Comparative Study on Pollen Viability of Camellia oleifera at Four Ploidy Levels

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The study carried out is aimed to study of correlation between the pollen size and ploidy level in Camellia oleifera on the base of pollen viability testing. Staining and in vitro germination methods were used to evaluate the fertility of some types of C. oleifera pollens at four ploidy levels. The results will shed light on the study of pollen fertility of new hybrids, and guide the hybrid breeding strategies of different varieties for C. oleifera, being Oil tea (Camellia oleifera) one of the most important woody edible oil tree species in China, with intraspecific polyploidy with the key problem in the cultivation, the acceleration and improving the selection and breeding of yield and quality of C. oleifera.

 

The manuscript is composed according to the requirements of “Agronomy”. The research methods applied are appropriate and sufficient to achieve the objectives of the study. The results are well presented and supported by tables and figures that are of good quality.

The following recommendations can be made:

Abstract:

The part below needs editing:

“The results showed that the pollen size of diploid, tetraploid, hexaploidy and octa-ploid were positively correlated with the ploidy level. Pollen viability of C. oleifera were determined by fluorescein diacetate (FDA) dye solution staining and medium containing 10% sucrose, 0.01% boric acid and 1% agar germination in vitro, which indicated that pollen viability and germination rate of the hexaploidy were relatively high among the four ploidy levels, were 79.69% and 71.78% respectively. For specific comparison, the pollen vigor of diploid NR-3, tetraploid DP43, hexaploidy CJ-12, and octoploid YNYC-1 are higher than other materials with the same ploidy level.”: The first sentence is quite long and for better understanding it can be divided into two sentences; and the second one would be better without the beginning: “The pollen vigor of diploid NR-3, tetraploid DP43, hexaploidy CJ-12, and octoploid YNYC-1 was higher than other materials with the same ploidy level.”

 

Introduction:

There is no formulated the aim of the study

 Materials and methods

 There are parts that need editing:

 

-         The last sentence in the paragraph 2.2. pollen ploidy “Each sample should be analyzed at least three times[22]” should be edited, as follow: “The analysis of each sample was done in a three time replication according to the metology requirements [22]”

 

-         The part “In this study, we used Fluorescein diacetate (FDA) fluorescence staining to evaluate pollen viability [27]. Under the fluorescence condition, the pollen that emits yellow-green fluorescence is considered to be active. Otherwise, it doesn't [28]. “in the paragraph “2.4. pollen viability”  needs editing, as follows:  “In this study, we used Fluorescein diacetate (FDA) staining to evaluate pollen viability [27]. According to the criteria for estimation of pollen efficiency given by Rotreklová,& Krahulcová[28], pollen that emits yellow-green fluorescence was considered as active.

 

 -  The last sentence in the paragraph 2.6. Statistical analysis “Use the origin pro8.5 software (origin lab company, Northampton, USA) to draw graphs

[31].” should be edited, as follow:”The origin pro8.5 software (origin lab company, Northampton, USA) was used to draw graphs

[31].”

 Results

 There was used “hexaploid” and “hexaploidy”: to be used the form “hexapod” like the others: diploid, tetraploid, octaploid, …

 

In the capture of Figure 5 was pointed that “1,2,3 and 4 in the ordinate represent diploid, tetraploid, hexaploid and octoploid respectively” , but . 1,2,3 and 4 are located on the abscissae

 Discussion

 

When authors are cited in the text, only the number of the cited literary source is given, without the year: cui et al. (2015) [40], Nico et al. (2013) [36], etc., must be: Cui et al [40], Nico et al. [36], …

 

In the second sentence of paragraph 4.1. Pollen size related to the ploidy level “In higher plants, polyploid plants generally have large organs compared with diploid plants …” large” have to be “larger”

 

At the line 11 of paragraph 4.1. Pollen size related to the ploidy level cui et al. (2015) – correct “c” with capital letter

 

In conclusion, this manuscript is recommended for publication in “Agronomy”.

 

Author Response

Response to Reviewer 1 Comments

Dear editors and reviewers,

Thank you very much for your careful review, constructive comments, and helpful suggestions with regard to our manuscript entitled “Comparative study on pollen viability of Camellia oleifera at four ploidy levels” (ID：agronomy-1978919). The comments are helpful not only for revising and improving our paper, but also for our future research. We have carefully revised the article according to the opinions of reviewers and editors, and marked it in yellow. We have revised the manuscript accordingly. The followings are specific responses:

Reviewer:

1. Abstract:

The first sentence is quite long and for better understanding it can be divided into two sentences; and the second one would be better without the beginning: “The pollen vigor of diploid NR-3, tetraploid DP43, hexaploidy CJ-12, and octoploid YNYC-1 was higher than other materials with the same ploidy level.”

