Distribution of cryl, cryll and cryV Genes within Bacillus thuringiensis Isolates from Spain

Ferrandis, María; Juárez-Pérez, Víctor; Frutos, Roger; Bel, Yolanda; Ferré, Juan

doi:10.1016/s0723-2020(99)80064-2

Cited by 55 publications

(47 citation statements)

References 31 publications

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“…The most abundant gene found in our collection was cry2, and the second most abundant was the cry1 gene family. Several reports show a high frequency of certain cry1 gene combinations, for example the linkage cry1C-cry1D (Bravo et al 1998, Ferrandis et al 1999. The cry1C-cry1D linkage may be explained by their common location on the same replicon (Sanchis et al 1988).…”

Section: Discussionmentioning

confidence: 99%

“…However, in this study cry1D gene was found alone at a relatively high frequency (24 strains) and only 7 strains presented such linkage. The absence of cry1C may be the result of a deletion or a negative selection of cry1C from an ancestral cry1C-cry1D linkage (Ferrandis et al 1999). One interesting observation was the high frequency of cry1D in combination with those of the subfamily cry1A.…”

Section: Discussionmentioning

confidence: 99%

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Diversity of Bacillus thuringiensis strains isolated from coffee plantations infested with the coffee berry borer Hypothenemus hampei

Arrieta

Hernández

Espinoza

2014

RBT

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Abstract:The coffee berry borer Hypothenemus hampei Ferrari (Coleoptera: Scolytidae) was first reported infecting Costa Rican coffee plantations in the year 2000. Due to the impact that this plague has in the economy of the country, we were interested in seeking new alternatives for the biological control of H. hampei, based on the entomopathogenic bacteria Bacillus thuringiensis. A total of 202 B. thuringiensis isolates obtained from Costa Rican coffee plantations infested with H. hampei were analyzed through crystal morphology of the crystal inclusions and SDS-PAGE of δ-endotoxins, while 105 strains were further evaluated by PCR for the presence cry, cyt and vip genes. Most of the Bt strains showed diverse crystal morphologies: pleomorphic (35%), oval (37%), bipyramidal (3%), bipyramidal and oval (12%), bipyramidal, oval and pleomorphic (10%) and bipyramidal, oval and cubic (3%). The SDS-PAGE analyses of the crystal preparations showed five strains with δ-endotoxin from 20 to 40 kDa, six from 40 to 50 kDa, seven from 50 to 60 kDa, 19 from 60 to 70 kDa, 29 from 70 to 100 kDa and 39 from 100-145 kDa. PCR analyses demonstrated that the collection showed diverse cry genes profiles having several genes per strain: 78 strains contained the vip3 gene, 82 the cry2 gene, 45 the cry1 and 29 strains harbored cry3-cry7 genes. A total of 13 strains did not amplified with any of the cry primers used: cry1, cry2, cry3-7, cry5, cry11, cry12 and cry14. Forty-three different genetic profiles were found, mainly due to the combination of cry1A genes with other cry and vip genes. The genetic characterization of the collection provides opportunities for the selection of strains to be tested in bioassays against H. hampei and other insect pests of agricultural importance. Rev. Biol. Trop. 52(3): 757-764. Epub 2004 Dic 15.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Diversity of Bacillus thuringiensis strains isolated from coffee plantations infested with the coffee berry borer Hypothenemus hampei

Arrieta

Hernández

Espinoza

2014

RBT

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“…Specific-primer PCR and multiplex PCR can directly detect known cry genes from B. thuringiensis (2,4,(6)(7)(8). Exclusive PCR and PCR-RFLP can identify not only known cry genes but also novel cry genes (3,5,11,15,17,23). We established a PCR-RFLP method to identify cry1I genes by using the conserved regions of four known cry1I genes as a basis for designing a pair of universal primers for cry1I-type genes which were different from the primers previously reported (18,19,22).…”

Section: Discussionmentioning

confidence: 99%

Identification of cry1I- Type Genes from Bacillus thuringiensis Strains and Characterization of a Novel cry1I- Type Gene

