Review Reports - Successive Harvesting Interval and Salinity Level Modulate Biomass Production and Nutritional Value in <i>Sarcocornia fruticosa</i> and <i>Arthrocaulon macrostachyum</i>

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript shows potential, but needs improvement. A clustering test and a Pearson correlation analysis (variables) could be included at the end of the discussion to determine which variables are most associated with biomass increase. The results should focus on the effect of salinity and the response of the evaluated ecotypes, enriching the discussion with more detailed explanations and updated references. The conclusion would benefit from a perspective of future work. Therefore, my opinion is that major revisions are needed. The file with the corrections is attached.

Comments for author File: Comments.pdf

Author Response

Response to the Reviewers' Comments

We thank the Reviewers for their comments and suggestions. We have responded to them all and feel that the revisions significantly improve the manuscript.

Reviewer: 1

Comments and Suggestions for Authors

Comment-1: The manuscript shows potential, but needs improvement. A clustering test and a Pearson correlation analysis (variables) could be included at the end of the discussion to determine which variables are most associated with biomass increase. The results should focus on the effect of salinity and the response of the evaluated ecotypes, enriching the discussion with more detailed explanations and updated references. The conclusion would benefit from a perspective on future work. Therefore, my opinion is that major revisions are needed. The file with the corrections is attached.

Response:

We have included a correlation matrix at the end of the discussion part and explanatory texts spread throughout the discussion section.

In 4.1. Growth responses

" Moderate salinity levels improve osmotic adjustment and ion compartmentalization, which support metabolic activity and plant growth, but excessive salinity causes oxidative stress, ion toxicity, and reduced nutrient uptake [39] [39,43]. In support of these notions, the accumulated yield (FW) exhibited high positive correlations with EC, TSS, and shoot diameter [Table 2, [44,45,46]".

In 4.2 Photosynthetic pigments.

Chlorophyll and carotenoids correlate positively (Table 2) due to their shared, interdependent roles in photosynthesis and photoprotection [47].

In 4.4 Antioxidant responses.

The current study shows that MDA displayed negative correlations with radical scavenging activity, polyphenols, flavonoids, proteins, chlorophylls, and carotenoids, further indicating that oxidative stress is inversely related to antioxidant capacity and photosynthetic pigments [Table 2, [45,47,74,75]]. …………..

Radical scavenging activity serves as a crucial defense mechanism under various stress conditions, and it plays a vital role in neutralizing reactive oxygen species that contribute to lipid peroxidation and abiotic stress effects [76]. Antioxidant-related traits, including total polyphenols, flavonoids, carotenoids, and total proteins, were significantly and positively correlated with radical scavenging activity (DPPH), highlighting their central role in antioxidative defense [Table 2, [47,48, 77].

Response: Table -2. Correlation analysis between different assays.

Variable

Correlations (Spreadsheet1) Marked correlations are significant at p < .05000 N=72 (Casewise deletion of missing data)

