Management of Water Supply in the Cultivation of Different Agaricus bisporus Strains
, Wagner Gonçalves Vieira Júnior 1, Douglas M. M. Soares 2, Lucas da Silva Alves 1, Bianca de Barros Nóbrega 2, Arturo Pardo-Giménez 3, Cassius V. Stevani 2 and Diego Cunha Zied 4,*Round 1
Reviewer 1 Report
Dear Authors, please rewrite the conclussions.
Author Response
Reviewer 1
Dear Authors, please rewrite the conclusions.
Answer: The conclusion was rewritten
Reviewer 2 Report
Agronomy-2622777 Paper Reviewed
The management of mushroom crops with less water use but with equal or greater productivity is a very important goal. But the reviewer has not well-understand some parts of the paper.
· Firstly, there are no clear relationship between the results of the genetic analysis and the cultivation trial. In fact, I understand them rather as two independent papers.
· The parameter “yield” has not been conventionally evaluated. Why is the reason because the authors have not used the standardised BE or the common value in kg m-2? In my opinion, the yield defined in “Variable analysed” section (with fresh weight of substrate), is not useful to compare treatments each other. This aspect must be clarified.
The methodology is not appropriate. The results are not sufficiently tested. More literature might be considered.
Title: It is ok
Abstract: The aim of the genetic analysis of the ITS 1+2 regions must be included. The results must be rewritten.
Key words: “irrigation” instead “water use”; yield; screening of mushroom quality; strain???; ..
Introduction: it is ok, but more literature might be considered.
Materials and methods
2.1. Are there any reasons because the ITS 1+ 2 regions have been studied? Are they related to some particular and defined growing parameter? This aspect must be explained.
2.4. There is ONLY one crop trial.
Moreover, I think the trial is not designed in a good way: 10, 5 kg m-2 is not the habitual packaging system, and 25 L m-2 is a very hard irrigation programme to this low compost weight. The height of the casing layer (3 cm) is also uncommon (so low).
How did this high water contribution affect to the temperature of the substrate? The temperature values for any time in the crop cycle must be shown.
I suppose that the authors used the oven dried measure of the compost to relativize the parameter “yield” along the crop cycle, but I think that is not the best methodology. The humidity sounding line would be able a better way to evaluate the water content of the substrates.
Quality parameters (apart from unitary weight) regarding water supply must be considered. [Noble et al, 2000; Barry et al., 2016]
Results
3.1. ITS sequencing of A. bisporus strain
What are the results of this section? If all used strains are closely related, how the authors could evaluate the relationship between irrigation system and genetic?
3.2. First flush results: Earliness has not been evaluated. I think it would help to draw final conclusions.
In the second flush, ….…. (Table 3), indicating a good recovery….. in the control treatments, isn’t it?
Figure 3. This figure was better understood if the three flushes were defined.
Figure 4 might be improved with the compost and casing temperature parameter evolution. What was the strain that was cultivated while moisture content was registered?
Discussion and Discussion
Some part of the Discussion section might be replaced on Introduction one [references 34 and 35].
The results were obtained after only one crop trial. The reviewer claims that is unsuitable for conclusions.
Author Response
Reviewer 2
The management of mushroom crops with less water use but with equal or greater productivity is a very important goal. But the reviewer has not well-understood some parts of the paper. Firstly, there are no clear relationship between the results of the genetic analysis and the cultivation trial. In fact, I understand them rather as two independent papers.
Answer: The authors are grateful for the comments on this manuscript and for all the Reviewer’s comments and suggestions. Although it may seem like our genetic analysis and our breeding trial are separate, they are interconnected. The genetic analysis provides valuable information about the genetic characteristics of the strains under study; the research assay allows us to observe how these characteristics manifest themselves in the real environment. This integrated approach helps us make meaningful connections between the genetic data and the behavior observed in the study. To make this clear, we propose to change the title to "Management of water supply in the cultivation of different Agaricus bisporus strains" in the new version of the manuscript. In mushroom science, unlike plant science, there are no manuscripts that associate the relationship between genotypic characteristics and phenotypic characteristics. This manuscript has this proposal.
The parameter “yield” has not been conventionally evaluated. Why is the reason because the authors have not used the standardized BE or the common value in kg m-2? In my opinion, the yield defined in “Variable analyzed” section (with fresh weight of substrate), is not useful to compare treatments to each other. This aspect must be clarified. The methodology is not appropriate. The results are not sufficiently tested. More literature might be considered.
Answer: The authors sincerely appreciate your constructive feedback on our manuscript. In response to your comment, in the new version of manuscript we have included an evaluation of yield using standardized biological efficiency (BE) values, alongside the yield parameter. We believe that this addition will provide a more comprehensive assessment and enable easier comparisons between them.