Song

Zhang

et al. 2003

Appl Environ Microbiol

109

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A PCR-restriction fragment length polymorphism method for identification of cry1I-type genes from Bacillus thuringiensis was established by designing a pair of universal primers based on the conserved regions of the genes to amplify 1,548-bp cry1I-type gene fragments. Amplification products were digested with the Bsp119I and BanI enzymes, and four kinds of known cry1I-type genes were successfully identified. The results showed that cry1I-type genes appeared in 95 of 115 B. thuringiensis isolates and 7 of 13 standard strains. A novel cry1I-type gene was found in one standard strain and six isolates. The novel cry1I gene was cloned from B. thuringiensis isolate Btc007 and subcloned into vector pET-21b. Then it was overexpressed in Escherichia coli BL21(DE3). The expressed product was shown to be toxic to the diamondback moth (Plutella xylostella), Asian corn borer (Ostrinia furnacalis), and soybean pod borer (Leguminivora glycinivorella). However, it was not toxic to the cotton bollworm (Helicoverpa armigera), beet armyworm (Spodoptera exigua), or elm leaf beetle (Pyrrhalta aenescens) in bioassays. Subsequently, the Cry protein encoded by this novel cry gene was designated Cry1Ie1 by the B. thuringiensis ␦-endotoxin nomenclature committee.Crystal proteins from the gram-positive spore-forming bacterium Bacillus thuringiensis are toxic to a wide variety of insects that are economically important as pests. Many different genes encoding the B. thuringiensis endotoxin have been isolated and characterized. The genes have been classified as cry1 to cry40, cyt1, and cyt2 and are ranked according to their homology (10, 21; see also the B. thuringiensis toxin nomenclature website at http://www.biols.susx.ac.uk/home/Neil_Crickmore /Bt/). Cry1 proteins that are active against lepidopteran insects are produced as crystalline parasporal inclusions during sporulation. Generally, the crystals are composed of protoxins of approximately 130 kDa, but cry1I-type genes are usually silent genes capable of encoding a protein of about 81 kDa in B. thuringiensis strains (9,13,21,24). We decided to screen B. thuringiensis isolates for cry1I genes with the aim of finding novel cry1I genes, which could encode insecticidal proteins toxic to insensitive or resistant insect pests.This screening approach included the development of an analysis protocol based on PCR-restriction fragment length polymorphism (RFLP). PCR-based methods have been developed to detect different cry genes from B. thuringiensis strains (1-8, 11, 13, 15, 17, 23). More than 80 primer pairs have been designed to identify entire groups and individual cry genes (19). Several specific primers and probes for Southern blotting were designed to detect cry1I-type genes. A wide distribution of cry1I-type genes among many different B. thuringiensis strains has been reported (13,22,25). However, the list of cry genes is increasing, and novel PCR primers are needed in order to identify some of the recently described genes (5).The present study establishes a PCR-RFLP method for identify...

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“…The lack of correspondence between δ-endotoxins genotype of Bt-S84-13a and its biological activity may be due to various factors, for example, the production of inactive crystal proteins whereby the gene that synthesize the crystal proteins could be under the control of a weak promoter (Masson et al, 1998;Ferrandis et al, 1999;Armengol et al, 2007). Some cry genes may be also expressed in low levels which may lead to the low expression of the respective Cry proteins.…”

Section: Discussionmentioning

confidence: 99%

Characterization of a native Bacillus thuringiensis isolates from Malaysia that produces exosporium-enclosed parasporal inclusion

Fun¹,

Rathinam²,

Ghazali³

et al. 2016

Emir. J. Food Agric

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Distribution of cryl, cryll and cryV Genes within Bacillus thuringiensis Isolates from Spain

Cited by 55 publications

References 31 publications

Diversity of <i>Bacillus thuringiensis</i> strains isolated from coffee plantations infested with the coffee berry borer <i>Hypothenemus hampei</i>

Diversity of <i>Bacillus thuringiensis</i> strains isolated from coffee plantations infested with the coffee berry borer <i>Hypothenemus hampei</i>

Identification of cry1I- Type Genes from Bacillus thuringiensis Strains and Characterization of a Novel cry1I- Type Gene

Characterization of a native Bacillus thuringiensis isolates from Malaysia that produces exosporium-enclosed parasporal inclusion

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