Campell B R, Town C D
Biology Department, Case Western Reserve University, Cleveland, Ohio 44106-7080.
Plant Physiol. 1991 Nov;97(3):1166-73. doi: 10.1104/pp.97.3.1166.
gamma-Radiation-induced tumors of Arabidopsis thaliana L. have been produced as a novel approach to isolation of genes that regulate plant development. Tumors excised from irradiated plants are hormone autonomous in culture and have been maintained on hormone-free medium for up to 4 years. Five tumor tissue lines having different morphologies and growth rates were analyzed for auxin, cytokinin, and 1-aminocyclopropane-1-carboxylic acid (ACC) content, ethylene production, and response to exogenous growth regulators. Normal tissues and two crown gall tissue lines were analyzed for comparison. Rosettes and whole seedlings each contained approximately 30 nanograms. (gram fresh weight)(-1) free indoleacetic acid (IAA), 150 nanograms. (gram fresh weight)(-1) ester-conjugated IAA, and 10 to 20 micrograms. (gram fresh weight)(-1) amide-conjugated IAA. The crown gall lines contained similar amounts of free and ester-conjugated IAA but less amide conjugates. Whereas three of the radiation-induced tumor lines had IAA profiles similar to normal tissues, one line had 10- to 100-fold more free IAA and three- to 10-fold less amide-conjugated IAA. The fifth line had normal free IAA levels but more conjugated IAA than control tissues. Whole seedlings contained approximately 2 nanograms. (gram fresh weight)(-1) of both zeatin riboside and isopentenyladenosine. The crown gall lines had 100- to 1000-fold higher levels of each cytokinin. In contrast, the three radiation-induced tumor lines analyzed contained cytokinin levels similar to the control tissue. The radiation-induced tumor tissues produced very little ethylene, although each contained relatively high levels of ACC. Normal callus contained similar amounts of ACC but produced several times more ethylene than the radiation-induced tumor lines. Each of the radiation-induced tumor tissues displayed a unique set of responses to exogenously supplied growth regulators. Only one tumor line showed the same response as normal callus to both auxin and cytokinin feeding. In some cases, one or more tumor lines showed increased sensitivity to certain growth substances. In other cases, growth regulator feeding had no significant effect on tumor tissue growth. Morphology of the radiation-induced tumor tissues generally did not correlate with auxin to cytokinin ratio in the expected manner. The results suggest that a different primary genetic event led to the formation of each tumor and that growth and differentiation in the tumor tissue lines are uncoupled from the normal hormonal controls.
通过伽马辐射诱导拟南芥产生肿瘤,这已成为一种分离调控植物发育基因的新方法。从受辐照植物上切除的肿瘤在培养中具有激素自主性,并且已在无激素培养基上维持了长达4年。分析了5个具有不同形态和生长速率的肿瘤组织系的生长素、细胞分裂素和1-氨基环丙烷-1-羧酸(ACC)含量、乙烯产生以及对外源生长调节剂的反应。分析了正常组织和两个冠瘿组织系以作比较。莲座叶和整株幼苗各自含有约30纳克/(克鲜重)的游离吲哚乙酸(IAA)、150纳克/(克鲜重)的酯结合型IAA以及10至20微克/(克鲜重)的酰胺结合型IAA。冠瘿系含有相似数量的游离和酯结合型IAA,但酰胺结合物较少。虽然三个辐射诱导的肿瘤系的IAA谱与正常组织相似,但有一个系的游离IAA多10至100倍,酰胺结合型IAA少3至10倍。第五个系的游离IAA水平正常,但结合型IAA比对照组织多。整株幼苗含有约2纳克/(克鲜重)的玉米素核苷和异戊烯基腺苷。冠瘿系的每种细胞分裂素水平高100至1000倍。相比之下,所分析的三个辐射诱导的肿瘤系的细胞分裂素水平与对照组织相似。辐射诱导的肿瘤组织产生的乙烯很少,尽管每个组织都含有相对较高水平的ACC。正常愈伤组织含有相似数量的ACC,但产生的乙烯比辐射诱导的肿瘤系多几倍。每个辐射诱导的肿瘤组织对外源供应的生长调节剂都表现出一组独特的反应。只有一个肿瘤系在生长素和细胞分裂素处理下与正常愈伤组织表现出相同的反应。在某些情况下,一个或多个肿瘤系对某些生长物质表现出更高的敏感性。在其他情况下,生长调节剂处理对肿瘤组织生长没有显著影响。辐射诱导的肿瘤组织的形态通常与生长素与细胞分裂素的比例没有预期的相关性。结果表明,不同的原发性基因事件导致了每个肿瘤的形成,并且肿瘤组织系中的生长和分化与正常激素控制脱钩。