Bt Cotton Bollgard I, II RoundUp in Pakistan Govt. Labs Tests 2012

Bio Chemical Testing Results – 2012 ABRI, CEMB, NIGAB& NIBGE

BIOTECH TEST REPORT OF CANDIDATE COTTON VARIETIRS FOR NCVT-2012 FROM

AGRICULTURAL BIOTECHNOLOGY RESEARCH INSTITUTE (ABRI), Faisalabad

TABLE‐1 Detailed results regarding purity test based on presence of cry1A/ cry2a/ RR determine through strip test using five plants/ entry are detailed below.

Code Variety Purity (%) Code Variety Purity (%) Cry 1A Cry 2A RR Cry 1A Cry 2A RR

V1 HBC‐SD‐134 100 100 100 V‐39 FH‐142 100 0 NT V2 HBC‐SB‐814 80 40 0 V‐40 CEMB‐33 100 0 NT V3 HSP‐3 100 0 NT V‐41 MNH‐456 100 0 NT V4 Kissan Early 100 100 0 V‐42 VH‐303 80 0 NT V5 HSP‐1 100 0 NT V‐43 BZU‐75 0 0 NT V6 Silver‐White 100 100 0 V‐44 VH‐282 60 0 NT V7 HSP‐2 100 0 NT V‐45 CRIS‐508 100 0 NT V8 Silver ‐Gold 100 100 100 V‐46 MM‐58 100 0 NT V9 AA‐904 100 0 0 V‐47 IR‐NIBGE‐5 100 0 NT V10 RS‐1 100 0 NT V‐48 SLH‐4 100 0 NT V11 Auriga‐213 100 0 NT V‐49 IUB‐222 100 0 NT V12 Tarzan‐3 100 0 NT V‐50 IR‐NIBGE‐4 100 0 NT V13 Sayban‐201 100 0 NT V‐51 CIM‐600 100 0 NT V14 A‐011 100 0 NT V‐52 BH‐180 100 0 NT V15 TARZAN‐2 100 0 NT V‐53 CIM‐602 100 0 NT V16 Sitara 10M 100 0 NT V‐54 CEMB‐55 40 0 NT V17 SB‐149 100 0 NT V‐55 CIM‐599 100 0 NT V18 Sitara‐12 100 0 NT V‐56 CEMB‐44 40 0 NT V19 Sun‐1 60 0 NT V‐57 Bt‐BH‐178 100 0 NT V20 Sitara 11M 100 0 NT V‐58 GS‐444 0 0 NT V21 KZ‐389 60 0 NT V‐59 DNH‐105 0 0 NT V22 JS‐1 80 0 NT V‐60 CIM‐612 0 0 NT V23 Leader‐1 100 0 NT V‐61 PB‐38 0 0 NT V24 BGC‐09 100 0 NT V‐62 IUB‐11 0 0 NT V25 Sayban‐202 40 0 NT V‐63 MPS‐II 0 0 NT V26 Silkee 100 0 NT V‐64 NIAB‐112 0 0 NT V27 RCA‐2 100 0 NT V‐65 Cyto‐124 0 0 NT V28 NS‐161 100 0 NT V‐66 JS‐212 40 0 NT V29 AGC‐777 100 0 NT V‐67 VH‐300 0 0 NT V30 AA‐919 100 0 NT V‐68 CRIS‐510 0 0 NT V31 RCA‐1 60 0 NT V‐69 NIA‐80 0 0 NT V32 BS‐52 100 0 NT V‐70 CIM‐591 0 0 NT V33 Trend‐1 100 0 NT V‐71 BH‐176 40 0 NT V34 A‐555 100 0 NT V‐72 CIM‐573 (Standard) 0 0 NT V35 NIAB‐Bt‐1 100 0 NT V‐73 CRIS‐342 (Standard) 0 0 NT V36 RH‐627 60 0 NT V‐74 CIM‐598 (standard) 100 0 NT V37 FH‐118 100 0 NT V‐75 MNH‐886 (Standard) 100 0 NT V38 GH‐142 60 0 NT

NT: Not Tested

TABLE‐2 Results obtained from event specific PCR for MON531 (Cry1A), MON 15985 (Cry2A) and MON 1445 (Roundup ready) are detailed below.

