OCCURRENCE OF 9.5 CELLULASE AND OTHER HYDROLASES IN FLOWER REPRODUCTIVE-ORGANS UNDERGOING MAJOR CELL-WALL DISRUPTION
OCCURRENCE OF 9.5 CELLULASE AND OTHER HYDROLASES IN FLOWER REPRODUCTIVE-ORGANS UNDERGOING MAJOR CELL-WALL DISRUPTION
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DOI：
10.1104/pp.99.3.1015
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发表时间：
1992-07-01
期刊：
影响因子：
7.4
通讯作者：
LEWIS, LN
中科院分区：
文献类型：
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作者：
DELCAMPILLO, E;LEWIS, LN
在菜豆（Phaseolus vulgaris）花生殖器官中研究了与豆叶脱落相关的酶的发生，其中分解代谢细胞壁事件在花药和雌蕊发育过程中至关重要。在开花前的豆花雌蕊和花药中检测到高水平的纤维素酶活性。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，随后用 9.5 纤维素酶抗体进行免疫印迹，鉴定出花药和雌蕊中的蛋白质具有相同的大小（51 千道尔顿），并且在血清学上与脱落纤维素酶密切相关。花药中 9.5 纤维素酶蛋白的积累受到发育调节，并从花药发育非常年轻阶段的不可检测水平增加到花药成熟时的高水平。在雌蕊中，9.5纤维素酶位于雌蕊的上部，柱头和花柱颈所在的位置，并且在所分析的最年轻的发育阶段中检测到。抗碱性几丁质酶的抗体在暴露于乙烯后在脱落区积累到高水平，在花药和雌蕊上部鉴定出具有相同大小（33千道尔顿）且血清学密切相关的蛋白质。相比之下，在豆类生殖器官中未检测到 45 千道尔顿的蛋白质和与脱落相关的碱性 β-1,3-葡聚糖酶。有趣的是，在幼豆花药中检测到 β-1,3-葡聚糖酶活性，并在开花时降低，但花药 β-1,3-葡聚糖酶在血清学上与碱性 β-1,3-葡聚糖酶无关。因此，似乎碱性纤维素酶和几丁质酶在许多需要主要细胞壁破坏的植物过程中组合出现，而半纤维素酶如β-1,3-葡聚糖酶是每个过程所特有的。
The occurrence of enzymes associated with bean leaf abscission was investigated in bean (Phaseolus vulgaris) flower reproductive organs in which catabolic cell wall events are essential during anther and pistil development. Cellulase activity was detected in high levels in both pistil and anthers of bean flowers before anthesis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by immunoblotting with 9.5 cellulase antibody identified a protein in anthers and pistil with the same size (51 kilodaltons) and serologically closely related to the abscission cellulase. The accumulation of 9.5 cellulase protein in the anther is developmentally regulated and increases from undetectable levels at very young stages of anther development to high levels as the anther matures. In the pistil, the 9.5 cellulase was localized in the upper part of the pistil where the stigma and the stylar neck reside and was detected in the youngest developmental stage analyzed. Antibodies against basic chitinase, which accumulates to high levels in abscission zones after exposure to ethylene, identified a protein with the same size (33 kilodaltons) and serologically closely related, in both anthers and upper portion of the pistil. In contrast, a 45-kilodalton protein and the basic beta-1,3-glucanase associated with abscission were undetected in bean reproductive organs. Interestingly, beta-1,3-glucanase activity was detected in young bean anthers and decreased at anthesis, but the anther beta-1,3-glucanase is serologically unrelated to the basic beta-1,3-glucanase. Thus, it appears that the basic cellulase and chitinase occur in combination in many plant processes that require major cell wall disruption, whereas hemicellulases such as beta-1,3-glucanase are specific to each process.