Table 2.
List of primers, gene targets, and amplicon lengths generated to detect the presence of each ACP-associated virus in all samples.
| Virus | Primers | Gene target | Length of amplicon |
|---|---|---|---|
| DcACV | F: 5' GCCGCACGAAACTAGTGATAAACGCA 3' R: 5' GGATCGGTGGTGCACGAGTATGTAAGTA 3' |
RNA1¹ segment | 473 bp |
| DcFLV | F: 5' AGGCGAGTACTCCCATCGGATACATT 3' R: 5' GAGGGCCGCTAAGTCTGTAGGACATATT 3' |
RdRp² | ~1.4 kb |
| DcDNV | F: 5' AGTCGGTGAGACTGATATCTTCGAGACC 3' R: 5' GTTTAGTTCGCTTGTCGGTTACACAGG 3' |
RdRp³ | ~1 kb |
| DcRV | F: 5' TTTTCCCAGGTACATCGA 3' R: 5' ACCATTCAGCCAGTCCTA 3' |
p8⁴ | 900 bp |
| DcPLV | F: 5' TAGGTGAACGTGATAATCCTGGTAT 3' R: 5' CAGAACGTCTGTTATGAATCGGAC 3' |
Polyprotein⁵ | 698 bp |
| DcCLV 5'-end | F: 5' AACACCCATGCTTCCAAAAC 3' R: 5' TCGTTTCACTAACGCCAATTT 3' |
Predicted RdRp6 | 492 bp |
| DcCLV 3'-end | F: 5' ATTTAGGGCCATGTGCAAAG 3' R: 5' CCAACACACCGAGCATACAC 3' |
Predicted RdRp6 | 526 bp |
| Internal control for integrity of ACP cDNA | F: 5' TCCAGAGTGATGGTCAGTAA 3' R: 5' GATCTCCTGTGTTCTGTAGC 3' |
ACP (Dc) wingless gene7 | 273 bp |
1Accession number: KX235518.1; 2Accession number: KX267823.1; 3Accession number: KX165268; 4 Nouri et al, 2016a; 5Accession number: KT698837.1; 6PCR primers designed for novel candidate virus from high throughput sequencing (HTS) results and used for screening of ACP populations in Florida; 7Accession number: XM_008486571.2. DcACV, Diaphorina citri associated C virus; DcFLV, Diaphorina citri flavi-like virus; DcDNV, Diaphorina citri densovirus; DcRV, Diaphorina citri reovirus; DcPLV, Diaphorina citri picorna-like virus; DcCLV, Diaphorina citri cimodo-like virus; F, Forward Primer; R, Reverse Primer; RdRp, RNA-dependent RNA polymerase; bp, base pair; kb, kilobase. An internal control was also used to verify the integrity of RNA extraction and cDNA synthesis steps.