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.

中文翻译：

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α-AI1抑制剂在咖啡颗粒中的种子特异性稳定表达及其对咖啡浆果Bor的发展的体内意义。

对咖啡（Coffea spp。）（全球第二大交易商品）的遗传转化是引入传统杂交不能融合的特征的替代方法。在表达α-淀粉酶抑制剂-1（α-AI1）基因的转基因阿拉伯芥中表征了菜豆种子特异性启动子植物血凝素（PHA-L）的转基因稳定性，遗传力以及定量和空间表达模式。α-Al1抑制剂对咖啡bore（CBB）的次生猪嗜血杆菌的消化酶表现出相当大的活​​性。这种害虫几乎完全在咖啡浆果中延长其生命周期。水果中的转基因遏制对于满足食品和环境安全要求以释放转基因作物非常重要。T2咖啡植物的PCR分析显示孟德尔单拷贝分离模式。异位转基因表达仅在咖啡粒中检测到，这通过不同植物组织的逆转录-PCR分析得以证实。与α-AI1蛋白表达相关的强烈免疫细胞化学信号定位于胚乳细胞。此外，在用昆虫攻击转基因咖啡种子后，观察到CBB幼虫发育延迟。这些结果表明，PHA-L启动子可能是咖啡中有用的工具，可用于与咖啡豆生产力相关的基因的种子特异性表达，质量和害虫保护。生物技术的适用性还讨论了控制CBB的α-AI1基因。这项工作是第一个报告，显示了咖啡植物中种子特异性转基因的表达。