Code Variety Event Detection Code Variety Event DetectionMON531 (Cry 1A)

MON15985 (Cry 2A)

MON1445(RR)

MON531 (Cry 1A)

MON15985(Cry 2A)

MON1445(RR)

V1 HBC‐SD‐134 + + + V‐39 FH‐142 + NT NT V2 HBC‐SB‐814 + ‐ NT V‐40 CEMB‐33 + NT NT V3 HSP‐3 + NT NT V‐41 MNH‐456 + NT NT V4 Kissan Early + + NT V‐42 VH‐303 + NT NT V5 HSP‐1 + NT NT V‐43 BZU‐75 NT NT NT V6 Silver‐White + + NT V‐44 VH‐282 + NT NT V7 HSP‐2 + NT NT V‐45 CRIS‐508 + NT NT V8 Silver ‐Gold + + NT V‐46 MM‐58 + NT NT V9 AA‐904 + NT NT V‐47 IR‐NIBGE‐5 + NT NT V10 RS‐1 + NT NT V‐48 SLH‐4 + NT NT V11 Auriga‐213 + NT NT V‐49 IUB‐222 + NT NT V12 Tarzan‐3 + NT NT V‐50 IR‐NIBGE‐4 + NT NT V13 Sayban‐201 + NT NT V‐51 CIM‐600 + NT NT V14 A‐011 + NT NT V‐52 BH‐180 + NT NT V15 TARZAN‐2 + NT NT V‐53 CIM‐602 + NT NT V16 Sitara 10M + NT NT V‐54 CEMB‐55 + NT NT V17 SB‐149 + NT NT V‐55 CIM‐599 + NT NT V18 Sitara‐12 + NT NT V‐56 CEMB‐44 + NT NT V19 Sun‐1 + NT NT V‐57 Bt‐BH‐178 + NT NT V20 Sitara 11M + NT NT V‐58 GS‐444 NT NT NT V21 KZ‐389 + NT NT V‐59 DNH‐105 NT NT NT V22 JS‐1 + NT NT V‐60 CIM‐612 NT NT NT V23 Leader‐1 + NT NT V‐61 PB‐38 NT NT NT V24 BGC‐09 + NT NT V‐62 IUB‐11 NT NT NT V25 Sayban‐202 + NT NT V‐63 MPS‐II NT NT NT V26 Silkee + NT NT V‐64 NIAB‐112 NT NT NT V27 RCA‐2 + NT NT V‐65 Cyto‐124 NT NT NT V28 NS‐161 + NT NT V‐66 JS‐212 + NT NT V29 AGC‐777 + NT NT V‐67 VH‐300 NT NT NT V30 AA‐919 + NT NT V‐68 CRIS‐510 NT NT NT V31 RCA‐1 + NT NT V‐69 NIA‐80 NT NT NT V32 BS‐52 + NT NT V‐70 CIM‐591 NT NT NT V33 Trend‐1 + NT NT V‐71 BH‐176 + NT NT V34 A‐555 + NT NT V‐72 CIM‐573 (Std) NT NT NT V35 NIAB‐Bt‐1 + NT NT V‐73 CRIS‐342 (Std) NT NT NT V36 RH‐627 + NT NT V‐74 CIM‐598 (std) + NT NT V37 FH‐118 + NT NT V‐75 MNH‐886 (Std) + NT NT V38 GH‐142 + NT NT

NT: Not Tested

TABLE‐3 Results on quantification of Cry1Ac are described below.

