Seed Physiological Potential of Capsicum annuum var. glabriusculum Genotypes and Their Answers to Pre-Germination Treatments

Round  1

Reviewer 1 Report

Abstract can be more compact. More compaction is recommended.

In Table 2, hot water, ‘at 83C let cool for 24hrs’ means ‘dipping the seeds in 83C hot water and let them naturally cool down for 24 hrs? Please make it clear.

Line 78, ‘The seeds were disinfected with chlorine at 1% during 30 seconds’, here did you use elemental chlorine or sodium hypochlorite or any other chlorine-generating compounds? Please make it clear.

Line 81, any sentence must begin with alphabetic words, not number. Petri dishes (100mm x 15mm) --- is recommended.

Line 82-84, two fungicides were used in rotation. No mention about the concentration and method of application and how long?

Line 99, essay? Not assay?

Line 149-153, in Fig. 1, PC1 and PC2 are given but you are citing CP1 and CP2. It is confusing. Which one is correct? Confusing thereafter in the text too.

According to Table 3, genotype, treatments, and genotype x treatment interaction was highly significant. It means that effect of treatments varying depending upon the genotypes. In that case, comparison of simple effects of treatment and genotype as in Table 4 and 5 does not give accurate information. Comparison of treatments by genotype or vice versa would be right.

Line 240, what does AG mean? There’s no explain. Not GA?

Author Response

Response to Reviewer 1 Comments

Point 1: Abstract can be more compact. More compaction is recommended.

Response 1: The summary was more compact by eliminating 26 words.

Point 2: In Table 2, hot water, ‘at 83C let cool for 24hrs’ means ‘dipping the seeds in 83C hot water and let them naturally cool down for 24 hrs? Please make it clear.

Response 2: It was clarified in the explanation of Table 2, that the seeds were immersed in hot water at 83 ° C and allowed to cool naturally for 24 hours.

Point 3: Line 78, ‘The seeds were disinfected with chlorine at 1% during 30 seconds’, here did you use elemental chlorine or sodium hypochlorite or any other chlorine-generating compounds? Please make it clear.

Response 3: Sodium hypochlorite was used.

Point 4: Line 81, any sentence must begin with alphabetic words, not number. Petri dishes (100mm x 15mm) --- is recommended.

Response 4: We correct this part for the recommended.

Point 5: Line 82-84, two fungicides were used in rotation. No mention about the concentration and method of application and how long?

Response 5: Concentration, method of application and time were included.

Point 6: Line 99, essay? Not assay?

Response 6: The word was corrected and "assay" was added.

Point 7: Line 149-153, in Fig. 1, PC1 and PC2 are given but you are citing CP1 and CP2. It is confusing. Which one is correct? Confusing thereafter in the text too.

Response 7: It was corrected and managed as PC1 and PC2.

Point 8: According to Table 3, genotype, treatments, and genotype x treatment interaction was highly significant. It means that effect of treatments varying depending upon the genotypes. In that case, comparison of simple effects of treatment and genotype as in Table 4 and 5 does not give accurate information. Comparison of treatments by genotype or vice versa would be right.

Response 8: We agree that the interaction provides us with more accurate information. It was decided to manage genotypes and treatments individually in order to make a complete analysis and see specifically its effect, for the case of genotypes the purpose was to know which of them has the greatest potential. On the other hand, the information of the treatments was for practical purposes in its use to promote germination.

Point 9: Line 240, what does AG mean? There’s no explain. Not GA?

Response 9: It was corrected we wanted to say GA, the meaning comes in table 2.

Author Response File: Author Response.pdf

Reviewer 2 Report

This work has as principal aim the study of the seed physiological potential of an important wild crop for the Mexican rural households. Reading the paper, the first things that I would like to know are the conditions of seeds storage at INIFAP, because the seed germination responses (including the sees dormancy state) may vary in function of the temperature and humidity in which they were preserved, as well as the duration in time (calculated from the date of the harvesting) of the seed storage. So, I suggest to include these important (in my opinion) information in the Material and Methods.

I give some suggestions which may help to clarify some points of the manuscript:

Page 2 LL 54: I suggest to add the references; see comment below.

Materials and Methods

Table 1: I suggest to add at least the coordinates of the different seed harvesting sites or the origin of the piquin pepper genotypes; in addition, in order to help the readers, I suggest to write in full (at least the first time) the State names.

Page 2 LLs 74-75: I suggest to change “… in order to promote germination in piquin…” in “… in order to evaluate the germination response in piquin…”.

In my personal opinion, if you write “in order to promote germination”, I understand that you already know the germination behaviour of piquin seeds in all 11 pre-treatments proposed in table 2, and all of them promote the germination. In addition, you should justify the choice of these eleven pre-treatments; without these pieces of information, any seed ecologist would like to know why you have excluded other important pre-treatments, like cold and/or warm stratification, in order to “solve the dormancy”. Probably, the authors can solve it by citing the previous works with similar topic published about the same species (i.e. Cano-Vázquez et al., 2015) on page 2 LL 54.

