Genetic Characterization and Fruit Quality Analysis of New Mandarin Hybrids

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

-Please specify exactly how many TRAPS and SSRs were used? The information is in the table but introduce it in the material and method.

-Please justify the use of those markers instead of sequencing given the low depth sequencing is very economical, cost? Allelic variations?

-I can not find data on color, shapes nor peel adhered. Please provide the rest of the data

 

#22-English needs to clarified

#45-state why they would not be suitable for the foreign market

#51-reference for 78 descriptors

#54-56-check English

#56-reference for the location

#58-Clearly define how many trees were studied.

#72-15 fruits per each tree? Not clear

#77-give in a table, mean, SD, and median of the measurements

#148-some? Not all 78?

-the resolution of the pictures of the leaf in (figure 1) could be improved-caption needs to be fixed

-figure 2 imaged need to be cleaned up. You can still the outline from the cropping process

#229-230-not readible

 -275-reference

-Figure 7, 8 please increase the resolution of the figures

#315-I do not see Rangpur in the Figure 7. 

Comments on the Quality of English Language

There are some sentences that could be improved so the readers can better understand what the authors are trying to convey. 

Author Response

COMMENTS AN AND SUGGESTIONS FOR AUTHORS

 

REVIEW REPORT1

Please specify exactly how many TRAPS and SSRs were used? The information is in the table but introduce it in the material and method.

We agree with this comment. Therefore, we have added the requested information between the lines 135-139.

Please justify the use of those markers instead of sequencing given the low depth sequencing is very economical, cost? Allelic variations?

We agree with this comment. SSR and Traps markers are used in plant genetic improvement programs and are important tools for faster and more efficient production of plants with advantages for agricultural production. So, we added a comment about this at the end of the Introduction item.

I cannot find data on color, shapes nor peel adhered. Please provide the rest of the data

Indeed, the color evaluation results were based on descriptors and not on the use of a colorimeter. Therefore, we removed it from the methodology item.

 

#22-English needs to clarify

The English language has been revised.

#45-state why they would not be suitable for the foreign market

They are not suitable mainly because they are not seedless, among other characteristics. Information was added in line 45.

#51-reference for 78 descriptors

This information was included in the methodology item.

#54-56-check English

The English language has been revised.

#56-reference for the location

We agree with this comment. We have added the location of the Centro de Citricultura SM/IAC.

 

#58-Clearly define how many trees were studied.

We agree with this comment. Therefore, we have added the information in the methodology item.

 

#72-15 fruits per each tree? Not clear

We agree with this comment. Therefore, we have added the information in the methodology item.

 

#77-give in a table, mean, SD, and median of the measurements

We appreciate the suggestion, but we have chosen to present the main features in the figures 2, 3 and 4.

#148-some? Not all 78?

We agree with this comment. We have revised the information in the methodology item.
the resolution of the pictures of the leaf in (figure 1) could be improved-caption needs to be fixed

we will send the original figures.

figure 2 imaged need to be cleaned up. You can still the outline from the cropping process

 we will send the original figures.

#229-230-not readable        

we will send the original figures.

-275-reference

The sentence in the text was modified.

-Figure 7, 8 please increase the resolution of the figures

we will send the original figures.

#315-I do not see Rangpur in the Figure 7.

The correct is Cravo Mandarin. It was changed in the text.

Comments on the Quality of English Language
There are some sentences that could be improved so the readers can better understand what the authors are trying to convey

The English language has been revised.

Reviewer 2 Report

Comments and Suggestions for Authors

Dear authors,

I think that your work is suitable for this journal, and have been carried out competently. The manuscript is well written and presented.

However, I have some suggestions and  several corrections, most of them are editorial ones, so MS can be published after minor revision.

Abstract.   It seems to me that more information about the results of this work can be given in the abstract. This article will benefit from this. In its current form it mainly contains introductory information.