Code Variety Cry1Ac 80 days after sowing (µg/g of the fresh weight)

S. No. Variety Cry1Ac 80 days after sowing(µg/g of the fresh weight)

V1 HBC‐SD‐134 6.19 V‐39 FH‐142 2.50 V2 HBC‐SB‐814 4.67 V‐40 CEMB‐33 1.25 V3 HSP‐3 3.85 V‐41 MNH‐456 1.08 V4 Kissan Early 2.62 V‐42 VH‐303 5.00 V5 HSP‐1 3.50 V‐43 BZU‐75 NT V6 Silver‐White 3.74 V‐44 VH‐282 0.98 V7 HSP‐2 3.11 V‐45 CRIS‐508 0.97 V8 Silver ‐Gold 1.72 V‐46 MM‐58 2.58 V9 AA‐904 5.73 V‐47 IR‐NIBGE‐5 1.15 V10 RS‐1 5.03 V‐48 SLH‐4 4.35 V11 Auriga‐213 4.56 V‐49 IUB‐222 5.03 V12 Tarzan‐3 4.87 V‐50 IR‐NIBGE‐4 4.35 V13 Sayban‐201 6.13 V‐51 CIM‐600 3.28 V14 A‐011 3.10 V‐52 BH‐180 0.45 V15 TARZAN‐2 7.45 V‐53 CIM‐602 3.53 V16 Sitara 10M 1.97 V‐54 CEMB‐55 4.99 V17 SB‐149 3.32 V‐55 CIM‐599 2.42 V18 Sitara‐12 0.99 V‐56 CEMB‐44 1.55 V19 Sun‐1 2.62 V‐57 Bt‐BH‐178 2.47 V20 Sitara 11M 0.01 V‐58 GS‐444 NT V21 KZ‐389 5.11 V‐59 DNH‐105 NT V22 JS‐1 4.85 V‐60 CIM‐612 NT V23 Leader‐1 4.34 V‐61 PB‐38 NT V24 BGC‐09 5.20 V‐62 IUB‐11 NT V25 Sayban‐202 0.65 V‐63 MPS‐II NT V26 Silkee 0.67 V‐64 NIAB‐112 NT V27 RCA‐2 0.03 V‐65 Cyto‐124 NT V28 NS‐161 3.18 V‐66 JS‐212 3.30 V29 AGC‐777 2.06 V‐67 VH‐300 NT V30 AA‐919 0.43 V‐68 CRIS‐510 NT V31 RCA‐1 3.50 V‐69 NIA‐80 NT V32 BS‐52 0.90 V‐70 CIM‐591 NT V33 Trend‐1 4.80 V‐71 BH‐176 3.21 V34 A‐555 2.33 V‐72 CIM‐573 (Standard) NT V35 NIAB‐Bt‐1 0.50 V‐73 CRIS‐342 (Standard) NT V36 RH‐627 2.53 V‐74 CIM‐598 (standard) 0.38 V37 FH‐118 1.06 V‐75 MNH‐886 (Standard) 1.64 V38 GH‐142 0.80

DNA AND PROTEIN BASED DETECTION OF BT GENE (CRY1AC AND CRY2A) IN COTTON VARIETIES AT NATIONAL CENTRE OF EXCELLENCE IN MOLECULAR BIOLOGY (CEMB), Lahore

