Immunogold localization of nodule-specific uricase in developing soybean root nodules
Immunogold localization of nodule-specific uricase in developing soybean root nodules
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DOI：
10.1007/bf00391217
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发表时间：
1986-04
期刊：
影响因子：
4.3
通讯作者：
K. Bosch;E. Newcomb
中科院分区：
文献类型：
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作者：
K. Bosch;E. Newcomb
用免疫金标记法研究了大豆根瘤特异性尿酸酶（EC 1.7.3.3）在大豆根瘤发育过程中的出现时间和分布。梅尔）接种慢生根瘤菌。通过采用抗大豆根瘤尿酸酶亚基的多克隆抗体制剂，在包埋于LR白色丙烯酸树脂或Spurr环氧树脂中的组织薄切片中检测到该酶。L R白色树脂保存抗原性较好，Spurr树脂保存超微结构较好。尿酸酶是第一次检测到与蛋白质A-金在年轻的，发展过氧化物酶体在未感染的细胞，符合释放的慢生根瘤菌类杆菌从感染线程在相邻的感染细胞。随着过氧化物酶体的扩大，密集的过氧化物酶体基质的标记增加。然而，在过氧化物酶体的次晶内含物上从未观察到金颗粒。尽管扩大过氧化物酶体和管状内质网之间的密切联系，尿酸酶在后者中检测不到。在成熟的根瘤中，尿酸酶的标记仅限于未感染细胞中的大过氧化物酶体。感染细胞中存在的小过氧化物酶体样小体没有被标记。
Immunogold labeling was used to study the time of appearance and distribution of a nodule-specific form of uricase (EC 1.7.3.3) in developing nodules of soybean (Glycine max(L.) Merr.) inoculated withBradyrhizobium japonicum. The enzyme was detected in thin sections of tissue embedded in either L R White acrylic resin or Spurr's epoxy resin, by employing a polyclonal antibody preparation active against a subunit of soybean nodule uricase. Antigenicity was better preserved in L R White resin, but ultrastructure was better maintained in Spurr's. Uricase was first detectable with protein A-gold in young, developing peroxisomes in uninfected cells, coincident with the release ofBradyrhizobiumbacteroids from infection threads in adjacent infected cells. As the peroxisomes enlarged, labeling of the dense peroxisomal matrix increased. Gold particles were never observed over the paracrystalline inclusions of peroxisomes, however. Despite a close association between enlarging peroxisomes and tubular endoplasmic reticulum, uricase was not detectable in the latter. In mature nodules, labeling of uricase was limited to the large peroxisomes in uninfected cells. Small peroxisome-like bodies present in infected cells did not become labeled.