Response：

Thank you very much for your good advice. We have made the adjustment according to your suggestion.

2. Introduction:

There is no formulated the aim of the study

Response：

Thank you very much for your good advice. We have made the additions according to your suggestion.

3. Materials and methods

The last sentence in the paragraph 2.2. pollen ploidy “Each sample should be analyzed at least three times[22]” should be edited, as follow: “The analysis of each sample was done in a three time replication according to the metology requirements [22]”

Response：

Thank you very much for your good advice. We have made the adjustment according to your suggestion.

The part “In this study, we used Fluorescein diacetate (FDA) fluorescence staining to evaluate pollen viability [27]. Under the fluorescence condition, the pollen that emits yellow-green fluorescence is considered to be active. Otherwise, it doesn't [28]. “in the paragraph “2.4. pollen viability”  needs editing, as follows:  “In this study, we used Fluorescein diacetate (FDA) staining to evaluate pollen viability [27]. According to the criteria for estimation of pollen efficiency given by Rotreklová,& Krahulcová[28], pollen that emits yellow-green fluorescence was considered as active.

Response：

Thank you very much for your good advice. We have made the adjustment according to your suggestion.

The last sentence in the paragraph 2.6. Statistical analysis “Use the origin pro8.5 software (origin lab company, Northampton, USA) to draw graphs[31].” should be edited, as follow:”The origin pro8.5 software (origin lab company, Northampton, USA) was used to draw graphs[31].”

Response：

Thank you very much for your good advice. We have made the adjustment according to your suggestion.

Results

here was used “hexaploid” and “hexaploidy”: to be used the form “hexapod” like the others: diploid, tetraploid, octaploid, …

Response：

Thank you very much for your good advice. We've replaced "hexaploid" and "hexaploidy" with "hexaploid".

In the capture of Figure 5 was pointed that “1,2,3 and 4 in the ordinate represent diploid, tetraploid, hexaploid and octoploid respectively” , but 1,2,3 and 4 are located on the abscissae.

Response：

Thank you very much for your careful discovery. We have made the modification.

Discussion

When authors are cited in the text, only the number of the cited literary source is given, without the year: cui et al. (2015) [40], Nico et al. (2013) [36], etc., must be: Cui et al [40], Nico et al. [36], …

Response：

Thank you very much for your good advice. We have made the adjustment according to your suggestion.

In the second sentence of paragraph 4.1. Pollen size related to the ploidy level “In higher plants, polyploid plants generally have large organs compared with diploid plants …” large” have to be “larger”

Response：

Thank you very much for your good advice. We have made the adjustment according to your suggestion.

At the line 11 of paragraph 4.1. Pollen size related to the ploidy level cui et al. (2015) – correct “c” with capital letter

Response：

Thank you very much for your careful discovery. We have made the modification.

We appreciate the efforts made by the Editors/Reviewers team and hope that our corrections and revisions are clear and sufficient. We will be happy to revise our manuscript if there are more questions and comments. We also look forward to our manuscript being accepted as soon as possible.

Sincerely,

Rui Zhao

[email protected]

 

 

Reviewer 2 Report

In the current manuscript titled “Comparative study on pollen viability of Camellia oleifera at four ploidy levels” authors compare pollen morphology and viability with the ploidy level of Camellia oleifera. The study is very streamline, very well written and studied. Authors studied 32 different types of pollens with different ploidy levels diploid, tetraploid, hexaploidy and octaploid. Authors observed These is a direct correlation between ploidy level and pollen size which is studied in various past research articles also so there is no novality here. Authors also added other morphological differences which is good for the study. Further they checked pollen viability. The current study is good and have some novality in Camellia oleifera and ploidy comparison. Overall the manuscript can be accepted with some correction in grammatical mistakes. 

