Overview

2021.12

Set up the 2022-SZU-China team.

2021.12-2022.2

Experimental skills training and literature reading training.

2022.2-2022.4

Brainstorming.

Project discussion and literature research were carried out in groups.

The projects of each group are summarized and finally the topics to be explored by the team are determined.

2022.5

The preliminary design of the complete project system and a large number of background research and literature research were completed.

The first new parts were designed and built for the project.

Some experiments were started.

2022.6-2022.9

The second batch of new elements was constructed following suggestions from interviews with professionals and professors.

Feedback from experiments and human practice is used to continuously improve our project (click here to jump to the Experiments and Human practice section).

The results of the vast majority of wet versus dry experiments covered by our project were obtained.

Write the first draft of the wiki.

2022.10

Complete the final wiki version and upload it.

Complete the poster production.

All experiments were completed.

2022 iGEM Giant Jamboree!

Teamlog

Brainstorming and meeting

3.10

SZU-China Project Concept Conference

Time: March 10 20:59-23:30.
Venue: Room 202, College of Life and Marine Science Building, Lihu Campus, Shenzhen University.
Minutes of the meeting: Xinying Zhang.

Highlights:

1. Discuss and determine the complete process of team brainstorming and establish the "three-step" strategy:

• Step 1. Release your thoughts and record them.
• Step 2. Search and organize a lot of data.
• Step 3. Initially select the project with the most development potential for further advancement, and finally select the most suitable one as our project after reviewing each pre-project from multiple perspectives.

2. Created a brainstorming summary sheet to record our interesting ideas.

3.25

SZU-China Preliminary Topic Selection Conference

Time: March 25, 21:00-23:00.
Venue: Room 202, College of Life and Marine Science Building, Lihu Campus, Shenzhen University
Minutes of the meeting: Xinying Zhang.

Highlights:

1. All members of SZU-China2022 introduced the project content and the specific design and investigation in turn.

• Bingru Feng: Early detection of plant related diseases.
• Bingru Feng: Oral diseases: prevention and adjuvant treatment of dental caries and periodontitis.
• Ruoyu Chen: Phage drug therapy for trachoma.
• Ruoyu Chen: Diagnosis of β-thalassemia.
• Yi Peng: Detection and degradation of microplastics in mangroves.
• Yi Peng: Rapid diagnosis and management of yellow Dragon disease in citrus crops.
• Xinying Zhang: Bacterial resistance and antibiotic resistance management.
• Xinying Zhang: Bioenrichment and degradation of industrial dust.
• Yingfeng Wu: Optimizing DNA-based data storage methods.
• Linzhen Bao: Detection of heartworm infection in pets.
• Xihe Wang: Treatment of cassava brown streak disease (CBSD).
• Minxi Zeng: Management of pine wood nematodes.
• Zhiying Zhang: The treatment of banana wilt disease.

2. After wonderful speeches and heated discussions, we finally confirmed the following four projects for further promotion:

Project 1: Detection of plant pathogens

Person in charge: Bingru Feng, Ruoyu Chen, Xihe Wang
Optimization direction: Technical research should be placed at the second-best level, and specific application scenarios (pathogens of cabbage, banana and other plants) should be prioritized.

Project 2: Bacterial resistance and antibiotic resistance

Person in charge: Bao Linzhen, Zhang Xinying
Optimization direction: Application scenarios such as phage therapy

Project 3: Detection and degradation of microplastics in mangroves

Person in charge: Yi Peng, Yingfeng Wu
Optimization direction: To find a better technical route.

Project 4: Management of pine wood nematodes

Person in charge: Minxi Zeng, Zhiying Zhang, Yixi Wang
Optimization direction: Continue to deepen according to the original planning plan

4.9

SZU-China Project Approval Conference

Time: 20:00-23:50, April 9
Venue: Room 202, Life and Marine Science Building, Lihu Campus, Shenzhen University
Minutes of the meeting: Xinying Zhang

Highlights:

1. The members of the four teams submitted the project planning plan that they were responsible for promoting, and gave project speeches in front of Advisor and PI in turn.

2. Under the joint decision of team members, Advisors and PI, we initially selected the new project "Early detection, prevention and treatment of rice sheath blighty", which was extended from the original plant pathogen detection project, as our preferred project. It will be further advanced in the coming week.

