Morphological and Phylogenetic Evidences Reveal Lasiodiplodia chonburiensis and L. theobromae Associated with Leaf Blight in Hevea brasiliensis in Southern Thailand
Round 1
Reviewer 1 Report
In the literature it is known that temperature and humidity condition a greater occurrence of fungi, mainly in tropical forests. However, I believe that the authors could better explore how abiotic factors such as temperature, precipitation and humidity can favor the appearance of an unprecedented form of fungal pathogen (Lasiodiplodia) in rubber trees in Thailand.
Author Response
In the literature it is known that temperature and humidity condition a greater occurrence of fungi, mainly in tropical forests. However, I believe that the authors could better explore how abiotic factors such as temperature, precipitation and humidity can favor the appearance of an unprecedented form of fungal pathogen (Lasiodiplodia) in rubber trees in Thailand.
Answer: Thank you very much for your suggestion and we added these data to the introduction (Lines 49-88).
Reviewer 2 Report
In this study, morphological characteristics and phylogenetic analysis were used to identify Lasiodiplodia chonburiensis and L. theobromae as pathogens causing leaf blight of rubber leaves. The genus Lasiodiplodia belong to the family Botryosphaeriaceae and can cause a variety of plant diseases. The Lasiodiplodia species do not form appressorium or other structures to penetrate plant tissues, but are pathogenic to host plants. Lasiodiplodia chonburiensis was first described as a saprophyte, and L. theobromae has been documented to cause disease in several plant species in different regions, but L. theobromae has not been reported to also cause leaf blight in Thailand rubber leaves. This study is the first to report L. theobromae, which causes rubber leaf blight. The study revealed that both L. chonburiensis and L. theobromae act as plant pathogens associated with leaf-blight disease in rubber trees. The paper cites many previous research results, and the overall idea is relatively clear and well-structured. All in all, I am very impressed with this paper and it is worth publishing after a little revision. I have included my comments in the attached PDF file.
Comments for author File: 
Comments.pdf
The quality of English language is good.
Author Response
In this study, morphological characteristics and phylogenetic analysis were used to identify Lasiodiplodia chonburiensis and L. theobromae as pathogens causing leaf blight of rubber leaves. The genus Lasiodiplodia belong to the family Botryosphaeriaceae and can cause a variety of plant diseases. The Lasiodiplodia species do not form appressorium or other structures to penetrate plant tissues, but are pathogenic to host plants. Lasiodiplodia chonburiensis was first described as a saprophyte, and L. theobromae has been documented to cause disease in several plant species in different regions, but L. theobromae has not been reported to also cause leaf blight in Thailand rubber leaves. This study is the first to report L. theobromae, which causes rubber leaf blight. The study revealed that both L. chonburiensis and L. theobromae act as plant pathogens associated with leaf-blight disease in rubber trees. The paper cites many previous research results, and the overall idea is relatively clear and well-structured. All in all, I am very impressed with this paper and it is worth publishing after a little revision. I have included my comments in the attached PDF file.
Answer: Thank you for your review and gave some valuable comments, we have addressed all your suggestion to improve this manuscript.
Have the authors used other solid/liquid media for their cultures?
Answer: We have tried in liquid media, but the fungus developed only mycelia, for solid culture, the fungus also rarely developed pycnidia on substrate.
The title of this paper is "Morphology and Phylogeny Evidence Reveal Lasiodiplodia chonburiensis and L. theobromae are Associated with Leaf Blight in Hevea brasiliensis in Southern Thailand", but in the introduction, the author used a lot of space to introduce the rubber tree and its importance as well as the rubber leaf drop disease, but we didn't find the author's introduction of the genus Lasiodiplodia, such as the disease characteristics, the host type, and the scope of influence, etc. It is suggested that the author can add.
Answer: Thank you for your suggestion and we added more data in the last paragraph in the introduction (Lines 66-82).
Reviewer 3 Report
I recommend revising the title as the current one is quite long.
I would suggest changing the introduction especially as it needs to be rewritten in scientific writing. I suggest getting the help of a native English speaker to check the whole paper.
Line 56 should be deleted as it has been mentioned above already.
Line 57 mentions the isolation of Neopestalotiopsis species from the hosts, why these were not also included in the current study? How did the authors confirm that the Neopestalotiopsis were also not responsible for the leaf-blight disease?
For the pathogenicity text, the authors mention reasons for not using spore suspension but it would have been better to include spore suspension even at a lower concentration as discussed in several papers.
In the tree, several strains of different species did not cluster together. The sampling is also strange as to why there are up to 5 isolates of some species that are not closely related to the isolates from this study. I recommend running the tree again.
see above
Author Response
Recommend revising the title as the current one is quite long.
Answer: We revised and corrected “Morphological and Phylogenetic Evidence Reveal Lasiodiplodia chonburiensis and L. theobromae Associated with Leaf Blight in Hevea brasiliensis in Southern Thailand” (Lines 1-5)
I would suggest changing the introduction especially as it needs to be rewritten in scientific writing. I suggest getting the help of a native English speaker to check the whole paper.
Answer: We have revise by adding more detail on introduction part and use MDPI English editing service to improve grammar.
Line 56 should be deleted as it has been mentioned above already.
Answer: We have deleted this sentence.
Line 57 mentions the isolation of Neopestalotiopsis species from the hosts, why these were not also included in the current study? How did the authors confirm that the Neopestalotiopsis were also not responsible for the leaf-blight disease?
Answer: We reported the leaf fall disease by Neopestalotiopsis in the rubber tree already (Pornsuriya et al., 2020.). The symptom on the rubber leaf in the current study is leaf blight and different from leaf spot by Neopestalotiopsis.
For the pathogenicity text, the authors mention reasons for not using spore suspension but it would have been better to include spore suspension even at a lower concentration as discussed in several papers.
Answer: It has been known that inoculation by mycelium plug was also showed the symptom on the wounded leaf similar with this study. Therefore, we inoculated only mycelial plug into wounded leaves and the result showed similar result with in nature.
In the tree, several strains of different species did not cluster together. The sampling is also strange as to why there are up to 5 isolates of some species that are not closely related to the isolates from this study. I recommend running the tree again.
Answer: Thank you for your nice suggestion and we have not reconstructed the phylogenetic tree because I would like to confirm that my isolates were not clustered in the other clades of the genus Lasiodiplodia.
