Search

Cultivation of genetically modified maize (Zea mays L.) expressing the Cry1Ab protein from Bacillus thuringiensis (Bt) has increased worldwide since Bt maize was first commercialised. However, the cultivation and return of Bt maize has been shown to affect nontarget symbiotic soil-borne microbes such as arbuscular mycorrhizal fungi (AMF). In this study, we compared the diversity and composition of the AMF communities between two Bt maize plants (5422Bt1 [event Bt 11] and 5422CBCL [event MON 810]) and their conventional (non-Bt) isoline (5422) after cultivation for five seasons and return of straws by using molecular approaches, including terminal restriction fragment length polymorphism and DNA sequencing. Our data revealed that the diversities of AMF communities did not consistently differ significantly in soils and roots of subsequently planted conventional maize (SCM 5422) grown with Bt maize straw at three sampling stages (seedling, large bell, and maturity stages). DNA sequencing showed that typical AMF communities included Glomus, Paraglomus, Diversispora, Acaulospora, and Rhizophagus, of which Glomus was the most abundant. Funneliformis was detected only in bulk and rhizospheric soils and in roots of maize at the seedling stage. Rhizophagus was detected only in rhizospheric soils and only at the maturity stage. No significant effects related to the presence of Bt maize straw (5422Bt1 or 5422CBCL) were found by general linear analysis. However, plant growth stage had a greater influence on AMF diversity than Bt traits. In conclusion, cultivation of non-Bt maize on soils previously cultivated with Bt maize for five seasons had minor effects on AMF communities.

Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) in combination with sequencing of amplified partial 18S ribosomal DNA was used to assess the effects of transgenic Bt corn 5422Bt1 (event Bt 11) and 5422CBCL (event MON810) on the community structure of a non-target microorganism, namely the arbuscular mycorrhizal fungi (AMF) Glomus in corn roots and rhizosphere soils, relative to their non-Bt isolines 5422 (conventional parent) and 5422wx (conventional hybrid). AMF colonization in roots of different corn genotypes was also assessed using microscopic visualization. No adverse effect was detected on the indigenous AMF colonization of the roots of Bt hybrids 5422Bt1 and 5422CBCL. Two-way indicator species analysis (TWINSPAN®) and detrended correspondence analysis (DCA) of the DGGE data from corn roots presented differences between Bt and non-Bt corn isolines (5422Bt1 vs. 5422wx, and 5422CBCL vs. 5422wx). However, differences were also recognized between the two non-Bt corn cultivars (5422 vs. 5422wx), and between the two Bt corn lines (5422Bt1 vs. 5422CBCL) in roots. Our results suggest that corn genotypes may have a greater influence on the AMF community structure of plant roots and rhizosphere soils than other factors, such as the age of the growing plants.

Earthworms Eisenia fetida, bred in substances with stover of two genetically-engineered Bacillus thuringiensis (Bt) corns (5422Bt1 (Event Bt11) and 5422CBCL (MON810)) expressing Cry1Ab and their near-isogenic non-Bt corn (5422), were used to investigate the non-target effects of Bt corn on soil-dwelling organisms. Cry1Ab concentrations in substances, casts and guts of E. fetida were also investigated by Enzyme-linked immunosorbent assay (ELISA). More than 90% individuals of E. fetida survived over a period of 30 d, irrespective of whether they received Bt corn or non-Bt corn. Compared to 5422 treatments, significantly higher relative growth rate and more number of new offspring and cocoons of E. fetida were found in 5422Bt1 and 5422CBCL treatments. These results were unlikely to be directly caused by Cry1Ab released from Bt corns but rather by differences in other factors of plants such as plant components (soluble sugar, total organic carbon, total protein and available phosphorus of Bt corns were more than 5422). ELISA results indicated immunoreactive Cry1Ab was detectable in substances, and the casts, guts of E. fetida from Bt corns treatments, of which the highest levels were detected in substances under the corresponding experimental conditions. With the increase of treated time, a strong decline was observed in Cry1Ab from substances and casts of E. fetida, whereas Cry1Ab in guts of E. fetida from 5422Bt1 treatments gradually increased and that from 5422CBCL treatments increased between 14 and 30 d. Therefore, the presence of Cry1Ab in E. fetida had no deleterious effects on their growth and reproduction.

We investigated the effects of soil temperature (15 °C, 25 °C, 35 °C), water content (20%, 33%, 50%) and pH (4.5, 7.0, 9.0) on the degradation of Cry1Ab protein released from the straw of Bt corn varieties 34B24 and 1246 × 1482 both expressing Cry1Ab protein. Our results showed that Cry1Ab protein released from both 34B24 and 1246 × 1482 straw was degraded in a similar way in all treatments, which demonstrated a rapid decline in the early stage but a slow decline in the middle and late stages. In the late stage (180 days after the experiment commenced) 0.03%–1.51% and 0.02%–0.91% of initial Cry1Ab protein released from 34B24 and 1246 × 1482 straw was detected in soil. In addition, degradation dynamics of Cry1Ab protein under different environmental conditions was well described by the shift-log model. DT 50 of Cry1Ab protein released from 34B24 and 1246 × 1482 straw was 0.97–9.97 d and 0.75–10.89 d, respectively, and DT 90 was 4.66–162.45 d and 6.44–57.46 d, respectively. The results suggested that soil temperature had significant effects on the degradation of Cry1Ab protein, with a higher degradation rate at higher temperature, but soil water content and pH had no obvious effects on the degradation of Cry1Ab protein.

Polymerase chain reaction–denaturing gradient gel electrophoresis (PCR–DGGE) and sequences of the 16S and 18S rRNA genes were used to access the effects of actively growing Bacillus thuringiensis (Bt) corn Pioneer 34B24 and Nongda 1246*1482, and plant straw (leaves plus stalks) of Bt hybrid Pioneer 34B24 and Nongda 61 on soil bacterial and fungal communities. Two-way indicator species analysis (TWINSPAN®), and detrended correspondence analysis (DCA) of the DGGE data indicated that neither the actively growing Bt corn nor its straw had any constant apparent effect on the soil bacteria and fungi community structure. The age of the growing plants, or the timing of plant straw decomposition may have more effect on the microbial community than other factors, i.e., the presence of Cry protein, plant hybrid and variety.

Bt corn is one of the top three large-scale commercialized transgenic crops around the world. It is increasingly clear that the complementary durable approaches for pest control, which combine the endogenous defense of the crop with the introduced foreign genes, are promising alternative strategies for pest resistance management and the next generation of insect-resistant transgenic crops. In the present study, we tested the inducible effects of exogenous jasmonic acid (JA) on direct-defense chemical content, Bt protein concentration, and related gene expression in the leaves of Bt corn cultivar 34B24 and non-Bt cultivar 34B23 by chemical analysis, ELISA, and RT-PCR. The results show that the expression of LOX, PR-2α, MPI, and PR-1 genes in the treated leaf (the first leaf) was promoted by exogenous JA both in 34B24 and 34B23. As compared with the control, the concentration of DIMBOA in the treated leaf was significantly increased by 63 and 18% for 34B24 and 34B23, respectively. The total phenolic acid was also increased by 24 and 12% for both 34B24 and 34B23. The Bt protein content of 34B24 in the treated leaf was increased by 13% but decreased significantly by 27% in the second leaf. The induced response of 34B24 was in a systemic way and was much stronger than that of 34B23. Those findings indicated that there is a synergistic interaction between Bt gene and internally induced chemical defense system triggered by externally applied JA in Bt corn.