4.16

Project Planning Meeting

Time: 9:00-12:00, April 16th
Venue: Room 202, Life and Marine Science Building, Lihu Campus, Shenzhen University
Minutes of the meeting: Xinying Zhang

Highlights:

1. Project Discussion

• (1) Suggestions and preferences for the current project.
• (2) Rice sheath blight project of Rhizoctonia solani.

2. Discussion of the rules of the Synthetic Biology card game

3. Assignment of research tasks to various parts in the early stage of the project

• (1) Find ways to buy R.solani and order it; Find culture Protocol：Zhiying Zhang.
• (2) Vector, synthetic gene and Protocol in Trichoderma vermiliformis：Bingru Feng, Minxi Zeng, Linzhen Bao.
• (3) RNAi design and related implementation scheme：Ruoyu Chen, Yi Peng.
• (4) Problems related to electronic nose system：Yingfeng Wu, Xinying Zhang, Zhiying Zhang.
• (5) Whether the application scenario needs to be changed：Zhiying Zhang, Xinying Zhang, Yixi Wang.
• (6) Life history and appropriate action time points of Rhizoctonia solani in rice field (secondary emphasis)：Xihe Wang, Yixi Wang.
• (7) How to design the spraying time of RNAi and Trichoderma lucidum (whether there is mutual influence and how to maximize the effect)：Wang Xihe, Bao Linzhen.

4.30

Preliminary Survey Summary and Experiment, HP Work Arrangement Meeting

Time: 9:00-12:00, April 30th
Venue: Room 202, Science of Life and Marine Sciences Building, Lihu Campus, Shenzhen University
Minutes of the meeting: Xinying Zhang

Highlights:

1. Coordinate and arrange follow-up work with PI: start to cultivate all kinds of bacteria needed in May, synthesize part of the designed components and biobrick, and define the complete technical route.

2. Early investigation and summary of the project.

• (1）Plasmid Design：The initial version has been completed and will continue to evolve iteratively.
• (2）RNAi research：Plants, bacteria RNAi uptake rate, spray modeling.
• (3）How to design the spraying time of RNAi and Trichoderma atroviride：no literature.
• (4）Experimental simulation of ethyl acetate volatilization：Explosion concentration test.
• (5）Toxicity (to rice), degradation and removal of ethyl acetate.

3.HP event planning:

• （1）iHP scheme: Construct IHP related framework of RNA part.
• （2）Rice greenhouses visit: Establish the time arrangement and the specific contents to be investigated.
• （3）To solve the problems related to the publication of the synthetic biology booklet prepared in cooperation with the team of Southern University of Science and Technology.

5.4

Conference on Plasmid Design

Time: 14:30-16:30, May 4th
Location: Room 202, College of Life and Marine Sciences Building
Minutes of the meeting: Xinying Zhang

Highlights:

1. Discussion on plasmid design:

• Overexpression of PRB1 (protease PRB1 gene).
• A protease Prb1 derived from T. Harzianum (IMI 206040) has been shown to be an effective biocontrol agent against a variety of pathogens. The transformation of this gene PRB1 in other Trichoderma strains increased the biocontrol potential of the transformed strain against R. Solani by many times compared with the wild-type strain.
• Overexpression of EPL1 (surface hydrophobic protein gene).
• Expression of antimicrobial peptide Snakin-1 (SN1) in Trichoderma atroviride.
• RNAi kills candidate genes of Rhizoctonia ruthenicum:
  (1) PMK1 homolog (RPMK1-1 / RPMK1-2) [two and four alternative shRNA molecules, respectively].
  Host-delivered RNA interference (HD-Rnai) technology was used to target two pathogenic MAP kinase 1 (PMK 1) homologs from R. solani, RPMK 1-1 and RPMK 1-2, using a hybrid RNAi construct. Homologs of PMK1 play an important role in other fungal pathogens for episome formation, fungal infection structures required to penetrate plant cuticle, and invasive growth once inside plant tissues and overall pathogen viability within plants.
  (2) PG (AG1IA_04727) [There are two alternative shRNA molecules]
  Polygalacturonic acid (endonuclease) enzyme (PG) is a cell wall structural protein that catalyzes the cleavage of pectin molecule poly α-(1,4) -polygalacturonic acid, participates in the degradation of pectin, and disintegrates the cell wall structure. The synthesis of PG enzymes by pathogenic fungi is key to initiating the host infection process.

