Difference between revisions of "Team:FAFU-CHINA/Proof"

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= <p><b><span style="font-size:12.0pt;font-family:&quot;:,serif">2.Material, Condition and Method</span></b></p> =
 
= <p><b><span style="font-size:12.0pt;font-family:&quot;:,serif">2.Material, Condition and Method</span></b></p> =
 
<span>&nbsp; &nbsp;&nbsp;<span><p><span style="text-indent:36pt;font-size:12pt;font-family:&quot">The larvae of <i>Ae. albopictus</i> are feeding in the Key Laboratory of Biopesticideand Chemical Biology. The generation of <i>Ae.albopictus </i>is 11. The <i>LLP229</i>strain of <i>Bacillus thuringiensis</i> isprovided by Key Laboratory of Biopesticide and Chemical Biology.</span><span style="text-indent:36pt;font-family:&quot"> The concentration of </span><i style="text-indent:36pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">LLP229</span></i><span style="text-indent:36pt;font-size:12pt;font-family:&quot">is recorded in eachtable.</span></p>
 
<span>&nbsp; &nbsp;&nbsp;<span><p><span style="text-indent:36pt;font-size:12pt;font-family:&quot">The larvae of <i>Ae. albopictus</i> are feeding in the Key Laboratory of Biopesticideand Chemical Biology. The generation of <i>Ae.albopictus </i>is 11. The <i>LLP229</i>strain of <i>Bacillus thuringiensis</i> isprovided by Key Laboratory of Biopesticide and Chemical Biology.</span><span style="text-indent:36pt;font-family:&quot"> The concentration of </span><i style="text-indent:36pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">LLP229</span></i><span style="text-indent:36pt;font-size:12pt;font-family:&quot">is recorded in eachtable.</span></p>
<p style="text-indent:36.0pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">The CC530 and FA479 strains of <i>Chlamydomonas reinhardtii </i>and TAP mediumare bought from the Aquatic Species Research Institute in Wuhan, Hubei.</span><span style="font-family:&quot;:,serif"> The concentration of </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif">Chlamydomonasreinhardtii </span></i><span style="font-size:12.0pt;font-family:&quot;:,serif">is recorded ineach table.</span></p>
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<p style="text-indent:36.0pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">The CC530 and FA479 strains of <i><em>Chlamydomonas reinhardtii</em> </i>and TAP mediumare bought from the Aquatic Species Research Institute in Wuhan, Hubei.</span><span style="font-family:&quot;:,serif"> The concentration of </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif"><em>Chlamydomonas reinhardtii</em> </span></i><span style="font-size:12.0pt;font-family:&quot;:,serif">is recorded ineach table.</span></p>
<p style="text-indent:36.0pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">We contrasted various toxins groupexpression vectors and transform them into <i>Chlamydomonasreinhardtii </i>as following groups: <i>Cry4Aa+2A+Cyt1,Cry4Aa+2A+Cyt2, Cry10Aa+2A+Cyt1, Cry10Aa+2A+Cyt2, Cry11Aa+2A+Cyt1,Cry11Aa+2A+Cyt2</i>.</span></p>
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<p style="text-indent:36.0pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">We contrasted various toxins groupexpression vectors and transform them into <i><em>Chlamydomonas reinhardtii</em> </i>as following groups: <i>Cry4Aa+2A+Cyt1,Cry4Aa+2A+Cyt2, Cry10Aa+2A+Cyt1, Cry10Aa+2A+Cyt2, Cry11Aa+2A+Cyt1,Cry11Aa+2A+Cyt2</i>.</span></p>
 
<p style="text-indent:36.0pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">The toxicity analysis condition is in anenvironmentally controlled room (</span><em><span>T</span></em><span style="font-family:&quot;:,serif">=28±1&nbsp;°C, RH=85%±5%, L: D=14:10).</span><span style="font-size:12.0pt;font-family:&quot;:,serif"></span></p>
 
<p style="text-indent:36.0pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">The toxicity analysis condition is in anenvironmentally controlled room (</span><em><span>T</span></em><span style="font-family:&quot;:,serif">=28±1&nbsp;°C, RH=85%±5%, L: D=14:10).</span><span style="font-size:12.0pt;font-family:&quot;:,serif"></span></p>
 
<p style="text-indent:36.0pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">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 (LC<sub>50</sub>) was calculated.</span><span style="font-size:12.0pt;font-family:&quot;:,serif"></span></p>
 