 Line 12    The ease of crosses among citrus species has been used

I think it would be better to write “among citrus taxa”

 I know that the systematic of citruses is very complicated, but not all readers of the "Agronomy" are experts in this field. That’s why  it is advisable to avoid discrepansies within the same text.

 For example:

Lines  14, 50, 60   Ponkan tangerine    Line  92  Ponkan mandarin

Line 32    Cravo mandarin              Line 153   Cravo tangerine

Line 64 Rangpur lime                Line 315  Rangpur mandarin It is not clear to all readers the difference between tangerine and mandarine. So it is better to explain it, and to use  one word  for the same  sample.

Line 30  The state of São Paulo is the largest Brazilian producer of tangerines, primarily the Ponkan (Citrus reticulata Blanco) and Murcott tangor (C. sinensis L Osbeck x C. reticulata) varieties, followed by tangerines (C. delicious Tenore) and Cravo mandarin (C. reticulata) 32 [1,2].

It is unclear whether all of listed items are tangerines, as is stated in the beginning of this phrase, or only one.

 Line 44  Another factor to considered is that most

Another factor to be considered is that most

Line 55 The have the potential to emerge as new commercial varieties.

They  have  the potential to emerge as new commercial varieties

Line 153   Additionally, individuals TMxTP 4 and TM x TP 8, along with the Murcott tangor, exhibited obtuse leaves (Figure 1).

However,  unmentioned individual TMxTP2 is shown on Figure 1 as representing obtuse leave.

Line 186 Among the hybrids most like Murcott tangor,  and       Line 196 However, for Ponkan “like” hybrids,

It is better to use a single format for the word “like”, with or without quotes

 Line 188 when compared to the other individuals

Line 198  in relation to the other  accessions

Line 202 Therefore, for the Ponkan group, there are also hybrids  with the potential to extend the period of fruit supply.

Do you mean that “Therefore, for the Ponkan group,  hybrids  with the potential to extend the period of fruit supply are needed and important»?  Lines 229-230          Some kind of glitch when printing the opening version of MS.  Line 250 compared to the other hybrids.  Line 251    showing statistical differences  across the three evaluated times.

It is not clear, what is meant by “evaluated times”.

Line 255  After 72 hours, four individuals (26.6%) were asymptomatic, while eleven were  symptomatic (73.3%),

Again it is not clear. Are eleven individuals are hybrids and four are parents?  It should be written more clearly.

 Figure 2  The format of the names of hybrids (TMXTP) in the legend differs from that in the text and in other legends (TM xTP)

Table 3    Number of allelos          should be:  Number of alleles

Line 312   Figure 7 …. depicted the presence of the male parent tangerine Ponkan and its hybrids TMxTP 13 and TMxTP 12 in group 1. In 313 the second group, the female parent tangor Murcott was grouped with the remaining hybrids from the cross,

The  topology on Figure 7 (one clade is Ponkan + TM x TP12 + TM x TP13,  the other clade is Murkott + TM x TP 1-11)  differs from that  on  Figure 8 (Ponkan + TM x TP 12-13 + TM x TP 1,5,3,11  and Murkott  + TM x TP2 + TM x TP 2, 7, 8, 9) .  But this pattern is not discussed in MS.

Line 315   the Rangpur mandarin was not closely associated with any hybrid, demonstrating a similarity of (0.47) in 316 comparison to the other individuals.

The Rangpur mandarin is not depicted on Figure 7.

Line 327          individual TM x TP 2, has a neck  that differs significantly from the other hybrids (Figure 7).

It is not clear, what does mean “has a neck”.

Line 345  In group 1, the Murcott tangor female parent was present, along with the same hy brids observed previously, namely TMxTP (2, 4, 7, 8, 9).

It is not clear to what Figure refers this phrase.

References.  The names of the  journals should be given in abbreviated form, this needs editing.

         Comments on the Quality of English Language

Minor editing of English language required.

Author Response

 

Reply review report2

Line 12 The ease of crosses among citrus species has been used I think it would be better to write “among citrus taxa”

We appreciate the suggestion. We made the modification.