GMO Detection of Bt Cotton

S. No.

Sample No. PCR ELISA Cry1Ac Cry2A Cry1Ac

µg/g Cry2A µg/g

V1 HBC‐SD‐134 +ve NC 0.352 NC V2 HBC‐SB‐814 +ve NC 0.354 NC V3 HSP‐3 +ve NC 0.343 NC V4 Kissan Early +ve NC 0.615 NC V5 HSP‐1 +ve NC 0.646 NC V6 Silver‐White +ve NC 0.652 NC V7 HSP‐2 +ve NC 0.567 NC V8 Silver ‐Gold +ve NC 0.582 NC V9 AA‐904 +ve +ve 0.691 0.365 V10 RS‐1 +ve NC 0.584 NC V11 Auriga‐213 +ve NC 0.615 NC V12 Tarzan‐3 +ve NC 0.567 NC V13 Sayban‐201 +ve NC 0.535 NC V14 A‐011 +ve NC 0.352 NC V15 TARZAN‐2 +ve NC 0.354 NC V16 Sitara 10M +ve NC 0.343 NC V17 SB‐149 +ve NC 0.535 NC V18 Sitara‐12 +ve NC 0.573 NC V19 Sun‐1 +ve NC 0.612 NC V20 Sitara 11M +ve NC 0.553 NC V21 KZ‐389 +ve NC 0.571 NC V22 JS‐1 +ve NC 0.683 NC V23 Leader‐1 +ve NC 0.332 NC V24 BGC‐09 +ve NC 0.595 NC V25 Sayban‐202 +ve NC 0.612 NC V26 Silkee +ve NC 0.593 NC V27 RCA‐2 +ve NC 0.323 NC V28 NS‐161 +ve NC 0.346 NC V29 AGC‐777 +ve NC 0.451 NC V30 AA‐919 +ve +ve 0.643 0.543 V31 RCA‐1 +ve NC 0.621 NC V32 BS‐52 +ve NC 0.362 NC V33 Trend‐1 +ve NC 0.682 NC V34 A‐555 +ve NC 0.572 NC V35 NIAB‐Bt‐1 +ve NC 0.675 NC V36 RH‐627 +ve NC 0.496 NC V37 FH‐118 +ve NC 0.332 NC V38 GH‐142 +ve NC 0.512 NC V39 FH‐142 +ve NC 0.625 NC V40 CEMB‐33 +ve +ve 0.673 0.632 V41 MNH‐456 +ve NC 0.632 NC V42 VH‐303 +ve NC 0.628 NC

V43 BZU‐75 +ve NC 0.621 NC V44 VH‐282 +ve NC 0.559 NC V45 CRIS‐508 +ve NC 0.650 NC V46 MM‐58 +ve NC 0.582 NC V47 IR‐NIBGE‐5 +ve NC 0.671 NC V48 SLH‐4 +ve NC 0.576 NC V49 IUB‐222 +ve NC 0.346 NC V50 IR‐NIBGE‐4 +ve NC 0.451 NC V51 CIM‐600 +ve NC 0.671 NC V52 BH‐180 +ve NC 0.576 NC V53 CIM‐602 +ve NC 0.683 NC V54 CEMB‐55 +ve +ve 0.652 0.468 V55 CIM‐599 +ve NC 0.595 NC V56 CEMB‐44 +ve +ve 0.612 0.673 V57 Bt‐BH‐178 +ve NC 0.562 NC V58 GS‐444 +ve NC 0.516 NC V59 DNH‐105 +ve NC 0.463 NC V60 CIM‐612 +ve NC 0.524 NC V61 PB‐38 +ve NC 0.489 NC V62 IUB‐11 +ve NC 0.561 NC V63 MPS‐II +ve NC 0.513 NC V64 NIAB‐112 +ve NC 0.646 NC V65 Cyto‐124 +ve NC 0.652 NC V66 JS‐212 +ve NC 0.567 NC V67 VH‐300 +ve NC 0.582 NC V68 CRIS‐510 +ve NC 0.691 NC V69 NIA‐80 +ve NC 0.584 NC V70 CIM‐591 +ve NC 0.615 NC V71 BH‐176 +ve NC 0.567 NC V72 CIM‐573 (Standard) +ve NC 0.535 NC V73 CRIS‐342 (Standard) +ve NC 0.573 NC V74 CIM‐598 (standard) +ve NC 0.612 NC V75 MNH‐886 (Standard) +ve NC 0.553 NC

RESULTS OF NATIONAL BT COTTON TRIAL-2012 CONDUCTED AT

NATIONAL INSTITUTE FOR GENOMICS & ADVANCED BIOTECHNOLOGY (NIGAB), NARC-ISLAMABAD

TABLE‐1: Quantitative ELISA for quantification of Cry 1 Ac toxin level.

