Expression of Storage Protein in Somatic Embryos of Plants

在第一阶段制备了针对紫花苜蓿11S种子贮藏蛋白和生菜14.5S种子贮藏蛋白的抗体。在第二阶段，PI建议利用这些抗体来确定这两种作物体细胞胚胎的发育阶段。在紫花苜蓿中，11S贮藏蛋白在合子种子成熟和萌发过程中被检测到。这将决定一个可以与体细胞胚胎发育进行比较的时间表。本文介绍了紫花苜蓿体细胞胚培养条件，可获得极高的胚萌发率。将对来自这些处理的体细胞胚胎进行测试，并确定胚胎萌发和11S种子蛋白质积累之间的相关性。将对控制体细胞胚胎早熟萌发的条件进行试验，以检测紫花苜蓿11S贮藏蛋白的损失。在生菜中，合子胚胎也将建立类似的发育时间表。将启动生菜细胞培养，为胚胎繁殖创造条件。胚胎成熟的条件将根据14.5S蛋白进行研究。该提案的总体目标是利用体细胞胚胎发生来改善商业植物的繁殖。具体目标是确定体细胞胚胎中的储存蛋白水平是否与胚胎发育和活力相关。根据这一假设，PI预期通过增加内部蛋白质储备来改善体细胞胚胎的发育。

Antibody to the 11S seed storage protein of alfalfa and to the 14.5S seed storage protein of lettuce were prepared during Phase I. In Phase II, the PIs propose to use these antibodies to determine the developmental stage of somatic embryos of both crops. In alfalfa, 11S storage protein will be measured during zygotic seed maturation and during germination. This will determine a timetable against which the development of somatic embryos can be compared. Conditions of alfalfa somatic embryo culture are presented which result in very high rates of embryo germination. Somatic embryos from these treatments will be tested and a correlation determined between embryo germination and 11S seed protein accumulation. Conditions which control precocious germination of somatic embryos will be tested for the loss of 11S storage protein in alfalfa. In lettuce, a similar developmental timetable for zygotic embryos will be established. Lettuce cell cultures will be initiated in which conditions for embryo propagation will be established. Conditions for embryo maturation will be studied with respect to 14.5S protein. The overall objective of the proposal is to improve commercial plant propagation using somatic embryogenesis. The specific objective is to determine whether storage protein levels in somatic embryos correlate with embryo development and vigor. On this assumption, the PIs anticipate improving somatic embryo development by increasing internal protein storage reserves.