Effect of Transgenic Bt Rice on the Survival of Three Nontarget Stored Product Insect Pests

Home , Chilo suppressalis, Rice weevil

TRANSGENIC PLANTS AND INSECTS Effect of Transgenic Bt Rice on the Survival of Three Nontarget Stored Product Insect Pests

1,2 1 1 3 JORDI RIUDAVETS, ROSA GABARRA, M. JOSE´ PONS, AND JOAQUIMA MESSEGUER

Environ. Entomol. 35(5): 1432Ð1438 (2006)

ABSTRACT The effects of Bacillus thuringiensis (Bt) transgenic Ariete and Senia rice lines devel- Downloaded from https://academic.oup.com/ee/article/35/5/1432/341097 by guest on 23 September 2021 oped for the control of Chilo suppressalis Walker were evaluated on three nontarget stored product insect pests: the lepidopteran Plodia interpunctella (Hu¨ bner), the coleopteran Sitophilus oryzae L., and the psocid Liposcelis bostrichophila (Badonnel). Two different cry genes (cry1B and cry1Aa) under the control of a constitutive plant promoter (ubi) and a wound inducible promoter (mpi) were tested. Plodia interpunctella was unable to survive in rice semolina obtained from the two Ariete and Senia lines, which contained the ubi:cry1B gene. A reduction in S. oryzae adult progeny was observed in cultivar Senia rice containing the ubi:cry1B gene but not in cultivar Ariete rice. Moreover, a sublethal effect on S. oryzae adult progeny weight was observed in both of the rice cultivar treatments that constitutively expressed this gene. The number of L. bostrichophila emerging from Ariete rice containing the ubi:cry1B gene also was reduced. Both a reduction in the number of emerged adults and a delay in the development of P. interpunctella were observed for Ariete rice semolina containing the mpi:cry1B gene. However, no lethal or sublethal effects were observed for P. interpunctella reared on a transformed Senia rice line expressing the ubi:cry1Aa gene, suggesting possible tolerance to the toxin present in the rice grains. Although no increase in mortality was observed with S. oryzae fed ubi:cry1Aa rice, there was a sublethal effect on adult progeny weight.

KEY WORDS Bacillus thuringiensis, transgenic rice, Plodia interpunctella, Sitophilus oryzae, Lipo- scelis bostrychophila

Bacillus thuringiensis (Bt) produces a variety of insec- pollinators, decomposers, and natural enemies (Losey ticide crystal proteins on sporulation that are encoded et al. 2004, OÕCallaghan et al. 2005). by a number of cry genes. These proteins are highly With regard to rice crops, transgenic Bt lines have toxic to lepidopteran, dipteran, and coleopteran in- been developed as alternative tools for preventing sects, interacting with the larval midgut epithelium losses caused by the incidence of the striped stem and causing a disruption in membrane integrity, ulti- borer, Chilo suppressalis Walker (Lepidoptera: Pyrali- mately leading to insect death (Gill et al. 1992). Bt cry dae) (Dirie et al. 2000, Breitler et al. 2004). Transgenic genes have been successfully engineered into a num- rice lines have been obtained with high expression of ber of crops. Several insect-resistant transgenic plant genes encoding insecticidal cry proteins through the cultivars of maize, cotton, and potato have been use of constitutive promoters. An alternative to the widely commercialized (James 2004). In these crops, continuous expression of insecticidal proteins through Bt toxins produced by plant tissues reduce attacks all of a plantÕs tissues is a conditional expression di- from target pest species such as the European corn rected by a wound inducible promoter, which could borer, Ostrinia nubilalis (Hu¨ bner) (Lepidoptera: direct toxin production both spatially and temporally Pyralidae), the noctuids Heliothis spp. and Helicoverpa (Breitler et al. 2004). spp. (Lepidoptera: Noctuidae), and the Colorado po- Several stored product pest species such as Plodia tato beetle, Leptinotarsa decemlineata (Say) (Co- interpunctella (Hu¨ bner) (Lepidoptera: Pyralidae) leoptera: Chrysomelidae). A beneÞt of Bt crops is the (Indianmeal moth), Sitophilus oryzae L. (Coleoptera: reduced reliance on broad-spectrum insecticide treat- Curculionidae) (Rice weevil), and Liposcelis ments (Shelton et al. 2002). Bt toxins also may have a bostrychophila (Badonnel) (Psocoptera: Liposcelidi- negative effect on other nontarget insect herbivores, dae) (Booklouse psocid) are common in rice and cause high economic losses during the postharvest period (Cogburn and Vick 1981, Riudavets et al. 2002). 1 Departament de Proteccio´ Vegetal, IRTA, Centre de Cabrils, Ctra. All three species are polyphagous, and P. interpunctella Cabrils s/n, 08348 Cabrils (Barcelona), Spain. and L. bostrychophila are pests of cereals and semi- 2 Corresponding author, e-mail: [email protected]. processed products, whereas S. oryzae is a pest of 3 Consorci laboratori CSICÐIRTA de Gene`tica Molecular Vegetal, Departament de Gene`tica Vegetal, IRTA, Centre de Cabrils, Ctra. whole grain cereals (Gorham 1991). Brown rice, with Cabrils s/n, 08348 Cabrils (Barcelona), Spain. its external grain layers and embryo, is more favorable

