Changes in Phytohormone and Antioxidant Enzyme Activities during the Healing Process of Different Crabapple Rootstock–Scion Combinations
Round 1
Reviewer 1 Report
I have received a manuscript submitted by Fenhou Shi et al., entitled „Changes in phytohormone and antioxidant enzyme activities during the healing process of different begonia rootstock-scion combinations.
Manuscript presents data on grafting success of a Malus „huabiao“ used as scion grafted on three different crab apple species as rootstock (Malus hupehensis, M. baccata and M. robusta, correlating grafting success assesed with activity of POD, PPO and PAL enzymes and presence of phytohormones IAA, Me-Ja and ZR,
Manuscript is a valuable contribution but it needs some corrections . Most but not all problems are related with translation and English editing. Translation in general has been very well done except in some technical terms, but these mistakes are also faults made by authors who failed to detect this inconsistencies in the text. There are also some comments which authors should keep in mind for their future studies as they can probably not be addressed right now.
Material
- The first and at the same time very large problem is connected with the type of used material. I believe that M.“huabiao“ is an apple, same as the three rootstocks. But it can not be both an apple and a begonia at the same time. I believe that letter M in the name M“huabiao“ comes from Malus (latin for apple) and that there is no place for mentioning Begonia. In my country none of the available Begonia species are trees and maybe I am wrong? Corrections need to be made in the text in the one or the other direction, but it can not stay written as it is.
Phytohormones
- phytohormone that you mention as zeatin nucleotide (ZR) is probably zeatin riboside, being perhaps a mixture of several different zeatin riboside forms (cis, trans, dihidro….) nowadays easily distinguished by LC-MS techniques. I don‘t mind if your analytical methods are not state of art, if they are standard or older ones, but please elaborate a bit more on the detection methods and their separation accuracy and fidelity.
Phytohormone content in plant tissues is subject to drastic daily changes with large amplitude differences between daily highest an lowest values. Having this in mind, it woud be good to state at what time of day sampling was done, how the sampled material was handled and how much time passed before it was procesed and stored at -80 0C. It would be nice to have non grafted controls but that is perhaps too much to ask the authors.
Although you graphics appear very dramatic, which perhaps forced you to describe direction of change between subsequent days, this differences are actually small, smaller than expected. Also statistical differences and their significance are more than good. Is it posssible that your material following sampling, was stored in common refrigerator giving sufficient time to phytohormones to diffuse from vascular into other tissue type which will later not be represented after scraping the bast? To elaborate, differences between days are fairly small and statistic is better than expected.
ZR and MeJA are translocated by phloem upwards (acropetally) and they should be present in bast during the day. IAA is mainly translocated via xylem and less in lateral direction, which makes it less likely to have much IAA in your samples after the bast scraping,
Apart from graphics you could perhaps provide a table with percentage changes in relation to the starting value, one day after the grafting was done. From this percentage values your readers can make their own conclusions if this are big or small changes.
Table with statistics for both phytohormones and isozymes should be provided after and not before the graphics are presented.
Enzymes
-it is well known that peroxidases, polyphenol oxidases, and other apple oxidases are all just color reactions of a same, albeit large group of related isozymes. In this case a single enzyme molecule, manifests color responses characteristic for diverse isozyme groups, In this sense data obtained by screening activity of oxidases are not specific and trustworthy enough and results of this analysis can not be accepted as valid until you provide a very detailed screening procedure which is missing in the paper. Connecting your data simply to procedures done by some other scientist is not good enough. Methods need to be presented in detail in your own manuscript.
Oxidative enzymes in apples and related fruit species have been studied for many years, and it is believed that their changes are not sufficiently specific to make correlations with their roles. Gaspar et al (1992) in Agronomie 12: 757-765 and his other publications claimed evidence for some specific roles of peroxidases in rooting but this data was later not sufficiently confirmed. To get the proper insight in the problem please get the paper by Passardi et al (2005) which appeared in Plant Cell Rep 24:255-265. Paper is entitled „ Peroxidases have more functions than a Swiss army knife“.
So extensive presentation of changes in isozymes after grafting is more or less indicative, and it is also up to the reader to decide ii it is a sufficient proof for them. How about some photographs of successfull and failed grafting.
- page 3, line 99 - kinds of grafted rooting seedlings, probably „seedlings intended for grafting were planted in rows 10cm apart and with 40 cm spacing between the rows“
- page 3, line 101 I don‘t know what is „root fixing water“ - this is some kind of misunderstanding in the translation
- page 3, line 120 – you can store apple branches in a refrigerator at + 3-5 0C for 2-3 months and then take them out and use for grafting. But if the same material is stored at -800C, damage is irreversible and material can further be used only for biochemical and mol. biology type analysis. So material at -800C can not be classified as backup bur rather as storage material.
Page 3, line 125 Bast of the healing grafting site is not a very good description of what you done to collect samples, You have scraped (with a knife or scalpel blade) stem bark at the site of grafting, and bast obtained in this way, comprising living periferal shoot tissues, functional phloem and cambium, was collected ad used for analysis. Xylem was probably not sraped of. Bast is a good material for isozyme studies, providing good results all around the year, but for phytohormone studies there are some restrictions which migh affect the results.
Finally, I hope that authors will correct this paper as much as possible giving the opportunity to Editor to further process.
Reviewer
Some technical terms have been improperly translated, they should be rechecked by authors.
Author Response
Please see the attachment.
Reviewer 2 Report
The manuscript is interesting, it contains some valuable results and suitable discussion. However, some aspects should be clarified.
It is really disturbing for an “average” reader to call begonia a tree (for the majority of gardeners, begonia is an ornamental flowering plant). Malus is an apple tree. Begonia and Malus are different plants, belonging to different families. If the Authors are writing about Malus, why “begonia” is in the title? Please check what “begonia” means for the majority of people (it is enough to check Wikipedia), explain what plant you are writing about, insert the proper taxonomy of this plant. A photograph would also be useful.
Abstract is too long, it should be shortened to the required size.
Methods for the determination of physiological indicators should be described more precisely, these analyses are the important part of the manuscript, so it is not enough just to cite the reference, not always easily accesible (besides, one of the reference is M.Sc. thesis, not the published paper, so these methods were not reviewed in any journal.
Please correct all expression as “From Fig. 2, it can be learned” or It can be learned from Table 3. Correct also expressions like “As can be seen from Table 1”, etc. Such expressions are awkward and better to avoid them. Please describe the phenomenon, conclusion etc. and just add (Table 1) or (Fig. 3) in the end of the sentence.
Author Response
Please see the attachment.
Author Response File: 
Author Response.docx
Round 2
Reviewer 1 Report
Manuscript has been significantly improved and I will recommend it for publishing. Do me/and yourself a favor and check daily variability of your parameters in morning, noon, late afternoon, dusk and midnight. It might pay our for some of your future studies.
Author Response
We appreciate for you directions and will consider it in future experiments. Once again, thank you very much for your comments and suggestions.
Reviewer 2 Report
The Authors have improved their manuscript according to my suggestions. I have no more comments.
Author Response
Thank you again for taking the time to review our manuscript.