<p style="text-indent:36.0pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">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 (LC<sub>50</sub>) was calculated.</span><span style="font-size:12.0pt;font-family:&quot;:,serif"></span></p>
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Note.</span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> Thenconcentration of <i>LLP29 </i>is <span>2.16 x 10<sup>9</sup> (OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750</span></sub><span><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">of 1.0 = 8 x 10<sup>8</sup><span>&nbsp;</span>cells/ml,OD</span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750</span></sub><sub><span style="font-size:12.0pt;font-family:&quot;:,serif"> </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">=2.7A)</span><span style="font-size:12.0pt;font-family:&quot;:,serif"></span></p>
 
Note.</span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> Thenconcentration of <i>LLP29 </i>is <span>2.16 x 10<sup>9</sup> (OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750</span></sub><span><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">of 1.0 = 8 x 10<sup>8</sup><span>&nbsp;</span>cells/ml,OD</span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750</span></sub><sub><span style="font-size:12.0pt;font-family:&quot;:,serif"> </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">=2.7A)</span><span style="font-size:12.0pt;font-family:&quot;:,serif"></span></p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid"><span style="font-size:12pt;font-family:&quot;color:windowtext">Table 2.1 Toxicity of </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Chlamydomonas reinhardtii</span><span style="font-size:12pt;font-family:&quot;color:windowtext">(</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Cry4Aa+2A+Cyt1</span><span style="font-size:12pt;font-family:&quot;color:windowtext">)</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"> </span><span style="font-size:12pt;font-family:&quot;color:windowtext">to</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">A. albopictus</span></p>
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<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid"><span style="font-size:12pt;font-family:&quot;color:windowtext">Table 2.1 Toxicity of </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"><em>Chlamydomonas reinhardtii</em></span><span style="font-size:12pt;font-family:&quot;color:windowtext">(</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Cry4Aa+2A+Cyt1</span><span style="font-size:12pt;font-family:&quot;color:windowtext">)</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"> </span><span style="font-size:12pt;font-family:&quot;color:windowtext"> to </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">A. albopictus</span></p>
  
 
<table border="1" cellspacing="0" cellpadding="0" align="left" width="567" style="border-collapse:collapse;border:none;margin-left:6.75pt;margin-right:6.75pt">
 
<table border="1" cellspacing="0" cellpadding="0" align="left" width="567" style="border-collapse:collapse;border:none;margin-left:6.75pt;margin-right:6.75pt">
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&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;Note.1. The <span>OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750 </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">of <i>Chlamydomonas reinhardtii is </i></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.559</span></p>
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&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;Note.1. The <span>OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750 </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">of <i><em>Chlamydomonas reinhardtii</em> is </i></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.559</span></p>
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2. The Control 1 group is </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif">Chlamydomonas reinhardtii</span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> (notransformation,<span> OD=</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.629</span><span style="font-size:12.0pt;font-family:&quot;:,serif">)</span></p>
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<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2. The Control 1 group is </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif"><em>Chlamydomonas reinhardtii</em></span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> (notransformation,<span> OD=</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.629</span><span style="font-size:12.0pt;font-family:&quot;:,serif">)</span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3. The Control 2 group is TAP medium </span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3. The Control 2 group is TAP medium </span></p>
 
<p><span style="font-family:&quot;:,serif">&nbsp;</span></p>
 
<p><span style="font-family:&quot;:,serif">&nbsp;</span></p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid"><span style="font-size:12pt;font-family:&quot;color:windowtext">Table 2.2 Toxicity of </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Chlamydomonas reinhardtii</span><span style="font-size:12pt;font-family:&quot;color:windowtext">(</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Cry4Aa+2A+Cyt2</span><span style="font-size:12pt;font-family:&quot;color:windowtext">)</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"> </span><span style="font-size:12pt;font-family:&quot;color:windowtext">to</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">A. albopictus</span></p>
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<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid"><span style="font-size:12pt;font-family:&quot;color:windowtext">Table 2.2 Toxicity of </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"><em>Chlamydomonas reinhardtii</em></span><span style="font-size:12pt;font-family:&quot;color:windowtext">(</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Cry4Aa+2A+Cyt2</span><span style="font-size:12pt;font-family:&quot;color:windowtext">)</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"> </span><span style="font-size:12pt;font-family:&quot;color:windowtext"> to </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">A. albopictus</span></p>
  
 
<table border="1" cellspacing="0" cellpadding="0" align="left" width="567" style="border-collapse:collapse;border:none;margin-left:6.75pt;margin-right:6.75pt">
 
