Ahad Abdul, Wolf Jochen, Nick Peter
Institut für Biologie II, Schänzlestr 1, D-79104 Freiburg, Germany.
Transgenic Res. 2003 Oct;12(5):615-29. doi: 10.1023/a:1025814814823.
T-DNA activation tagging was used to generate tobacco mutants with increased tolerance to antimicrotubular herbicides and chilling stress. After transformation, protoplast-derived calli were screened for tolerance to treatments that affect microtubule assembly. In one screen mutants with tolerance to aryl carbamates (a blocker of microtubule assembly) were selected, the second screen was targeted to chilling-tolerant mutants that could survive for several months at 3 degrees C, a third screen combined both factors. The resistance of these mutants to aryl carbamates or chilling was accompanied by resistance of microtubules to these factors. The carbamate tolerant mutants were cross-resistant to chilling stress. This was mirrored by an adaptive reorganization of microtubules and a reduction of microtubule dynamics in response to chilling. The analysis of these mutants suggests (1) that microtubule dynamics limit the tolerance to chilling and EPC, and (2) that the cold sensitivity of microtubules limits chilling tolerance in tobacco.
采用T-DNA激活标签法来生成对抗微管除草剂和冷胁迫耐受性增强的烟草突变体。转化后,对原生质体来源的愈伤组织进行筛选,以检测其对影响微管组装处理的耐受性。在一次筛选中,选择了对芳基氨基甲酸酯(一种微管组装阻断剂)具有耐受性的突变体,第二次筛选针对的是能够在3摄氏度下存活数月的耐冷突变体,第三次筛选则综合了这两个因素。这些突变体对芳基氨基甲酸酯或冷胁迫的抗性伴随着微管对这些因素的抗性。氨基甲酸酯耐受性突变体对冷胁迫具有交叉抗性。这反映在微管的适应性重组以及冷胁迫下微管动态性的降低。对这些突变体的分析表明:(1)微管动态性限制了对冷胁迫和EPC的耐受性;(2)微管的冷敏感性限制了烟草的耐冷性。