During the past decade the pesticidal bacterium Bacillus thuringiensis has been the subject of intensive research. These efforts have yielded considerable data about the complex relationships between the structure, mechanism of action, and genetics of the organism’s pesticidal crystal proteins, and a coherent picture of these relationships is beginning to emerge. Other studies have focused on the ecological role of the B. thuringiensis crystal proteins, their performance in agricultural and other natural settings, and the evolution of resistance mechanisms in target pests. Armed with this knowledge base and with the tools of modern biotechnology, researchers are now reporting promising results in engineering more-useful toxins and formulations, in creating transgenic plants that express pesticidal activity, and in constructing integrated management strategies to insure that these products are utilized with maximum efficiency and benefit.

Bacillus thuringiensis and Its Pesticidal Crystal Proteins

Insecticidal crystal proteins of Bacillus thuringiensis.

characterized by its ability to produce crystalline inclusions during spor-ulation. TIhese inclusions consist of proteins exhibiting a highly specific insecticidal activity (reviewed in references 4 and 97). Many B. thuriuigiensis strains with different insect host spectra have been identified (9). TIhey ar-e classified into different serotypes or subspecies based on their flagellar antigens. Most strains are active against larvae of certain members of the Lepidoptera, but some show toxicity against dipterian (reviewed in refer-ence 22) or coleopteran (53) species. For several cryst.al-producing strains.

Insecticidal crystal proteins of Bacilllus thuringiensis

Pectin structure and biosynthesis

The structure, function, and biosynthesis of plant cell wall pectic polysaccharides.

Soybean soluble polysaccharides (SSPS) extracted from soybean cotyledons are acidic polysaccharides and have a pectin-like structure. The results of a structural analysis of SSPS by using polygalacturonase (PGase) and rhamnogalacturonase (RGase) clarified that the main backbone consisted of galacturonan (GN) and rhamnogalacturonan (RG), which were composed of the diglycosyl repeating unit, -4)-α-D-GalpA-(1→2)-α-L-Rhap-(1-. The side chains of β-1,4-galactans, branched with fucose and arabinose residues, were linked to the C-4 side of rhamnose residues in the RG regions. The degree of polymerization (dps) of GN, which linked the RG regions together, was estimated to be about 4-10 residues, and some were modified with xylose residues on the C-3 side of the galacturonates. The dps of GN at the reducing end of SSPS was estimated to be about 7-9 residues. Moreover, the fragment of the basic structure of the RG region, -[4)-α-D-GalpA-(1→2)-α-L-Rhap- (1-]2-, some of which had long-chain β-1,4-galactans branched o...

Structural studies by stepwise enzymatic degradation of the main backbone of soybean soluble polysaccharides consisting of galacturonan and rhamnogalacturonan.

https://febs.onlinelibrary.wiley.com/doi/pdf/10.1016/S0014-5793%2899%2901327-7

The cadherin-like protein is essential to specificity determination and cytotoxic action of the Bacillus thuringiensis insecticidal CryIAa toxin

Soybean soluble polysaccharides (SSPS) extracted from soybean cotyledons have a pectin-like structure. The core polysaccharides after treatments with four kinds of hemicellulases and a pectinase contained approximately equal numbers of L-rhamnose and D-galacturonate residues, suggesting the presence of the rhamnogalacturonan (RG) I structure consisting of the diglycosyl repeating unit, -4)-alpha-D-GalpA-(1-->2)-alpha-L-Rhap-(1-. The lengths of RG chains were calculated as approximately 15, 28, and 100 diglycosyl repeats. The RG components linked to each other by intervention of galacturonan (GN) chains, constituting the backbone of SSPS. All arabinose residues, which constitute 21% of total SSPS sugars, were found to be in side chains from RG regions, and this was also true for galactose residues, which constitute 50% of total sugars. Of arabinose residues, 94% are present as alpha-1,3- or alpha-1,5-arabinans, and 89% of galactose residues were present as beta-1,4-galactans. Galactan chains are modified with arabinose, xylose, fucose, and glucose at the sites close to the RG regions.

Analysis of structural components and molecular construction of soybean soluble polysaccharides by stepwise enzymatic degradation.

Bacillus thuringiensis strains produce insect-specific Bt toxins. Bt CryIA(a) toxin binds to a 175-kDa glycoprotein (BtR175) on the microvillus membranes of columnar cells in the Bombyx mori midgut and causes lysis of the cells. BtR175 was purified, and its cDNA was cloned. The cDNA encodes a newly identified 193.3-kDa preproprotein form of BtR175 that includes nine extracellular cadherin repeats, a 23.5-kDa membrane-proximal domain, a membrane-spanning region, and a 13.6-kDa cytoplasmic domain. Spodoptera frugiperda cells transfected with a recombinant baculovirus DNA carrying the cDNA produced a 175-kDa protein that reacted with anti-BtR antibodies and the Bt CryIA(a) toxin.

Yasunori Nagamatsu

Papers

Structural studies by stepwise enzymatic degradation of the main backbone of soybean soluble polysaccharides consisting of galacturonan and rhamnogalacturonan.

The cadherin-like protein is essential to specificity determination and cytotoxic action of the Bacillus thuringiensis insecticidal CryIAa toxin

Analysis of structural components and molecular construction of soybean soluble polysaccharides by stepwise enzymatic degradation.

Cloning, Sequencing, and Expression of the Bombyx mori Receptor for Bacillus thuringiensis Insecticidal CryIA(a) Toxin

Effect of soybean soluble polysaccharides on the stability of milk protein under acidic conditions