Midgut–Transgenic Crops Expressing<i>Bacillus thuringiensis</i>Cry Proteins

Rie, Jeroen Van; Jansens, Stefan

doi:10.1002/9783527644179.ch30

Cited by 1 publication

(1 citation statement)

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“…Currently, more than 800 sequences of Cry proteins are registered, which are grouped into 78 different classes and are specifically active mostly against some insects and nematodes [12]. The genes encoding the insecticidal proteins in some Bt microbial products have been successfully cloned, integrated, and expressed in genetically modified plants [13,14,15,16,17,18,19] to confer resistance against insect damage. Bt-protected crops such as corn, cotton, soybean, and potato have demonstrated significant benefits since their introduction in 1996.…”

Section: Introductionmentioning

confidence: 99%

Development of an Efficient Protocol to Obtain Transgenic Coffee, Coffea arabica L., Expressing the Cry10Aa Toxin of Bacillus thuringiensis

Valencia-Lozano

Cabrera-Ponce

Gómez-Lim

et al. 2019

IJMS

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This report presents an efficient protocol of the stable genetic transformation of coffee plants expressing the Cry10Aa protein of Bacillus thuringiensis. Embryogenic cell lines with a high potential of propagation, somatic embryo maturation, and germination were used. Gene expression analysis of cytokinin signaling, homedomains, auxin responsive factor, and the master regulators of somatic embryogenesis genes involved in somatic embryo maturation were evaluated. Plasmid pMDC85 containing the cry10Aa gene was introduced into a Typica cultivar of C. arabica L. by biobalistic transformation. Transformation efficiency of 16.7% was achieved, according to the number of embryogenic aggregates and transgenic lines developed. Stable transformation was proven by hygromycin-resistant embryogenic lines, green fluorescent protein (GFP) expression, quantitative analyses of Cry10Aa by mass spectrometry, Western blot, ELISA, and Southern blot analyses. Cry10Aa showed variable expression levels in somatic embryos and the leaf tissue of transgenic plants, ranging from 76% to 90% of coverage of the protein by mass spectrometry and from 3.25 to 13.88 μg/g fresh tissue, with ELISA. qPCR-based 2−ΔΔCt trials revealed high transcription levels of cry10Aa in somatic embryos and leaf tissue. This is the first report about the stable transformation and expression of the Cry10Aa protein in coffee plants with the potential for controlling the coffee berry borer.

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