Honee G, Buitink J, Jabs T, Sijbolts F, Apotheker M, Weide R, Sijen T, Stuiver M
Plant Physiol. 1998 Jul;117(3):809-20. doi: 10.1104/pp.117.3.809.
The AVR9 elicitor from the fungal pathogen Cladosporium fulvum induces defense-related responses, including cell death, specifically in tomato (Lycopersicon esculentum Mill.) plants that carry the Cf-9 resistance gene. To study biochemical mechanisms of resistance in detail, suspension cultures of tomato cells that carry the Cf-9 resistance gene were initiated. Treatment of cells with various elicitors, except AVR9, induced an oxidative burst, ion fluxes, and expression of defense-related genes. Agrobacterium tumefaciens-mediated transformation of Cf9 tomato leaf discs with Avr9-containing constructs resulted efficiently in transgenic callus formation. Although transgenic callus tissue showed normal regeneration capacity, transgenic plants expressing both the Cf-9 and the Avr9 genes were never obtained. Transgenic F1 seedlings that were generated from crosses between tomato plants expressing the Avr9 gene and wild-type Cf9 plants died within a few weeks. However, callus cultures that were initiated on cotyledons from these seedlings could be maintained for at least 3 months and developed similarly to callus cultures that contained only the Cf-9 or the Avr9 gene. It is concluded, therefore, that induction of defense responses in Cf9 tomato cells by the AVR9 elicitor is developmentally regulated and is absent in callus tissue and cell-suspension cultures, which consists of undifferentiated cells. These results are significant for the use of suspension-cultured cells to investigate signal transduction cascades.
来自真菌病原体番茄叶霉病菌的AVR9激发子可诱导防御相关反应,包括细胞死亡,这种情况特别发生在携带Cf-9抗性基因的番茄(Lycopersicon esculentum Mill.)植株中。为了详细研究抗性的生化机制,启动了携带Cf-9抗性基因的番茄细胞悬浮培养。用除AVR9之外的各种激发子处理细胞,可诱导氧化爆发、离子通量以及防御相关基因的表达。根癌农杆菌介导用含Avr9的构建体转化Cf9番茄叶盘,高效导致转基因愈伤组织形成。尽管转基因愈伤组织表现出正常的再生能力,但从未获得同时表达Cf-9和Avr9基因的转基因植株。由表达Avr9基因的番茄植株与野生型Cf9植株杂交产生的转基因F1幼苗在几周内死亡。然而,从这些幼苗的子叶上起始的愈伤组织培养物可维持至少3个月,并且其发育与仅含有Cf-9或Avr9基因的愈伤组织培养物相似。因此得出结论,AVR9激发子在Cf9番茄细胞中诱导的防御反应受到发育调控,并且在由未分化细胞组成的愈伤组织和细胞悬浮培养物中不存在。这些结果对于利用悬浮培养细胞研究信号转导级联具有重要意义。