Article Figures & Data
Figures
- Fig. 1.
Distribution of cry-type genes obtained from 496 field-collected strains of B. thuringiensis and identified by PCR analysis with general primers and ELISAs with polyclonal antisera.
- Fig. 2.
Transmission electron microscopy of representative Mexican B. thuringiensis strains that did not react with any PCR primers. Strain IB5 produced a small bipyramidal crystal. Strain IB7 had a square crystal. Strain IB183 had a bipyramidal crystal. Strain IB358 had a round crystal enclosed in a multilayered envelope.
- Fig. 3.
SDS-PAGE of B. thuringiensis crystal proteins. Lane M, molecular mass markers; the numbers beside the gel indicate molecular masses of standard marker proteins. Lane 1, strain IB7; lane 2, strain IB5; lane 3, strain IB183; lane 4, strain IB358.
- Fig. 4.
Distribution of putative novel cry genes in the different regions of Mexico. PN, Pacific-North region; N, North region. These two regions correspond to semiarid steppes. C, Central High Plateau region, corresponding to temperate and cold climates. GM, Gulf of Mexico region; PS, South-Pacific region. The last two regions correspond to tropical rainy climates.
Tables
- Table 1.
Characteristics of general and specific primers forcry1, cry5, cry8, cry9,cry11, cry12, cry13, cry14,cry21, and cyt genes
Primer pair Positionsa Gene(s) recognized Product size (bp) Annealing temp (°C) Sequenceb Accession no.c gral-cry1 1472–2029 cry1Aa,cry1Ad 558 52 5′ CTGGATTTACAGGTGGGGATAT (d) M11250 , M73250 1475–2032 cry1Ab, cry1Ae 558 5′ TGAGTCGCTTCGCATATTTGACT (r) M13898 , M65252 1472–2035 cry1Ac 564 M11068 1636–2194 cry1Af 558 U82003 1559–2116 cry1Ba 558 X06711 1577–2131 cry1Bb 555 L32020 2181–2723 cry1Bc 555 Z46442 1463–2056 cry1Ca 594 X07518 1463–2017 cry1Cb 555 M97880 1442–1984 cry1Da,cry1Db 543 X54160 , 1454–2011 cry1Ea,cry1Fa 558 X53985 , M63897 1451–2005 cry1Eb 555 M73253 1936–2490 cry1Fb 555 Z22512 1430–1987 cry1G 558 Z22510 1457–2005 cry1Ha 549 Z22513 2181–2723 cry1Hb 543 U35780 1583–2140 cry1Ia,cry1Ib 558 X62821 , U07642 1439–1993 cry1Ja 555 L32019 1615–2172 cry1Jb 558 U31527 2012–2569 cry1K 558 U28801 gral-cry8 1–373 cry8A 373 49 5′ ATGAGTCCAAATAATCTAAATG (d) U04364 1–376 cry8B 376 5′ TTTGATTAATGAGTTCTTCCACTCG (r) U04365 1–376 cry8C 376 U04366 gral-cry11 1553–1857 cry11A 305 51 5′ TTAGAAGATACGCCAGATCAAGC (d) M31737 1588–1892 cry11B 305 5′ CATTTGTACTTGAAGTTGTAATCCC (r) X86902 gral-nem 2848–3321 cry5Aa 474 50 5′ TTACGTAAATTGGTCAATCAAGCAAA (d) L07025 2560–3033 cry5Ab 474 5′ AAGACCAAATTCAATACCAGGGTT (r) L07026 2353–2826 cry5Ac 474 I34543 2428–2901 cry5B 474 U19725 2401–2877 cry12A 477 L07027 2326–2808 cry14A 483 U13955 2263–2751 cry21Aa 489 I32932 gral-cyt 193–714 cyt1Aa 522 51 5′ AACCCCTCAATCAACAGCAAGG (d) X03182 81–605 cyt1Ab 525 5′ GGTACACAATACATAACGCCACC (r) X98793 spe-cry9A 2103–2673 cry9A 571 51 5′ GTTGATACCCGAGGCACA (d) X58120 5′ CCGCTTCCAATAACATCTTTT (r) spe-cry9B 1770–2171 cry9B 402 51 5′ TCATTGGTATAAGAGTTGGTGATAGAC (d) X75019 5′ CCGCTTCCAATAACATCTTTT (r) spe-cry9C 1853–2158 cry9C 306 51 5′ CTGGTCCGTTCAATCC (d) Z37527 5′ CCGCTTCCAATAACATCTTTT (r) spe-cry8A 1–338 cry8A 338 49 5′ ATGAGTCCAAATAATCTAAATG (d) U04364 5′ TCTCCCCATATATCTACGCTC (r) spe-cry8B 1–510 cry8B 510 49 5′ ATGAGTCCAAATAATCTAAATG (d) U04365 5′ GAACATCTCGTAAGGCTC (r) spe-cry8C 1–963 cry8C 963 49 5′ ATGAGTCCAAATAATCTAAATG (d) U04366 5′ GGTACTCGATTGTCCAGT (r) spe-cry13 225–537 cry13 313 50 5′ CTTTGATTATTTAGGTTTAGTTCAA (d) L07023 5′ TTGTAGTACAGGCTTGTGATTC (r) - Table 2.