Table 2: I suggest “Treatments used to evaluate the germination response of piquin pepper seeds” instead to “Treatments used to promote the germination of piquin pepper seed”.

Page 3 LL 89-90: What do you mean “normal seedlings”? Among seedlings or germinated seeds? Probably you refer to “germinated seeds”.

Results

Page 5 LL150-151; Figure 1, Page 9 LLs 231, 232, 236: I suggest to standardize “CP1” and “CP2” as reported in figure 1 (PC1 and PC2) or vice versa.

Page 6 LL 154: “Figure 1” should be written as “Figure 1 GS” and “(GS)” should be deleted.

Page 6 LLs 181-182: double check this sentence.

Page 6 LL 185: “as well as G9 applying HPGA and AV”… very hard to understand it in Fig.1 HS. See comment below.

Figure 1: It is very hard to read the abbreviations inside the figures. Is it possible to modify it in order to increase the readability?

Discussion

Page 10 LL 240: GA instead to AG?

Page 10 LLs 262-263: I suggest to modify the sentence with “These results confirm that endogenous plant hormones like GA, play an important role in seed dormancy release”.

Author Response

Response to Reviewer 2 Comments

Point 1: This work has as principal aim the study of the seed physiological potential of an important wild crop for the Mexican rural households. Reading the paper, the first things that I would like to know are the conditions of seeds storage at INIFAP, because the seed germination responses (including the sees dormancy state) may vary in function of the temperature and humidity in which they were preserved, as well as the duration in time (calculated from the date of the harvesting) of the seed storage. So, I suggest to include these important (in my opinion) information in the Material and Methods

Response 1: The conditions in which the seed was stored and the time were included in materials and methods. The percentage of humidity was not measured, but it was kept low to keep the fruits dry and prevent the spread of fungi.

Point 2: I give some suggestions which may help to clarify some points of the manuscript:

Page 2 LL 54: I suggest to add the references; see comment below.

Response 2: The reference was added.

Point 3: Table 1: I suggest to add at least the coordinates of the different seed harvesting sites or the origin of the piquin pepper genotypes; in addition, in order to help the readers, I suggest to write in full (at least the first time) the State names.

Response 3: The coordinates of the different seed collection sites were added and the states were completely written.

Point 4: Page 2 LLs 74-75: I suggest to change “… in order to promote germination in piquin…” in “… in order to evaluate the germination response in piquin…”.

Response 4: The prayer was changed to the one suggested.

Point 5: In my personal opinion, if you write “in order to promote germination”, I understand that you already know the germination behaviour of piquin seeds in all 11 pre-treatments proposed in table 2, and all of them promote the germination. In addition, you should justify the choice of these eleven pre-treatments; without these pieces of information, any seed ecologist would like to know why you have excluded other important pre-treatments, like cold and/or warm stratification, in order to “solve the dormancy”. Probably, the authors can solve it by citing the previous works with similar topic published about the same species (i.e. Cano-Vázquez et al., 2015) on page 2 LL 54.

Response 5: The choice of treatments was justified by citing previous works.

Point 6: Table 2: I suggest “Treatments used to evaluate the germination response of piquin pepper seeds” instead to “Treatments used to promote the germination of piquin pepper seed”.

Response 6: We accept and add the suggestion.

Point 7: Page 3 LL 89-90: What do you mean “normal seedlings”? Among seedlings or germinated seeds? Probably you refer to “germinated seeds”

Response 7: We add germinated seeds instead of normal seedlings.

Point 8: Page 5 LL150-151; Figure 1, Page 9 LLs 231, 232, 236: I suggest to standardize “CP1” and “CP2” as reported in figure 1 (PC1 and PC2) or vice versa.

Response 8: It was standardized as PC1 and PC2.

Point 9: Page 6 LL 154: “Figure 1” should be written as “Figure 1 GS” and “(GS)” should be deleted.

 Response 9: We eliminated (GS) and added the suggestion "Figure 1 GS".

Point 10: Page 6 LLs 181-182: double check this sentence.

Response 10: The sentence was revised twice and we agreed that the most suitable treatments for genotypes are those that are closest. This condition was presented by the G6 genotype with the MS treatment, in the case of the G7 genotype, it presented a positive response in several treatments, but the best response was with the HPGA treatment.

Point 11: Page 6 LL 185: “as well as G9 applying HPGA and AV”… very hard to understand it in Fig.1 HS. See comment below.

Figure 1: It is very hard to read the abbreviations inside the figures. Is it possible to modify it in order to increase the readability?

Response 11: We increase readability.

Point 12: Page 10 LL 240: GA instead to AG?

Response 12: It was corrected we wanted to say GA.

Point 13: Page 10 LLs 262-263: I suggest to modify the sentence with “These results confirm that endogenous plant hormones like GA, play an important role in seed dormancy release”.

Response 13: Suggested sentence added.

Author Response File: Author Response.pdf

Reviewer 3 Report

This topic is a very interesting one since there is a interest in domesticating the plant. However, I have some concerns about the paper:

There is not enough information about the methodology of how some of the experiments are conducted. There should be enough information for someone to repeat the experiments. For example, the authors talked about sanding the seeds softly, but with what kind of sand (grain size)? Is it rubbing with sand paper or vortexing the seeds with the sand? For how long? A few seconds or  minutes? 

