1st question: Yes, the synthesized 7-CP was purified by column chromatography and the
product were subjected to structural analysis by GC-MS and NMR to confirm that there is no
interference from precursors. Dilutions of the purified product were made to get the working
concentrations for the assay. This information was added to the manuscript as the reviewer
suggested.
2nd question: The paragraph related to the substrate synthesis was moved to method section
as was suggested.
3rd question: L182-186: Additional information about the primers and PASA assay were added
to method section as was suggested by the reviewer.
The explanation about how is shown the presence/absence of the mutations was added to
method section.
4th question: The missing n from table 2 was an error and was corrected. The values from both
tables are means of individual assays.
5th The P-450 activity of Ti-R1 type measured in previous papers was done on first instar
nymphs. There are not previous studies that measure P450 activity in developing eggs of T.
infestans as was done in this work.
6th KDR: Yes, the 476bp fragment amplification appears in S and ER/CL meaning that the
populations have homozygous susceptible individuals, or heterozygous individuals.
The ADN used for the PASA assay was pooled from many individuals.
Yes, the reference to the figure 2 is incorrect and was corrected. Text was improved as
suggested.
7th Lane 224, 248-250 was improved.
Lanes 257 and 260 were corrected.
Paragraph 259-264 was improved.
Lane 279 reference was added.
8th L292-298 We tested the presence of the L925I mutation in Campo Largo and Entre Ríos
because the mutation was not previously studied in nymphs or eggs of these populations. The
L9251 mutation was previously identified in Madrejones, a small locality close to Campo Largo
and Entre Ríos. It is important to notice that other populations of this area were tested
negative. As was described for other species, both mutations can be present at the same time.
L302-305 Results showed that the 476bp band is present in the susceptible and both resistant
populations, suggesting that the presence of the susceptible allele in heterozygous resistant
individuals. The higher resistant values of Campo Largo can be explained as a higher
proportion of the resistant allele in resistant individuals (e.g. higher proportion of homozygote
resistant individuals).
Language and phrasing have been improved throughout the manuscript.