Plant Biotechnology Institute, National Research Council of Canada, 110 Gymnasium Place, S7N 0W9, Saskatoon, SK, Canada.
Plant Cell Rep. 1993 Nov;13(1):17-23. doi: 10.1007/BF00232308.
Cotyledonary somatic embryos of white spruce [Picea glauca (Moench) Voss] were subjected to microprojectile bombardment with a gene construct containing a gus::nptll fusion gene. Somatic embryos were used to re-induce the embryogenic tissue after bombardments. Histochemical assay using X-gluc as a substrate showed that all the embryos (100%) were GUS positive 48 h after bombardment. However, only thirteen out of 605 embryos (2.2%) remained GUS positive after two months in culture. Three of those thirteen (23%) embryo-derived tissues consistently showed GUS activity for eight months in culture. These putatively transfomed embryogenic tissues were subjected to Southern blot analysis and the results suggested integration of the gus::nptll gene expression cassette in the white spruce genome.
白松 [松科(Pinaceae)云杉属(Picea)] 的子叶体胚经过含有 Gus::nptll 融合基因的基因构建体的微弹丸轰击。体胚在轰击后用于重新诱导胚胎发生组织。使用 X-gluc 作为底物的组织化学分析表明,轰击后 48 小时,所有胚胎(100%)均为 GUS 阳性。然而,在培养两个月后,只有 605 个胚胎中的 13 个(2.2%)仍保持 GUS 阳性。这 13 个胚胎中有 3 个(23%)胚胎衍生组织在培养 8 个月内始终表现出 GUS 活性。这些推测的转化胚胎发生组织进行了 Southern blot 分析,结果表明 Gus::nptll 基因表达盒已整合到白松基因组中。