Thompson W F, Cleland R
Department of Botany, University of Washington, Seattle, Washington 98105.
Plant Physiol. 1971 Dec;48(6):663-70. doi: 10.1104/pp.48.6.663.
The ability of auxin to alter gene transcription in pea (Pisum sativum L.) stem tissues has been investigated by means of DNA/RNA hybridization-competition techniques. In order to obtain reproducible hybridization with total nucleic acid preparations from plants it was found necessary to remove interfering substances, probably polysaccharides; this was accomplished by methoxyethanol extraction and precipitation with cetyltrimethylammonium bromide. When purified in this fashion, plant nucleic acids could be made to form hybrids which showed both species specificity and high thermal stability.No change in hybridizable RNA of stem sections in response to auxin could be detected over a 2- to 24-hour period, regardless of the auxin level employed. In contrast, when large doses of auxin were applied to intact pea seedlings, definite changes in the hybridizable RNA of stem tissue were detected both 8 and 24 hours after treatment. Many of the 2,4-D-induced species of RNA which were present at 24 hours were also present at 8 hours. Sections apparently lack a factor needed for the manifestation of the auxin effect on RNA synthesis.Since the hybridization assay employed does not measure all cellular RNA, it is still possible that certain RNA species may be synthesized in sections in response to auxin. However, the auxin promotion of cell elongation in such sections is clearly not associated with changes in hybridizable RNA such as have been reported for several hormonal responses in animal systems.
利用DNA/RNA杂交竞争技术研究了生长素改变豌豆(Pisum sativum L.)茎组织中基因转录的能力。为了使植物总核酸制剂获得可重复的杂交结果,发现有必要去除可能是多糖的干扰物质;这通过甲氧基乙醇提取并用十六烷基三甲基溴化铵沉淀来完成。以这种方式纯化后,植物核酸可形成具有物种特异性和高热稳定性的杂交体。在2至24小时内,无论使用何种生长素水平,均未检测到茎段可杂交RNA因生长素而发生变化。相反,当向完整的豌豆幼苗施用大剂量生长素时,在处理后8小时和24小时均检测到茎组织可杂交RNA有明显变化。24小时时存在的许多2,4 - D诱导的RNA种类在8小时时也存在。切片显然缺乏生长素对RNA合成产生作用所需的一种因子。由于所采用的杂交测定法并未测量所有细胞RNA,因此仍有可能某些RNA种类在切片中因生长素而合成。然而,生长素对这类切片中细胞伸长的促进作用显然与可杂交RNA的变化无关,而动物系统中的几种激素反应已有此类变化的报道。