2. Discussion on the project experimental design scheme

• (1) The culture of Rhizoctonia orbicularis was started.
• (2) R-body (electron microscope +OD value), which was verified separately from RNAi.

3. Rice plant acquisition, field investigation and electronic nose system arrangement:

• (1) Rice greenhouses can be built in the base, after successful verification on rice plants.
• (2) Difficulty: It is difficult to grow rice from scratch in the culture room with constant conditions.

5.8

Weekly team meeting and South China Exchange Conference to host seminars

Meeting time: 9:00-12:00, May 8th
Venue: Room 202, College of Life and Ocean Sciences, Shenzhen University
Minutes of the meeting: Zhang Xinying

Highlights:

1. Weekly work report of each member.

2. Human practice, meeting up and Collaboration:

• (1) Liaise with other teams: Bingru Feng, Zhiying Zhang.
• (2) Social account operation and contact: Xihe Wang.
• (3) Human Practice: divide the work according to the type of activities and participation.

3. Discuss the overall issues of holding the Meeting up:

• (1) Confirm attendance list.
• (2) Exchange meeting process arrangement.
• (3) Invite guests to participate in comments related matters.
• (4) Making and planning of the 2022 conference manual of South China Exchange Conference.
• (5) Confirm whether it will be held offline.
• (6) The specific time will be finalized.

Experiment

5.28-5.31

—Yi Peng: Amplification and culture of Escherichia coli containing pPIC-P-T empty plasmid; Small amount extraction and verification of pPIC-P-T plasmid; Preparation and transformation of Trichoderma protoplasm.
—Xinying Zhang: Incubation and planting of the first batch of rice seeds.

6.1-6.4

—Bingru Feng: PCR of Rhizoctonia solani colony (preliminary experiment), failed. After investigation and analysis, we planned to use TPS simple method and CTAB method to extract DNA of T. paraquat, and then perform colony PCR.

6.6-6.9

—Bingru Feng: pCAMBIA1302 was transferred into Escherichia coli BL21(DE3) and verified. It failed due to the failure of kanamycin.
—Xinying Zhang & Yi Peng & Yingfeng Wu: The second batch of rice sowing.

6.10-6.12

—Yi Peng & Ruoyu Chen: Determination of the growth curve of Rhizoctonia solani.
—Ruoyu Chen: Plate inoculation of Rhizoctonia Aquarius (Rotary plate method with hole punch).
—Ruoyu Chen: A culture of Rhizoctonia solani was inoculated into isolated rice leaves.
—Yi Peng & Xinying Zhang: Protoplast transformation of Trichoderma atroviride. (empty plasmid pPIC-P-T)

6.14-6.15

—Xinying Zhang: Trichoderma protoplast transformation (all processes were changed to ice).
—Xinying Zhang: The transformed Trichoderma suspension was inoculated onto resistant plates for screening.

6.16-6.18

—Xinying Zhang: Sowing the third batch of rice.
—Xinying Zhang: Activation and cultivation of the deep green Trichoderma preservation collection.
—Xinying Zhang: Observation of Trichoderma pseudotransformants on PDA plates containing hygromycin resistance.

7.7

—Xinying Zhang: Purification of Trichoderma atroviride, single colonies of Trichoderma atroviride were obtained by dilution of colonies on bevel medium.

7.16-7.19

—Xinying Zhang: Plasmid Pan-7 and PPIC were extracted and prepared for the second transformation.
—Xinying Zhang: Preparation of Trichoderma protoplasts.
—Xinying Zhang: PDA plate inoculation of Trichoderma dark green.
—Ruoyu Chen: RNAi-associated plasmids were transformed into Escherichia coli DH5a and selected using a medium containing kanamycin resistance.
—Ruoyu Chen: Rhizoctonia ligulatum culture infected isolated rice leaves (optimized infection device).
—Ruoyu Chen: PDA plate inoculation of Rhizoctoromyces erectus.

7.20-7.21

—Xinying Zhang: Three plasmids designed for engineering Trichoderma were transformed into Escherichia coli DH5a for verification.