S. No.

Variety 80 *DAS µg/g fresh leaf weight

S. No. Variety 80 *DASµg/g fresh leaf weight

V1 HBC‐SD‐134 0.633138686 V‐39 FH‐142 0.609820174 V2 HBC‐SB‐814 0.672189781 V‐40 CEMB‐33 0.627311042 V3 HSP‐3 0.516532847 V‐41 MNH‐456 0.367810219 V4 Kissan Early 0.477664234 V‐42 VH‐303 0.60229927 V5 HSP‐1 0.567080292 V‐43 BZU‐75 0.499744526 V6 Silver‐White 0.475225487 V‐44 VH‐282 0.616350365 V7 HSP‐2 0.542992701 V‐45 CRIS‐508 0.725302051 V8 Silver ‐Gold 0.579124088 V‐46 MM‐58 0.45850365 V9 AA‐904 0.800109489 V‐47 IR‐NIBGE‐5 0.385145985 V10 RS‐1 0.702822229 V‐48 SLH‐4 0.813430657 V11 Auriga‐213 0.75959854 V‐49 IUB‐222 0.330036496 V12 Tarzan‐3 0.318375948 V‐50 IR‐NIBGE‐4 0.392554088 V13 Sayban‐201 0.332043796 V‐51 CIM‐600 0.63879562 V14 A‐011 0.530433518 V‐52 BH‐180 0.476934307 V15 TARZAN‐2 0.786240876 V‐53 CIM‐602 0.507773723 V16 Sitara 10M 0.461970803 V‐54 CEMB‐55 0.626569343 V17 SB‐149 0.822737226 V‐55 CIM‐599 0.224744526 V18 Sitara‐12 0.455352627 V‐56 CEMB‐44 0.385328467 V19 Sun‐1 0.729971303 V‐57 Bt‐BH‐178 0.242627737 V20 Sitara 11M 0.389603821 V‐58 GS‐444 0.067580145 V21 KZ‐389 0.621094891 V‐59 DNH‐105 NEGETIVE V22 JS‐1 0.861681699 V‐60 CIM‐612 0.095687036 V23 Leader‐1 0.412883212 V‐61 PB‐38 0.049014599 V24 BGC‐09 0.697007299 V‐62 IUB‐11 0.761085249 V25 Sayban‐202 0.676934307 V‐63 MPS‐II 0.07065755 V26 Silkee 0.462616339 V‐64 NIAB‐112 0.067063082 V27 RCA‐2 0.71700611 V‐65 Cyto‐124 0.720182482 V28 NS‐161 0.533534701 V‐66 JS‐212 0.583868613 V29 AGC‐777 0.583740876 V‐67 VH‐300 0.568722628 V30 AA‐919 0.286765945 V‐68 CRIS‐510 0.609051095 V31 RCA‐1 0.08179938 V‐69 NIA‐80 0.773086994 V32 BS‐52 0.444270073 V‐70 CIM‐591 0.070941055 V33 Trend‐1 0.355766423 V‐71 BH‐176 NEGETIVE V34 A‐555 0.305218978 V‐72 CIM‐573 (Standard) NEGETIVE V35 NIAB‐Bt‐1 0.392554292 V‐73 CRIS‐342 (Standard) 0.093661377 V36 RH‐627 0.419270073 V‐74 CIM‐598 (standard) 0.280578993 V37 FH‐118 0.647554745 V‐75 MNH‐886 (Standard) 0.587169043 V38 GH‐142 0.414708029

* DAS‐Days After Sowing. Tabulated values are the mean of three plants tested for each entry.

Results revealed that Cry1Ac expression in all the tested lines was found to be lower than the expression level recommended by USDA (1.5 µg/g fresh leaf weight) is alarming as it will result in developing resistance for Cry1Ac in the target insect, thus may deprive the farmer of the benefit of Bt technology.

TABLE‐2: Purity test for different transgenes.