<table border="1" cellspacing="0" cellpadding="0" align="left" width="567" style="border-collapse:collapse;border:none;margin-left:6.75pt;margin-right:6.75pt">
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&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;Note1. The <span>OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750 </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">of <i>Chlamydomonas reinhardtii is </i></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.559</span></p>
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&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;Note1. The <span>OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750 </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">of <i><em>Chlamydomonas reinhardtii</em> is </i></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.559</span></p>
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2. The Control 1 group is </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif">Chlamydomonas reinhardtii</span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> (notransformation,<span> OD=</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.629</span><span style="font-size:12.0pt;font-family:&quot;:,serif">)</span></p>
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<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2. The Control 1 group is </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif"><em>Chlamydomonas reinhardtii</em></span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> (notransformation,<span> OD=</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.629</span><span style="font-size:12.0pt;font-family:&quot;:,serif">)</span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3. The Control 2 group is TAP medium</span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3. The Control 2 group is TAP medium</span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;</span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;</span></p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid"><span style="font-size:12pt;font-family:&quot;color:windowtext">Table 2.3 Toxicity of </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Chlamydomonas reinhardtii</span><span style="font-size:12pt;font-family:&quot;color:windowtext">(</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Cry10Aa+2A+Cyt1</span><span style="font-size:12pt;font-family:&quot;color:windowtext">)</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"> </span><span style="font-size:12pt;font-family:&quot;color:windowtext">to</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">A. albopictus</span></p>
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<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid"><span style="font-size:12pt;font-family:&quot;color:windowtext">Table 2.3 Toxicity of </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"><em>Chlamydomonas reinhardtii</em></span><span style="font-size:12pt;font-family:&quot;color:windowtext">(</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Cry10Aa+2A+Cyt1</span><span style="font-size:12pt;font-family:&quot;color:windowtext">)</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"> </span><span style="font-size:12pt;font-family:&quot;color:windowtext"> to </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">A. albopictus</span></p>
  
 
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<table border="1" cellspacing="0" cellpadding="0" align="left" width="567" style="border-collapse:collapse;border:none;margin-left:6.75pt;margin-right:6.75pt">
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&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;Note1.The <span>OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750 </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">of <i>Chlamydomonas reinhardtii is </i></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.559</span><span style="font-size:12.0pt;font-family:&quot;:,serif"></span></p>
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&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;Note1.The <span>OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750 </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">of <i><em>Chlamydomonas reinhardtii</em> is </i></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.559</span><span style="font-size:12.0pt;font-family:&quot;:,serif"></span></p>
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2. The Control 1 group is </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif">Chlamydomonas reinhardtii</span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> (notransformation,<span> OD=</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.629</span><span style="font-size:12.0pt;font-family:&quot;:,serif">)</span></p>
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<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2. The Control 1 group is </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif"><em>Chlamydomonas reinhardtii</em></span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> (notransformation,<span> OD=</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.629</span><span style="font-size:12.0pt;font-family:&quot;:,serif">)</span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3. The Control 2 group is TAP medium </span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3. The Control 2 group is TAP medium </span></p>
 
<p><span style="font-family:&quot;:,serif">&nbsp;</span></p>
 
<p><span style="font-family:&quot;:,serif">&nbsp;</span></p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid"><span style="font-size:12pt;font-family:&quot;color:windowtext">Table 2.4 Toxicity of </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Chlamydomonas reinhardtii</span><span style="font-size:12pt;font-family:&quot;color:windowtext">(</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Cry10Aa+2A+Cyt2</span><span style="font-size:12pt;font-family:&quot;color:windowtext">)</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"> </span><span style="font-size:12pt;font-family:&quot;color:windowtext">to</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">A. albopictus</span></p>
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<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid"><span style="font-size:12pt;font-family:&quot;color:windowtext">Table 2.4 Toxicity of </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"><em>Chlamydomonas reinhardtii</em></span><span style="font-size:12pt;font-family:&quot;color:windowtext">(</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Cry10Aa+2A+Cyt2</span><span style="font-size:12pt;font-family:&quot;color:windowtext">)</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"> </span><span style="font-size:12pt;font-family:&quot;color:windowtext"> to </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">A. albopictus</span></p>
  
