毛地黄皂苷诱导的水稻悬浮细胞防卫基因表达的信号传导  

SIGNAL TRANSDUCTION PATHWAYS OF DEFENSE GENE EXPRESSION IN RICE SUSPENSION CELLS INDUCED BY DIGITONIN

作　　者：王钧[1] 朱群 Christopher J.Lamb 

机构地区：[1]中国科学院上海植物生理研究所 [2]Salk Institute for Biological Studies,La Jolla,CA 92037,USA

出　　处：《实验生物学报》1997年第2期133-145,共13页Acta Biologiae Experimentalis Sinica

摘　　要：用带水稻苯丙氨酸解氨酶PAL 2启动子和GUS读码框嵌合基因的水稻悬浮细胞证明毛地黄皂苷能诱导水稻PAL 2的转录。NAD-PH、NADH、过氧化氢、超氧歧化酶、过氧化氢酶、甘露醇、1,2-羟基苯-3,5-二磺酸(Tiron)、paraquat、精氨酸、还原型和氧化型的谷胱甘肽、N-乙酰-L-半胱氨酸、二硫苏糖醇等都不影响毛地黄皂苷诱导的PAL2转导。只有在超氧歧化酶和过氧化氢酶或超氧歧化酶和Tiron一起时才微弱地抑制这种诱导。毛地黄皂苷能在缺钙培养基中诱导胞外介质碱性化,但它只在含钙离子的条件下才诱导PAL 2转录。钙离子载体A 23187及Stauros-poline不诱导PAL 2基因转录和胞外介质碱性化。钙调素的拮抗剂N-(6-Aminohexy1)-5-chloro-1-naphthalenesulfonamide(W7)和trifluoperazine(TFP)能抑制毛地黄皂苷诱导的PAL 2转录,但一些蛋白激酶的激活剂和抑制剂包括phorbol-12-myristate(TPA)、1-(5-Isoquinoline sulfony1)-2-methylpiperaz-ine(H7)、staurosporine及genestein不干扰它对PAL 2转录的诱导,G蛋白激活剂GTP-γS和抑制剂百日咳毒素也不影响这种诱导。毛地黄皂苷不诱导几丁酶转录。二硫苏糖醇能抑制几丁酶转录。伴刀豆球蛋白及麦胚凝集素能微弱地促进几丁酶基因转录,但对PAL 2表达没有影响。看来毛地黄皂苷诱导的水稻悬浮细胞PAL 2转录与活性氧激活没有简单和直接的关系。但它依赖于胞外介质碱性化和钙-钙调素,它的信号传导途径与诱导几丁酶转录的信号传导途径是不同的。A transgenic rice suspension cell cultures expressing rice phenylalinine am-monia-lyase PAL 2 promoter-GUS fusion gene was established. The GUS activity and PAL 2 transcription of this cultures could be induced by digitonin. The PAL 2 transcription induced by digitonin was not affected by treated with NADPH, NADH, hydrogen peroxide, superoxide di-smutase (SOD), 1,2-dihydroxybenzene-3, 5-disulfonic acid (Tiron), catalase, man-nitol, paraquat, arginine, glutathione reduced (GSH), glutathione oxidative (GS-SG), N-acetyl-L-cysteine (NAC), and dithiothreitol (DTT) alone. However, it was weakly inhibited by the combination between SOD and catalase or SOD and Tiron. Digitonin could induce extracellular alkalization in rice suspension culture even in calcium free medium, but it could not induce PAL 2 transcription in this condition. Calcium ionophone A 23187 and staurospoline could not induce PAL 2 transcription, nor extracellular alkalization. The antagonists of calmodulin, trif-luoperazine (TFP) and (6-aminohexyl)-5 -chloro-1-naphthalenesulfonamide (W 7) could weakly inhibit digitonin-induced PAL 2 transcription, but the stimulator and inhibitors of some protein kinases such as phorbol-12-myristate (TPA), st-aurosporine, 1-(5-isoquinoline sulfonyl)-2-methylpiperazine (H 7) and genistein could not affect this induction. The stimulator and inhibitor of G protein, GTP-γS and pertussis toxin could not affect this induction either. Digitonin could not induce chitinase transcription. Dithiothreitol (DTT) could inhibit chitinase transcription. Concanavalin A (Con A) and wheat germ agglutinin (WGA) could weakly induce transcription of chitinase, but not PAL gene expression. It seems that the PAL 2 transcription induced by digitonin is not simply and directly related to oxidative burst, but depends on extracellular calcium ion, extracellular alkalization and calmodulin, its signal transduc-tion pathway is different from the induction of chitinase gene in rice suspension cultures.

关 键 词：毛地黄皂苷 水稻 基因表达 信号传导 悬浮细胞 

分 类 号：S511.01[农业科学—作物学]