Meyer B, Houlné G, Pozueta-Romero J, Schantz M L, Schantz R
Institut de Biologie Moléculaire des Plants, Université Louis Pasteur, Strasbourg, France.
Plant Physiol. 1996 Oct;112(2):615-22. doi: 10.1104/pp.112.2.615.
We have isolated a 454-bp cDNA that encodes a novel fruit specific defensin from bell pepper (Capsicum annuum). The encoded 75-amino-acid polypeptide contains an N-terminal domain characteristic of a signal peptide and a 48-amino-acid mature domain named J1. The mature protein, from which the N-terminal amino acid sequence was determined, contains eight cysteines that from four intramolecular disulfide bridges, suggesting a monomeric form for J1. In healthy fruits J1 is undetectable at the green stage but high levels accumulate during ripening. In wound areas of the green fruit the accumulation of J1 dramatically increased, suggesting a role for J1 in the plant's defense response. Moreover, we have demonstrated that J1 possesses an antifungal activity. We have isolated and characterized the corresponding two homologous genes (j1-1 and j1-2) that exist in the bell pepper genome. Both genes are interrupted by the insertion, at the same position, of one intron of 853 bp for j1-1 and 4900 bp for j1-2. Northern blot and reverse transcriptase-polymerase chain reaction and restriction fragment length polymorphism analyses revealed that j1-1 transcripts are present only in fruits, only in trace amounts in mature green fruits, and that they accumulate to high levels in fully ripe fruits, whereas no j1-2 transcripts were detected in the samples monitored.
我们从甜椒(辣椒属)中分离出一个454碱基对的cDNA,它编码一种新的果实特异性防御素。所编码的75个氨基酸的多肽包含一个具有信号肽特征的N端结构域和一个名为J1的48个氨基酸的成熟结构域。确定了N端氨基酸序列的成熟蛋白含有八个半胱氨酸,形成四个分子内二硫键,表明J1为单体形式。在健康果实中,J1在绿色阶段不可检测,但在成熟过程中大量积累。在绿色果实的伤口部位,J1的积累显著增加,表明J1在植物防御反应中起作用。此外,我们已证明J1具有抗真菌活性。我们分离并鉴定了甜椒基因组中存在的两个相应的同源基因(j1-1和j1-2)。两个基因都在相同位置被一个内含子插入打断,j1-1的内含子为853碱基对,j1-2的内含子为4900碱基对。Northern印迹、逆转录-聚合酶链反应和限制性片段长度多态性分析表明,j1-1转录本仅存在于果实中,在成熟绿色果实中含量极少,在完全成熟的果实中积累到高水平,而在所监测的样品中未检测到j1-2转录本。