 
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<table border="1" cellspacing="0" cellpadding="0" align="left" width="567" style="border-collapse:collapse;border:none;margin-left:6.75pt;margin-right:6.75pt">
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&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;Note1. The <span>OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750 </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">of <i>Chlamydomonas reinhardtii is </i></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.559</span></p>
+
&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;Note1. The <span>OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750 </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">of <i><em>Chlamydomonas reinhardtii</em> is </i></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.559</span></p>
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2. The Control 1 group is </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif">Chlamydomonas reinhardtii</span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> (notransformation,<span> OD=</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.629</span><span style="font-size:12.0pt;font-family:&quot;:,serif">)</span></p>
+
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2. The Control 1 group is </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif"><em>Chlamydomonas reinhardtii</em></span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> (notransformation,<span> OD=</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.629</span><span style="font-size:12.0pt;font-family:&quot;:,serif">)</span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3. The Control 2 group is TAP medium </span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3. The Control 2 group is TAP medium </span></p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid"><span style="font-size:12pt;font-family:&quot;color:windowtext">Table 2.5 Toxicity of </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Chlamydomonas reinhardtii</span><span style="font-size:12pt;font-family:&quot;color:windowtext">(</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Cry11Aa+2A+Cyt1</span><span style="font-size:12pt;font-family:&quot;color:windowtext">)</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"> </span><span style="font-size:12pt;font-family:&quot;color:windowtext">to</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">A. albopictus</span></p>
+
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid"><span style="font-size:12pt;font-family:&quot;color:windowtext">Table 2.5 Toxicity of </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"><em>Chlamydomonas reinhardtii</em></span><span style="font-size:12pt;font-family:&quot;color:windowtext">(</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Cry11Aa+2A+Cyt1</span><span style="font-size:12pt;font-family:&quot;color:windowtext">)</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"> </span><span style="font-size:12pt;font-family:&quot;color:windowtext"> to </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">A. albopictus</span></p>
  
 
<table border="1" cellspacing="0" cellpadding="0" align="left" width="567" style="border-collapse:collapse;border:none;margin-left:6.75pt;margin-right:6.75pt">
 
<table border="1" cellspacing="0" cellpadding="0" align="left" width="567" style="border-collapse:collapse;border:none;margin-left:6.75pt;margin-right:6.75pt">
Line 427: Line 427:
 
<br/>
 
<br/>
 
<br/>
 
<br/>
&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;Note1.The <span>OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750 </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">of <i>Chlamydomonas reinhardtii is </i></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.559</span></p>
+
&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;Note1.The <span>OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750 </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">of <i><em>Chlamydomonas reinhardtii</em> is </i></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.559</span></p>
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2. The Control 1 group is </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif">Chlamydomonas reinhardtii</span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> (notransformation,<span> OD=</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.629</span><span style="font-size:12.0pt;font-family:&quot;:,serif">)</span></p>
+
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2. The Control 1 group is </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif"><em>Chlamydomonas reinhardtii</em></span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> (notransformation,<span> OD=</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.629</span><span style="font-size:12.0pt;font-family:&quot;:,serif">)</span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3. The Control 2 group is TAP medium</span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3. The Control 2 group is TAP medium</span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;</span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;</span></p>
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid"><span style="font-size:12pt;font-family:&quot;color:windowtext">Table 2.5 Toxicity of </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Chlamydomonas reinhardtii</span><span style="font-size:12pt;font-family:&quot;color:windowtext">(</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Cry11Aa+2A+Cyt2</span><span style="font-size:12pt;font-family:&quot;color:windowtext">)</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"> </span><span style="font-size:12pt;font-family:&quot;color:windowtext">to</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">A. albopictus</span></p>
+
<p align="center" style="text-align:center;text-indent:36.0pt;page-break-after:avoid"><span style="font-size:12pt;font-family:&quot;color:windowtext">Table 2.5 Toxicity of </span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"><em>Chlamydomonas reinhardtii</em></span><span style="font-size:12pt;font-family:&quot;color:windowtext">(</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">Cry11Aa+2A+Cyt2</span><span style="font-size:12pt;font-family:&quot;color:windowtext">)</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext"> </span><span style="font-size:12pt;font-family:&quot;color:windowtext">to</span><span style="font-size:12.0pt;font-family:&quot;:,serif;color:windowtext">A. albopictus</span></p>
  
 
<table border="1" cellspacing="0" cellpadding="0" align="left" width="567" style="border-collapse:collapse;border:none;margin-left:6.75pt;margin-right:6.75pt">
 