Title: It is ok
Abstract: The aim of the genetic analysis of the ITS 1+2 regions must be included. The results must be rewritten.
Answer: ITS 1+2 analysis are commonly used for fungal taxonomy, and we analyzed this DNA fragment in order to assess genetic variations among the strains evaluated in this study. We revised the abstract clarifying this purpose.
Key words: “irrigation” instead “water use”; yield; screening of mushroom quality; strain???; ..
Answer: As requested by the Reviewer, the mentioned modification was made in the revised manuscript.
Introduction: it is ok, but more literature might be considered.
Answer: More literatures were considered.
Materials and methods
2.1. Are there any reasons because the ITS 1+ 2 regions have been studied? Are they related to some particular and defined growing parameter? This aspect must be explained.
Answer: DNA sequencing of ITS 1+2 regions is adopted as a primary fungal barcode (Schoch et al., 2012; Xu et al., 2016) and we performed this analysis to investigate genetic variations among the ABI 19/03, ABI 18/04, ABI 18/02, and ABI 11/19 A. bisporus strains. Phylogenetic analysis suggests that additional molecular markers are required to properly characterize these strains at a genetic level and investigate the relationships between genotypic variations and differences in agronomic traits.
2.4. There is ONLY one crop trial.
Moreover, I think the trial is not designed in a good way: 10, 5 kg m-2 is not the habitual packaging system, and 25 L m-2 is a very hard irrigation programme to this low compost weight. The height of the casing layer (3 cm) is also uncommon (so low).
Answer: Firstly, we acknowledge the limitations imposed by the use of plastic boxes with dimensions of 40 x 30 cm in this experiment. This choice of container size did indeed result in a more confined substrate space, and we understand that it differs from the more common, larger packaging systems. We selected this size due to practical constraints and the need for control in our experimental design. The aqueous layer was previously subjected to experimentation (Marinsek, 2019), revealing that the application of 3.2 liters (per 0.9 m²) to A. bisporus colonized compost exhibited a significant augmentation in yield relative to the alternative treatments involving 0.2 liters, 0.4 liters, 0.8 liters, and 1.6 liters per 0.9 m².
With regard to the casing layer, we understand that conventional practices, mentioned in Gómez (2017) [https://doi.org/10.1002/9781119149446.ch10], normally involve the application of a casing layer 4 cm high over the compost. However, recent research, such as the study by Wang et al. (2023) [https://doi.org/10.3389/fmicb.2023.1154903], introduced the use of a thinner casing layer, specifically 2 cm high. In our study, we believe that 3cm of casing layer is adequate for mushroom harvesting.
Reference: Marinsek, B.P. Diferentes lâminas de irrigação em compost colonizado por Agaricus bisporus antes da adição da camada de cobertura. Undergraduate thesis, Universidade Estadual Paulista “Júlio de Mesquita Filho”, Campus Dracena, 2019, 35p.
How did this high water contribution affect to the temperature of the substrate? The temperature values for any time in the crop cycle must be shown.
I suppose that the authors used the oven dried measure of the compost to relativize the parameter “yield” along the crop cycle, but I think that is not the best methodology. The humidity sounding line would be able a better way to evaluate the water content of the substrates. Quality parameters (apart from unitary weight) regarding water supply must be considered. [Noble et al, 2000; Barry et al., 2016].
Answer: We appreciate the points raised in our manuscript. Unfortunately, it was not possible to evaluate temperature and other qualitative parameters to consider water supply. The compost drying method measured moisture monitoring.
Results
3.1. ITS sequencing of A. bisporus strain
What are the results of this section? If all used strains are closely related, how the authors could evaluate the relationship between irrigation system and genetic?
Answer: ITS analysis was performed as a first attempt to genetically characterize the A. bisporus strains investigated in this study. We clarified the discussion of these results to rectify that additional molecular markers and QTL analyzes, for instance, are required to suitably evaluate the relationship between genetic variations and superior agronomic traits for those strains.
3.2. First flush results: Earliness has not been evaluated. I think it would help to draw final conclusions.
Answer: We appreciate the points raised in our manuscript. Additional insights regarding precocity (earliness) were incorporated into the revised manuscript. However, no significant differences were observed, prompting its inclusion as supplementary material within the work.
In the second flush, ….…. (Table 3), indicating a good recovery….. in the control treatments, isn’t it? Figure 3. This figure was better understood if the three flushes were defined.
Answer: As requested by the Reviewer, the mentioned modification was made in the revised manuscript.
Figure 4 might be improved with the compost and casing temperature parameter evolution. What was the strain that was cultivated while moisture content was registered?
