Bonghi C, Rascio N, Ramina A, Casadoro G
Institute of Pomology, University of Padua, Italy.
Plant Mol Biol. 1992 Dec;20(5):839-48. doi: 10.1007/BF00027155.
Ethylene-induced abscission in leaf and fruit explants of peach involves different enzymes. In leaves abscission is accompanied by increased occurrence of cellulase forms differing in isoelectric point (pI 6.5 and 9.5). A polypeptide with a molecular mass of 51 kDa gives in a western blot a strong cross-reaction with an antibody raised against a maturation cellulase from avocado fruit. Cellulase activity is also found in abscising fruit explants but the amount is very low compared to that of the leaf explants. A northern analysis with a cellulase clone from avocado reveals the presence of two hybridizing mRNAs with a size of 2.2 kb and 1.8 kb, respectively. The steady-state level of the 2.2 kb mRNA is significantly increased by treatment with ethylene. Polygalacturonases are not detected in abscising leaves, but are strongly induced by ethylene in fruit explants. Of the three forms found, two are exopolygalacturonases while the third is an endoenzyme. Ethylene activates preferentially the endoenzyme and the basic exoenzyme but depresses the acid exopolygalacturonases. A northern analysis carried out with a cDNA coding for tomato endopolygalacturonase shows hybridization only with one endopolygalacturonase mRNA form in the fruit abscission zone. Treatment with ethylene causes an increase in the steady-state level of this mRNA. The differences in the enzyme patterns observed in fruit and leaf abscission zones and a differential enzyme induction suggest the feasibility to regulate fruit abscission in peach with the aid of antisense RNA genes.
乙烯诱导的桃树叶和果实外植体脱落涉及不同的酶。在叶片中,脱落伴随着等电点不同(pI 6.5和9.5)的纤维素酶形式出现频率的增加。一种分子量为51 kDa的多肽在蛋白质免疫印迹中与针对鳄梨果实成熟纤维素酶产生的抗体发生强烈的交叉反应。在脱落的果实外植体中也发现了纤维素酶活性,但与叶片外植体相比,其含量非常低。用鳄梨的纤维素酶克隆进行的Northern分析显示存在两种杂交mRNA,大小分别为2.2 kb和1.8 kb。用乙烯处理可使2.2 kb mRNA的稳态水平显著增加。在脱落的叶片中未检测到多聚半乳糖醛酸酶,但在果实外植体中乙烯可强烈诱导其产生。在所发现的三种形式中,两种是外多聚半乳糖醛酸酶,第三种是内切酶。乙烯优先激活内切酶和碱性外多聚半乳糖醛酸酶,但抑制酸性外多聚半乳糖醛酸酶。用编码番茄内切多聚半乳糖醛酸酶的cDNA进行的Northern分析表明,在果实脱落区仅与一种内切多聚半乳糖醛酸酶mRNA形式杂交。用乙烯处理会导致这种mRNA的稳态水平增加。在果实和叶片脱落区观察到的酶模式差异以及不同的酶诱导作用表明,借助反义RNA基因调控桃果实脱落是可行的。