Articles
ELISA detection of Apple chlorotic leafspot virus (ACLSV) and Apple mosaic virus (ApMV) in comparison to RT-PCR detection and the determination of viral variation in South Africa
Article number
1297_65
Pages
499 – 506
Language
English
Abstract
Apple chlorotic leaf spot virus (ACLSV) and Apple mosaic virus (ApMV) are responsible for reduced yield in stone- and pome-fruit.
The detection limits of DAS-ELISA and RT-PCR for ApMV and ACLSV were compared.
The RT-PCR was more sensitive than DAS-ELISA by a factor of about 5 to 100 times in the case of ACLSV and 100 to 1000 times for ApMV. Differences in the detection levels of DAS-ELISA versus RT-PCR of different isolates were additionally found.
Certain isolates of ApMV was not detected by DAS-ELISA in apples and one peach isolate, which were detected by RT-PCR. The genetic variation of ApMV and ACLSV isolates from South Africa was investigated.
Extracted RNA was used for RT-PCR of the coat protein genes and then sequenced.
Phylogenetic trees were constructed using these sequences and others deposited on GenBank.
Phylogenetic analysis revealed that South African isolates of ACLSV grouped into 3 of 4 possible clades and were similar to the isolates from the rest of the world.
It also indicated that the isolates are not restricted to a particular fruit group indicating that cross-infection between pome- and stone-fruit is a strong possibility.
Phylogenetic analysis of ApMV isolates indicated that two major groups occurred in South Africa.
The first group consisted of only 4 isolates, grouped together with isolates from India, while the second large group, consisting of 33 isolates, grouped together with isolates from the Czech Republic and India.
It was concluded that ApMV was also not restricted to a particular fruit group indicating the potential risk of cross-infection.
The detection limits of DAS-ELISA and RT-PCR for ApMV and ACLSV were compared.
The RT-PCR was more sensitive than DAS-ELISA by a factor of about 5 to 100 times in the case of ACLSV and 100 to 1000 times for ApMV. Differences in the detection levels of DAS-ELISA versus RT-PCR of different isolates were additionally found.
Certain isolates of ApMV was not detected by DAS-ELISA in apples and one peach isolate, which were detected by RT-PCR. The genetic variation of ApMV and ACLSV isolates from South Africa was investigated.
Extracted RNA was used for RT-PCR of the coat protein genes and then sequenced.
Phylogenetic trees were constructed using these sequences and others deposited on GenBank.
Phylogenetic analysis revealed that South African isolates of ACLSV grouped into 3 of 4 possible clades and were similar to the isolates from the rest of the world.
It also indicated that the isolates are not restricted to a particular fruit group indicating that cross-infection between pome- and stone-fruit is a strong possibility.
Phylogenetic analysis of ApMV isolates indicated that two major groups occurred in South Africa.
The first group consisted of only 4 isolates, grouped together with isolates from India, while the second large group, consisting of 33 isolates, grouped together with isolates from the Czech Republic and India.
It was concluded that ApMV was also not restricted to a particular fruit group indicating the potential risk of cross-infection.
Authors
S.S. Malan, L. Mostert, D.U. Bellsteadt
Keywords
Apple chlorotic leaf spot virus (ACLSV), Apple mosaic virus (ApMV), enzyme-linked immune sorbent assay (ELISA), reverse transcriptase-polymerase chain reaction (RT-PCR)
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