Content of review 1, reviewed on September 19, 2018

This study explores the relationship between leaf reflectance and leaf biochemistry in the context of insect attack and plant-plant communication. The question is quite important and very appealing. The experimental design seems to be well conceived and structured, especially with acquisition of time series for both hyperspectral data and biochemistry. Nevertheless, the results do not mirror the nice design and I think that these results, as shown in this version of the MS at least, are not convincing and do not support well the main conclusion on plant-plant communication reflected by a change in leaf reflectance in the NIR range. I develop these concerns below: 1- The text reports at several places "a highly significant and consistent reflectance response to insect herbivory in 10 spectral bands from 886 – 1009" (line 319 for instance). I don't see this at all in the figure 4b, in which the lines look all the same in this waveband range. The Table 2 reports the statistical relationships but it fails to give any insight into comparisons among the various treatments. Was it possible to discriminate between the (spectral) responses of non-infested plants that communicated with infested plants from individual infested plants alone? Moreover, the fig 4b seems to provide averaged reflectance spectra, ie averaged over the replicates but also across time. Why this? the biochemistry of the leaf clearly changes across time (from fig 3), so why not trying to relate reflectance and biochemistry at each time step? This would reinforce the study a lot! 2- The statistical approach is not appropriate to me. A pairwise comparison test (a glm ?) was done but it is not mentioned if a correction was provided for testing multiple comparisons (ie, across multiple time steps, and among treatments). This statistical design does not make advantage of having these wonderful time series for both hyperspectral data and biochemistry. From what I understand of the data, I suggest a RM-ANOVA (ANOVA with repeated measures) to take into account for the time effect and also to obtain the interaction terms, AND most importantly to be able to compare the treatments with each other appropriately. 3- While I think the experimental design is nice, I think it still lack important details. (A) The plastic cages used "to prevent interference communication across replications and among treatments" (line 172): please give more details: were they totally volatile-proof? how were they designed to be totally closed? What was the light level inside the cages? (B) Hyperspectral measurements: you need to specify how the plants were handled during the hyperspec measurements. Were they taken as fast as possible to the spectro room? Was this done at the same time of the day systematically to ensure that plants were photosynhtetizing at the same rate during measurement? How did you manage to avoid plant-plant communication during these spectral measurements? (C) Numbers of replicates are given, but I am confused as to the total number of plants used. For example, lines 202-203: 3 replicates x 6 treatments x 3 time period = 36 plants, am I correct? Minor issues: - line 135: not sure everyone will understand what is meant by 'subtle herbivory'. This is rather unusual term. - line 148: experimentally induced herbivory: delete experimentally because this term may be confusing as readers may think that you simulated herbivory mechanically for example instead of using a living insect. - line 168: "establishement" ? delete - line 365: delete "provides"

Source

    © 2018 the Reviewer (CC BY 4.0).

References

    Prado, R. L. d., Lidia, S. K. A., Americo, W. F. J., Aurelio, T. M., Almeida, T. M., Axel, M., Christian, N. 2018. Hyperspectral imaging to characterize plant-plant communication in response to insect herbivory. Plant Methods.