Author Response

Response to Reviewer 2 Comments

Reviewer:

In the current manuscript titled “Comparative study on pollen viability of Camellia oleifera at four ploidy levels” authors compare pollen morphology and viability with the ploidy level of Camellia oleifera. The study is very streamline, very well written and studied. Authors studied 32 different types of pollens with different ploidy levels diploid, tetraploid, hexaploidy and octaploid. Authors observed These is a direct correlation between ploidy level and pollen size which is studied in various past research articles also so there is no novality here. Authors also added other morphological differences which is good for the study. Further they checked pollen viability. The current study is good and have some novality in Camellia oleifera and ploidy comparison. Overall the manuscript can be accepted with some correction in grammatical mistakes. 

Response：

Dear reviewer,

Thank you very much for your careful review, constructive comments, and helpful suggestions with regard to our manuscript entitled “Comparative study on pollen viability of Camellia oleifera at four ploidy levels” (ID：agronomy-1978919). The comments are helpful not only for revising and improving our paper, but also for our future research. We have improved the article according to your suggestions. Thanks again for your careful review.

Reviewer 3 Report

Dear authors, please consider the following suggestions when revising your paper:

2.2. pollen ploidy - capital letter for pollen. 

2.3. pollen size - capital letter for pollen. 

2.4. pollen viability- capital letter for pollen. and so on....

Table 2: Different letters in the column of the table indicate significant differences. Based on which test?

Table 3: What are different letters showing? Is it within the ploidy magnitude group of genotypes or between all genotypes? 

Please be very specific here since it influences a results interpretation. Again according to which test were the differences? 

Table 4: the same issue. 

Figure 5 and Table 5: delete the surplus punctuation. 

 

It is very well known that ploidy had a significant effect on viability and pollen grain size, and pollen grain size increased with the increase of ploidy level. What is the considerable advancement from your study? Please discuss the applicability. How do your results affect the breeding methods, can they shorten or simplify the pollen choice prior to the pollination?

 

Author Response

Response to Reviewer 3 Comments

Dear editors and reviewers,

Thank you very much for your careful review, constructive comments, and helpful suggestions with regard to our manuscript entitled “Comparative study on pollen viability of Camellia oleifera at four ploidy levels” (ID：agronomy-1978919). The comments are helpful not only for revising and improving our paper, but also for our future research. We have carefully revised the article according to the opinions of reviewers and editors, and marked it in yellow. We have revised the manuscript accordingly. The followings are specific responses:

Reviewer:

2.2. pollen ploidy - capital letter for pollen.

2.3. pollen size - capital letter for pollen. 

2.4. pollen viability- capital letter for pollen. and so on....

Response：

Thank you very much for your good advice. We have made the adjustment according to your suggestion.

Table 2: Different letters in the column of the table indicate significant differences. Based on which test?

Response：

SPSS 19.0 software (IBM, New York, USA) was used for data analysis, and one-way analysis of variance (ANOVA) was used to test the differences in average polar and equatorial lengths of Camellia oleifera. When P≤0.05, Duncan's multiple comparison method was used to evaluate the significant differences between means.

Table 3: What are different letters showing? Is it within the ploidy magnitude group of genotypes or between all genotypes? 

Response：

Different letters in a column indicate a significant difference, it is within the ploidy magnitude group of genotypes.

Table 4: the same issue. 

Response：

Different letters in a column indicate a significant difference. It is a comparison of pollen from a total of 32 species of diploid, tetraploid, hexaploidy and octaploid.

Figure 5 and Table 5: delete the surplus punctuation. 

Response：

Thank you very much for your careful advice. We have made the adjustment according to your suggestion.

It is very well known that ploidy had a significant effect on viability and pollen grain size, and pollen grain size increased with the increase of ploidy level. What is the considerable advancement from your study? Please discuss the applicability. How do your results affect the breeding methods, can they shorten or simplify the pollen choice prior to the pollination?

Response：

At present, studies on the correlation between pollen and ploidy have been reported in some plants, while studies on pollen viability and ploidy in Camellia oleifera have not been reported. This study showed the correlation between pollen size and ploidy level of C. oleifera. The fertility of different types of C. oleifera pollen at four ploidy levels was evaluated by staining and in vitro germination. The results will be helpful for the study of pollen fertility of new camellia varieties and guide the hybrid breeding strategy of different camellia varieties. C. oleifera is one of the most important woody edible oil tree species in China. Intraspecies polyploidy is a key problem in C. oleifera cultivation. Our study will help accelerate and improve the breeding of C. oleifera for yield and quality.