 

I know that the systematic of citruses is very complicated, butnot all readers of the "Agronomy" are experts in this field. That’swhy it is advisable to avoid discrepansies within the same text.

For example:Lines 14, 50, 60 Ponkan tangerine Line 92 Ponkan mandarin

 

We appreciate the suggestion. We made the modification.

 

Line 32 Cravo mandarin Line 153 Cravo tangerineLine 64 Rangpur lime Line 315 Rangpur mandarin It isnot clear to all readers the difference between tangerine andmandarine. So it is better to explain it, and to use one word forthe same sample.

 

We appreciate the suggestion. We made the modification.

 

Line 30 The state of São Paulo is the largest Brazilian producerof tangerines, primarily the Ponkan (Citrus reticulata Blanco) andMurcott tangor (C. sinensis L Osbeck x C. reticulata) varieties,followed by tangerines (C. delicious Tenore) and Cravo mandarin(C. reticulata) 32 [1,2].It is nlyr whether all of listed items are tangerines, as isstated in the beginning of this nly, or nly one.

 

We appreciate the suggestion. We made the modification: The São Paulo state is the largest Brazilian producer of mandarins, mainly Ponkan (Citrus reticulata Blanco) and Murcott tangor (C. sinensis L Osbeck x C. reticulata), followed by Willow Leaf mandarin  (C. delicious Tenore) and Cravo mandarin (C. reticulata) [1].

 

 

Line 44 Another factor to considered is that most. Another factor to be considered is that most

We appreciate the suggestion. We made the modification.

 

 

Line 55 The have the potential to emerge as new comercial varieties. They have the potential to emerge as new commercial varieties

We appreciate the suggestion. We made the modification.

 

Line 153 Additionally, individuals TMxTP 4 and TM x TP 8,along with the Murcott tangor, exhibited obtuse leaves (Figure 1).

However, unmentioned individual TMxTP2 is shown on Figure 1as representing obtuse leave.

 

We appreciate the suggestion. We made the modification. Additionally, individuals TMxTP 2, TMxTP 4 and TM x TP 8, along with the Murcott tangor, exhibited obtuse leaves (Figure 1).

 

Line 186 Among the hybrids most like Murcott tangor, and Line 196 However, for Ponkan “like” hybrids, It is better to use a single format for the word “like”, with orwithout quotes

We removed the quotation marks.

 

 

Line 188 when compared to the other individuals

Line 198 in relation to the other accessions

Line 202 Therefore, for the Ponkan group, there are also hybrids with the potential to extend the period of fruit supply.Do you mean that “Therefore, for the Ponkan group, hybrids with the potential to extend the period of fruit supply are neededand important»

 

We appreciate the suggestion. We made the modification: Therefore, for the Ponkan group, there are also hybrids with the potential to extend the period of fruit supply which is an important and necessary characteristic.

 

? Lines 229-230 Some kind of glitch whenprinting the opening version of MS. Line 250 compared to theother hybrids.

We appreciate the suggestion. We made the modification.

 

Line 251 showing statistical differences acrossthe three evaluated times.

It is not clear, what is meant by “evaluated times”.

We appreciate the suggestion. We made the modification: Moreover, these values exceeded those of Ponkan and Murcott, showing statistical differences across the hours after inoculation.

 

Line 255 After 72 hours, four individuals (26.6%) wereasymptomatic, while eleven were symptomatic (73.3%). Again it is not clear. Are eleven individuals are hybrids and fourare parents? It should be written more clearly.

We made the correction in the text: After 72 hours after inoculation, four individuals (26.6%) were asymptomatic (the hybrids TM x TP 1, TM x TP 3, TM x TP11 and the parental Pera Sweet orange), while eleven genotypes were symptomatic (73.3%), eight hybrids  TM x TP 2, TM x TP 4, TM x TP 5, TM x TP 7, TM x TP 8, TM x TP 9, TM x TP 12 and TM x TP 13, the controls (Cravo and Ponkan mandarins) and the parental Murcott tangor, exhibiting varying levels of disease severity (Table 2).

