Molecular Characterization and Functional Analysis of Wheat TtLOX Gene Involved in Aphid Resistance
Round 1
Reviewer 1 Report
minor suggestions:
1) Page 3, L103-111: tell the resistance or susceptible for Lanmei and Poland 305.
2) P3, L117-1221: how many plants per cultivar were used, replications and experimental design for EGA infestation, not very clear.
3) Figure 2, why wheat was not included in (a)?
4) Fig. 6, the graph text contents are not consistent and uniform
Author Response
Response to Reviewer 1 Comments
Dear Editors and Reviewers,
Thank you for your replies and comments concerning our manuscript entitled “Molecular characterization and Functional Analysis of wheat TtLOX gene involved in aphid Resistance” These comments were all valuable and very helpful for revising and improving our paper, as well as providing important guidance for our future research. We have studied the comments carefully and made appropriate corrections, which we hope meet with your approval. The main corrections in the paper and the responses to the reviewers’ comments are as follows:
Respond to Reviewer
Major points:
1) Page 3, L103-111: tell the resistance or susceptible for Lanmei and Poland 305.
Response: We accept this comment. We have revised it line 119-127.
Line 103-105 “Seeds of tetraploid wheat cultivars, Lanmai (Shaanxi, Zhashui)(T. turgidum) and Poland 305 (T. polonicum), common wheats cultivar Xiaoyan 6 (T. aestivum), tobacco cultivar Nicotiana benthamiana (Nicotiana tabacum) were obtained from Northwest A&F University.” has been replaced by “Three tetraploid wheat varieties differed in EGA resistance levels were used. Variety Lanmai (Shaanxi, Zhashui)(T. turgidum) was highly resistance to EGA , variety Poland 305 (T. polonicum) was moderately susceptible to EGA and variety and variety Xiaoyan 6 (T. aestivum) was highly susceptible to EGA [10]. Seeds of the three varieties and tobacco cultivar Nicotiana benthamiana (Nicotiana tabacum) were obtained from Northwest A&F University.”.
2) P3, L117-1221: how many plants per cultivar were used, replications and experimental design for EGA infestation, not very clear.
Response: We accept this comment. We have revised it and improve the methods description line 139-147.
3) Figure 2, why wheat was not included in (a)?
Response: We accept this comment. We have revised it.
Figure 3. Phylogenetic analysis of TtLOX in plants. The phylogenetic tree included 24 plant LOXs and TtLOX, among them TaLOX-B1 and TaLOX3 from wheat.
4) Fig. 6, the graph text contents are not consistent and uniform
Response: We accept this comment. We have revised it.
Figure 7. Functional validation of TtLOX with BSMV-VIGS. The graph text contents are consistent and uniform.
With best wishes,
Yours sincerely,
Wanquan Ji and Xinlun Liu.
Author Response File: 
Author Response.docx
Reviewer 2 Report
The manuscript reports the isolation and molecular characterization of a gene (TtLOX) encoding a lipoxygenase (LOX) enzyme involved in the resistance to the English grain aphid (EGA) in durum wheat. The Authors investigated the conserved domains, the phylogenetic relationship and subcellular localization of the protein encoded by TtLOX, the expression of TtLOX in durum wheat by EGA infestation, and changes in TtLOX expression levels and EGA numbers under methyl jasmine treatment. The role of TtLOX in wheat resistance against EGA was investigated by evaluating the aphid reproduction and the transcript level of TtLOX in both TtLOX-silenced and non-silenced plants. The Author concluded that TtLOX contributes to EGA resistance in wheat regulating aphid feeding behavior and population growth by the jasmonic acid biosynthesis.
The title adequately describes the subject of the manuscript and the abstract briefly tell what was done and summarize the main results and conclusions. The author’s contribution is placed in its proper perspective in relation to the state of knowledge. The conclusions are adequate and supported by the data. The information is presented in a relatively simple and straightforward manner to be understood by a competent scientist or reader. The subject falls within the scope of the Agronomy Journal.
Main comment:
- One of the main objectives of the manuscript is to report the cloning of a lipoxygenase gene involved to EGA resistance in durum wheat, as reported in the abstract (lines 21-22) and elsewhere (lines 95, 211 and 211). However, in the manuscript there are few or missing details on the isolation and characterization of the TtLOX gene sequence. The authors should report more details on the isolation of the nucleotide sequence in the aphid-resistant wheat variety Lanmai and in the aphid-sensitive wheat variety Poland 305, the comparison with the sequences reported in the sequenced genomes of T. turgidum, T. dicoccoides and T. aestivum, its genetic mapping and physical position in the AABB genomes, the designation of the TtLOX gene in the sequenced durum wheat genome to easily identify it, etc.
- According to the international nomenclature, the scientific name should be always written in italics.