0046-225X/06/1432Ð1438$04.00/0 ᭧ 2006 Entomological Society of America October 2006 RIUDAVETS ET AL.: EFFECT OF TRANSGENIC Bt RICE ON STORED PRODUCT PESTS 1433 for S. oryzae preimaginal development, survival, and Table 1. List of Ariete and Senia Bt-transformed lines tested reproduction than is white polished rice (Lucas and against P. interpunctella and L. bostrichophila on semolina and Riudavets 2000). S. oryzae on white rice In the United States, Bt has been commercially Line Treatment Gene Promoter applied for stored-product insect control since the Ariete AControl Non-Bt 1970s (Golob et al. 2002), while high levels of insec- A3.4 cry1B ubi ticidal activity have been observed in laboratory stud- A3.4E cry1B ubi ies involving the use of different cry protoxins and A9.1 cry1B mpi toxins against P. interpunctella (Herrero et al. 2001). A9.1E cry1B mpi Senia SControl Non-Bt Nevertheless, the resistance of some P. interpunctella S98.9 cry1B ubi strains to sprayable Bt proteins has been shown (Mc- S05 cry1Aa ubi Gaughey 1985, McGaughey and Johnson 1994). Cry

proteins present in transgenic maize kernels from hy- Brown rice semolina treatments (A3.4E and A9.1E) containing the Downloaded from https://academic.oup.com/ee/article/35/5/1432/341097 by guest on 23 September 2021 brids with the CaMV/35s promoter expressing cry1Ab embryo and endosperm only were tested against P. interpunctella. and cry9C genes reduced the survival and reproduction of P. interpunctella, and developmental time also ler et al. 2000) or through Agrobacterium, following was delayed (Giles et al. 2000, Hanley et al. 2004). the procedures detailed in Sallaud et al. (2003) and Similarly, Sedlacek et al. (2001) found that emergence Pons et al. (2000) for Ariete and Senia, respectively. and fecundity of P. interpunctella were lower and de- Synthetic cry1B and cry1Aa genes are Bt ␦-endo- velopmental times longer for individuals reared on Bt toxin genes. The synthetic cry1B gene corresponds to transgenic maize hybrids expressing cry1Ab genes. In the 1Ð1,944 region of the cry1Ba1 gene originally iso- contrast, no differences in P. interpunctella develop- lated from Bt strain HD2 by Brizzard and Whiteley mental time and survival were observed between (1988) and encodes the active toxin plus 29 amino transgenic maize hybrids with the phosphoenolpyru- acids at the N-terminal end. The synthetic cry1Aa vate carboxylase promoter expressing the same gene (M. Royer, unpublished data) corresponds to the cry1Ab genes or transgenic hybrids expressing cry1Ac 1Ð1,854 region of the cry1Aa gene isolated from Bt versus their isolines (Giles et al. 2000). strain FU2Ð7 by Udayasuriyan et al. (1994). In an artiÞcial maize seed bioassay, Bt cry1A and Homozygous Ariete line A3.4 and Senia S05 and cry1B toxins had no effect on S. oryzae mortality (Pit- S98.9 integrated the cry gene under the control of the tendrigh et al. 1997). However, transgenic rice ex- maize ubiquitin (ubi) promoters Þrst intron and Þrst pressing the trypsin inhibitor Itr1 gene encoding the exon (Christensen and Quail 1996), whereas Ariete BTI-CMe toxin reduced the survival rate of S. oryzae line A9.1 integrated the cry gene under the control of (Alfonso-Rubõ´ et al. 2003). The effects of Bt toxins the wound inducible (mpi) promoter (Breitler et al. produced by transgenic rice on stored-product rice 2001). Details of the materials used from each one of pests have not been reported. these lines are provided in Table 1. Transgenic commercial rice lines will be approved Insect Cultures. All colonies were originally estab- in the near future in several countries. One of the Þrst lished from individuals collected in 1997 from storage candidates likely will be rice lines resistant to attack by areas located in the northeast of Spain. P. interpunc- stem borers. The objective of this study was to eval- tella was reared on a diet based on wheat bran and uate the extent to which harvested grain from trans- glycerin. S. oryzae was reared on brown rice (cultivar genic rice expressing the cry1B and cry1Aa genes, Tebre). L. bostrichophila was reared on a diet of wheat especially designed for controlling C. suppressalis, ßour and yeast. Rearing and experiments were con- could affect nontarget pests present in stored rice such ducted in a climatic chamber at 25 Ϯ 1ЊC, 75 Ϯ 15% as the lepidopteran P. interpunctella, the coleopteran RH, and at a photoperiod of 16:8 (L:D). S. oryzae, and the psocid L. bostrichophila. We eval- Testing Method. The experimental arenas were uated the effect of transgenic rice lines harboring the ventilated glass jars (110 ml) containing 30 g of rice cry1B gene with constitutive and wound inducible kernels or 20 g of rice semolina. Brown rice (with promoters and the cry1A gene with a constitutive embryo and external layers) was polished to obtain promoter on adult weight and survival rates. Addi- white rice (only endosperm) using a laboratory pol- tionally, the effects of the cry1B gene under the con- isher (Univeral Brevetto 65378, vertical type; A. trol of both promoters in brown and white rice on GuidÕetti, Italy), and white rice was milled to obtain P. interpunctella life parameters were studied. semolina in a standard kitchen blender. Insects were added to arenas in doses of 100 P. interpunctella eggs or 30 L. bostrichophila adults in rice semolina and 30 Materials and Methods S. oryzae adults in rice kernels. Survival rates for each Plant Material. Control and transgenic Bt seeds of species were assessed for the Þrst generation progeny. the Mediterranean rice lines Ariete and Senia were In weeks 6, 8, and 10 after P. interpunctella infestation, used in this study. Ariete is grown in La Camargue in emerging adult progeny were counted, removed, and the south of France, and Senia is grown in the Ebro frozen. One week after S. oryzae infestation, adults in Delta in the northeast of Spain. The cry genes were the arenas were removed. Seven weeks after S. oryzae introduced into seed embryoÐderived embryogenic infestation, adult progeny were counted, removed, calli either after microprojectile bombardment (Breit- and frozen. Nine weeks after L. bostrichophila infes- 1434 ENVIRONMENTAL ENTOMOLOGY Vol. 35, no. 5

Table 2. Total P. interpunctella and S. oryzae progeny emerged (means ؎ SEM) and total no. (means ؎ SEM) of L. bostrichophila adults present 9 wk after infestation from each Bt and non-Bt rice semolina treatment

