Centre de Recherche on Horticulture, pavillon de l'Envirotron, Université Lavai, G1K 7P4, Québec, Canada.
Plant Cell Rep. 1995 Jun;14(9):585-8. doi: 10.1007/BF00231943.
Transformation of potato (Solanum tuberosum L.) with cysteine proteinase inhibitor (PI) genes represents a potential way of controlling the major insect pest Colorado potato beetle (CPB; Leptinotarsa decemlineata Say). The present study describes the Agrobacterium-mediated transformation of potato (cv. Kennebec) with an oryzacystatin I (OCI) cDNA clone linked to a CaMV 35S promoter. The transgenic plants accumulated active OCI in potato leaves, as demonstrated by the papain-inhibitory activity of transgenic plant leaf extracts. In addition to their anti-papain activity, the extracts also caused a partial but significant inhibition of CPB digestive proteinases, similar to that observed with pure inhibitors. Recombinant OCI did not alter the activity of the major potato leaf endogenous proteinases, which seemed to be of the serine-type. Therefore we suggest that the OCI cDNA can be used for the production of CPB-resistant transgenic potato plants without interfering with endogenous proteinases of these plants.
利用半胱氨酸蛋白酶抑制剂（PI）基因转化马铃薯（Solanum tuberosum L.）是控制主要害虫——马铃薯甲虫（CPB；Leptinotarsa decemlineata Say）的一种潜在方法。本研究描述了利用携带玉米蛋白酶抑制剂 I（OCI） cDNA 克隆的根癌农杆菌介导转化马铃薯（品系 Kennebec），该克隆与 CaMV 35S 启动子相连。转基因植物在马铃薯叶片中积累了有活性的 OCI，这可以通过木瓜蛋白酶抑制活性的转基因植物叶片提取物来证明。除了抗木瓜蛋白酶活性外，提取物还对 CPB 消化蛋白酶有部分但显著的抑制作用，类似于纯抑制剂的作用。重组 OCI 不会改变主要马铃薯叶片内源性蛋白酶的活性，这些蛋白酶似乎属于丝氨酸型。因此，我们建议可以利用 OCI cDNA 生产抗 CPB 的转基因马铃薯植株，而不会干扰这些植物的内源性蛋白酶。