Distribution of cry1 gene profiles present in the B. thuringiensis Mexican strain collection ( n = 246)a
No. of strains cry gene profile % 5 cry1Aa 2.0 3 cry1Ab 1.2 3 cry1Ac 1.2 29 cry1Aa,cry1Ab, cry1Ac 11.7 4 cry1Aa,cry1Ab, cry1B 1.6 2 cry1Ac,cry1B 0.8 7 cry1Aa, cry1B,cry1D 2.8 9 cry1Aa, cry1Ab,cry1Ac, cry1B, cry1D 3.6 6 cry1Aa, cry1C, cry1D 2.4 15 cry1Aa, cry1Ab, cry1C,cry1D 6.1 2 cry1Aa, cry1Ab,cry1Ac, cry1C, cry1D 0.8 17 cry1Aa, cry1D 6.9 1 cry1Ac,cry1D 0.4 6 cry1Aa, cry1Ab,cry1Ac, cry1D 2.4 3 cry1Ac,cry1E 1.2 3 cry1Aa, cry1D,cry1F 1.2 2 cry1Ab, cry1D,cry1F 0.8 1 cry1Aa, cry1Ab,cry1B, cry1F 0.4 1 cry1Aa,cry1C, cry1D, cry1F 0.4 2 cry1Ac, cry1F 0.8 19 cry1B 7.7 1 cry1B, cry1D 0.4 1 cry1B,cry1E 0.4 4 cry1C, cry1D 1.6 6 cry1D 2.4 2 cry1D, cry1F 0.8 2 cry1Aa, cry1Ab, cry1Ac,cry3Ba 0.8 1 cry1Ab, cry3Ba 0.4 1 cry1Ac, cry1D, cry1Bb 0.4 1 cry1Aa, cry1Ab, cry1Ac,cry3A 0.4 1 cry1Aa, cry1C,cry1D, cry1F, cry3A 0.4 1 cry1Ac, cry7A 0.4 1 cry1D,cry3Ba 0.4 19 Did not react with any cry1 specific primer 7.7 65 Produced different PCR products 26.4 ↵a PCR analysis was done with the 246cry1-containing strains identified with the general primer pair gral-cry1 (Table 1). The analysis shown here was done with the cry1 specific primers (CJ1 to CJ19) reported by Cerón et al. (7, 8).
- Table 3.
Dose-response activities of B. thuringiensisisolates against S. frugiperda, S. exigua, andT. ni larvaea
Strain cry gene profile identified by PCR analysis LC50, ng/cm2 (95% confidence interval) for: S. frugiperda S. exigua T. ni IB4 cry1Aa,cry1D 34 (23–52) 32 (23–45) 80 (50–112) IB87 cry1Aa, cry1Ab, cry1Ac,cry1D 177 (119–261) 72 (51–101) 6 (4–9) IB126 cry1C,cry1D 22 (15–30) 22 (16–31) 121 (80–190) IB23 cry1Aa, cry1C,cry1D 106 (71–157) 62 (44–86) 62 (40–95) IB33 cry1Aa, cry1Ab,cry1Ac 698 (480–977) 579 (413–811) 20 (14–32) IB104 cry1B 843 (558–1,273) 654 (436–981) NDc IB109 cry1Aa, cry1Ab, cry1C,cry1D 47 (30–71) 24 (17–33) 170 (121–255) IB132 cry1Aa, cry1Ab, cry1Ac,cry1B 997 (651–1,525) 656 (452–918) 23 (16–32) IB217 cry1Aa, cry1C, cry1D,cry1F 154 (100–235) ND ND IB304 cry1Aa, cry1Ac,cry1D 31 (20–47) ND ND HD1 cry1Aa, cry1Ab,cry1Ac 969 (651–1,363) 1,330 (950–1, 862) 25 (17–35) HD73 cry1Ac 1,124 (802–1,573) 1,202 (828–1, 742) ND HD137 cry1Aa, cry1B, cry1C,cry1D 43 (32–71) 25 (17–35) 350 (250–495) ↵a Bioassays were done with the spore-crystal complex. LC50, 50% lethal concentration; ND, not determined.
- Table 4.
Distribution of cry3, cry7, andcry8 gene profiles present in the B. thuringiensis Mexican strain collection ( n = 117)a
No. of strains cry gene profile % 6 cry3A 5.1 16 cry3Ba 13.6 6 cry3Bb 5.1 2 cry3C 1.7 3 cry7A 2.5 3 cry8B 2.5 3 cry8C 2.5 53 Did not react with any cry3, cry8, or cry7 specific primer 45.2 25 Produced different PCR products 21.3 ↵a PCR analysis was done with the 117 strains that were previously identified with the general primers CJ20 and CJ21 (8). The analysis shown here was done with the cry3 specific primers (CJ22 to CJ27) reported by Cerón et al. (8) and novel spe-cry8 primers (Table 1).
