PhoGV and potato moth geographic data in the north andean region

Description

1) Insect data (datosFeromonas.csv). A total of 1216 pheromone trap samples from 106 localities were obtained across Northern Andes. 2) Virus occurrence data (virusecofor.txt). A total of 517 larval samples from 15 localities in Ecuador and Colombia were PCR amplified for teh detection of PhoGV

Steps to reproduce

1) T. solanivora densities were evaluated using pheromone baited traps located along altitudinal gradients of larger amplitude than for virus prevalence in Ecuador, Colombia and Venezuela. Five different surveys, representing different regions and/or periods of time, were carried out. A total of 1216 pheromone trap samples from 106 localities were obtained across Northern Andes 2) PhoGV entomovirus data on T. solanivora larvae were evaluated by larvae sampling and phoGV PCR detection. T. solanivora larvae (517) were collected from infested tubers from 15 localities dispatched along 5 altitudinal gradients, three located in Colombia from 2800 to 2940 masl Subachoque, Siachoque and Zipaquira and three in Ecuador from (Montufar-Carchi and Riobamba ) (distribution of localities in presented in figure 1). In these localities, the granulovirus had never been applied for biological control purposes. Tubers were collected and dissected in the laboratory. Healthy larvae and/or larvae with signs of granulovirus disease were individually collected and conserved in alcohol for DNA analysis. Larval tissues were homogenized in 50 μL bidistilled H2O and centrifuged at 1000rpm for 5 min at 4ºC. To recover OBs, supernatants were centrifuged at 15000 rpm for 15 min and pellets resuspended in 100 μL bidistilled H2O. Suspensions were incubated with 100 μl vol. of 0,5M Na2CO3, 50 μL SDS 10% for 10 min at 60ºC and centrifuged at 5000 rpm for 5 min. The supernatants were digested with 25 μL proteinase K (20mg/mL) at 56ºC for one hour and DNA was extracted using a phenol/chloroform/isoamyl alcohol protocol and then precipitated with ethanol, as described in previous works [37]. DNA was extracted using proteinase K digestion, The presence of the PhopGV was detected by PCR using primer pair, 83.2 5'-CCGCGCCGATTACCAACAGCAGC-3' and 84.1 5'-GAACTGTTAAACGGCTTGAGTGAGCG-3' designed over the GenBank sequence JX170206.1. on 517 T. solanivora collected larvae, amplifying a 241 bp region encompassing part of gene 83 and 84 of the PhoGV genome. Amplification conditions were: 94ºC for 1min, 30 cycles (94ºC for 1min, 50ºC for 1min, 72ºC for 1 min), and a final extension of 7 min. Conditions were optimized in Colombia and Ecuador separately to reduce false negatives. Colombian samples were amplified in Corpoica-Tibaitata, Colombia using the following 10X mix: dNTPs 200 μM (Pharmacia Biotech 27-1850), 0.5 μM of both primers, MgCl2 2.0 mM, buffer 10X (50 mM KCl, 10 mM Tris HCl pH 9.0 0.1% Nonidet), 2U Taq polymerase (Promega M1665). Ecuadorian samples were amplified in Santa Catalina: PCR mix contained dNTPs 250 μM (Invitrogen, reference?), 0.5 μM of both primers, MgCl2 1.6 mM, buffer 10X (50 mM KCl, 20 mM Tris Hcl, pH 8.4), 0.075U/ul Platinium Taq polymerase (Invitrogen).

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