6][7][8][9][10][11][12][13][14][15], RT-PCR using genus degenerate primers and multiplex primers [16][17][18][19][20][21][22][23][24][25][26][27][28][29][30], RT-PCR dot-blot hybridization (RT-PCR-DBH) [31], real time RT-PCR [32][33][34][35][36][37], molecular hybridization [38][39][40], reverse transcription loop-mediated isothermal amplification (RT-LAMP) [41,42] and microarrays [43][44][45][46][47].…”