Pollen Variability of Alnus glutinosa (L.) Gaerth. (Betulaceae) from Southern Range Edge Populations in Northern Morocco

Round 1

Reviewer 1 Report

The paper is interesting as introduce the study of polyploid Alnus glutinosa populations in Morocco that is not a well known case study. The samples for analyses on pollen morphology are many, although not systematic observations on all the parameters of the pollen have been done, and measurements were carried out on fresh pollen. The parameters selected for this study may be enough to infer general conclusions.

However, methods must be improved by explaining better the way measurements were taken, and the discussion must include more detailed comparisons with other studies also including clearly if the other studies were made on fresh or acetolysed pollen. In my opinion, the paper can be considered for publication on IJPlant Biology after reformulating the discussion section. Below, some minor remarks along the text:

 

Betulaceae = not in italics

Use ‘pollen’ or ‘pollen grains’ (not ‘pollens’) in. all the text

 

Introduction

“including [6,7], and [8] “ = including Erdtman [6], Blackmore et al. [7], and Leopold et al. [8]

Methods 2.2
“following the protocol outlined by [7]. = following the protocol outlined by Blackmore et al. [7].”
I suggest to mention the names of authors when their methods or ideas are referred in the text (and not in the case of usual references)
Add the microscope magnification of measurements.

Results
“as shown in Figures 2 and 3.” = as shown in Figures 3 and 4.
About exine description: “The exine surface of the pollen grains is both striated and granulated” = maybe, this is not clear in fresh pollen but usually the exine is described as ‘scabrate’.
Figure 3 = the quality of figure should be improved, especially the image ‘e’

Figure 4 legend = please, don’t use ‘pollens’ but ‘pollen’ or ‘pollen grains’

Figure 5 seems not so important, but if the Authors want to maintain it, the graph should and could be reduced in dimension

Table 1 legend = explain ‘Code P’

Discussion
avellana (not ‘avellena’)
4.1 = check the description of ‘striate exine’ because usually it is ‘scabrate’
4.2 = there are several repetitions with things already said in the text
“This suggests that the variability observed at the level of the pollen diameters mainly corresponds to an expression of phenotypic plasticity. Therefore, most of the variability within the analyzed samples may be the result of irregular pollen development.” = therefore, is it a phenomenon due to phenotypic plasticity (environmental trigger) or irregular microgametogenesis? Arguments supporting links between the two aspects in alder are not reported. The discussion should be articulated in a quite different way posing the different possibilities causing the observed results.

Author Response

We thank the reviewer for his interest in our manuscript. The suggested corrections and comments are pertinent and help to improve the quality and readability of the manuscript. We have incorporated all suggested corrections. For comments aimed at improving the measurements section of the methodology and discussions, we have revised and expanded the relevant sections (see modifications on the file in change-tracking mode).

 

Comments	Response
1. However, methods must be improved by explaining better the way measurements were taken.	Details of the measures have been added to the text in section 2.2. Changes are visible in the track changes mode.
2. The discussion must include more detailed comparisons with other studies also including clearly if the other studies were made on fresh or acetolysed pollen.	Section 4.2 has been revised and expanded to take account of the commentary. Changes are visible in the track changes mode.
3. Betulaceae = not in italics.	Corrected
4. Use ‘pollen’ or ‘pollen grains’ (not ‘pollens’) in all the text.	Corrected
5. Introduction: “including [6,7], and [8] “ = including Erdtman [6], Blackmore et al. [7], and Leopold et al. [8].	Corrected
6. Methods 2.2: I suggest to mention the names of authors when their methods or ideas are referred in the text (and not in the case of usual references).	Ok, done.
7. Add the microscope magnification of measurements.	Added as suggested in section 2.2
8. Results: as shown in Figures 2 and 3.	Corrected
9. About exine description.
10. Figure 3 = the quality of figure should be improved, especially the image ‘e’.	Improved ad suggested
11. Figure 4 legend = please, don’t use ‘pollens’ but ‘pollen’ or ‘pollen grains’.	Corrected as suggested
12. Figure 5 seems not so important, but if the Authors want to maintain it, the graph should and could be reduced in dimension.	Graph dimensions reduced as suggested
13. Table 1 legend = explain ‘Code P’.	Done
14. Discussion: avellana (not ‘avellena’).	Corrected.
15. 4.1 = check the description of ‘striate exine’ because usually it is ‘scabrate’.	Corrected.

 

We hope these corrections meet your expectations. We remain at your disposal for any further explanation.

Sincerely yours,

Author Response File: Author Response.pdf

Reviewer 2 Report

The study of pollen grains morphology are important for the systematic botany, aerobiology and environmental medicine (allergology), geobotany. Therefore, the study undertaken by authors of the ms is justified.

In fact it is good paper, properly designed and described. The methods are appreciate.