 

Figure 2 The format of the names of hybrids (TMXTP) in thelegend differs from that in the text and in other legends (TM xTP)

We appreciate the suggestion. We made the modification.

 

 

Table 3 Number of allelos should be: Number of alleles

 

We appreciate the suggestion. We made the modification.

 

Line 312 Figure 7 …. depicted the presence of the male parentt angerine Ponkan and its hybrids TMxTP 13 and TMxTP 12 in group 1.  In 313 the second group, the female parent tangorMurcott was grouped with the remaining hybrids from the cross,

The topology on Figure 7 (one clade is Ponkan + TM x TP12 +TM x TP13, the other clade is Murkott + TM x TP 1-11) differsfrom that on Figure 8 (Ponkan + TM x TP 12-13 + TM x TP1,5,3,11 and Murkott + TM x TP2 + TM x TP 2, 7, 8, 9) . Butthis pattern is not discussed in MS.

We believe that this paragraph can explain:

In group 2, the Murcott tangor female parent was present, along with the same hybrids observed previously, namely TMxTP (2, 4, 7, 8, 9). Within this clade, the similarities varied, indicating that no hybrid was identical to another. Such variation can be attributed to the type of marker used, as TRAP markers generate numerous polymorphic bands compared to microsatellite markers, rendering them more effective for detecting polymorphisms and distinguishing between individuals.

 

 

Line 315 the Rangpur mandarin was not closely associated withany hybrid, demonstrating a similarity of (0.47) in 316comparison to the other individuals.

The Rangpur mandarin is not depicted on Figure 7.

We made the modification to Cravo mandarin.

 

Line 327 individual TM x TP 2, has a neck that differssignificantly from the other hybrids (Figure 7). It is not clear, what does mean “has a neck”.

This fruit characteristic, the neck, can be observed in figure 3. We made the correction in the text.

 

Line 345 In group 1, the Murcott tangor female parent was present, along with the same hy brids observed previously,namely TMxTP (2, 4, 7, 8, 9).

It is not clear to what Figure refers this phrase.

 

We made the correction in the text. In group 2, the Murcott tangor female parent was present, along with the same hybrids observed previously, namely TMxTP (2, 4, 7, 8, 9).

 

 

 

 

 

Reviewer 3 Report

Comments and Suggestions for Authors

The manuscript entitled "Genetic characterization and fruit quality analysis of new mandarin hybrids" by Mariângela Cristofani-Yaly et al., genetically and phenotypically evaluates eleven hybrids of the cross between Murcott tangor and Ponkan tangerine for the selection of varieties with commercial potential. This manuscript was clearly written down and the results look promising; however, from my point of view, it lacks marketable storage or shelf-life conditions which are very important and meaningful. So, this manuscript has a major revision, and details are listed below.

·       General comment: The manuscript does not consider the order. For example, the title mentions the genetic evaluation before fruit quality, even though the manuscript addressed fruit quality first, then genetic assessment. Also, would be better if the authors had drawn all steps of the manuscript in a pictorial form.

·       Lines 13−15 "In previous studies, eleven hybrids of the cross between Murcott tangor and Ponkan tangerine were obtained and identified. Plants of these hybrids were grafted onto Rangpur lime rootstock and genetically and phenotypically evaluated for the selection of varieties with commercial potential." This point should be indicated in detail in the materials and methods (M&M) section. Also, the importance of this rootstock should be indicated. e.g. In the M&M section, the reader will wonder if these are hybrids that are normally generated by seed...why they are grafted?

·       Line 67: A reference or website of this descriptor must be included. This descriptor is ambiguous throughout the manuscript and needs clarification, e.g. Line 148.

·       Line 99: Is this a scientific way to confirm the target A. alternata strain? please add ref.

