Resistance to Leaf and Yellow Rust in a Collection of Spanish Bread Wheat Landraces and Association with Ecogeographical Variables
Round 1
Reviewer 1 Report
Please find my comments on the manuscript.
Page 3 line 97: what are the PR experiments?
Page 3 line 108: What type of inoculum were used? Fresh or stored spores?
Page 3 line 110: What is the advantage of fixing leaves on soil?“ Leaves were laid and fixed on the soil with metallic hairpins”
Page 3 line 112: what method was used to saturation of humidity in the incubation compartment and how is it controlled?
Page 3 line 114: What are the conditions in the greenhouse?
Page 3 line 120: The necrotic reaction is dead tissue around the infection spot NOT (yellowish spot), Yellowing around the spot call chlorotic.
Page 3 line 130: Is the isolate JC13 of stripe rust the only used? What about leaf rust?
Page 3 line 136: What is the method was followed to count pustules? Please describe.
Page 4 line 138: Is just only the leaf rust evaluation was performed in the field?
Page 4 line 143: The author says, “Field infection was natural”? Does this isolate has been raced typed? Because different virulent/avirulent races can give different virulent in tested materials, the data of the seedling stage will not be matching with field data because it is not the same virulent pattern for the race that used in the greenhouse test. ((The same concern about stripe rust))
Page 4 line 229: Figure 2. Is not successful to represent the data, it's not clear and confusing for the reader. I prefer to change it to a table.
Page 5 line 234: How Relations between seedling resistance to leaf rust and agronomic traits has been done and there is no evidence that the natural infection in the field for rust race did not confirm the virulent pattern is the same one to the race used in a greenhouse test? ((Natural infection may be occurring by more than one race of rust)) ((The same concern about stripe rust))
Page 10 line 378: With the concern of field screening of stripe rust in the field. The author needs to mention the field condition (Temperature and humidity) because it’s very important especially for stripe rust, because the infection can stop when the temperature rises more than 27C˚, and in this case, the resistance data will be false.
Author Response
Response to reviewer 1
Point 1. Page 3 line 97: what are the PR experiments?
Response 1: I added the point where PR experiments are described: ‘see 2.4.’.
Point 2. Page 3 line 108: What type of inoculum were used? Fresh or stored spores?
Response 2: I wrote ’uredospores were stored in an ultra-freezer at -80 °C. They were taken out given a heat treatment in a 40 °C water bath for 5 minutes to break cold-induced dormancy of the spores’.
Point 3. Page 3 line 110: What is the advantage of fixing leaves on soil? “Leaves were laid and fixed on the soil with metallic hairpins”
Response 3: It was changed to ‘Leaves were laid with the adaxial side up and fixed on the soil with metallic hairpins to get a more uniform inoculation’.
Point 4. Page 3 line 112: what method was used to saturation of humidity in the incubation compartment and how is it controlled?
Response 4: It was changed to ‘…and humidity at saturation (by means of ultrasonic humidifiers) for 15 hours.’
Point 5. Page 3 line 114: What are the conditions in the greenhouse?
Response 5: It as changed to ‘The next day, hairpins were removed from the leaves and plants were transferred back to their greenhouse compartment, where temperature ranged 15-25 °C.’
Point 6. Page 3 line 120: The necrotic reaction is dead tissue around the infection spot NOT (yellowish spot), Yellowing around the spot call chlorotic.
Response 6: Necrosis is macroscopically seen as yellowish spots, chlorosis as pale green spots. I changed the sentence to ‘Hypersensitive resistance is characterized by necrotic foliar tissue surrounded the pustule and macroscopically seen as yellowish spots.’
Point 7. Page 3 line 130: Is the isolate JC13 of stripe rust the only used? What about leaf rust?
Response 7: Partial resistance was only assessed to the leaf rust isolate JC13. Anyway, I renamed the title to ‘partial resistance to leaf rust…’.
Point 8. Page 3 line 136: What is the method was followed to count pustules? Please describe.