My main concern is related to (i)  the use of the phrase ‘pollen’ used throughout the text instead of ‘pollen grains’. To be precise the morphology  (size, shape) is related to pollen grains. Pollen is broader expression.  This comment refers to the entire text.   (ii) As the authors measured P/E ratio(0.69- 0.73) , therefore the shape can be named as oblate. Please add this information to the pollen grains morphological description.

 

Key words – the key words should not repeat the phrases form the ms title. Therefore, remove Alnus glutinosa, range edge populations.

I suggest to add – pollen grains apertures, P axis diameter, E axis diameter

Page 6/7 Table 1 add the information on the units of the measurements. I guess that you should add µm

Table 3 add explanation for P4-P 7

P 5 Fiigure 4 add the word ‘grains” i.e. it should be Observation of Alnus glutinosa pollen grains with a scanning electron microscope: a: general view of pollen grains; b: pollen grains with 5 apertures; c: pollen grains with 4 apertures; d: annulus and arcus; e:aperture with f1: ectoaperture; f2: endoaperture.

P 8 Fig 6 Please add the explanation for P1-P7

Author Response

First, we would like to thank you for this positive assessment of the work and the interest you showed in it. So, we would like to express our gratitude for the time dedicated to the review and constructive remarks which allowed us to improve the quality of our article.

In this regard, we approached every comment, hopefully, in a proper and exhaustive manner. The table below presents in detail the way of addressing each comment;

The text was marked using the “Track changes” function.

 

 

Comments	Response
1. (i)the use of the phrase ‘pollen’ used throughout the text instead of ‘pollen grains’.	Done as suggested
ii) As the authors measured P/E ratio(0.69- 0.73) , therefore the shape can be named as oblate. Please add this information to the pollen grains morphological description.	Added as suggested
Key words – the key words should not repeat the phrases form the ms title. Therefore, remove Alnus glutinosa, range edge populations.	Removed as suggested
I suggest to add – pollen grains apertures, P axis diameter, E axis diameter.	Added as suggested
Page 6/7 Table 1 add the information on the units of the measurements. I guess that you should add µm.	Added as suggested
Table 3 add explanation for P4-P 7.	Added as suggested
P 5 Figure 4 add the word ‘grains” i.e. it should be Observation of Alnus glutinosa pollen grains with a scanning electron microscope: a: general view of pollen grains; b: pollen grains with 5 apertures; c: pollen grains with 4 apertures; d: annulus and arcus; e: aperture with f1: ectoaperture; f2: endoaperture.	Added as suggested
P 8 Fig 6 Please add the explanation for P1-P7.	Added as suggested

 

We hope these corrections meet your expectations. We remain at your disposal for any further explanation.

Sincerely yours,

The Authors.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

you have made several improvements but there are still minor remarks to be corrected. In particular,  the change of some keywords and the use ' pollen grains' instead of 'pollen' are wrong, and I have to invite now the authors to return back to their previous version on this point. I send you a pdf with red evidence on what must be deleted.

Keywords:
pollen grains apertures = pollen apertures
the Paxis and E axis are not ‘key words’: you should use ‘pollen morphology’ or ‘morphopalynology’

Introduction:
including [Erdtman [6], Blackmore et al. [7], and Leopold et al. [8], = including Erdtman [6], Blackmore et al. [7], and Leopold et al. [8].

I am aware that the other referee has possibly recommended the use of ‘pollen grains’ BUT this is not the common use in palynology. Therefore, I recommended the use of ‘pollen’ as a colletive word meaning ‘many grains of pollen’. Please, delete all the ‘grains’ you added in this version.

Methods:
please remove the sentence you added about ‘oblate’ shape because it is repeated.

Pag 11 = Although you have not added the specification to the method section, I understand that you measured fresh pollen by considering that you cite Wodehouse, and by the photos.
Please, specify in the pertinent parts of the discussion section, if the comments and data from other papers were made of fresh or acetolysed pollen.

‘cytotypes’ is not in italics (?)

Comments for author File: Comments.pdf

Author Response

Cover letter to Reviewer 2

 

Thank you for the proofreading and for the corrections and suggestions.

We are pleased that the changes made to the manuscripts have at least partially met the reviewer's expectations.

All minor corrections, such as revising key words, replacing "pollen grains" by "pollen", deleting a sentence, "cytotypes" not in italics,.......; were made directly on the text following the reviewer's recommendations.

For the question relating to the precision of the use of fresh non-acetolysed pollen. In the methods section (2.2), p3, L87, we have replaced "pollen" with "fresh pollen", and in the discussions section (4.2), p11, L264.

We hope that the corrections validated and made to the manuscript are satisfactory.

Thank you

 

Sincerly yours

 

The authors

Author Response File: Author Response.pdf