Line Treatment Construct P. interpunctella S. oryzae L. bostrichophila Ariete AControl Non-Bt 22 Ϯ 0.7a 400 Ϯ 21.1a 249 Ϯ 17.0a A3.4 ubi:cry1B 0c 430 Ϯ 23.3a 128 Ϯ 9.8b A9.1 mpi:cry1B 6 Ϯ 0.8b 360 Ϯ 29.9a 274 Ϯ 11.6a Senia SControl Non-Bt 10 Ϯ 0.6b 403 Ϯ 21.0a 248 Ϯ 11.1a S98.9 ubi:cry1B 0c 321 Ϯ 22.6b 218 Ϯ 15.5a S05 ubi:cry1Aa 17 Ϯ 1.1a 374 Ϯ 23.8ab 257 Ϯ 7.4a

Means followed by the same letter indicates no signiÞcant difference between treatments within the same line tested (Tukey, n ϭ 10, ␣ ϭ 0.05). Downloaded from https://academic.oup.com/ee/article/35/5/1432/341097 by guest on 23 September 2021 tation, the total number of adults present in each trol treatment (F ϭ 35.11; df ϭ 2,29; P Ͻ 0.001). No repetition was counted. The total adult progeny of differences were observed in the number of adults at P. interpunctella and S. oryzae from each repetition was the ninth week for the Senia treatments (F ϭ 3.00; df ϭ weighed using a digital balance, and individual 2,29; P ϭ 0.07). A signiÞcant weight reduction was weights for fresh specimens were calculated. Ten rep- observed for S. oryzae adults reared on both lines with etitions of each treatment were conducted, except in this Bt gene and promoter combination (F ϭ 27.07; the case of lines A3.4E and A9.1E, with three repeti- df ϭ 2,29; P Ͻ 0.001 for Ariete, F ϭ 19.66; df ϭ 2,29; P Ͻ tions per treatment. 0.001 for Senia; Table 3). Statistical Analyses. The mean number of adults and Effect of the Cry1B Gene Under Control of the Mpi individual weights were compared between treat- Promoter. The emergence of P. interpunctella in Ari- ments of the same rice line (Ariete and Senia) using ete rice semolina containing the mpi:cry1B gene (line a one-way analysis of variance (ANOVA) procedure A9.1) was signiÞcantly higher than from A3.4 with the and a subsequent Tukey test (PROC GLM; SAS In- constitutive promoter ubi, yet Ͼ70% lower than in the stitute 2000). A Bartlett test was applied before Ariete control treatment (Table 2). There was also a ANOVA to test variance homogeneity, and data were delay in the preimaginal development of P. interpunc- log-transformed when necessary. tella in line A9.1 (Fig. 1). This construct had no ap- parent effect on total adult progeny for S. oryzae and Results L. bostrichophila. P. interpunctella adults fed on rice line A9.1 had signiÞcantly greater weights than adults Effect of the Cry1B Gene Under Control of the Ubi that emerged from the rice control treatment (F ϭ Promoter. The moth P. interpunctella did not complete 4.96; df ϭ 1,14; P Ͻ 0.05; Table 3). The weights of its preimaginal development in rice semolina obtained S. oryzae adults that emerged in line A9.1 were sig- from either Ariete or Senia lines containing the syn- niÞcantly