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Article
Peer-Review Record

Arbuscular Mycorrhizal Fungi Associated with Tree Species in a Planted Forest of Eastern China

Forests 2019, 10(5), 424; https://doi.org/10.3390/f10050424
by Jinping Wang 1,2, G. Geoff Wang 2, Bo Zhang 3, Zhongming Yuan 1, Zhiyuan Fu 1, Yingdan Yuan 1, Lingjun Zhu 1, Shilin Ma 1 and Jinchi Zhang 1,*
Reviewer 1: Anonymous
Reviewer 2: Anonymous
Forests 2019, 10(5), 424; https://doi.org/10.3390/f10050424
Submission received: 17 April 2019 / Revised: 9 May 2019 / Accepted: 15 May 2019 / Published: 16 May 2019
(This article belongs to the Special Issue Forest Microbial Communities and Processes)

Round  1

Reviewer 1 Report

The work presented entitled “Effects of edaphic factors on arbuscular mycorrhizal fungi in a planted forest of eastern China” is an interesting work that needs to be better explained before acceptance in Forests. 

Few insights for your convenience: 

Introduction is Ok, and there you introduce all the important issues you want to discuss.

Material and Methods

You said that your study was undertaken in 2017 but the plants were installed in 1990-1998, which means they have 21-27 years old. However you previously said that the forest was established in 1959. What forest? And since you say that in spite of the efforts afforestation was not properly accomplished, you need to characterise better the stands where you were working. And this means, average height, differences between stands, “healthy” aspect. Did the plants mycorrhizae in the field or did they have been previously mycorrhized before planting? Apparently the stands have a mixed tree species but you didn’t refer in how many stands did you collect your samples. On the other hand what is the influence of a shelter forest for all your stands!? Please give a better detailed description of all your sampling site. 

Results/Discussion

It is better to reduce the information you provide, particularly I refer figure 1, which is not quite informative since you have already table 2, although not properly with the same data. Relationship between soil factors and AMF community (Table 5) are important but you also said in your objectives that you want to evaluate the influence of abiotic factors on fungal and species distribution. Under this, why did you not establish correlations between AMF colonisation and tree species in order to understand the role of AMF on sustainable tree growth. On the other hand you show that AMF can establish - with different degrees of association - with the studied species. 

Of course you found important relationships between AMF and soil characteristics but you said nothing about the variations you observed in terms of soil properties and tree species. Are these due to soil heterogeneity or have been influenced by tree species? This is important to clarify for a better understanding of your discussions. Some aspects, for example P and OM, are repeatedly justified in your discussions. You should avoid those repetitions. Under all these data I do not understand how your study can provide insights on the utilization and management of AMF to maintain sustainable management of forests. What were thus your suggestions to fulfil the initially aim of this study and give an added value for afforestation?

Author Response

Dear reviewer:

 Thank you for your constructive comments of our manuscript. Your suggestion is  helpful for improving quality of the manuscript. We have revised the manuscript  using *Track Changes* function according to your comments which we hope meet with  approval. The responses to your comments are showed below: 

Point 1: The work presented entitled “Effects of edaphic factors on arbuscular mycorrhizal fungi in a planted forest of eastern China” is an interesting work that needs to be better explained before acceptance in Forests.  Introduction is Ok, and there you introduce all the important issues you want to discuss.

Response 1: Thank you for your favourable comments on our manuscript.

Point 2: Material and Methods: (1) You said that your study was undertaken in 2017 but the plants were installed in 1990-1998, which means they have 21-27 years old. However you previously said that the forest was established in 1959. What forest? And since you say that in spite of the efforts afforestation was not properly accomplished, you need to characterise better the stands where you were working. And this means, average height, differences between stands, “healthy” aspect. (2) Did the plants mycorrhizae in the field or did they have been previously mycorrhized before planting? Apparently the stands have a mixed tree species but you didn’t refer in how many stands did you collect your samples. On the other hand what is the influence of a shelter forest for all your stands!? Please give a better detailed description of all your sampling site. 

 Response 2: (1) We apologize that we indeed did not say clearly in the paper before. Chinese government had been trying to develop forestry in the Jiangsu coastal area. Dafeng forest farm was established to developing plantations in the 1950s, and different tree species was transplanted to the farm at different time. Eight ecological and economic tree species in the present study, C. illinoensis, P. lasiocarpa, M. glyptostroboides, Z. serrata, T. ‘zhongshansha’, E. ulmoides, G. biloba, and E. pungens, were imported from local tree nurseries in 1990–1998. We have revised it in study sites and sampling section to make it clearly, please see line 90-99. (2) Generally, seedlings growth in field will colonized by AMF. The trees in our study were imported from local tree nurseries, so despite the roots of transplanted trees might be companied with some AMF, but the AMF species were local AMF. Moreover, many years after the transplant, the trees and the local AMF species would be form a more favourable symbiotic relationship in order to adapt to local environment. We have re-declared the stands and sample number in line104-108. The soil properties influenced by shelter forest were presented in table 4. I have made a better detailed description of sampling site. Please reread it.

 Point 3: Results: (1) It is better to reduce the information you provide, particularly I refer figure 1, which is not quite informative since you have already table 2, although not properly with the same data. (2) Relationship between soil factors and AMF community (Table 5) are important but you also said in your objectives that you want to evaluate the influence of abiotic factors on fungal and species distribution. Under this, why did you not establish correlations between AMF colonisation and tree species in order to understand the role of AMF on sustainable tree growth. On the other hand you show that AMF can establish - with different degrees of association - with the studied species. 

