Susceptibility of Cry1Ab maize-resistant and -susceptible strains of sugarcane borer (Lepidoptera: Crambidae) to four individual Cry proteins.

Sugarcane borer, Diatraea saccharalis (F.), is a major target of Bt maize in South America and many areas of the US mid-south region. Six laboratory strains of D. saccharalis were established from six single-pair F(2) families possessing major resistance alleles to Cry1Ab maize hybrids. Susceptibility of the six strains was evaluated on diet treated with each of four purified trypsin-activated Bt proteins, Cry1Ab, Cry1Aa, Cry1Ac and Cry1F. Bt susceptibility of the six strains was compared with that of known Cry1Ab-susceptible and -resistant strains of D. saccharalis. At least two of the six strains demonstrated a similar level (>526-fold) of resistance to Cry1Ab as shown in the known Cry1Ab-resistant strain, while resistance levels were relatively lower for other strains (116- to 129-fold). All the six strains were highly cross-resistant to Cry1Aa (71- to 292-fold) and Cry1Ac (30- to 248-fold), but only with a low level to Cry1F (<7-fold). Larval growth of all six strains was also inhibited on Bt-treated diet, but, except for Cry1F, the growth inhibition of the six strains was considerably less than that of the Cry1Ab-susceptible larvae. The results provide clear evidence that the observed resistance to Cry1Ab maize in the six strains is a result of resistance to the Cry1Ab protein in the plants. The low level of cross-resistance between Cry1A and Cry1F suggests that pyramiding these two types of Bt proteins into a plant could be a good strategy for managing D. saccharalis.

Inheritance of resistance to Bacillus thuringiensis Cry1Ab protein in the sugarcane borer (Lepidoptera: Crambidae).

Inheritance traits of a Cry1Ab-resistant strain of the sugarcane borer, Diatraea saccharalis (F.) were analyzed using various genetic crosses. Reciprocal parental crosses between Cry1Ab-susceptible and Cry1Ab-resistant populations, F(1) by F(1) crosses, and backcrosses of F(1) with the Cry1Ab-resistant population were successfully completed. Larval mortality of the parental and cross-populations were assayed on Cry1Ab diet and Bacillus thuringiensis (Bt)-corn leaf tissue. Maternal effects and sex linkage were examined by comparing the larval mortality between the two F(1) populations. Dominance levels of resistance were measured by comparing the larval mortality of the Cry1Ab-resistant, -susceptible, and -heterozygous populations. Number of genes associated with the resistance was evaluated by fitting the observed mortality of F(2) and backcross populations with a Mendelian monogenic inheritance model. Cry1Ab resistance in D. saccharalis was likely inherited as a single or a few tightly linked autosomal genes. The resistance was incompletely recessive on Bt corn leaf tissue, while the effective dominance levels (D(ML)) of resistance increased as Cry1Ab concentrations decreased with Cry1Ab-treated diet. D(ML) estimated based on larval mortality on intact Bt corn plants reported in a previous study ranged from 0.08 to 0.26. This variability in D(ML) levels of Cry1Ab resistance in D. saccharalis suggests that Bt corn hybrids must express a sufficient dose of Bt proteins to make the resistance genes functionally recessive. Thus, Bt resistant heterozygous individuals can be killed as desired in the "high/dose refuge" resistance management strategy for Bt corn.

Susceptibility of Cry1Ab-resistant and -susceptible sugarcane borer (Lepidoptera: Crambidae) to four Bacillus thuringiensis toxins.

Sugarcane borer, Diatraea saccharalis (F.), is a primary corn stalk borer pest targeted by transgenic corn expressing Bacillus thuringiensis (Bt) proteins in many areas of the mid-southern region of the United States. Recently, genes encoding for Cry1A.105 and Cry2Ab2 Bt proteins were transferred into corn plants (event MON 89034) for controlling lepidopteran pests. This new generation of Bt corn with stacked-genes of Cry1A.105 and Cry2Ab2 will become commercially available in 2009. Susceptibility of Cry1Ab-susceptible and -resistant strains of D. saccharalis were evaluated on four selected Bt proteins including Cry1Aa, Cry1Ac, Cry1A.105, and Cry2Ab2. The Cry1Ab-resistant strain is capable of completing its larval development on commercial Cry1Ab-expressing corn plants. Neonates of D. saccharalis were assayed on a meridic diet containing one of the four Cry proteins. Larval mortality, body weight, and number of surviving larvae that did not gain significant weight (<0.1mg per larva) were recorded after 7 days. Cry1Aa was the most toxic protein against both insect strains, followed in decreasing potency by Cry1A.105, Cry1Ac, and Cry2Ab2. Using practical mortality (larvae either died or no significant weight gain after 7 days), the median lethal concentration (LC(50)) of the Cry1Ab-resistant strain was estimated to be >80-, 45-, 4.1-, and -0.5-fold greater than that of the susceptible strain to Cry1Aa, Cry1Ac, Cry1A.105 and Cry2Ab2 proteins, respectively. This information should be useful to support the commercialization of the new Bt corn event MON 89034 for managing D. saccharalis in the mid-southern region of the United States.