7.22

—Xinying Zhang: Validation of Escherichia coli DH5a transformants by colony PCR.
—Xinying Zhang: Washing and germination of spores of Trichoderma deep green.
—Linzhen Bao: Colony PCR for snakin-1 plasmid and plasmid PCR after plasmid extraction.

7.23

—Xinying Zhang: Three plasmids designed for engineering Trichotilbe were transformed into Escherichia coli DH5a for verification (the second batch of plate).
—Xinying Zhang: Trichoderma spore count.

7.24

—Xinying Zhang: The spores germinated overnight were observed under microscope, and the germinated Trichoderma atroviride were inoculated on PDA plates.
—Xinying Zhang: Determination of the relationship curve between spore OD550 and spore concentration of Trichoderma atroviride.
—Ruoyu Chen: Rhizoctonia litmus culture infected isolated rice leaves (infection observation device was optimized again).

7.25

—Xinying Zhang: Colony PCR validation of Escherichia coli DH5a recombinant (epl1, prB1, snakin-1).

7.26-7.27

—Xinying Zhang: Expanded culture of Escherichia coli DH5a recombinant (epl1, prB1, snakin-1).

7.28

—Xinying Zhang: Small volume extraction of engineered Trichoderma Plasmids.
—Xinying Zhang & Yi Peng: Agarose gel electrophoresis verification of small amount of plasmid extract.
—Xinying Zhang & Yi Peng: Preparation of reagents for mass plasmid extraction.
—Xinying Zhang & Yi Peng: Mass extraction of Plasmids from engineered Trichoderma atroviride.
—Xinying Zhang & Yi Peng: Validation of large amount of plasmid extract by agarose gel electrophoresis.
—Xinying Zhang & Yi Peng: Verification of double digestion of a large amount of plasmid extract.
—Linzhen Bao: Verification of double digestion of small amounts of plasmid extracts.
—Ruoyu Chen: Transformation of E. coli HT115(DE3) (RNAi partial plasmids: PG2, RPMK1-1-2, RPMK1-2-2).

7.29

—Ruoyu Chen: Retransformation of HT115(DE3) : RPMK1-1-1, RPMK1-2-1.
—Ruoyu Chen: Mix 50% glycerol with equal volume of bacterial solution and store at -80°C refrigerator.
—Xinying Zhang & Yi Peng: Mass extraction of plasmids from engineered Trichoderma fungi (Repeat 7.28 to prepare subsequent plasmids for transformation) .
—Xinying Zhang & Yi Peng: PCR verification and double digestion verification of a large number of plasmid extracts.
—Xinying Zhang: Inoculate Trichoderma atroviride.

7.30-7.31

—Ruoyu Chen: Colony PCR of HT115 (DE3) transformants.
—Ruoyu Chen: IPTG induces microRNA production in Escherichia coli.
—Ruoyu Chen: Trizol method was used to extract Escherichia coli RNA.
—Xinying Zhang: Transformation of Trichoderma verticillum by carbon nanotube delivery method; Transformation of Trichoderma atroviride by transmembrane peptide method.

8.1

—Ruoyu Chen: Colony PCR validation of HT115 (DE3) transformants transferred into RNAi-associated plasmids.
—Ruoyu Chen: IPTG induces a large number of micrornas in HT115 (DE3).
—Ruoyu Chen & Yi Peng: Large volume extraction of Escherichia coli RNA.
—Linzhen Bao & Wang Xihe: Small amount extraction of snakin-1 plasmid.
—Xinying Zhang & Linzhen Bao & Xihe Wang: Plasmid transformation into Trichoderma dark green by nanomaterial transformation method.

8.2

—RuoYu Chen & Yi Peng: IPTG induces HT115 (DE3) to produce a large number of micrornas.
—RuoYu Chen & Yi Peng: Large scale extraction of E.coli RNA.
—Xinying Zhang: Verification of Snakin-1 plasmid digestion.

8.3

—Xinying Zhang: Transformation of Trichoderma atroviride by carbon nanotube delivery method; Transformation of Trichoderma atroviride by transmembrane peptide method.

8.5

—Ruoyu Chen: Chemical transformation of HT115 (DE3).
—Ruoyu Chen: Experiment on the infection of rice leaves in vitro by Rhizoctonia solani culture.
—Xinying Zhang: Transformation of snakin-1 plasmid into BL21 (DE3).

