Department of Biochemistry and Biophysics, University of California, Davis, California 95616.
Plant Physiol. 1985 Mar;77(3):630-4. doi: 10.1104/pp.77.3.630.
Callus cultures have been produced from the epicotyl and leaves, hypocotyl, and roots of germinating Dolichos biflorus seeds. These cultures were initiated on media containing 2,4-dichlorophenoxyacetic acid and kinetin, transferred to media with increased amounts of these hormones, and then maintained on hormone-free media. Extracts of these cultures were examined by radioimmunoassays specific for the lectin from the seeds of this plant and for a lectin that is present only in the stems and leaves of the intact plant. Although the seed lectin was not detected in any cultures, the stem and leaf lectin was produced in those cultures grown on the hormone free media. Lectin isolated from these cultures had subunits identical in electrophoretic mobilities to the subunits from the lectin isolated from intact stems and leaves. Levels of this lectin decreased when the cells were transferred back to media containing hormones and increased again upon transfer to the hormone-free media. The absence of exogenous hormones and the production of lectin were also correlated with the rapid growth and greening of the cells. Immunofluorescence and immunocytochemical studies on sections of cultured cells indicated that the stem and leaf lectin is associated with the cytoplasm as well as the cell wall as has been found in previous studies on the subcellular localization of this lectin in the intact plant.
已从萌发的豇豆种子的上胚轴和叶片、下胚轴和根中产生愈伤组织培养物。这些培养物最初在含有 2,4-二氯苯氧乙酸和激动素的培养基上启动,转移到含有这些激素的增加量的培养基上,然后在无激素培养基上维持。通过针对该植物种子中的凝集素和仅存在于完整植物的茎和叶中的凝集素的放射免疫测定法检查这些培养物的提取物。尽管在任何培养物中均未检测到种子凝集素,但在无激素培养基上生长的培养物中产生了茎和叶凝集素。从这些培养物中分离出的凝集素有与从完整茎和叶中分离出的凝集素相同的亚基在电泳迁移率上相同。当将细胞转移回含有激素的培养基中时,该凝集素的水平降低,而当再次转移到无激素培养基时,其水平又升高。细胞快速生长和变绿也与缺乏外源激素和产生凝集素有关。对培养细胞切片的免疫荧光和免疫细胞化学研究表明,与先前在完整植物中对该凝集素的亚细胞定位的研究一样,茎和叶凝集素与细胞质以及细胞壁有关。