Vogelsang R, Barz W
Westfälische Wilhelms-Universität, Institut für Biochemie und Biotechnologie der Pflanzen, Münster, FRG.
Planta. 1993 Jan;189(1):60-9. doi: 10.1007/BF00201344.
Chickpea (Cicer arietinum L.) cell-suspension cultures were used to isolate one beta-1,3-glucanase (EC 3.2.1.29) and two chitinases (EC 3.2.1.14). The beta-1,3-glucanase (M(r) = 36 kDa) and one of the chitinases (M(r) = 32 kDa) belong to class I hydrolases with basic isoelectric points (10.5 and 8.5, respectively) and were located intracellularly. The basic chitinase (BC) was also found in the culture medium. The second chitinase (M(r) = 28 kDa), with an acidic isoelectric point of 5.7, showed homology to N-terminal sequences of class III chitinases and represented the main protein accumulating in the culture medium. Polyclonal antibodies raised against the basic beta-1,3-glucanase (BG) and the acidic chitinase (AC) were shown to be monospecific. The anti-AC antiserum failed to recognize the BC on immune blots, confirming the structural diversity between class I and class III chitinases. Neither chitinase exhibited lysozyme activity. All hydrolases were endo in action on appropriate substrates. The BC inhibited the hyphal growth of several test fungi, whereas the AC failed to show any inhibitory activity. Expression of BG activity appeared to be regulated by auxin in the cell culture and in the intact plant. In contrast, the expression of neither chitinase was apparently influenced by auxin, indicating a differential hormonal regulation of beta-1,3-glucanase and chitinase activities in chickpea. After elicitation of cell cultures or infection of chickpea plants with Ascochyta rabiei, both system were found to have hydrolase patterns which were qualitatively and quantitatively comparable.(ABSTRACT TRUNCATED AT 250 WORDS)
鹰嘴豆（Cicer arietinum L.）细胞悬浮培养物被用于分离一种β-1,3-葡聚糖酶（EC 3.2.1.29）和两种几丁质酶（EC 3.2.1.14）。β-1,3-葡聚糖酶（分子量 = 36 kDa）和其中一种几丁质酶（分子量 = 32 kDa）属于具有碱性等电点（分别为10.5和8.5）的I类水解酶，且位于细胞内。碱性几丁质酶（BC）也存在于培养基中。第二种几丁质酶（分子量 = 28 kDa），等电点为酸性的5.7，与III类几丁质酶的N端序列具有同源性，是培养基中积累的主要蛋白质。针对碱性β-1,3-葡聚糖酶（BG）和酸性几丁质酶（AC）产生的多克隆抗体显示为单特异性。抗AC抗血清在免疫印迹中未能识别BC，证实了I类和III类几丁质酶之间的结构差异。两种几丁质酶均未表现出溶菌酶活性。所有水解酶对合适的底物均起内切作用。BC抑制了几种测试真菌的菌丝生长，而AC未表现出任何抑制活性。BG活性的表达似乎在细胞培养物和完整植株中受生长素调节。相比之下，两种几丁质酶的表达显然均不受生长素影响，表明鹰嘴豆中β-1,3-葡聚糖酶和几丁质酶活性存在不同的激素调节。在用拉宾斯壳二孢菌诱导细胞培养物或感染鹰嘴豆植株后，发现两个系统的水解酶模式在定性和定量上具有可比性。（摘要截短于250词）
