Anderson M A., Van Heeswijck R, West J, Bateman K, Lee M, Christeller J T., McDonald G, Heath R L.
The Plant Cell Biology Research Centre, School of Botany, University of Melbourne, Parkville, Australia
J Insect Physiol. 1997 Sep;43(9):833-842. doi: 10.1016/s0022-1910(97)00026-7.
The ornamental tobacco (Nicotiana alata) produces one 6-kDa chymotrypsin inhibitor and four 6-kDa trypsin inhibitors from a single 40.3-kDa precursor protein. Three different approaches have been used to assess the potential of these proteinase inhibitors (PIs) in insect control. The first was an in-vitro approach in which all five inhibitors, the single chymotrypsin inhibitor or three of the four trypsin inhibitors were tested for their ability to inhibit gut protease activity in insects from four orders. The second approach was to incorporate the N. alata PIs in the artificial diet of the native budworm (Helicoverpa punctigera) and the black field cricket (Teleogryllus commodus). H. punctigera larvae and T. commodus nymphs had a significant (P<0.01) reduction in growth after ingestion of the PI and were more lethargic than insects on the control diet. Several of the H. punctigera larvae also failed to complete moulting at the third or fourth instar. The third approach was to express the N. alata PIs in transgenic tobacco under the control of the 35S CaMV promoter. When H. punctigera larvae were fed tobacco leaves expressing the N. alata PIs at 0.2% soluble protein, significant (P<0.01) differences in mortality and/or growth rate were observed.
观赏烟草（Nicotiana alata）从一个40.3 kDa的前体蛋白中产生一种6 kDa的胰凝乳蛋白酶抑制剂和四种6 kDa的胰蛋白酶抑制剂。已采用三种不同方法来评估这些蛋白酶抑制剂（PIs）在昆虫防治中的潜力。第一种是体外方法，其中测试了所有五种抑制剂，即单一的胰凝乳蛋白酶抑制剂或四种胰蛋白酶抑制剂中的三种，以评估它们抑制来自四个目的昆虫肠道蛋白酶活性的能力。第二种方法是将N. alata PIs掺入本地棉铃虫（Helicoverpa punctigera）和黑田蟋（Teleogryllus commodus）的人工饲料中。棉铃虫幼虫和黑田蟋若虫在摄入PI后生长显著（P<0.01）减缓，并且比食用对照饲料的昆虫更慵懒。一些棉铃虫幼虫在三龄或四龄时也未能完成蜕皮。第三种方法是在35S CaMV启动子的控制下在转基因烟草中表达N. alata PIs。当用表达0.2%可溶性蛋白的N. alata PIs的烟草叶喂养棉铃虫幼虫时，观察到死亡率和/或生长速率有显著（P<0.01）差异。