Answer: Unfortunately, we do not have data on the evolution of temperature. Figure 4 illustrates average moisture trends across all mushroom strains.
Discussion
Some part of the Discussion section might be replaced on Introduction one [references 34 and 35].
Answer: As requested by the Reviewer, the mentioned modification was made in the revised manuscript.
The results were obtained after only one crop trial. The reviewer claims that is unsuitable for conclusions
Answer: We appreciate the points raised in our manuscript. In the new version of manuscript, the conclusion has been formulated, considering the comments.
Reviewer 3 Report
The idea of addition of 25L of water per squere meter, to the copost before the addition of the casing layer, is problematic and even unrealistic in modern mushroom cultivation, due to several aspects. The authors must add a paragraph that will indicate that this experiment was done in smal scale' and can not serve as an example for mushroom growers before upscaling study using cultivation beds or shelves,
The calculation of the yields as percents of mushrooms weight of the fresh wieght the substrate (compost + casing) can lead to mistakes. The commonly used presentation is as "Biologicl efficiency" where the yield is calculated as percents of the substrate dry weight. I recomend that the authors will check the possibility to present the results as "Biologiclal efficiency", or as yield per squere meter, as mushroom yield results are presented commonly in mushroom research publucations.
Moderate editing of English language is requiered, in several paragraphs .
Author Response
Reviewer 3
The idea of addition of 25L of water per square meter, to the compost before the addition of the casing layer, is problematic and even unrealistic in modern mushroom cultivation, due to several aspects. The authors must add a paragraph that will indicate that this experiment was done in smal scale' and can not serve as an example for mushroom growers before upscaling study using cultivation beds or shelves,
Answer: The authors are grateful for the comments on this manuscript and for all the Reviewer’s comments and suggestions. As requested by the Reviewer, the mentioned modification was made in the revised manuscript.
The calculation of the yields as percents of mushrooms weight of the fresh wieght the substrate (compost + casing) can lead to mistakes. The commonly used presentation is as "Biologicl efficiency" where the yield is calculated as percents of the substrate dry weight. I recomend that the authors will check the possibility to present the results as "Biologiclal efficiency", or as yield per squere meter, as mushroom yield results are presented commonly in mushroom research publucations.
Answer: We appreciate the points raised in our manuscript. In the new version of manuscript, the Biological Efficiency (BE) has been added, considering the comments.
Author Response File: 
Author Response.pdf
Round 2
Reviewer 2 Report
Paper has been substantially improved, although some aspect regarding materials and methods section have not been able to concluded (only one trial, substrates temperature measures, other quality parameters considered, etc.). Authors have avoided them by the inclusion of the term “pilot”. I hope that they could consider these aspects in future trials. Otherwise, there are only some clarifications to consider.
Abstract: the second treatment of water added to the crop might be included in the Abstract section.
Materials and Methods
· The inclusion of BE parameter has made better the understanding of the result section, but the BE definition must be clarified:
“To calculate the biological efficiency (BE), the weight of harvested mushrooms was divided by the dry weight of the substrate…….. (compost and casing or only compost dry weight?
· The earliness parameter must be reconsidered
“The precocity was determined by dividing the harvest time into two periods (yield in the first period/total yield harvested), and the result was multiplied by 100 to obtain a percentage”
Reviewer suggests the earliness defined as the number of days between casing and the beginning of harvest of the first flush.
Author Response
Reviewer 2
Paper has been substantially improved, although some aspect regarding materials and methods section have not been able to concluded (only one trial, substrates temperature measures, other quality parameters considered, etc.). Authors have avoided them by the inclusion of the term “pilot”. I hope that they could consider these aspects in future trials. Otherwise, there are only some clarifications to consider.
The authors are gratefully for the contributions and comments about the new version the paper. We will consider the aspects cited.
Abstract: the second treatment of water added to the crop might be included in the Abstract section.
Answer: This information is already included in the abstract.
Materials and Methods
The inclusion of BE parameter has made better the understanding of the result section, but the BE definition must be clarified:
“To calculate the biological efficiency (BE), the weight of harvested mushrooms was divided by the dry weight of the substrate…….. (compost and casing or only compost dry weight?
Answer: The BE was calculated by the compost dry weight only.
The earliness parameter must be reconsidered
“The precocity was determined by dividing the harvest time into two periods (yield in the first period/total yield harvested), and the result was multiplied by 100 to obtain a percentage”
Reviewer suggests the earliness defined as the number of days between casing and the beginning of harvest of the first flush.
Answer: The days for the first flush are provided in the manuscript (page 7, first line). The term “earliness” has been added into the text. Precocity is employed to divide the cultivation timing and determine when the highest crop production occurs, which is distinct from “earliness”.