greater than for those in A3.4 and the Ariete thetic ubi: cry1B gene (F ϭ 341.04; df ϭ 2,29; P Ͻ 0.001 control. for Ariete and F ϭ 150.43; df ϭ 2,29; P Ͻ 0.001 for Senia; Effect of Grain Components. SigniÞcantly fewer Table 2). In the case of the weevil S. oryzae, no sig- P. interpunctella adults emerged from transgenic Ari- niÞcant differences in the number of adult progeny ete white rice semolina (A9.1) that contained only were observed between the control and line A3.4 endosperm than from Ariete transgenic brown rice treatments in the Ariete cultivar (F ϭ 1.96; df ϭ 2,29; semolina that contained endosperm, external layers, P ϭ 0.16). Nevertheless, a signiÞcantly lower number and embryo (A9.1E; F ϭ 240.62; df ϭ 4,35; P Ͻ 0.001; of weevil progeny emerged from Senia transgenic rice Fig. 2). However, no signiÞcant differences in adult with this construct (S98.9; F ϭ 3.45; df ϭ 2,29; P Ͻ progeny were observed between line A9.1E and the 0.05). The number of L. bostrichophila adults that had Ariete control with endosperm only. Ariete brown emerged by the ninth week after inoculation was sig- rice semolina from line A3.4E and white rice semolina niÞcantly lower for line A3.4 than for the Ariete con- containing only endosperm (A3.4) both produced 100% mortality. The weights of emerged adults fed on Table 3. Individual weight (mg; mean ؎ SEM) of P. interpunc- semolina A9.1 were signiÞcantly higher than those of tella and S. oryzae adult progeny that emerged from each Bt and adults fed on semolina A9.1E (F ϭ 4.34; df ϭ 2,17; P Ͻ non-Bt rice semolina treatment 0.05; Fig. 2). However, no signiÞcant differences were observed between the transgenic A9.1 lines and Ariete Line Treatment Construct P. interpunctella S. oryzae control. Ariete AControl Non-Bt 3.45 Ϯ 0.130b 2.55 Ϯ 0.030b Effect of the Cry1Aa Gene Under Control of the A3.4 ubi:cry1B Ñ 2.39 Ϯ 0.018c A9.1 mpi:cry1B 4.40 Ϯ 0.552a 2.66 Ϯ 0.028a Ubi Promoter. The total number of emerged adults of Senia SControl Non-Bt 3.61 Ϯ 0.323b 2.81 Ϯ 0.035a P. interpunctella and adult weight were signiÞcantly S98.9 ubi:cry1B Ñ 2.56 Ϯ 0.024b higher in the cry1Aa-transformed Senia rice treat- S05 ubi:cry1Aa 4.52 Ϯ 0.246a 2.61 Ϯ 0.031b ment (line S05) than in treatment S98.9 and the Senia control treatment (emergence: F ϭ 150.43; df ϭ 2,29; Means followed by the same letter indicates no signiÞcant differ- Ͻ ϭ ϭ Ͻ ence between treatments within the same line tested (Tukey, n ϭ 10, P 0.001; adult weight: F 5.40; df 1,14; P 0.05; ␣ ϭ 0.05). Table 2). No signiÞcant differences in the number of October 2006 RIUDAVETS ET AL.: EFFECT OF TRANSGENIC Bt RICE ON STORED PRODUCT PESTS 1435 Downloaded from https://academic.oup.com/ee/article/35/5/1432/341097 by guest on 23 September 2021