8.6

—Ruoyu Chen: Colony PCR validation of RNAi partial transformants.
—Yi Peng: Bulk extraction of PG-1 microRNA.
—Yi Peng: Stability experiments of complexes obtained from CNT coated micrornas.

8.7

—Ruoyu Chen: Bulk extraction of RNA from Escherichia coli.
—Xinying Zhang: Resuscitation and expanded culture of transformants stored in a refrigerator at -80°C (prb 1 and epl 1).
—Xinying Zhang: Germination of spores of Trichoderma atroviride.

8.8-8.10

—Xinying Zhang: Bulk extraction of epl 1, prb 1 plasmids.
—Xinying Zhang: Transformation of protoplasts from Trichoderma atroviride.
—Xinying Zhang: Washing dark green Trichoderma spores.
—Yi Peng: Time and stability of CNT encapsulated RNA experiment.

8.11

—Yi Peng: Investigation on the duration and encapsulation rate of RNA wrapped by CNT.
—Xinying Zhang: Confrontation experiments between Trichoderma atroviride and Rhizoctonia solani at different concentrations on plates.
—Ruoyu Chen: Repeated experiments on the infection of rice leaves by Rhizoctonia solani.
—Ruoyu Chen: Transformation of GFP plasmid in HT115(DE3).
—Xinying Zhang: Preparation of Trichoderma atroviride protoplasts.

8.13

—Ruoyu Chen: IPTG induces a large number of micrornas in HT115 (DE3).
—Ruoyu Chen: Mass extraction of Escherichia coli RNA.
—Ruoyu Chen: Repeated experiments on the infection of rice leaves by Rhizoctonia solani.
—Xinying Zhang: Preparation of magnesium silicate nanoclay.

8.14

—Xinying Zhang: Transformation of Trichoderma deep green by magnesium silicate nanoclay method.
—Ruoyu Chen & Yi Peng: Large-scale extraction of Escherichia coli RNA and verification by agarose gel electrophoresis.
—Xinying Zhang: Germination and washing of spores of Trichoderma atroviride.

8.15

—Ruoyu Chen: Re-transformation of GFP plasmid in Escherichia coli HT115 (DE3).
—Ruoyu Chen: Repeated experiments on the infection of rice leaves by Rhizoctonia solani.
—Xinying Zhang: Antifungal experiment of Trichoderma atroviride.

8.16-8.18

—Ruoyu Chen & Yi Peng: Large-scale RNA extraction from Escherichia coli (Plasmid：GFP, PG-1, PG-2, RPMK1-1-1, RPMK1-1-2, RPMK1-2-1，RPMK1-2-2).
—Xinying Zhang: Transformation of Trichoderma deep green by magnesium silicate nanoclay method.
—Xinying Zhang: Preparation and transformation of protoplasts from Trichoderma atroviride.
—Xinying Zhang: Antifungal experiment of Trichoderma atroviride.
—Yi Peng: Encapsulation assay of LDH-RNA complex.

8.19

—Xinying Zhang: Screening of resistance genes in protoplast transformants.
—Ruoyu Chen: Repeated experiments on the infection of rice leaves by Rhizoctonia solani.

8.21

—Ruoyu Chen: Transformation of the second batch of RNAi partial plasmids.
—Xinying Zhang: Resuscitation and expansion culture of glycerol bacteria containing Snakin-1 plasmid.

8.22

—Xinying Zhang: A small amount of plasmid was extracted from the resuscitation solution of glycerin preserving bacteria containing snakin-1 plasmid.
—Xinying Zhang: Snakin-1 transformants were verified by PCR and double restriction enzyme digestion.

8.23-8.24

—Ruoyu Chen: IPTG induced Escherichia coli HT115 (DE3) to produce a large amount of target RNA (plasmids: C6, E3, Psm1, β1).
—Ruoyu Chen & Yi Peng: Large scale extraction of RNA from Escherichia coli.
—Xinying Zhang: Protoplast transformation of Trichoderma atroviride.
—Xinying Zhang: Subculture of Trichoderma atroviride.

8.25

—Ruoyu Chen: Effect of RNA on the morphology of dead cells under the microscope.
—Yi Peng: Preparation and transformation of Trichoderma atroviride protoplasm.

