Büchter R, Strömberg A, Schmelzer E, Kombrink E
Max-Planck-Institut für Züchtungsforschung, Abteilung Biochemie, Köln, Germany.
Plant Mol Biol. 1997 Dec;35(6):749-61. doi: 10.1023/a:1005830706507.
Infection of potato (Solanum tuberosum) leaves by the late blight fungus Phytophthora infestans or treatment with fungal elicitor leads to a strong increase in chitinase activity. We isolated cDNAs encoding acidic (class II) chitinases (ChtA) from potato leaves and determined their structures and expression patterns in healthy and stressed plants. From the total number of cDNAs and the complexity of genomic DNA blots we conclude that acidic chitinase in potato is encoded by a gene family which is considerably smaller than that encoding basic (class I) chitinase (ChtB). The deduced amino acid sequences show 78 to 96% identity to class II chitinases from related plant species tomato, tobacco) whereas the identity to basic chitinases of potato is in the range of 60%. RNA blot analysis revealed that both acidic and basic chitinases were strongly induced by infection or elicitor treatment and that the induction occurred both locally at the site of infection and systemically in upper uninfected leaves. In contrast, a differential response to other types of stress was observed. Acidic chitinase mRNA was strongly induced by salicylic acid, whereas basic chitinase mRNA was induced by ethylene or wounding. In healthy, untreated plants, acidic chitinase mRNA accumulated also in an organ-, cell-type- and development-specific manner as revealed by RNA blot analysis and in situ RNA hybridization. Relatively high transcript levels were observed in old leaves and young internodes as well as in vascular tissue and cells constituting the stomatal complex in leaves and petioles. Lower, but appreciable mRNA levels were also detectable in roots and various flower organs, particularly in sepals and stamens. The possible implications of these findings in pathogen defense, development and growth processes are discussed.
晚疫病菌(致病疫霉)感染马铃薯(茄属)叶片或用真菌激发子处理会导致几丁质酶活性大幅增加。我们从马铃薯叶片中分离出编码酸性(II类)几丁质酶(ChtA)的cDNA,并确定了它们在健康和受胁迫植物中的结构及表达模式。根据cDNA总数和基因组DNA印迹的复杂性,我们得出结论:马铃薯中的酸性几丁质酶由一个基因家族编码,该家族比编码碱性(I类)几丁质酶(ChtB)的基因家族小得多。推导的氨基酸序列与相关植物物种番茄、烟草的II类几丁质酶有78%至96%的同一性,而与马铃薯碱性几丁质酶的同一性在60%左右。RNA印迹分析表明,酸性和碱性几丁质酶均受感染或激发子处理强烈诱导,且诱导在感染部位局部发生,并在未感染的上部叶片中系统性发生。相比之下,观察到对其他类型胁迫的差异反应。酸性几丁质酶mRNA受水杨酸强烈诱导,而碱性几丁质酶mRNA受乙烯或创伤诱导。如RNA印迹分析和原位RNA杂交所示,在健康、未处理的植物中,酸性几丁质酶mRNA也以器官、细胞类型和发育特异性方式积累。在老叶和幼嫩节间以及维管组织和构成叶片及叶柄气孔复合体的细胞中观察到相对较高的转录水平。在根和各种花器官中也可检测到较低但可观的mRNA水平,特别是在萼片和雄蕊中。讨论了这些发现对病原体防御、发育和生长过程的可能影响。