S I R E N S ntelligent .coli ynthetic ematode anger Agianst - PowerPoint PPT Presentation

S I R E N S ntelligent .coli ynthetic ematode anger Agianst

1 Economic damage Environment hazard lack of weariness background Meloisogyne spp. [1] Meloisogyne incoginta [2] Heterodera spp. [1] Bursaphelenchus xylophilus [1] [1] wikipidia [2] Beijing Agricultural database

1 Economic damage Environment hazard lack of weariness background HALF of CHINA! 3rd announcement of China’s State Forestry Administration (SAF) in 2015

1 Environment hazard Economic damage lack of weariness background Common Nematicides Nematicides Targets Disadvantages High toxicity. Do harm to the soil, the environment even the health. The using period is limited. Nematodes in the soil Ethylene dibromide Other small pests It is probably do harm to the crops. Preventive nematicides. The operation is very complex. Apelenchoides fragariae Have toxicity. Fenamiphos Most of the nematodes The underground water will be polluted. Meloidogyne; Pratylenchus; Abamectin Radopholus; Tylenchulus; Heterodera glycines Expensive Meloidogyne; Heterodera; Biological agent DiT Pratyleachus spp.; Trichodorus spp.; Belonolaimus spp.

1 Lack of awareness Economic damage Environment hazard background Investigation in Hebei Province Officials and local pesticide retailer knew little about nematodes Villagers only care about visible pests instead of invisible nematodes

1 Lack of awareness Economic damage Environment hazard background Investigation in Hebei Province Sample analysis - Soil nematode screening

1 Lack of awareness Economic damage Environment hazard background Communication With Other Teams The abuse of pesticide have brought about severe agricultural diseases all over the world!

1 Lack of awareness Economic damage Environment hazard background An interview with Dr. Yan Houfu (Beijing Normal University) Lack of bills on bio-safety , Lack of protection of farmers’ rights!

Lack of protection of farmers’ rights The abuse of pesticide have brought about severe agricultural diseases all over the world! How to solve these problems ? We aim to solve these problems ! ? Villagers only care about visible pests instead of invisible nematodes Economic damage of nematodes Officials and local pesticide retailer knew little about nematodes Environmental hazard Lack of bills on bio-safety

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The !Project

2 Overview Photoinduced System BDT System Bait & Killer System Suicide Project Photo-induced System Bidirectional Transcription System Bait & Killer System Suicide

2 Photoinduced System Overview BDT System Bait & Killer System Suicide Project Cph8 The construction result of light regulation system

2 BDT System Overview Photoinduced System Bait & Killer System Suicide Project Biobrick 1 Biobrick 2 Detection experiment result Biobrick 1 & 2

2 Project

2 Bait & Killer System Overview Photoinduced System BDT System Suicide Project Limonene terpenoid attracts nematodes such as Tylenchulus semipenetrans d-limonene l-limonene

2 Bait & Killer System Overview Photoinduced System BDT System Suicide Project Biosysthesis Pathway Engineered pathway for l-limonene biosynthesis in E. coli Reference : Du et al. Bioresources and Bioprocessing,2014,1:10 .

2 Bait & Killer System Overview Photoinduced System BDT System Suicide Project Vector : pGEX-4T-1 plasmid C hassis : E. coli BL21 (DE3) RBS GPPS LS Promoter GPPS gene : Abies grandis (GenBank : AF513112) BBa_J23100 BBa_B0032 d-LS gene : Citrus unshiu (GenBank : AB110636.1) Strong constitutive promoter l-LS gene : Mentha spicata (GenBank : L13459)

2 Bait & Killer System Overview Photoinduced System BDT System Suicide Project Limonene LS : 89kDa GPPS: 49kDa Production Agarose gel electrophoresis SDS-PAGE of l-/d- limonene synthase and GPPS d-limonene GC-MS

2 Bait & Killer System Overview Photoinduced System BDT System Suicide Project Limonene attracts C.elegans significantly The distribution of C. elegans Proportion of nematodes' distribution

2 Bait & Killer System Overview Photoinduced System BDT System Suicide Project Consulting with Prof. Liu Xingzhong (PhD, CAS)

2 Bait & Killer System Overview Photoinduced System BDT System Suicide Project Vector : pSB1C3 plasmid Toxins: Bace16 & rMpL C hassis : E. coli BL21(DE3) C hassis : E. coli BW25113 Promoter Promoter RBS bace16 / rMpL bace16 gene : Bacillus nematocida BBa_J23100 BBa_K206000 BBa_B0032 (GenBank : AY708655) Constitutive promoter pBAD promoter rMpL gene : Macrolepiota procera (GenBank : HQ449739)

2 Bait & Killer System Overview Photoinduced System BDT System Suicide Project rMpL successfully expressed SDS-PAGE of rMpL Lane 1, supernatant of pSB1C3-BL21(DE3) with constitutive promoter; Lane 2, supernatant of pSB1C3-BW25113 with pBAD promoter after 0uM L-Ara induction; Lane 3, supernatant of pSB1C3.

2 Bait & Killer System Overview Photoinduced System BDT System Suicide Project rMpL stops the C.elegans growth rMpL PSB1C3 empty plasmid

2 Suicide Overview Photoinduced System BDT System Bait & Killer System Project LuxR AHL LuxI LuxR-AHL

Device&Modeling

3 Device Modeling Device&Modeling Light Controlling Chamber CO 2 Generating Chamber E.coli Culturing Chamber Cuboid Outer Shell outer shell slide glasses separating funnel red LED light test tube

3 Device Modeling Device&Modeling http://2015.igem.org/wiki/images/0/00/BNU-MODELING-DEVICE-VIDEO.mp4

3 Modeling Device Device&Modeling Nematodes’ Diffusion movement velocity, coefficient of attraction time limonene √ L = V × T AB = 2 2 EF V: nematodes’ crawl C 0 x [1] Limm W, Begley T H, 2 (1 − erf ) speed √ Fick’s Law: 2 Dt Lickly T, et al. Diffusion of gas diffusion limonene inpolyethylene[J]. 18 The longest distance = C 0 To Find The Best Position For X ( − 1) i 2 (1 − model Food additives and T: the half ∂ 2 C ( t, x ) ∂ C ( t, x ) between nematodes = D A contaminants, 2006, 23(7): i =0 time of which ∂ x 2 ∂ t and the equipment 738-746. x 2 i +1 The Device To Be Placed In Farmland nematodes grow from Dt ) 2 i +1 ) √ i !(2 i + 1)(2 eggs to larvae Distance between two devices dichotomy the concentration cellular automaton simulation of every device

4 Public engagement Database extension Voluntary education in GuiZhou Province Synthetic biology Plant diseases and insect pests SIRENS

4 Public engagement Database extension Open day of BNU-CHINA Interviewed by BNU TV station Synthetic biology Our innovative idea iGEM competition How iGEM impact us SIRENS Our achievements

4 Database Public engagement extension

5 Team Modeling Wet lab Wiki Art Policy & Practice

6 Ackonwledgement

Thanks ！

BNU CHINA