Code Variety Cry1Ac/Ab Cry 2A Cry 1F CP4 Code Variety Cry1Ac Cry 2A Cry 1F CP4

V1 HBC‐SD‐134 YES No No No V‐39 FH‐142 YES No No No V2 HBC‐SB‐814 YES No No No V‐40 CEMB‐33 YES No No No V3 HSP‐3 YES No No No V‐41 MNH‐456 YES No No No V4 Kissan Early YES YES No No V‐42 VH‐303 YES No No No V5 HSP‐1 YES No No No V‐43 BZU‐75 YES No No No V6 Silver‐White YES No No No V‐44 VH‐282 YES No No No V7 HSP‐2 YES No No No V‐45 CRIS‐508 YES No No No V8 Silver ‐Gold YES No No No V‐46 MM‐58 YES No No No V9 AA‐904 YES No No No V‐47 IR‐NIBGE‐5 YES No No No V10 RS‐1 YES No No No V‐48 SLH‐4 YES No No No V11 Auriga‐213 YES No No No V‐49 IUB‐222 YES No No No V12 Tarzan‐3 YES No No No V‐50 IR‐NIBGE‐4 YES No No No V13 Sayban‐201 YES No No No V‐51 CIM‐600 YES No No No V14 A‐011 YES No No No V‐52 BH‐180 YES No No No V15 TARZAN‐2 YES No No No V‐53 CIM‐602 YES No No No V16 Sitara 10M YES No No No V‐54 CEMB‐55 YES No No No V17 SB‐149 YES No No No V‐55 CIM‐599 YES No No No V18 Sitara‐12 YES No No No V‐56 CEMB‐44 YES No No No V19 Sun‐1 YES No No No V‐57 Bt‐BH‐178 YES No No No V20 Sitara 11M YES No No No V‐58 GS‐444 YES No No No V21 KZ‐389 YES No No No V‐59 DNH‐105 No No No No V22 JS‐1 YES No No No V‐60 CIM‐612 YES No No No V23 Leader‐1 YES No No No V‐61 PB‐38 YES No No No V24 BGC‐09 YES No No No V‐62 IUB‐11 YES No No No V25 Sayban‐202 YES No No No V‐63 MPS‐II YES No No No V26 Silkee YES No No No V‐64 NIAB‐112 YES No No No V27 RCA‐2 YES No No No V‐65 Cyto‐124 YES No No No V28 NS‐161 YES No No No V‐66 JS‐212 YES No No No V29 AGC‐777 YES No No No V‐67 VH‐300 YES No No No V30 AA‐919 YES No No No V‐68 CRIS‐510 YES No No No V31 RCA‐1 YES No No No V‐69 NIA‐80 YES No No No V32 BS‐52 YES No No No V‐70 CIM‐591 YES No No No V33 Trend‐1 YES No No No V‐71 BH‐176 No No No No V34 A‐555 YES No No No V‐72 CIM‐573 (Standard) No No No No V35 NIAB‐Bt‐1 YES No No No V‐73 CRIS‐342 (Standard) YES No No No V36 RH‐627 YES No No No V‐74 CIM‐598 (standard) YES No No No V37 FH‐118 YES No No No V‐75 MNH‐886 (Standard) YES No No No V38 GH‐142 YES No No No

Note: Three plants were tested for each entry.

TABLE‐3: Very low level of Bt toxin (0.049-0.093 µg/g)

Code Variety toxin µg/g V31 RCA‐1 0.081799 V58 GS‐444 0.06758 V60 CIM‐612 0.095687 V61 PB‐38 0.049015 V63 MPS‐II 0.070658 V64 NIAB‐112 0.067063 V70 CIM‐591 0.070941 V73 CRIS‐342 (Standard) 0.093661

TABLE‐4: Low level of Bttoxin (0.224‐0.392 µg/g)

Code Variety toxin µg/g V12 Tarzan‐3 0.318376 V13 Sayban‐201 0.332044 V20 Sitara 11M 0.389604 V30 AA‐919 0.286766 V33 Trend‐1 0.355766 V34 A‐555 0.305219 V35 NIAB‐Bt‐1 0.392554 V41 MNH‐456 0.36781 V47 IR‐NIBGE‐5 0.385146 V49 IUB‐222 0.330036 V50 IR‐NIBGE‐4 0.392554 V55 CIM‐599 0.224745 V56 CEMB‐44 0.385328 V57 Bt‐BH‐178 0.242628 V74 CIM‐598 (standard) 0.280579

