Wagner Lucena

Brazilian Agricultural Research Corporation (EMBRAPA) | Embrapa · Embrapa Cotton

Figure S1 Template determination for drafting the Cry10Aa in silico three‐dimensional model. The Cry10Aa sequence (Thorne et al., 1986; Journal of Bacteriology 166, 801‐811; Aguiar et al., 2012, Bt Research 3, 20‐28) and the crystal structure of Cry1Ac (PDB ID: 4W8J; solved at 2.78 Å) were used to generate the Cry10Aa three‐dimensional structure...

Figure S4 Phenotypic comparison between T0 Cry10Aa cotton plants and nontransgenic plants. The images compare the macroscopic phenotypes of Cry10Aa cotton P#008 with wild type (WT—BRS 372), both at 4 months of age. In (a) is shown the view of the whole aerial part of transgenic (T) and non‐trangenic (WT) plants. In (b) and (c), the floral structure...

Table S2 Summary of qPCR experiments—cry10Aa transcript quantification.

Table S5 Commercially approved Bt cotton plants worldwide.

Figure S5 Leaf area damaged by CBW in T0 GM cotton plants. The graph shows the percentage of leaf area damaged by CBW in leaves of eleven T0 GM cotton plants expressing the toxin Cry10Aa and WT (control), all collected every three days during the bioassays. The rate (damaged leaf area/total leaf area) was obtained using ImageJ software (Schneider e...

Table S3 Mortality rate (%) of cotton boll weevil adults fed tissues from T0 GM and non‐GM cotton plants.

Table S6 Summary of used primers.

Figure S2 Prediction of Cry10Aa toxin secondary structure. Representation of Cry10Aa (accession number AAA22614.1) secondary structure obtained from in silico modelling using Cry1Ac crystal (PDB ID: 4W8J) as a template. The 66 first residues present in the Cry10Aa protein do not show structural similarity with Cry1Ac crystal. Predicted helixes are...

Figure S3 Analyses of candidate promoters supporting the design of the Cry10Aa expression cassette for cotton transformation. The cotton ubiquitin‐conjugating enzyme (uceA) constitutive promoter, rather than CaMV35S, was used to design the transformation vector. This choice was based on comparative results of the strength and tissue specificity of...

Figure S6 Flower bud drop rate in T0 GM cotton plants infested with CBW. The graph shows the percentage of dropped flower buds in eleven T0 GM cotton plants expressing the toxin Cry10Aa and WT (control) evaluated during the bioassays. (*) Asterisks represent the level of statistical significance when compared to the same rate in WT plants (Student'...

Table S4 Mortality rate (%) of cotton boll weevil adults fed tissues from T1 GM and non‐GM cotton plants.

Figure S7 Leaf area damaged by CBW in T1 GM cotton plants. The graph shows the percentage of leaf area damaged by CBW in leaves of eleven T1 GM cotton plants expressing the toxin Cry10Aa and WT (control), all collected every three days during the bioassays. The rate (damaged leaf area/total leaf area) was obtained using ImageJ software (Schneider e...

Table S1 LC50 values of recombinant Cry10Aa protein against Anthonomus grandis

Genetically modified (GM) cotton events that effectively control cotton boll weevil (CBW), which is the most destructive cotton insect pest in South America, are reported here for the first time. This work presents the successful development of new GM cotton with high resistance to CBW conferred by Cry10Aa toxin, a protein encoded by entomopathogen...

Gossypium hirsutum (commercial cooton) is one of the most economically important fibers sources and a commodity crop highly affected by insect pests and pathogens. Several transgenic approaches have been developed to improve cotton resistance to insect pests, through the transgenic expression of different factors, including Cry toxins, proteinase i...

α-Amylases are common enzymes responsible for hydrolyzing starch. Insect-pests, whose larvae develop in seeds, rely obligatorily on α-amylase activity to digest starch, as their major food source. Considering the relevance of insect α-amylases and the natural α-amylase inhibitors present in seeds to protect from insect damage, we report here the mo...

