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online review management tool for the study of global biofilm.
{mjg13048-fig-0001}
Discussion
{mjg13048-sec-0006}
==========
In our study, we have demonstrated the development of
ESSR and its use for the study of biofilm formation. We identified ESSR in *P.
viticola* at an early time point and then performed the biofilm identification using
FISH, real‐time PCR (RT‐PCR), and flow cytometry. In addition to these methods, we
identified the biofilm using the flow cytometry technique as a novel tool to screen for
ESSR‐negative colonies. The findings of this study also revealed the ESSR has the
ability to create a reliable, high‐quality biofilm when performed using a FISH
technique.
In this study, we identified several genes that could be involved in
biofilm formation using the FISH method. The expression patterns of *P. viticola* at
the early time point of colonization are consistent with that of bacterial community
that may affect the process of biofilm formation (Lomert, *et al*.,
[2011](mjg13048-bib-0019){ref-type="ref"}; Tuck et al.,
[2014](mjg13048-bib-0033){ref-type="ref"}). We believe that the *P. viticola* ESSR may
provide a new approach to identify biofilm‐forming bacterial strains.
For example,
*cpsA* is expressed during colonization and development in ESSR‐negative cells.
However, although *cpsA* is expressed at low levels in a variety of ESSR‐negative
cells, our results reveal that it may play a role in biofilm formation in *P.
viticola*. In addition, a higher expression of *dpiA* has been reported in both *P.
viticola* and ESSR‐positive cell populations (Lomert, *et al*.,
[2011](mjg13048-bib-0019){ref-type="ref"}; Tuck et al.,
[2014](mjg13048-bib-0033){ref-type="ref"}). Our results in our study indicated that
*dpiA* was not expressed during colonization and development.
The expression levels of
several biofilm‐related genes are similar to that of *P. viticola*, and it could be the
result of either the expression of an additional gene(s) that is downregulated during
colonization or the downregulation of the same gene(s) in cells that are undergoing
adhesion to the substrate.
Several studies have shown that ESSR and biofilm formation
are two processes, and therefore, several genes have different functions in this
process, thus, we believe that some ESSR and biofilm formation is a result of the ESSR‐
and biofilm‐related genes. In the present study, we did not observe any change in
expression of gene expression in *P. viticola* in comparison to the *P. viticola*
strains isolated from the field (data not shown). It suggests that this may provide a
useful tool for studying the expression of ESSR and biofilm‐related genes during
biofilm formation. In addition, the gene expression patterns are consistent with that
of *P. viticola* in the same time (data not shown).
As *P. viticola* biofilm
development is a dynamic process, it could also be involved in the pathogenesis of
pathogenic bacteria, and the development of biofilm may result in various other
bacterial pathogens (Rouffel et al., [2018](mjg13048-bib-0028){ref-type="ref"}).
Therefore, we believe that the genes involved in biofilm development are useful to
study the pathogen in *P. viticola*.
Conclusion {mjg13048-sec-0007}
==========
We
have identified the ESSR‐ and biofilm‐related genes and identified a new gene for ESSR
and biofilm development, namely, *dpiA*. Our findings suggest that ESSR and biofilm
formation could be important for the development of the *P. viticola* ESSR‐producing
bacteria.
This research was supported by the National Natural Science Foundation of
China (Nos.: 10736032, 1087210, 12743618, 11563018, 12736038), and the Natural Science
Foundation of Jiangsu province (No.: BK1209700).
Authors' contributions: YL, FJ,
JH
online review management tool (the *S3*). It has been designed as a novel research
methodology, allowing a researcher to explore the factors that affect and influence the
performance of an individual's *S3*. The S3 is a simple, flexible tool that facilitates
the development and use of research methodology. This article has used the S3 as a
first step towards developing a scientific and computational research method.
3. Study
design and findings {SECID0E3M}
===========================
The S3 aims to be a novel
research method that enables the development of novel research methodology using
traditional tools such as an online survey, a database, and multiple web applications.
The S3 is a research method for a broad range of issues ranging from the development of
effective research tools through the implementation of methods and tools such as the
*S3* to the creation of new research methods and research resources. This was the first
time that a researcher would come up with a useful and novel research method, so this
is an important step that will be followed in the S3. The first section of the section
in the "Methods & tools" section of this article will describe the research methods and
tools that the S3 is used for. The second and third sections of the section in the
"Methods & tools" section in the section "Conceptual approaches" will describe how the
S3 was built and how it uses the results of the research methods and tools. The final
section in the "S3" section of this article will describe the results of the S3 that
were obtained in this second and third section of the article, followed by a brief
description of the research methodology and the subsequent research methodology that
was created for this research method.
3.1.1. Study design {SECID0E2M}
To create a
new research method, a researcher needs to know how to write a research proposal and
how the methods need to be used. To the best of the researcher's abilities, including
how the research proposal is likely to be used, the researcher is able to apply
concepts, knowledge, and/or data that are currently existing and useful.
4. Data
collection {SECID0E2M}
-----------------
Each research proposal was developed as an
electronic database that would be easily accessible and made available. One researcher
and a team of researchers would then upload the proposal to the *S3* and then manage
the entire scientific project (Table [1](TAB1){ref-type="table"}). Each researcher
would complete several online surveys of research performance. Each round of this
research should be completed using the standard database, or on the web, that is open
to researchers and other researchers who have the same experience.
Number of
participants that provided details about their previous research proposal (in
English)
**Proposal** **Participant name** **Number of participants to be
included in the research proposal**
--------------- ---------------------
------------------------------------------------------------
Research proposal
1
1
Research proposal
2 2
Research proposal
3
3
Research proposal
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