Department of Plant Sciences, University of Western Ontario, N6A 5B7, London, Ontario, Canada.
Theor Appl Genet. 1990 Oct;80(4):497-502. doi: 10.1007/BF00226751.
A leaf disc method is described to permit the localized incorporation of (35)S-L-methionine into polypeptides synthesized in individual leaves of maize plants grown in the field. The method of incorporation employs minimal external manipulation of the intact leaf, is simple, repeatable, and may be used at any plant age after leaf emergence. Incorporation (cpm/μg protein) in 12 leaves per plant was compared among three inbred (Oh43, W23, M14) and three F1 hybrid (Oh43/M14, W23/M14, Oh43/W23) genotypes. The incorporation was 40% higher (hybrid versus inbred) in 9 of the 12 leaves studied. Samples from leaf 07 (7th leaf numbered from base of plant) for four inbreds (Oh43, M14, B73, Mol 7) and two pairs of reciprocal F1 hybrids (Oh43/M14, M14/Oh43; B73/Mo17, Mo17/B73) were labelled in situ using the leaf disc method. Each cultivar was sampled at three different ages in each of 1985, 1986, and 1987. High-resolution, two-dimensional isoelectric focusing sodium-dodecyl-sulfate polyacrylamide gel electrophoresis and fluorography were used to display the polypeptides synthesized in the samples. Multivariate methods - Principal Coordinate Analysis, Cluster Analysis, and Standard Deviation Distance - were used to analyze variation and to identify trends in the variation for year, genotype, and age sampled. Our analyses disclose a hierarchy to polypeptide synthesis variation in maize leaves: differences in polypeptide synthesis are greater for year-to-year comparisons than differences due to sample age, which in turn are greater than differences for inbred versus hybrid comparisons.
描述了一种叶圆片方法,可使在田间生长的玉米植株的单个叶片中合成的多肽进行局部掺入(35)S-L-甲硫氨酸。该掺入方法对完整叶片的外部操作最少,简单、可重复,并且可以在叶片出现后任何植物年龄使用。比较了三种自交系(Oh43、W23、M14)和三种 F1 杂种(Oh43/M14、W23/M14、Oh43/W23)基因型的每株 12 片叶片的掺入(cpm/μg 蛋白)。在研究的 12 片叶片中的 9 片中,杂交种的掺入率(杂交种与自交系)高 40%。对来自四个自交系(Oh43、M14、B73、Mol 7)和两个正反交 F1 杂种(Oh43/M14、M14/Oh43;B73/Mo17、Mo17/B73)的叶 07 号叶片(从植株基部算起的第 7 片叶片)进行原位标记使用叶圆片方法。在 1985 年、1986 年和 1987 年的每个时期,对每个品种进行了三个不同年龄的采样。使用高分辨率、二维等电聚焦十二烷基硫酸钠聚丙烯酰胺凝胶电泳和荧光显影来显示样品中合成的多肽。使用主坐标分析、聚类分析和标准差距离等多变量方法来分析变化并确定年度、基因型和采样年龄的变化趋势。我们的分析揭示了玉米叶片中多肽合成变化的层次结构:与样本年龄引起的差异相比,年际差异引起的多肽合成差异更大,而与自交系与杂种比较引起的差异相比,样本年龄引起的差异更大。