8.26

—Ruoyu Chen: The effect of shRNA against dryness at the micro level.
—Ruoyu Chen: The effect of shRNA on the growth of vegetative cells on plates.
—Xinying Zhang: Transformation and screening of protoplasts from Trichoderma atroviride.
—Xinying Zhang: Antifungal experiment of Trichoderma atroviride.

8.28

—Yi Peng: Transformation of Agrobacterium GV3101 (plasmid: epl 1 and prb 1).
—Ruoyu Chen: Large scale RNA extraction from Escherichia coli (plasmid: PG-1, PG-2, Psm1, β1, GFP).

8.30

—Yi Peng: Colony PCR validation of Agrobacterium GV3101 transformants.
—Yi Peng: Amplification and culture of Agrobacterium GV3101 transformants.

8.31-9.2

—Yi Peng: Trichoderma deep green spores were transformed using Agrobacterium GV3101 transformants.
—Ruoyu Chen: Bulk extraction of RNA from Escherichia coli.
—Ruoyu Chen: Stability verification of shRNA.
—Ruoyu Chen: Verification of shRNA stability after binding LDH.
—Xinying Zhang: Protoplast transformation of Trichoderma atroviride.
—Ruoyu Chen & Xinying Zhang: An experimental study on the inhibitory effect of shRNA on Rhizoctonia solani on plates.
—Xinying Zhang: Resistance plate screening for protoplast transformation of Trichoderma atroviride.
—Xinying Zhang: Transformation of Agrobacterium tumefaciens GV3101 (plasmid: snakin-1).
—Ruoyu Chen: Microscopic examination showed that shRNA could enter the cells of Trichoderma atroviride.

9.3

—Xinying Zhang: Agrobacterium containing PRB 1 and epl 1 plasmids on culture plates was resuscitated and expanded.
—Xinying Zhang & Yi Peng: Transformation of Trichoderma atroviride spores using Agrobacterium Tumefaciens GV3101 transformants.

9.5-9.12

At this stage, the previous agrobacterium transformation experiment and in vitro rice leaf infection experiment were repeated continuously, so no duplicate records were made.

—Xinying Zhang & Yi Peng: Transformation of Trichoderma atroviride spores or protoplasts using Agrobacterium Tumefaciens GV3101 transformants.
—Ruoyu Chen: Observation of rice leaves under stereomicroscope after mycelium inoculation.
—Ruoyu Chen: RNA extraction from Rhizoctonia solani.

9.13-9.14

To our great excitement, we successfully obtained the first Trichoderma atroviride pseudotransformants transferred into the prb 1 plasmid on September 13th! Moreover, the transformants transferred into epl 1 and snakin-1 plasmids also showed signs of growth.

—Xinying Zhang & Yi Peng: Screening of the secondary rotation plate of transformants by Trichoderma atroviride.
—Ruoyu Chen: RNA was extracted from Rhizoctonia solani and cDNA was obtained by reverse transcription.
—Ruoyu Chen & Bingru Feng: Screening assay for standard primers (18S, GADPH).

9.15

—Ruoyu Chen & Bingru Feng: Re-extraction of cDNA from Rhizoctonia solani.
—Ruoyu Chen & Bingru Feng: Revalidation of standard primers.

9.18

—Xinying Zhang & Yi Peng: DNA extraction and PCR of the pseudotransformants of Trichoderma atroviride.
—Xinying Zhang & Yi Peng: The proposed transformants were transferred to PDA plates without resistance for expanded culture.

9.22

—Yi Peng: The proposed transformants were transferred into PDB liquid medium containing hygromycin B resistance for expanded culture.

9.24

—Yi Peng & Xinying Zhang: Extraction of whole protein from the transformants of Trichoderma deep green. The target protein containing His-tag was purified using an affinity purification kit.
—Yi Peng：The purified target protein was verified by SDS-PAGE.

9.26-10.1

—Yi Peng: LC-MS technology was used to further verify the target protein and relative quantitative analysis of its overexpression.

Products

7.25-8.3

—Minxi Zeng: Preliminary experimental research on inhaling powder.
—Minxi Zeng: Spore production induced by Trichoderma atroviride.
—Minxi Zeng: Drawing of standard curve of absorbance and spore number.
—Bingru Feng: Simple method was used to extract the DNA of Rhizoctonia solani and verify it by PCR.