Total

chlorophyll

Total carotenoid

Total polyphenol

Total flavonoid

DPPH

Total protein

Shoot diameter

TSS

RWC

Anthocyanin

MDA

Total chlorophyll

1.000

0.908*

0.166

0.337*

0.313*

0.179

0.397*

0.411*

0.548*

0.319*

0.333*

0.257*

-0.645*

Total carotenoid

0.908*

1.000

0.151

0.432*

0.364*

0.317*

0.367*

0.475*

0.468*

0.212

0.365*

0.330*

-0.496*

Total polyphenol

0.166

0.151

1.000

0.645*

0.763*

0.333*

0.286*

-0.115

0.077

-0.410*

-0.388*

-0.064

-0.182

Total flavonoid

0.337*

0.432*

0.645*

1.000

0.919*

0.732*

0.538*

0.025

0.215

-0.329*

-0.102

0.285*

-0.137

DPPH

0.313*

0.364*

0.763*

0.919*

1.000

0.565*

0.517*

-0.015

0.320*

-0.379*

-0.293*

0.144

-0.183

Total protein

0.179

0.317*

0.333*

0.732*

0.565*

1.000

0.114

-0.117

0.087

-0.207

-0.098

-0.031

0.205

Shoot diameter

0.397*

0.367*

0.286*

0.538*

0.517*

0.114

1.000

0.417*

0.172

0.063

0.366*

0.774*

-0.532*

TSS

0.411*

0.475*

-0.115

0.025

-0.015

-0.117

0.417*

1.000

0.186

0.213

0.719*

0.601*

-0.354*

RWC

0.548*

0.468*

0.077

0.215

0.320*

0.087

0.172

0.186

1.000

0.278*

-0.084

-0.070

-0.333*

Anthocyanin

0.319*

0.212

-0.410*

-0.329*

-0.379*

-0.207

0.063

0.213

0.278*

1.000

0.402*

0.193

-0.342*

0.333*

0.365*

-0.388*

-0.102

-0.293*

-0.098

0.366*

0.719*

-0.084

0.402*

1.000

0.703*

-0.312*

0.257*

0.330*

-0.064

0.285*

0.144

-0.031

0.774*

0.601*

-0.070

0.193

0.703*

1.000

-0.410*

MDA

-0.645*

-0.496*

-0.182

-0.137

-0.183

0.205

-0.532*

-0.354*

-0.333*

-0.342*

-0.312*

-0.410*

1.000

References (added for Reviewers 1 and 2)

[33] R. Ksouri et al., “Influence of biological, environmental and technical factors on phenolic content and antioxidant activities of Tunisian halophytes,” C. R. Biologies, vol. 331, no. 11, pp. 865–873, 2008, doi: 10.1016/j.crvi.2008.07.024.

[34] D. Katschnig, R. Broekman, and J. Rozema, “Salt tolerance in the halophyte salicornia dolichostachya moss: Growth, morphology and physiology,” Environ Exp Bot, vol. 92, pp. 32–42, Aug. 2013, doi: 10.1016/j.envexpbot.2012.04.002.

[35] S. K. Polutchko, J. J. Stewart, W. W. Adams, and B. Demmig-Adams, “Photosynthesis and foliar vascular adjustments to growth light intensity in summer annual species with symplastic and apoplastic phloem loading,” J Plant Physiol, vol. 267, Dec. 2021, doi: 10.1016/j.jplph.2021.153532.

[66] H. Homayouni et al., “Temporal Changes in Biochemical Responses to Salt Stress in Three Salicornia Species,” Plants, vol. 13, no. 7, Apr. 2024, doi: 10.3390/plants13070979.

[67] G. Lee, R. N. Carrow, and R. R. Duncan, “Growth and water relation responses to salinity stress in halophytic seashore paspalum ecotypes,” Sci Hortic, vol. 104, no. 2, pp. 221–236, Mar. 2005, doi: 10.1016/j.scienta.2004.08.011.

[68] H. Kumar, S. B. Lal, and A. M. Wani, “Correlation Studies for Morphological and Biomass Traits in Half Sib Families of Terminalia Arjuna (L.),” Current World Environment, vol. 12, no. 2, pp. 345–354, Aug. 2017, doi: 10.12944/cwe.12.2.18.

[69] I. Mansinhos, S. Gonçalves, R. Rodríguez-Solana, G. Pereira-Caro, J. M. Moreno-Rojas, and A. Romano, “Nutrient Deficiency-Induced Stress Improves Skincare Effects and Phytochemical Content of Green Extracts from Lamiaceae In Vitro Cultures,” Horticulturae, vol. 10, no. 9, Sep. 2024, doi: 10.3390/horticulturae10090947.

[70] S. Saadati, A. Borzouei, M. R. Rahemi, and B. Naserian Khiabani, “Alteration of physiological and biochemical properties in leaves and fruits of pomegranate in response to gamma irradiation,” Sci Rep, vol. 12, no. 1, Dec. 2022, doi: 10.1038/s41598-022-08285-y.