<table border="1" cellspacing="0" cellpadding="0" align="left" width="567" style="border-collapse:collapse;border:none;margin-left:6.75pt;margin-right:6.75pt">
Line 470: Line 470:
 
<br/>
 
<br/>
 
<br/>
 
<br/>
&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;Note1. The <span>OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750 </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">of <i>Chlamydomonas reinhardtii is </i></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.559</span><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></p>
+
&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;Note1. The <span>OD</span></span><sub><span style="font-size:12.0pt;font-family:&quot;:,serif">750 </span></sub><span style="font-size:12.0pt;font-family:&quot;:,serif">of <i><em>Chlamydomonas reinhardtii</em> is </i></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.559</span><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></p>
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;2. The Control 1 group is </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif">Chlamydomonas reinhardtii</span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> (notransformation,<span> OD=</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.629</span><span style="font-size:12.0pt;font-family:&quot;:,serif">)</span></p>
+
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;2. The Control 1 group is </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif"><em>Chlamydomonas reinhardtii</em></span></i><span style="font-size:12.0pt;font-family:&quot;:,serif"> (notransformation,<span> OD=</span></span><span style="font-size:12.0pt;font-family:&quot;:,serif">0.629</span><span style="font-size:12.0pt;font-family:&quot;:,serif">)</span></p>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3. The Control 2 group is TAP medium<br/>
 
<p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3. The Control 2 group is TAP medium<br/>
 
</span></p>
 
</span></p>

Revision as of 02:09, 20 October 2016

Team FAFU-CHINA

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.

T--FAFU-CHINA--proof2.png

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.

The CC530 and FA479 strains of Chlamydomonas reinhardtii and TAP mediumare bought from the Aquatic Species Research Institute in Wuhan, Hubei. The concentration of Chlamydomonas reinhardtii is recorded ineach table.

We contrasted various toxins groupexpression vectors and transform them into Chlamydomonas reinhardtii as following groups: Cry4Aa+2A+Cyt1,Cry4Aa+2A+Cyt2, Cry10Aa+2A+Cyt1, Cry10Aa+2A+Cyt2, Cry11Aa+2A+Cyt1,Cry11Aa+2A+Cyt2.

The toxicity analysis condition is in anenvironmentally controlled room (T=28±1 °C, RH=85%±5%, L: D=14:10).

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.

Table 1.1 Toxicity of LLP29 to A. albopictus

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











Note.
Thenconcentration of LLP29 is 2.16 x 109 (OD750 of 1.0 = 8 x 108 cells/ml,OD750 =2.7A.)

 

Table  1.2 Toxicity of LLP29 to Culex quinquefasciatus

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












Note.
Thenconcentration of LLP29 is 2.16 x 109 (OD750 of 1.0 = 8 x 108 cells/ml,OD750 =2.7A)

Table 2.1 Toxicity of Chlamydomonas reinhardtii(Cry4Aa+2A+Cyt1) to A. albopictus

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














           Note.1. The OD
750 of Chlamydomonas reinhardtii is 0.559

         2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629)

         3. The Control 2 group is TAP medium

 

Table 2.2 Toxicity of Chlamydomonas reinhardtii(Cry4Aa+2A+Cyt2) to A. albopictus

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














        Note1. The OD
750 of Chlamydomonas reinhardtii is 0.559

         2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629)

         3. The Control 2 group is TAP medium

 

Table 2.3 Toxicity of Chlamydomonas reinhardtii(Cry10Aa+2A+Cyt1) to A. albopictus

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













        Note1.The OD
750 of Chlamydomonas reinhardtii is 0.559

         2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629)

         3. The Control 2 group is TAP medium

 

Table 2.4 Toxicity of Chlamydomonas reinhardtii(Cry10Aa+2A+Cyt2) to A. albopictus

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












        Note1. The OD
750 of Chlamydomonas reinhardtii is 0.559

         2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629)

         3. The Control 2 group is TAP medium

Table 2.5 Toxicity of Chlamydomonas reinhardtii(Cry11Aa+2A+Cyt1) to A. albopictus

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













        Note1.The OD
750 of Chlamydomonas reinhardtii is 0.559

         2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629)

         3. The Control 2 group is TAP medium

 

Table 2.5 Toxicity of Chlamydomonas reinhardtii(Cry11Aa+2A+Cyt2) toA. albopictus

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














        Note1. The OD
750 of Chlamydomonas reinhardtii is 0.559     

         2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629)

         3. The Control 2 group is TAP medium

3.Reference

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