Response 3: (1) Indeed, Some information in figure 1 and table 2 were coincided, but they cannot be replaced by each other. Table 2 showed the occurrence of AMF species in rhizosphere soil of every trees species, and the occurrence frequency, relative abundance and important value of each AMF species, but figure 1 can showed the abundance of AMF species means percentage of AMF species in rhizosphere soil of every trees species. This information were very important for understanding the AMF community composition. So we thought figure 1 should not be deleted. (2) We are very sorry for that we did not say clearly in the paper before. Our objectives were to evaluate the influence of abiotic factors on root colonization and fungal community composition, but not distribution, we have revised it in introduction section, see line 69, line 77-80, line 288.

Point 4: Discussion: (1) Of course you found important relationships between AMF and soil characteristics but you said nothing about the variations you observed in terms of soil properties and tree species. Are these due to soil heterogeneity or have been influenced by tree species? This is important to clarify for a better understanding of your discussions. (2) Some aspects, for example P and OM, are repeatedly justified in your discussions. You should avoid those repetitions. (3) Under all these data I do not understand how your study can provide insights on the utilization and management of AMF to maintain sustainable management of forests. What were thus your suggestions to fulfil the initially aim of this study and give an added value for afforestation?

Response 4: (1) We have supplemented the instruction about the variations in terms of soil properties and tree species in discussion section, see 276-278. (2) The repeated OM in discussion has been deleted, see line 274-277. (3) About the insights on the utilization and management of AMF to maintain sustainable management of forests, we have made an revision in conclusion section. In the studied planted forest, the roots of most trees were colonized by AMF. Among them, the roots of C. illinoensis, Z. serrata, T. zhongshansha, E. ulmoides, and E. pungens were readily colonized by AMF which indicated that the application of AMF in afforestation of these tree species had a great potential. AMF colonization rate was significantly and negatively correlated with soil P which suggested that the application of AMF in afforestation was more suitable in low P soil. In the rhizosphere soil of all the examined tree species, abundant spore numbers and high diversity of AMF species were found. Glomus and Acaulospora were the dominant genera means they were the most promise genera for applying in afforestation. The AMF community composition and diversity were highly related to the host trees and edaphic factors which indicated that local edaphic factors should be considered when applying AMF in afforestation. In total, the results of this study could be beneficial for the sustainable management of planted forests.

Special thanks to you for your good comments! Hope to learn more from you!

Reviewer 2 Report

Authors studied the Arbuscular mycorrhizal fungi diversity in tree plantations. They studied AMF diversity in variety of tree species. I highly appreciate the authors effort to study the correlation between AMF diversity and edaphich factors. Authors used conventional methods such as AMF colonization and spore identification. It would have been nice to utilize molecular methods for the better understanding of AMF diversity and their correlation between edaphic factors. However, the current experimental results are still valid.

Generally, the manuscript is clear and written well. I have following comments,

Line 42: Soil microorganisms technology provides.....

I would modify the sentence as "Harnessing soil microbes provides.....

Line 101 to 104: The sampling methods are not clear. Please provide total number of samples taken for analysis. Also, please provide details about sampling of rhizosphere soil. Is the rhizosphere soils are soils adhere to roots? Please clarify it properly. 

Line 117: AP and total phosphorus.....

Please explain what is "AP"? 

Line 118: AK and total potassium...

Please explain what is "AK"?

Line 126: The clearing washed fine root....

I would modify the sentence as "The clearly washed fine roots..... 

Line 350 and 351: Glomus and Acaulospora are the genus, please modify the sentence to make it clear. 

Author Response

Dear reviewer:

 Thank you for your constructive comments of our manuscript. Your suggestion is  helpful for improving quality of the manuscript. We have revised the manuscript  using *Track Changes* function according to your comments which we hope meet with  approval. The responses to your comments are showed below:

Point 1: Authors studied the Arbuscular mycorrhizal fungi diversity in tree plantations. They studied AMF diversity in variety of tree species. I highly appreciate the authors effort to study the correlation between AMF diversity and edaphich factors. Authors used conventional methods such as AMF colonization and spore identification. It would have been nice to utilize molecular methods for the better understanding of AMF diversity and their correlation between edaphic factors. However, the current experimental results are still valid. Generally, the manuscript is clear and written well. I have following comments,

Response 1: Thank you for your favourable comments on our manuscript.

Point 2: Line 42: Soil microorganisms technology provides..... I would modify the sentence as "Harnessing soil microbes provides.....; Line 101 to 104: The sampling methods are not clear. Please provide total number of samples taken for analysis. Also, please provide details about sampling of rhizosphere soil. Is the rhizosphere soils are soils adhere to roots? Please clarify it properly; Line 117: AP and total phosphorus..... Please explain what is "AP"?; Line 118: AK and total potassium... Please explain what is "AK"?; Line 126: The clearing washed fine root.... I would modify the sentence as "The clearly washed fine roots.....;  Line 350 and 351: Glomus and Acaulospora are the genus, please modify the sentence to make it clear. 

Response 2: We are very sorry for our incorrect writing. We had corrected them: Soil microorganisms technology provides has been modified as Harnessing soil microbes provides, please see line 44; Total number of samples taken for analysis was provided in the sampling methods, please see line 107; Rhizosphere soil are soil adhere to roots, supplement instruction was added behind rhizosphere soil, please see line 106; AP and AK were explained, please see line 121 and line 123; The clearing washed fine root, has been modified as The clearly washed fine roots, please see line 130; Glomus and Acaulospora were the dominant genera, has been made clearly, please see line 357-358.

Special thanks to you for your good comments! Thanks again! Hope to learn more from you!

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