Bacillus thuringiensis (Bt) is a gram-positive spore-forming soil bacterium that is distributed worldwide. Originally recognized as a pathogen of the silkworm, several strains were found on epizootic events in insect pests. In the 1960s, Bt began to be successfully used to control insect pests in agriculture, particularly because of its specificity...

Numerous species of insect pests attack cotton plants, out of which the cotton boll weevil (Anthonomus grandis) is the main insect in Brazil and must be controlled to avert large economic losses. Like other insect pests, A. grandis secretes a high level of α-amylases in the midgut lumen, which are required for digestion of carbohydrates. Thus, α-am...

The cotton boll weevil (Anthonomus grandis) is a serious insect-pest in the Americas, particularly in Brazil. The use of chemical or biological insect control is not effective against the cotton boll weevil because of its endophytic life style. Therefore, the use of biotechnological tools to produce insect-resistant transgenic plants represents an...

Biotech crops expressing Bacillus thuringiensis Cry toxins present a valuable approach for insect control. Cry8Ka5, which is highly toxic to the cotton boll weevil (Anthonomus grandis), was used as a model to study toxin-ligand interactions. Three Cry-binding proteins were detected after toxin overlay assays. Following de novo sequencing, a heat-sh...

Sugarcane giant borer (Telchin licus licus) is a serious sugarcane pest in Americas whose endophytic lifestyle hampers effective chemical and biological controls. Therefore, development of alternative control methods is extremely important. Envisaging development of transgenic plants resistant to this pest, we investigated the effect of the Bacillu...

Bacillus thuringiensis (Bt) Berliner is a promising agent for microbial control of agriculturally and medically important insects. This study aimed at searching for Bt strains encoding Cry proteins that act more efficiently against fall armyworm. Thirty Bt strains were isolated from soil samples in Pernambuco State and evaluated through bioassays....

Twenty-five RAPD loci and 6 isozyme loci were studied to characterize the genetic variability of natural populations of Anthonomus grandis from two agroecosystems of Brazil. The random-amplified polymorphic DNA data disclosed a polymorphism that varied from 52 to 84% and a heterozygosity of 0.189 to 0.347. The index of genetic differentiation (GST)...

Many international forums have identified the need for comprehensive, scientific methods for the pre-release testing and post-release monitoring of transgenic plants to ensure their environmental safety and sustainable use. In response to this requirement, the International Project on GMO Environmental Risk Assessment Methodologies develops biosafe...

Many international forums have identified the need for comprehensive, scientific methods for the pre-release testing and post-release monitoring of transgenic plants to ensure their environmental safety and sustainable use. In response to this requirement, the International Project on GMO Environmental Risk Assessment Methodologies develops biosafe...

A sensitive and specific polymerase chain reaction (PCR) based on a highly repeated deoxyribonucleic acid (DNA) sequence (188 bp; SspI repeat) was tested for the detection of Wuchereria bancrofti DNA in blood and urine samples collected during the day from individuals in Coque, Recife, Brazil, an endemic area for W. bancrofti. All microfilaraemic i...

Cotton fiber is a tubular single cell trichomes derived from individual epidermal cells of the outer integument of the ovules in the developing cotton fruit. In addition to being the most important natural textile fiber crop, cotton fibers provide an excellent experimetal system to study cellular and development of cell wall. This work was concerne...

Gossypium hirsuntum is attacked by five crop-pest species, among them; Spodoptera frugiperda constitutes one of the most important. Bacillus thuringiensis is an entomopathogenic bacterium with high useful potential: as biolarvicide production as for obtaining insect-resistant transgenic plants, due the presence of the cry genes encoding species-spe...

Microsatelite markers or SSR (Simple Sequence Repeat) are short base pairs repeated sequences spreaded in the entire genome. Genotypic identification using genetic markers can to light out the polymorphism that exists through the genome, which will help to select important genes for specific agronomic characteristics. The objective of this work was...