Development and Optimization of Label-Free Quantitative Proteomics under Different Crossing Periods of Bottle Gourd

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Reviewer #1:

Is the research design appropriate? Can be improved

Needful done.

Are the methods adequately described?

Can be improved

The methods have been corrected as per the suggestions.

Kindly refer line no.  114-327.

Are the results clearly presented? Can be improved

The results have been improved as suggested.

Kindly refer line no.  328-628.

Are the conclusions supported by the results? Can be improved

Rewrote the conclusion to support the findings of the present study.

Kindly refer to line no.  722-743.

1

 In line 108, the experimental place should display the altitude.

The altitude has been added.

Kindly refer line no.  119.

2

Line 121, leave and seed were harvest at different period, why the author choose the leaves? Which type of leave you choose?

The leaves were chosen to determine the quantification of proteins for superior seed quality and to compare the seed protein quality under the source-sink relationship. Tender leaves present at the axillary buds were chosen for this study.

Kindly refer line no. 147-148.

3

Line 130, seeds were extract from each crossing periods? How to identified the crossing successful and protect from pollution (insects and other factor disturb)?

The fruits were harvested from the hybrid and parental lines at maturity, when colour turned from green to brown. Seeds were extracted manually by slicing the ripened fruits into small pieces and seeds were separated by squeezing the fruit pulp.

At the start of the crossing period in hybrid and parental lines seed production, the male flower was chosen in the parental lines and bagged with paper bags in the previous evening and pinched off in the next morning and kept carefully in a glass container.

Kindly refer line no. 134-142.

After bagging, waited for one week and observed female flower ovary growth. The increased size of the ovary confirmed the successful crossing after that it was tagged using different color threads as shown in the picture.

4

F4 period showed different from others? Why, dose the author think the real-time and other methods to prove this?

Based on the speed of germination studies, seed vigour indexes, seed test weight, and higher antioxidant potential of seeds, we proved that F4 period showed different from others (the data is not presented in the manuscript).

Kindly refer line no. 143-148.

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Reviewer #2:

Does the introduction provide sufficient background and include all relevant references? Can be improved

The introduction has been improved as per the suggestions.

Kindly refer to line no.  45-113.

Are the methods adequately described? Can be improved

The methods have been corrected as per the suggestions.

Kindly refer line no.  114-327.

Are the results clearly presented? Can be improved

The results have been improved as suggested.

Kindly refer line no.  328-628.

Are the conclusions supported by the results? Must be improved

Rewrote the conclusion to support the findings of the present study.

Kindly refer to line no.  722-743

1

The introduction is correctly described, I checked, but overall I am not convinced that this is the first report.

Rectified the mistake and removed the first report.

Kindly refer to line no. 104.

2

The results of the paper, according to the authors, will facilitate future scrutiny of protein related to quality traits and agronomic importance of bottle gourd through selective breeding programs. And in general in the world of Molecular Plant Sciences.  it is very important.   It is suggested to compare this species with other species in the future.

Thank you for this valuable suggestion. We will do it in further studies.

3

Figure 2 is not clear and needs further explanation.

Figure 2 explains the interaction plot of seed vigour potential of bottle gourd hybrid HBGH-35 and two parental lines and explained that hybrid HBGH-35 at F4 crossing periods had maximum seed vigour potential.

4

Due to the large number of tables, it makes reading challenging and requires careful editing of the text.

The text has been carefully revised as suggested throughout the manuscript.

5

In terms of written language, it needs some changes.

The English editing has been done throughout the manuscript.

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Reviewer #3:

1

Update references for introduction and discussion

The references have been updated for introduction and discussion.

2

Add recommendations for farming community

Generally, the farmers produce tender fruits for selling as vegetables, and in the last weeks of the crossing period they leave fruits for seed production, which results into poor quality of seeds with higher numbers of immature and unfilled seeds. According to the findings of the present study, it is recommended that if farmers keep the fruits of the initial crossing period for seed purposes, the subsequent seed setting won‘t be good enough. Due to higher source-sink strength in the older growing fruits, the seed setting in younger ovaries of subsequent developing fruits in the upcoming crossing period will be inhibited. The photosynthetic activities and limited transfer of assimilates in developing fruits and seeds do not provide enough energy to sustain their own development. The first developing fruit on the basal vine node utilizes a more considerable portion of photosynthates than subsequent fruit formed on the upper node. Thus, the earliest developed fruits are bigger in cucurbits due to better source-sink strength and have a suppressive impact on the subsequent fruits.

The farmers‘ income will be increased many folds if he/she sells the tender fruits from early crossing periods and leave the fruits set during the 4th week of the crossing period for seed production. Because during the 4th week of the crossing period, the overall growth of the vine is robust and higher assimilate transfer to the developing seeds and it yields higher quality of seeds. This seed production model, allows farmers to take the seed production and tender fruit production in the same season and on the same plant, without compromising the seed quality. Secondly, farmers get additional income from sale of tender fruits in market. The crossing in the 4th week after the anthesis of first flowers resulted in high-quality seed production without affecting tender fruit yield. Hybrid HBGH-35 gave maximum seed yield, better seed quality, and greater economic returns, which could be exploited in future sustainable seed production programs.

3

Format the references according to journal recommended

Needful done