·       Line 100: PAD and BOD should be mentioned in full form not abbreviated.

·       Line 103: Please mention the size of the Petri dishes that are used.

·       Line 113: ", respectively" is missing after A. alternata.

·       Line 117: Which software did you use for performing the statistics?

·       Line 123: The number of TRAP primers that were used should also be cleared.

·       Line 128: The whole recipe of the PCR reaction should be stated.

·       Line 132: The type of DNA ladder is missing since you use it for calculating the band sizes.

·       Line 132: The type of DNA ladder is missing since you use it for calculating the band sizes.

·       Line 154: Which leaf was relied upon for the description - was it the third one or what?

·       Line 207: The figures need quality improvement and should clarify the criteria for positioning the upper and lowercase letters, explaining the reason for not being collected in the M&M section.

·       Line 306: Table2: How could Trap2 seq be different? Please check. Also, check the Trap11 seq.

 

·       A combined analysis can be done using the data generated by both markers, and creating a single dendrogram, as well as a PCA analysis will be useful and helpful to understand the results in a greater way.

Author Response

COMMENTS AN AND SUGGESTIONS FOR AUTHORS

 

Reply review report3

 

 

Lines 13−15 "In previous studies, eleven hybrids of the cross between Murcott tangor and Ponkan tangerine were obtained and identified. Plants of these hybrids were grafted onto Rangpur lime rootstock and genetically and phenotypically evaluated for the selection of varieties with commercial potential." This point should be indicated in detail in the materials and methods (M&M) section. Also, the importance of this rootstock should be indicated. e.g. In the M&M section, the reader will wonder if these are hybrids that are normally generated by seed...why they are grafted?

 

We appreciate the suggestion.  This information was added to the methodology as suggested. The original plants, from seeds, are kept in pots, containing 60L, in a greenhouse.  For multiplication and evaluation in the field, the hybrids were grafted onto Rangpur lime (C. limonia Osbeck). The experimental design was a randomized blocks design, with one plant per plot and four replications, spaced 6 m between rows and 3 m between plants in the same row, totalizing 60 plants.

 

 Line 67: A reference or website of this descriptor must be included. This descriptor is ambiguous throughout the manuscript and needs clarification, e.g. Line 148.

The website has been included, and the text has been revised.

 

Line 99: Is this a scientific way to confirm the target A. alternata strain? please add ref. · Line 100: PAD and BOD should be mentioned in full form not abbreviated.

We appreciate the suggestion. Is this a scientific way to confirm A. alternata described in the cited literature (Peever et al 1999). Other suggestions were added to the text.

 

Line 103: Please mention the size of the Petri dishes that are used. ·

This information was added.

 Line 113: ", respectively" is missing after A. alternata. ·

This word was added.

 Line 117: Which software did you use for performing the statistics?

Fruit quality parameters as well as ABS leaf severity were evaluated using the statistical program SASM-AGRI [9]

 

Line 123: The number of TRAP primers that were used should also be cleared.

This information was added.

 

 Line 128: The whole recipe of the PCR reaction should be stated. ·

This information has been referenced.

 

Line 132: The type of DNA ladder is missing since you use it for calculating the band sizes. ·

 This information was added.

 

Line 154: Which leaf was relied upon for the description - was it the third one or what?

It was the third one on the selected branch, since the plants were mature.

 

Line 207: The figures need quality improvement and should clarify the criteria for positioning the upper and lowercase letters, explaining the reason for not being collected in the M&M section.

The quality of the figure has been improved. We will send the original figures.

 

Line 306: Table2: How could Trap2 seq be different? Please check. Also, check the Trap11 seq

A combined analysis can be done using the data generated by both markers, and creating a single dendrogram, as well as a PCA analysis will be useful and helpful to understand the results in a greater way

Thank you for the observation. We have made the correction.

 

 

 

Round 2

Reviewer 3 Report

Comments and Suggestions for Authors

Next time, it is preferable to put the line number in the revised version when responding to comments for easy tracking