TABLE‐5: Medium level of Bttoxin (0.412‐0.676 µg/g)

Code Variety toxin µg/g V1 HBC‐SD‐134 0.633139 V2 HBC‐SB‐814 0.67219 V3 HSP‐3 0.516533 V4 Kissan Early 0.477664 V5 HSP‐1 0.56708 V6 Silver‐White 0.475225 V7 HSP‐2 0.542993 V8 Silver ‐Gold 0.579124 V14 A‐011 0.530434 V16 Sitara 10M 0.461971 V18 Sitara‐12 0.455353 V21 KZ‐389 0.621095 V23 Leader‐1 0.412883 V24 BGC‐09 0.697007

V25 Sayban‐202 0.676934 V26 Silkee 0.462616 V28 NS‐161 0.533535 V29 AGC‐777 0.583741 V32 BS‐52 0.44427 V36 RH‐627 0.41927 V37 FH‐118 0.647555 V38 GH‐142 0.414708 V39 FH‐142 0.60982 V40 CEMB‐33 0.627311 V42 VH‐303 0.602299 V43 BZU‐75 0.499745 V44 VH‐282 0.61635 V46 MM‐58 0.458504 V51 CIM‐600 0.638796 V52 BH‐180 0.476934 V53 CIM‐602 0.507774 V54 CEMB‐55 0.626569 V66 JS‐212 0.583869 V67 VH‐300 0.568723 V68 CRIS‐510 0.509051 V75 MNH‐886 (Standard) 0.587169

TABLE‐6: High level of Bt toxin (0.702‐0.86 µg/g)

Code Variety toxin µg/g V9 AA‐904 0.800109 V10 RS‐1 0.702822 V11 Auriga‐213 0.759599 V15 TARZAN‐2 0.786241 V17 SB‐149 0.822737 V19 Sun‐1 0.729971 V22 JS‐1 0.861682 V27 RCA‐2 0.717006 V45 CRIS‐508 0.725302 V48 SLH‐4 0.813431 V62 IUB‐11 0.761085 V65 Cyto‐124 0.720182 V69 NIA‐80 0.773087

TESTING OF Bt COTTON LINESOF NCVT 2012AT

NATIONAL INSTITUTE FOR BIOTECHNOLOGY & GENETIC ENGINEERING (NIBGE)

Sr. No

Cotton Lines (Code)

Variety PCR Immuno Strip test for % purity

ELISA

Sad 1 npt II Mon 531 Cry 1Ab/Cry 1Ac Cry 1 Ac (µg/g or ppm)

