Team:FAFU-CHINA/Proof
Contents
Proof
1.Introduction
In our project, we transform theexpression vector of Cry and Cyt toxins. Based on the toxin groups,we construct six various toxins express devices as Figure1. To compare the toxicities, we also tested the toxicity of Bacillus thuringiensis strain LLP229.
2.Material, Condition and Method
The larvae of Ae. albopictus are feeding in the Key Laboratory of Biopesticideand Chemical Biology. The generation of Ae.albopictus is 11. The LLP229strain of Bacillus thuringiensis isprovided by Key Laboratory of Biopesticide and Chemical Biology. The concentration of LLP229is recorded in eachtable.</p>
<p style="text-indent:36.0pt">The CC530 and FA479 strains of Chlamydomonas reinhardtii and TAP mediumare bought from the Aquatic Species Research Institute in Wuhan, Hubei. The concentration of Chlamydomonasreinhardtii is recorded ineach table.</p>
<p style="text-indent:36.0pt">We contrasted various toxins groupexpression vectors and transform them into Chlamydomonasreinhardtii as following groups: Cry4Aa+2A+Cyt1,Cry4Aa+2A+Cyt2, Cry10Aa+2A+Cyt1, Cry10Aa+2A+Cyt2, Cry11Aa+2A+Cyt1,Cry11Aa+2A+Cyt2.</p>
<p style="text-indent:36.0pt">The toxicity analysis condition is in anenvironmentally controlled room (T=28±1 °C, RH=85%±5%, L: D=14:10).</p>
<p style="text-indent:36.0pt">Bioassay of variousconcentrations were assayed against 30 third-instar larvae in 20 ml of dechlorinatedwater. Each group was replicated 3 times. The level of mortality was recordedafter 48 hours. Then calculated the corrected mortality. The bioassay was thenrepeated such that a concentration range yielding 10–90% mortality was obtainedand the 50% lethal concentration (LC50) was calculated.</p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid">Table 1.1 Toxicity of LLP29 to A. albopictus</p>
Repeating group Number of insects Number of mortality Corrected mortality (%) 1 30 30 100 2 30 30 100 3 30 30 100 Control 30 0 0 <p align="center" style="text-align:center"> Repeating group Number of insects Number of mortality Corrected mortality (%) 1 30 30 100 2 30 30 100 3 30 30 100 Control 30 0 0 <p align="center" style="text-align:center"> Repeating group Number of insects Number of mortality Corrected mortality (%) 1 100 93 93% 2 100 89 89% 3 100 91 91% Control 1 Control 2 100 0 0 <p> Repeating group Number of insects Number of mortality Corrected mortality (%) 1 100 89 89% 2 100 90 90% 3 100 88 88% Control 1 Control 2 100 0 0 <p> Repeating group Number of insects Number of mortality Corrected mortality (%) 1 100 90 90% 2 100 92 92% 3 100 91 91% Control 1 Control 2 100 0 0 <p> Repeating group Number of insects Number of mortality Corrected mortality (%) 1 100 90 90% 2 100 90 90% 3 100 89 89% Control 1 Control 2 100 0 0 <p> Repeating group Number of insects Number of mortality Corrected mortality (%) 1 100 90 90% 2 100 95 95% 3 100 91 91% Control 1 Control 2 100 0 0 <p> Repeating group Number of insects Number of mortality Corrected mortality (%) 1 100 91 91% 2 100 90 90% 3 100 89 89% Control 1 Control 2 100 0 0 <p> <p>1.Zhang, L., Huang, E., Lin, J., Gelbič,I., Zhang, Q., Guan, Y., … Guan, X. (2010). A novel mosquitocidal strain LLP29isolated from the phylloplane of. Microbiological Research, 165(2), 133–141.doi: 10.1016/j.micres.2009.03.002 </p>
Note. Thenconcentration of LLP29 is 2.16 x 109 (OD750 of 1.0 = 8 x 108 cells/ml,OD750 =2.7A.)</p>
<p align="center" style="text-align:center"> </p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid">Table 1.2 Toxicity of LLP29 to Culex quinquefasciatus</p>
Note. Thenconcentration of LLP29 is 2.16 x 109 (OD750 of 1.0 = 8 x 108 cells/ml,OD750 =2.7A)</p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid">Table 2.1 Toxicity of Chlamydomonas reinhardtii(Cry4Aa+2A+Cyt1) toA. albopictus</p>
Note.1. The OD750 of Chlamydomonas reinhardtii is 0.559</p>
<p> 2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629)</p>
<p> 3. The Control 2 group is TAP medium </p>
<p> </p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid">Table 2.2 Toxicity of Chlamydomonas reinhardtii(Cry4Aa+2A+Cyt2) toA. albopictus</p>
Note1. The OD750 of Chlamydomonas reinhardtii is 0.559</p>
<p> 2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629)</p>
<p> 3. The Control 2 group is TAP medium</p>
<p> </p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid">Table 2.3 Toxicity of Chlamydomonas reinhardtii(Cry10Aa+2A+Cyt1) toA. albopictus</p>
Note1.The OD750 of Chlamydomonas reinhardtii is 0.559</p>
<p> 2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629)</p>
<p> 3. The Control 2 group is TAP medium </p>
<p> </p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid">Table 2.4 Toxicity of Chlamydomonas reinhardtii(Cry10Aa+2A+Cyt2) toA. albopictus</p>
Note1. The OD750 of Chlamydomonas reinhardtii is 0.559</p>
<p> 2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629)</p>
<p> 3. The Control 2 group is TAP medium </p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid">Table 2.5 Toxicity of Chlamydomonas reinhardtii(Cry11Aa+2A+Cyt1) toA. albopictus</p>
Note1.The OD750 of Chlamydomonas reinhardtii is 0.559</p>
<p> 2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629)</p>
<p> 3. The Control 2 group is TAP medium</p>
<p> </p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid">Table 2.5 Toxicity of Chlamydomonas reinhardtii(Cry11Aa+2A+Cyt2) toA. albopictus</p>
Note1. The OD750 of Chlamydomonas reinhardtii is 0.559 </p>
<p> 2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629)</p>
<p> 3. The Control 2 group is TAP medium
</p>
<p align="center" style="text-align:center">3.Reference</p>
