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火疫病病原菌韧皮部坏死杆菌(Erwinia amylovora)腺嘌呤转运蛋白 EaAdeP 的功能表征及其对苹果和梨中病害形成的影响。

Functional characterization of the adenine transporter EaAdeP from the fire blight pathogen Erwinia amylovora and its effect on disease establishment in apples and pears.

机构信息

Department of Biology, Purdue University Fort Wayne, 2101 East Coliseum Blvd., Fort Wayne, IN 46805, USA.

Department of Plant Pathology & Ecology, The Connecticut Agricultural Experiment Station, 123 Huntington Street, New Haven, CT 06511, USA.

出版信息

FEMS Microbiol Lett. 2020 Nov 5;367(20). doi: 10.1093/femsle/fnaa173.

Abstract

Erwinia amylovora is the causal agent of fire blight, an economically important disease of apples and pears. As part of the infection process, Er. amylovora propagates on different plant tissues each with distinct nutrient environments. Here, the biochemical properties of the Er. amylovora adenine permease (EaAdeP) are investigated. Heterologous expression of EaAdeP in nucleobase transporter-deficient Escherichia coli strains, coupled with radiolabel uptake studies, revealed that EaAdeP is a high affinity adenine transporter with a Km of 0.43 ± 0.09 μM. Both Es. coli and Er. amylovora carrying extra copies of EaAdeP are sensitive to growth on the toxic analog 8-azaadenine. EaAdeP is expressed during immature pear fruit infection. Immature pear and apple fruit virulence assays reveal that an E. amylovora ΔadeP::Camr mutant is still able to cause disease symptoms, however, with growth at a lower level, indicating that external adenine is utilized in disease establishment.

摘要

果胶杆菌是火疫病的病原体,火疫病是苹果和梨的一种重要经济病害。作为感染过程的一部分,果胶杆菌在不同的植物组织上繁殖,每个组织都有不同的营养环境。在这里,研究了果胶杆菌腺嘌呤通透酶(EaAdeP)的生化特性。将 EaAdeP 在核碱基转运蛋白缺陷型大肠杆菌菌株中的异源表达,结合放射性摄取研究,表明 EaAdeP 是一种高亲和力的腺嘌呤转运蛋白,Km 值为 0.43±0.09 μM。携带额外 EaAdeP 拷贝的大肠杆菌和果胶杆菌对毒性类似物 8-氮杂腺嘌呤的生长都很敏感。EaAdeP 在未成熟的梨果实感染过程中表达。未成熟的梨和苹果果实的致病性试验表明,E. amylovora ΔadeP::Camr 突变体仍然能够引起疾病症状,但生长水平较低,表明在疾病建立过程中利用了外部腺嘌呤。

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