[71] A. K. Esmaeili, R. M. Taha, S. Mohajer, and B. Banisalam, “Antioxidant Activity and Total Phenolic and Flavonoid Content of Various Solvent Extracts from in Vivo and in Vitro Grown Trifolium pratense L. (Red Clover),” Biomed Res Int, vol. 2015, 2015, doi: 10.1155/2015/643285.

Reviewer 2 Report

Comments and Suggestions for Authors

The paper: Successive Harvesting Intervals and Salinity Level Modulate 2 Biomass Production and Nutritional Value in 3 Sarcocornia fruticosa and Arthrocnemum macrostachyum. 4 by the following authors (Tesfaye Asmare Sisay 1,#, Jaykumar Patel 2, Kusum Khatri 2, Babita Choudhary 2, Dominic Standing 3, Zai Du Nja 1, 5 Muki Shpigel 4, Luísa Margarida Batista Custódio 5 and Ilya Gelfand 3 Moshe Sagi 3,6,7,) is valuable but some points and sections should be addressed to be accepted for publication in Agriculture.

INTRODUCTION

Species names should be written in full at first mention (Sarcocornia fruticosa, Arthrocnemum macrostachyum) and always italicized. Acronyms such as “AM” should be avoided in the text. If abbreviations are needed after the first mention, the correct form is A. macrostachyum.

The use of the term ecotype is unclear. If referring to distinct populations or accessions, this should be explained explicitly.

The objective of the study is relevant. Highlighting successive harvesting intervals, in addition to salinity effects, adds novelty compared with previous research. Nonetheless, I believe the introduction might follow a “funnel” structure to make it more appealing for the readers.

MATERIAL AND METHODS

The use of Sarcocornia fruticosa VM, which has been used before, Ventura et al., 2011a, is not well-explained. The authors should explain what “VM” refers to (population, accession, or locality) and provide precise information on the source of plant material. Without this clarification, the description is incomplete.

The description of Sarcocornia fruticosa “ecotypes” and A. macrostachyum seeds is confusing. The information should be reorganized more clearly, ideally on a table. Do the authors have problems to get high yield of germination percentages? Some authors refer in Salicornia (including former Sarcocornia) and Arthrocaulon (formerly Arthrocnemum) the necessity of seed priming before germination.

The term ecotype should be defined explicitly. Does it refer to morphological forms (prostrate versus shrubby growth) or to populations from distinct localities? Clarification is needed to avoid ambiguity.

A suggested table should include at least:

Species (in italics).
Ecotype designation (Ecotype 1, Ecotype 2, etc.).
Locality with precise geographic coordinates.
Date of collection.
Collectors or source reference.
Ecological/morphological description (e.g., coastal sandy habitat, saline marsh, prostrate vs. shrubby growth form, tidal influence vs. no tidal flooding, low, medium or high marsh, inland).

A high-quality map would be valuable as well.

This reorganization would make the meaning of “ecotypes” transparent and scientifically rigorous.

The description of pigment analysis (Section 2.4) lacks essential detail on the equipment used. Spectrophotometers (model, company, country) used for the measurements should be added.

Data Analysis (Section 2.14) is not sufficiently clear. The authors should explain:

Whether assumptions of normality and homogeneity of variance were verified before applying the Tukey-Kramer HSD test?

Why the number of replicates is not homogeneous (n = 4 to 9)?

RESULTS

Results should be limited to the data generated in the present study, while explanations, references, and implications belong in the Discussion.

Reorganization is highly recommended: concise reporting of findings and based on significant statistical support only, so that should be written in Results elegantly and concisely. With interpretation moved to the Discussion, would improve clarity, rigor, and readability.

Keep this subsection strictly to the data, please.

It would very valuable as well to add a Figure with a layout of different photographs showing the experiments and the best results after 21 days and 30 days of harvesting.

3.1. Fresh Biomass Production