8.4-8.10

—Minxi Zeng: To determine the effect of suction powder on different kinds of common solutions.
—Minxi Zeng: Determination of encapsulation rate and loading rate of suction powder microspheres.
—Minxi Zeng: Explore the balance process of suction and swelling powder content detachment.
—Minxi Zeng: To explore the degradation effect of suction powder.
—Minxi Zeng: Selection of gas producing materials.
—Bingru Feng: Various methods were used to extract DNA from Rhizoctonia solani and verified by PCR and electrophoresis.
—Bingru Feng：Preform LAMP reactions and verified by electrophoresis.

8.11-8.21

—Minxi Zeng: Determination of adsorption radius of adsorptive powder.
—Minxi Zeng: Short-term release of Trichoderma spore.
—Minxi Zeng: Determination of sedimentation rate of suction powder.
—Bgru Feng: Perform LAMP reaction and verified by electrophoresis.
—Bgru Feng: Use varieties of methods to extract DNA from R.solani, and the DNA was verified by PCR and electrophoresis.

8.23-8.31

—Minxi Zeng: T.ACE quality control.
—Minxi Zeng: Calculate the effective concentration of each recommended method.
—Bingru Feng: pre-experiment of LFD.
—Bgru Feng: Perform LAMP reaction and verified by electrophoresis.
—Bgru Feng: Use varieties of methods to extract DNA from R.solani, and the DNA was verified by PCR and electrophoresis.

9.1-9.10

—Minxi Zeng: Measurement and calculation of morphology, particle size and other parameters of T.ACE prescription II.
—Minxi Zeng: Determination of the release rate of T.ACE with different particle sizes.
—Minxi Zeng: T.ACE product effect monitoring.
—Minxi Zeng: Temporary photographic monitoring of the release process of T.ACE.
—Bingru Feng: Perform LAMP reaction and verified by electrophoresis.
—Bingru Feng: Use varieties of methods to extract DNA from R.solani, and the DNA was verified by PCR and electrophoresis.

9.10-9.20

—Bingru Feng: CTAB method and TPS method were used to extract DNA of R.solanum AG-1, respectively.
—Bingru Feng: PCR verification of DNA of Rhizoctonia solani.
—Bingru Feng: PCR gradient experiment of DNA extraction from R.solanum AG-1 by TPS.

Model，software and hardware

5.1-7.10

—Yingfeng Wu：Algorithm design and system architecture design of electronic nose.

7.11-7.23

—Yingfeng Wu：The appearance design of the electronic nose.
—Yingfeng Wu：The circuit principle design of electronic nose.
—Yingfeng Wu：Design of electronic nose supporting APP (version 1.0).
—Yingfeng Wu：Prediction model of co-culture concentration of R.solani and T.atroviride.

7.24-8.21

—Yingfeng Wu & Yi Peng：Write the user guide of electronic nose supporting APP (version 1.0).
—Yingfeng Wu：Modeling of disease spot detection and culture medium colony detection of rice infected by R.solani.
—Yingfeng Wu：Endurance curve modeling and pollen spore release modeling of fish suction.

8.24-9.17

—Yingfeng Wu:Determine the overall framework of the wiki.
—Yingfeng Wu & Yixi Wang：Design of the second generation electronic nose supporting APP.
—Yingfeng Wu & Yi Peng：Write the user guide of the second generation electronic nose supporting APP.
—Yingfeng Wu：Build the wiki template.
—Yingfeng Wu：Electronic nose hardware assembly and debugging.
—Yingfeng Wu：Electronic nose data acquisition experiment.
—Yingfeng Wu：Modification of the spore release model of T.atroviride.
—Yingfeng Wu：The algorithm development of the hardware part of the electronic nose.

9.18-9.30

—Yingfeng Wu:Refinement of all models.
—Yingfeng Wu：Electronic nose data acquisition experiment perfection.
—Yingfeng Wu：Analyze the differences in data collection between R.solani and Pirospora oryzae.

Wiki

After the initial confirmation of our wiki framework on August 24th, all of us started writing the manuscript for our respective pages. We started making demos of each page on September 10th, and the first phase of Wiki Freeze was launched on September 20th. Between September 20th and October 12th, we made further changes and content additions to each page, and finally completed the entire wiki.