1. V‐1 HBC‐SD‐134 + + + 80 0.49

2. V‐2 HBC‐SB‐814 + + + 100 0.45

3. V‐3 HSP‐3 + + + 60 0.34

4. V‐4 Kissan Early + + + 80 0.47

5. V‐5 HSP‐1 + + + 60 0.61

6. V‐6 Silver‐White + + + 60 1.19

7. V‐7 HSP‐2 + + + 80 0.44

8. V‐8 Silver ‐Gold + + + 80 0.38

9. V‐9 AA‐904 + + + 80 1.93

10. V‐10 RS‐1 + + + 80 0.57

11. V‐11 Auriga‐213 + + + 60 0.23

12. V‐12 Tarzan‐3 + + + 80 0.40

13. V‐13 Sayban‐201 + + + 60 0.67

14. V‐14 A‐011 + + + 80 0.53

15. V‐15 TARZAN‐2 + + + 80 1.09

16. V‐16 Sitara 10M + + + 80 0.28

17. V‐17 SB‐149 + + + 60 0.70

18. V‐18 Sitara‐12 + + + 60 0.25

19. V‐19 Sun‐1 + + + 60 0.10

20. V‐20 Sitara 11M + + + 80 0.18

21. V‐21 KZ‐389 + + + 80 0.82

22. V‐22 JS‐1 + + + 80 0.26

23. V‐23 Leader‐1 + + + 60 0.50

24. V‐24 BGC‐09 + + + 60 1.35

25. V‐25 Sayban‐202 + + + 60 0.55

26. V‐26 Silkee + + + 40 0.49

27. V‐27 RCA‐2 + + + 60 0.61

28. V‐28 NS‐161 + + + 80 0.98

29. V‐29 AGC‐777 + + + 80 0.33

30. V‐30 AA‐919* + + + 80 0.78

31. V‐31 RCA‐1 + + + 80 0.18

32. V‐32 BS‐52 + + + 60 0.71

33. V‐33 Trend‐1 + + + 60 0.42

34. V‐34 A‐555 + + + 80 0.41

35. V‐35 NIAB‐Bt‐1 + + + 60 0.62

36. V‐36 RH‐627 + + + 80 0.60

37. V‐37 FH‐118 + + + 60 0.60

38. V‐38 GH‐142 + + + 60 0.47

39. V‐39 FH‐142 + + + 80 0.32

40. V‐40 CEMB‐33 + + + 60 0.31

41. V‐41 MNH‐456 + + + 60 0.60

42. V‐42 VH‐303 + + + 60 0.57

Sr. No

Cotton Lines (Code)

Variety PCR Immuno Strip test for % purity

ELISA

Sad 1 npt II Mon 531 Cry 1Ab/Cry 1Ac Cry 1 Ac (µg/g or ppm)

43. V‐43 BZU‐75 + + + 80 0.10

44. V‐44 VH‐282 + + + 60 0.23

45. V‐45 CRIS‐508 + + + 80 0.43

46. V‐46 MM‐58 + + + 80 0.44

47. V‐47 IR‐NIBGE‐5 + + + 60 0.15

48. V‐48 SLH‐4 + + + 80 0.83

49. V‐49 IUB‐222 + + + 60 0.19

50. V‐50 IR‐NIBGE‐4 + + + 80 0.16

51. V‐51 CIM‐600 + + + 80 0.36

52. V‐52 BH‐180 + + + 60 0.31

53. V‐53 CIM‐602 + + + 80 0.40

54. V‐54 CEMB‐55 + + + 80 0.84

55. V‐55 CIM‐599 + + + 80 0.12

56. V‐56 CEMB‐44 + + + 80 0.19

57. V‐57 Bt‐BH‐178 + + + 60 0.43

58. V‐58 GS‐444 + + + 40 0.23

59. V‐59 DNH‐105 + + + 40 0.23

60. V‐60 CIM‐612 + + + 60 0.24

61. V‐61 PB‐38 + + + 80 0.26

62. V‐62 IUB‐11 + + + 40 0.22

63. V‐63 MPS‐II + ‐ ‐ 0 0.00

64. V‐64 NIAB‐112 + ‐ ‐ 0 0.00

65. V‐65 Cyto‐124 + ‐ ‐ 0 0.00

66. V‐66 JS‐212 + + + 60 0.40

67. V‐67 VH‐300 + ‐ ‐ 0 0.00

68. V‐68 CRIS‐510 + ‐ ‐ 0 0.00

69. V‐69 NIA‐80 + + + 60 0.19

70. V‐70 CIM‐591 + ‐ ‐ 0 0.00

71. V‐71 BH‐176 + ‐ ‐ 0 0.00

72. V‐72 CIM‐573 (Non Bt Standard)

+ ‐ ‐ 0 0.00

73. V‐73 CRIS‐342 (Non Bt Standard)

+ + + 80 0.12

74. V‐74 CIM‐598 (Btstandard)

+ + + 80 0.21

75. V‐75 MNH‐886 (BtStandard)

+ + + 60 0.40

Bt Cotton Bollgard I, II RoundUp in Pakistan Govt. Labs Tests 2012

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