It's not clear to me in table 2 if the seeds treated with chemicals were submerged or the authors sprayed the chemicals on the seeds, or they were agitated in a flask? Do the authors rinse off the chemicals after the treatment with dIH2O or they were placed on filter paper directly for germination? They also did not mention the solvents, I assume it's just dI water, but it may have been certain % of ethanol or other solvents, a paragraph to clarify the methodology would be helpful for readers.

Germination rate and dormancy has a lot to do with the age of the seeds and the storage condition. Are these seeds all harvest at the same time? If so, how old are they and what is their storage condition? If previous research show that this is not a factor for Piquin peppers, the authors must address that and cite the article properly. 

Germination rate and physical trait is also affected by the mother plant where the seeds come from. Were they grown in the same condition before the seeds were harvest? It is not clearly stated.

Table 2. I'm having trouble with some of the treatment controls, were there none that was done with no treatment? Simply straight germinated in water?

 Throughout the article the authors kept using the word "behavior" to describe the plants. Like in row 138, 148, or "best behavior" in row 155 and 160, "behaved well" in 166 and "positive behavior" in row 168. Behavior is define as something that human and animal would do, so I'm not sure it's a good word to use for plants. It maybe better to replace it with words like "desirable traits" or "response to xxx treatment" rather than using the word behavior. 

Row 187, the authors use the word "discriminating" treatments. Again, this is a word usually used to describe human behavior and not used in plants. So, maybe replace it with a better term. 

Overall, the result is very interesting and can be applied in future breeding work on domesticating the pepper. I'd recommend the authors work on the revisions.      

Author Response

Response to Reviewer 3 Comments

Point 1: This topic is a very interesting one since there is a interest in domesticating the plant. However, I have some concerns about the paper:

There is not enough information about the methodology of how some of the experiments are conducted. There should be enough information for someone to repeat the experiments. For example, the authors talked about sanding the seeds softly, but with what kind of sand (grain size)? Is it rubbing with sand paper or vortexing the seeds with the sand? For how long? A few seconds or minutes?

Response 1: We add the indicated information (With what is done, the way it is done, grain size and time).

Point 2: It's not clear to me in table 2 if the seeds treated with chemicals were submerged or the authors sprayed the chemicals on the seeds, or they were agitated in a flask? Do the authors rinse off the chemicals after the treatment with dIH2O or they were placed on filter paper directly for germination? They also did not mention the solvents, I assume it's just dI water, but it may have been certain % of ethanol or other solvents, a paragraph to clarify the methodology would be helpful for readers.

Response 2: This part was clarified, where we mentioned that the seed was submerged in each treatment and then rinsed with distilled water. It was also added that the distilled water was used as a solvent to obtain the desired concentration.

Point 3: Germination rate and dormancy has a lot to do with the age of the seeds and the storage condition. Are these seeds all harvest at the same time? If so, how old are they and what is their storage condition? If previous research show that this is not a factor for Piquin peppers, the authors must address that and cite the article properly.

Response 3: The conditions in which the seed was stored and the time were included in materials and methods. The percentage of humidity was not measured, but it was kept low to keep the fruits dry and prevent the spread of fungi.

Point 4: Germination rate and physical trait is also affected by the mother plant where the seeds come from. Were they grown in the same condition before the seeds were harvest? It is not clearly stated.

Response 4: The conditions of the mother plants from which the seed was obtained were added.

Point 5: Table 2. I'm having trouble with some of the treatment controls, were there none that was done with no treatment? Simply straight germinated in water?

Response 5: Yes, the WIT treatment was carried out without treatment and the planting was direct

Point 6: Throughout the article the authors kept using the word "behavior" to describe the plants. Like in row 138, 148, or "best behavior" in row 155 and 160, "behaved well" in 166 and "positive behavior" in row 168. Behavior is define as something that human and animal would do, so I'm not sure it's a good word to use for plants. It maybe better to replace it with words like "desirable traits" or "response to xxx treatment" rather than using the word behavior.

Response 6: The word "behavior" was replaced by the suggested words "desirable traits" or "response to treatment xxx".

Point 7: Row 187, the authors use the word "discriminating" treatments. Again, this is a word usually used to describe human behavior and not used in plants. So, maybe replace it with a better term.

Response 7: The word was reviewed and it was determined that the discriminatory term is also used in plants. The ability to discriminate is one of the most important parameters of the GGE biplot when assessing an environment (Kendal & Sener, 2015, Hagos & Abay, 2013, Muthoni et al., 2015, Gedif & Yigzaw, 2014).

Point 8: Overall, the result is very interesting and can be applied in future breeding work on domesticating the pepper. I'd recommend the authors work on the revisions.

Response 8: We worked on the revisions and added more references to give more support to work.

Author Response File: Author Response.pdf

Round  2

Reviewer 3 Report

The authors have addressed all of my concerns. 

Author Response

Point 1: The authors have addressed all of my concerns. 

Response 1: We welcome your comments