Seed-Specific Stable Expression of the α-AI1 Inhibitor in Coffee Grains and the In Vivo Implications for the Development of the Coffee Berry Borer

Abstract

Genetic transformation of coffee (Coffea spp.), the second most traded commodity worldwide, is an alternative approach to introducing features that cannot be introgressed by traditional crossings. The transgenic stability, heritability and quantitative and spatial expression patterns of the seed-specific promoter phytohemagglutinin (PHA-L) from Phaseolus vulgaris were characterized in genetically modified C. arabica expressing the α-amylase inhibitor-1 (α-AI1) gene. The α-AI1 inhibitor shows considerable activity toward digestive enzymes of the coffee berry borer (CBB) Hypothenemus hampei. This insect pest expends its life cycle almost entirely in coffee berries. Transgene containment in the fruit is important to meeting food and environmental safety requirements for releasing genetically modified (GM) crops. PCR analysis of T2 coffee plants showed a Mendelian single-copy segregation pattern. Ectopic transgene expression was only detected in coffee grains, as demonstrated by reverse transcription-PCR analysis of different plant tissues. An intense immunocytochemical signal associated with α-AI1 protein expression was localized to endospermic cells. In addition, a delay in the larval development of CBB was observed after challenging transgenic coffee seeds with the insect. These results indicate that the PHA-L promoter might be a useful tool in coffee for the seed-specific expression of genes related to coffee bean productivity, quality and pest protection. The biotechnological applicability of the α-AI1 gene for controlling CBB is also discussed. This work is the first report showing a seed-specific transgene expression in coffee plants.Electronic supplementary materialThe online version of this article (doi:10.1007/s12042-015-9153-0) contains supplementary material, which is available to authorized users.

Análise De Métodos De Transformação Do Café E Sua Possível Eficácia No Ramo Agrícola

“…Quanto a pesquisa feita por Albuquerque et al (2015), os pesquisadores concluíram que o promotor PHA-L pode ser utilizado em grãos de café com a função de guiar a expressão de transgene específica de sementes, tornando-se assim um recurso valioso para o controle de transgenes em análises direcionadas ao melhoramento de plantas.…”

CRISPR technology towards genome editing of the perennial and semi-perennial crops citrus, coffee and sugarcane

“…Fruits of genetically transformed plants inhibited the development and infestation capacity of CBB females ( Valencia-Lozano et al., 2021 ). Similarly, particle bombardment was used to transform embryogenic calli of C. arabica with the α-amylase-1 inhibitor gene ( α-AI1 ) that triggers resistance to CBB ( Albuquerque et al., 2015 ). The authors evaluated 54 plants and observed that the α-AI1 transgene was stably inherited in the T2 progeny and expressed in a tissue-specific manner in seeds by using the PHA-L promoter.…”