Response 8: Ok, counting was performed with a pocket lens.
Point 9. Page 4 line 138: Is just only the leaf rust evaluation was performed in the field?
Response 9: Yellow rust was also evaluated in one location (Cordoba). I rename the title.
Point 10. Page 4 line 143: The author says, “Field infection was natural”? Does this isolate have been raced typed? Because different virulent/avirulent races can give different virulent in tested materials, the data of the seedling stage will not be matching with field data because it is not the same virulent pattern for the race that used in the greenhouse test. ((The same concern about stripe rust))
Response 10: That comment is appropriate and true, but the virulent pattern of the isolate/s or race/s from the field in the season 1996/97 were not characterized.
Point 11. Page 4 line 229: Figure 2. Is not successful to represent the data, it's not clear and confusing for the reader. I prefer to change it to a table.
Response 11: The Figure 2 was removed and the data from that figure can be seen in Table S2 (MS Excel). The following Figures (and their citations in the text) were renamed accordingly.
Point 12. Page 5 line 234: How Relations between seedling resistance to leaf rust and agronomic traits has been done and there is no evidence that the natural infection in the field for rust race did not confirm the virulent pattern is the same one to the race used in a greenhouse test? ((Natural infection may be occurring by more than one race of rust)) ((The same concern about stripe rust))
Response 12: it is true. The data at seedling stage could not match with field data because the virulent pattern of the races could be different, but field data are from 1996/97 and race analysis was not performed at that time.
We indicate in the Materials and Methods section that the relationships between variables were examined using Pearson correlation coefficient (line 178) and a t-test for means (p-value < 0.05) were used to compare the resistant and susceptible groups (line 175). The agronomic data (days to heading and plant height) to study the correlations with resistance in the field were taken in the field experiments (see 2.6)
Point 13. Page 10 line 378: With the concern of field screening of stripe rust in the field. The author needs to mention the field condition (Temperature and humidity) because it’s very important especially for stripe rust, because the infection can stop when the temperature rises more than 27C˚, and in this case, the resistance data will be false.
Response 13: Yes, it is true, but the stripe rust arrives in late February-early March, and leaves by mid-April, and the severity was taken before yellow rust went off. The temperature was lower than 27 ˚C at that time in the field experiments. In this website you can check that maximum temperature in Cordoba in April is 21.1 ˚C (https://es.climate-data.org/europe/espana/andalucia/cordoba-121/t/abril-4/#monthly-weather-data).
Author Response File: 
Author Response.pdf
Reviewer 2 Report
The Authors screened a large number of bread wheat landraces of Spanish origin for their susceptibility to leaf and yellow rust. A very interesting and original part of this study was the discover of a correlation between the susceptibility level of landraces and some climatic parameters of the areas from which the wheat landraces originated. This study provides a huge amount of informationand is basical to breeding programs of wheat cultivars for resistance to rust.
The English style can be improved but is basically correct. I only introduced few suggestions and corrections (see notes in the text: attached file). See line 520 in the Discussion, where I added few sentences to expand the discussion and introduced three new references (add these references also in the section References and cite them with the proper numbers)
Comments for author File: Comments.pdf
Author Response
Response to reviewer 2
The Authors screened a large number of bread wheat landraces of Spanish origin for their susceptibility to leaf and yellow rust. A very interesting and original part of this study was the discovery of a correlation between the susceptibility level of landraces and some climatic parameters of the areas from which the wheat landraces originated. This study provides a huge amount of information and is basical to breeding programs of wheat cultivars for resistance to rust.
The English style can be improved but is basically correct. I only introduced few suggestions and corrections (see notes in the text: attached file). See line 520 in the Discussion, where I added few sentences to expand the discussion and introduced three new references (add these references also in the section References and cite them with the proper numbers).
Response: All corrections, suggestions and adding of new text performed by reviewer 2 were applied to the manuscript. Thank you.
Author Response File: Author Response.pdf