Fig. 1. Emergence (mean Ϯ SEM) of P. interpunctella adults collected during weeks 6, 8, and 10 after infestation from Ariete (Acontrol, A9.1; left) and Senia (SControl, S05; right) rice semolina treatments.

S. oryzae and L. bostrichophila adult progeny were Discussion observed between this transformed Senia rice and the The accumulation of cry1B toxin in the seed tissues Senia control. Nevertheless, weevils emerging from of lines A3.4 and S98.9 harboring the ubi promoter Senia line S05 were signiÞcantly lighter than those ϭ ϭ could explain the high mortality observed in P. inter- emerging from the Senia control (F 19.66; df 2,29; punctella (Table 2). Breitler et al. (2001) and Breitler Ͻ P 0.001; Table 3). There was no delay in the pre- et al. (2004) showed that these transformed Ariete and imaginal development of P. interpunctella in the pres- Senia lines provide stable, full protection against the ence of cry1Aa (Fig. 1). lepidoptera C. suppressalis in greenhouse and Þeld trials. Furthermore, Western blot analysis has shown a stable accumulation of Bt toxins in the vegetative and seed tissues of lines A3.4 and S98.9, constitutively expressing the cry1B gene (Breitler et al. 2004). Adult progeny from S. oryzae were affected by Senia line S98.9, and weevils emerging from A3.4 and S98.9 suffered a signiÞcant weight loss that may reduce their fecundity (Table 3). Hespenheide (1973) and Scriber and Slansky (1981) described a positive relationship between weight loss and reduction of adult fecundity for numerous insect species. Moreover, adult progeny from L. bostrichophila were reduced by the Ariete line A3.4. The stable accumulation of cry1B toxins in the seeds of the Ariete line under the control of the ubi promoter (Breitler et al. 2004) seems to have been responsible for these sublethal effects on S. oryzae and L. bostrichophila. However, Pittendrigh et al. (1997) found no effect on the number of emerged S. oryzae adults when cry1A/cry1B ␦-endotoxins were incorpo- rated into artiÞcial seeds at levels of up to 0.01% wt:wt. The effect of Bt toxins on L. bostrichophila has not been previously described. It is not possible to discern whether the decreases in the number of adult progeny and in adult weight observed in S. oryzae and L. bostrichophila were caused by the Bt toxin or whether Fig. 2. Number and individual weight (mg; mean Ϯ they were the result of differences in the nutritional SEM) of P. interpunctella adult progeny that emerged from quality of the plant material ingested by the tested Ariete brown rice semolina treatments that contained em- insects. Genetic manipulation or tissue culturing of ϭ bryos, external layers, and endosperm (A3.4E, A9.1E; n 3) plants may cause certain changes in plant character- compared with white rice semolina treatments that contained only the endosperm (A3.4, A9.1; n ϭ 10) and the istics, such as in their nitrogen or carbohydrate con- control treatment (n ϭ 10). Means with the same letters tents (Wei-xiang et al. 2003). indicate no signiÞcant difference between treatments The emergence of P. interpunctella was affected by (Tukey, n ϭ 10, ␣ ϭ 0.05). the presence of the mpi:cry1B gene. Both a reduction 1436 ENVIRONMENTAL ENTOMOLOGY Vol. 35, no. 5 in the number of emerged adults and a delay in de- 2001, Marfa` et al. 2002b), and Þeld conditions (Breit- velopment were observed in insects fed with the mpi: ler et al. 2004). In our experiment, rice grains con- cry1B Ariete line (Table 2; Fig. 1). Breitler et al. (2004) taining cry1Aa proteins were not toxic to P. interpunc- found that, in rice from this line, cry1B was not de- tella, which indicates a lack of susceptibility to the tected in unwounded tissues, but toxin accumulation cry1Aa toxins. In contrast, the transformation signiÞ- was detected around attack sites. Therefore, induction cantly improved P. interpunctella survival and adult of the transgene was in response to insect attack. weight. From our results, it is not possible to discern Moreover, cry1B was not detected in embryo or en- whether this effect was caused by changes in the plant dosperm. Nevertheless, Tu et al. (2000) showed that nutritional quality of this transgenic line or wheher Bt very low levels of toxin (0.002%) can be effective this Bt gene and promoter improved Lepidoptera per- in controlling some pests. It is plausible that line A9.1 formance (Wei-xiang et al. 2003). could have contained an undetectable level of cry1B Lepidopteran species with documented resistance toxin, yet enough to affect the development of the Downloaded from https://academic.oup.com/ee/article/35/5/1432/341097 by guest on 23 September 2021 P. interpunctella larvae. In rice transformed with a to Bt include P. interpunctella (McGaughey 1985), and proteinase inhibitor under the control of the same