Phenotypic and Molecular-Markers-Based Assessment of Jamun (Syzygium cumini) Genotypes from Pakistan

Suggestions

Changes made

 In the abstract, mention where the phenotypic characterization was carried out, and that it was carried out in situ. Include in general terms the genotype selection criteria. The same for molecular characterization, in which laboratory it was carried out.

Surveys were made in natural jamun inhabits across Punjab, Pakistan and genotypes were marked based on visual diversity of tree and fruits.

Phenotypic characterization was done in situ.  Properly preserved leaves (for DNA extraction) of phenotypically characterized plants were brought to Wheat Biotechnology Lab., University of Agriculture, Faisalabad, Pakistan for genetic characterization of Jamun genotypes.

In general the keywords are related to the nature of the study. The change of germplasm resources will continue, only leaving germplasm.

Change has been made accordingly

In the introduction it is suggested to complement the antecedent studies considering the international and regional context. Include studies 2019 onwards. If these studies do not exist, include those of related species.

Antecedent studies has been added throughout the introduction considering international and regional context.

Introduction has been updated using studies from 2019 and onward. Citations of older studies have also been strengthened by using recent studies with them.

In the materials and methods section, it is suggested that in the phenotypic characterization part, the information be complemented with the criteria used for the selection of genotypes, the type of sampling carried out, and the experimental units considered. It is important to mention here the age of the selected trees and the agronomic management. Also include that the phenotypic characterization was carried out in situ. In the table where the descriptors are mentioned, include the source from which they were taken or adapted.

Data for phenotypic characterization was obtined in situ from trees of aproximately same age (based on trunk size and height) between 8-12 years as mentioed earlier by Khan et al. (2010) and Sharif et al. (2019). Genotypes were randomly marked based of visual diversity of tree and fruits. Each individual tree was selected as an experimental unit witout replication and mature leaves and fruits were used for data collection.

Description of Pakistani Jamun genotypes were scored considering Jamun biodiversity descriptor by Agro Biodiversity, Plant Genetic Resources (Mahajan et al., 2002)

 In the molecular characterization, include the criteria taken into account for the selection of the markers. Include the DNA concentration value used in the PCR reactions, and some additional considerations in electrophoresis that are included directly in the manuscript.

Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) markers having more than 50 % GC conetent were employed on the basis of their ability to reveal high PIC values (>0.8) in previous studies on Jamun by Sharma et al. (2023); Gajera et al. (2018), Shakya et al. (2010), Khan et al. (2010).

DNA preps were diluted in molecular grade water making final concentration of 50 ng/µl and used as templates for subsequent polymerase chain reactions (PCRs).

After PCR resulting products were separated by standard agarose gel electrophoresis (1.2% w/v) in 1X TAE buffer at 70 volts and 250 mA for one and a half (1½) hours.

Correct the legend of table 3 and other aspects. Statistical analyzes are adjusted to the nature of the study

Changes has been made accordingly

The results are presented very well described, it is suggested to include the MDS analysis in this item and not in the discussion item.

MDS analysis has been shifted to result portion as suggested

The discussion of the results considers comparative studies, however, in some of them the contexts are not mentioned whether it is international or national. It is suggested that the authors delve into the aspects and impact at the biological, agronomic and genetic level of the results found; practical implications of the research findings

Comparative studies have been mentioned with the information regarding where they were carried out. Discussion related to biological, agronomic and genetic concerns has been mentioned with practical implications.

The conclusions are pertinent but could be improved by including some data on the results found, which effectively support the arguments presented.

 Results based data has been added in the conclusion

The bibliographic references are pertinent, it is suggested that they be more up-to-date.

Bibliographic references have been updated by using references from 2019 and onward. Older references have also been strengthen by using recent studies.

Suggestions

Changes made

The introduction would be enriched by an expanded discussion on the imperative nature of genetically and phenotypically characterizing Jamun. It should provide a more comprehensive context regarding its economic and ecological importance. Moreover, the current exposition does not sufficiently address the research's contribution to the existing body of literature on molecular markers for genetic diversity assessment in plants.

Introduction has been improved and economical and ecological concern of jamun plant has been added

In the Materials and Methods section, the utilization of three-tier subheadings is unnecessary and should be streamlined for clarity and consistency.

Change has been made accordingly

The presentation of results could be augmented by incorporating more nuanced statistical analyses, such as Analysis of Molecular Variance (AMOVA), to delineate genetic variance among the various groups more precisely.

Analysis of Molecular Variance (AMOVA) for computing genetic diversity within and among Jamun genotypes has been added in the study under the heading “3.2: Polymorphism and genetic differentiation among Jamun genotypes” on page no. 15.

It is recommended that the Discussion section provide a more profound comparative analysis with analogous studies, especially those from disparate geographical locales. The current section's reliance on restating findings without a robust contextualization within the existing literature is notable. Furthermore, it does not offer a clear vista on potential avenues for future research

Discussion section has been improved accordingly

A meticulous review and refinement of the manuscript's formatting and grammar are imperative throughout, including within figures and tables, to align with the journal's standards. For instance, the genus name "Syzygium" in the Introduction's opening sentence should be italicized for taxonomic accuracy. Additionally, Figure 1's resolution and quality require enhancement to meet the journal's publication criteria.

Manuscript has been reviewed carefully for corrections. Figure one has been changed with high resolution picture.