- Write title of the manuscript and titles of the paragraphs according to the Instructions for Authors
- Lines 46-48: Breeding of wheat for EGA resistance shows that there are few varieties of wheat resistant to aphid, there are still several resistance genes which inhibit the growth of aphid, yet [10]. Please, rewrite
- Line 76 : “In Barley, LOX2was highly up-regulated after Bird Cherry-Oat Aphid (BCA)” should be” In barley, LOX2 was highly up-regulated after bird cherry-oat aphid (BCA)”
- Line 82:” In wheat, LOX1 was upregulated significantly in a wheat landrace plants, carrying …..” Please, delete plants,
-Line 347: Spodoptera exigua, to be written in italic
- The English form and punctuation should be reviewed by a native English speaker
Author Response
Dear Editors and Reviewers,
Thank you for your replies and comments concerning our manuscript entitled “Molecular characterization and Functional Analysis of wheat TtLOX gene involved in aphid Resistance” These comments were all valuable and very helpful for revising and improving our paper, as well as providing important guidance for our future research. We have studied the comments carefully and made appropriate corrections, which we hope meet with your approval. Language revisions of the entire manuscript can be seen in manuscript with track changes from Prof. Zhaogui Yan and PhD. Amo Aduragbemi Emmanuel. The revised sections can be seen in manuscript with track changes.
The main corrections in the paper and the responses to the reviewers’ comments are as follows:
Respond to Reviewer
Major points:
- One of the main objectives of the manuscript is to report the cloning of a lipoxygenase gene involved to EGA resistance in durum wheat, as reported in the abstract (lines 21-22) and elsewhere (lines 95, 211 and 211). However, in the manuscript there are few or missing details on the isolation and characterization of the TtLOX gene sequence. The authors should report more details on the isolation of the nucleotide sequence in the aphid-resistant wheat variety Lanmai and in the aphid-sensitive wheat variety Poland 305, the comparison with the sequences reported in the sequenced genomes of T. turgidum, T. dicoccoides and T. aestivum, its genetic mapping and physical position in the AABB genomes, the designation of the TtLOX gene in the sequenced durum wheat genome to easily identify it, etc.
Response: We accept this comment.
Lines 248-262 describe the details on the isolation and characterization of the TtLOX gene sequence.
The EST sequence of LOX in Poland 305 is full-identical (100%) to compared to the full length sequence of TtLOX.
- According to the international nomenclature, the scientific name should be always written in italics.
Response: We accept this comment. We have and revised each one in the article.
- Write title of the manuscript and titles of the paragraphs according to the Instructions for Authors
Response: We accept this comment.
Lines 248: “3.1 Isolation and Phylogenetic Analysis of the TtLOX cDNA Sequence” has been replaced by “3.1 Isolation and Characteristic Analysis of the TtLOX cDNA Sequence”
Lines 352: “3.5. Gene silencing and the variation of aphid population” has been replaced by “3.5. Gene silencing and changes in aphid reproduction”
- Lines 46-48: Breeding of wheat for EGA resistance shows that there are few varieties of wheat resistant to aphid, there are still several resistance genes which inhibit the growth of aphid, yet [10]. Please, rewrite
Response: We accept this comment. We have revised it and deleted it
Lines 54-59: “Breeding of wheat for EGA resistance shows that there are few varieties of wheat resistant to aphid, there are still several resistance genes which inhibit the growth of aphid, yet [10]” has been replaced by “In our study, we found most wheat cultivars were susceptible to aphid attack although aphid resistant genes were present in tetraploid wheat cultivars [10] ”
- Line 76 : “In Barley, LOX2was highly up-regulated after Bird Cherry-Oat Aphid (BCA)” should be” In barley, LOX2 was highly up-regulated after bird cherry-oat aphid (BCA)”
Response: We accept this comment. We have revised it.
Line88-89 “In Barley, LOX2 was highly up-regulated after Bird Cherry-Oat Aphid (BCA) and GPA infestation” has been replaced by “In barley, LOX2 was highly up-regulated after bird cherry-oat aphid (BCA) and GPA infestation.”
- Line 82:” In wheat, LOX1 was upregulated significantly in a wheat landrace plants, carrying …..” Please, delete plants,
Response: We accept this comment. We have revised it line 97.
-Line 347: Spodoptera exigua, to be written in italic
Response: We accept this comment. We have revised it.
Line413: “Spodoptera exigua” has been replaced by “Spodoptera exigua”.
- The English form and punctuation should be reviewed by a native English speaker
Response: Language revisions of the entire manuscript can be seen in manuscript with track changes from Prof. Zhaogui Yan and PhD. Amo Aduragbemi Emmanuel. The revised sections can be seen in manuscript with track changes.
Zhaogui Yan is Prof. of Forest Ecology College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan, 430070, China; and Associate Editor，Ecology and Evolution, ISSN: 2045-7758; Associate Editor, Plant Ecology, ISSN: 1573-5052.
With best wishes,
Yours sincerely,
Wanquan Ji and Xinlun Liu.