cry1Aa is present in commercial insecticide formula- wound-inducible mpi, Vila et al. (2005) barely de- tions of Bt (Oppert et al. 2000). Although the moths tected the transgenic protein in the endosperm, but used in our experiment were reared for several years not in the embryo at the limit of detection by Western without insecticide treatments, it is possible that the blot analysis. observed lack of efÞcacy could have been caused by There were no negative effects of Bt toxin on resistance in the colony. McGaughey (1985) found P. interpunctella survival in the A9.1E treatment with that, after seven generations, a resistant strain did not brown rice semolina containing the embryo, external suffer any loss of resistance to Bt. This author also layers, and endosperm, which contrasted with the suggested that resistance was inherited as a recessive results obtained for the A9.1 treatment with white rice trait and may therefore be caused by a single major semolina that only contained the endosperm (Fig. 2). factor that occurs at high frequency. It is possible that the presence of the embryo and From the results obtained in this study, it can be external layers of the endosperm was beneÞcial for the concluded that transgenic Bt rice lines developed for survival of P. interpunctella. Furthermore, the pres- the control of a particular Þeld lepidopteran pest ence of the embryo and the external layers of the (C. suppressalis) produce lethal and sublethal ef- endosperm improved rice quality and was more con- fects not only on other lepidopteran species, such as ducive to the development of another rice pest, the P. interpunctella, but also on the coleopteran S. oryzae weevil S. oryzae, when brown and white rice were and the psocopteran L. bostrichophila. These results compared (Lucas and Riudavets 2000). Adult insect Bt weight also is related to the amount of food available. suggest potential beneÞts of rice events that express In our study, lower larval density in rice semolina Cry proteins in the grain caused by a reduction in the without embryo from the A9.1 event might have im- damage produced by P. interpunctella and L. bos- proved the adult weight of survivors (Fig. 2). In this trichophila and also in adult weight loss among study, rice semolina obtained from cultivar Ariete was S. oryzae, suggesting reduced Þtness of survivors. Sim- more suitable for preimaginal development of the ilar results for P. interpunctella with transgenic Bt corn moth P. interpunctella than rice semolina obtained have been observed (Giles et al. 2000, Sedlacek et al. from cultivar Senia; this was evident from survivor- 2001). However, P. interpunctella has developed re- ship in control treatments. Differences in nutrient sistance to spray Bt formulations (McGaughey and composition or in efÞciency of use of food energy for Johnson 1994). Therefore, sublethal doses of Cry tox- P. interpunctella could perhaps explain these differ- ins in cereals may promote rapid selection for resis- ences (Johnson et al. 1992). Similarly, cultivar Ariete tance in P. interpunctella. Further studies are needed was more sensitive to C. suppressalis than cultivar to determine if transgenic Bt grains will result in the Senia under Þeld conditions (Breitler et al. 2004). development of resistant populations of P. interpunc- When P. interpunctella larvae were reared on the tella and will serve to further characterize the beneÞts event S05, constitutively expressing the ubi:cry1Aa and durability of transgenic cereals in the control of gene, no detrimental effect on survival, larval devel- stored-product pests. opment time, or adult weight was detected. Moreover, P. interpunctella larvae fed this transgenic event ex- hibited greater larval survival and adult weight than moths fed on Senia control rice (Tables 2 and 3; Fig. 1). The ubi promoter is known to be highly active in rice Acknowledgments seeds, leading to the accumulation of Cry toxins in We thank E. Guiderdoni and M. Royer from CIRAD endosperm (Breitler et al. 2001, Wu et al. 2002, Breitler (Montpellier, France) for kindly providing Ariete transgenic et al. 2004). Western blot analysis has shown a stable lines and Bt constructs. We also thank R. Albajes from UdL- accumulation of cry1Aa toxin in leaf, stem, and seed IRTA (Lleida, Barcelona) and C. Castan˜ e´ from IRTA (Ca- tissues of line S05, and full protection against C. sup- brils, Barcelona) for critically reading the manuscript and pressalis also has been described under laboratory J. Artes for technical assistance. This study was supported by (Marfa` et al. 2002a), greenhouse (Breitler et al. 2000, project INIA RTA2005-00068-00-00. October 2006 RIUDAVETS ET AL.: EFFECT OF TRANSGENIC Bt RICE ON STORED PRODUCT PESTS 1437

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