Lawrence Susan D, Novak Nicole G
United States Department of Agriculture, Agricultural Research Service, Insect Biocontrol Laboratory, BARC-West, Bldg. 011A, Room 214, Beltsville, MD 20705, USA.
Biotechnol Lett. 2006 Apr;28(8):593-9. doi: 10.1007/s10529-006-0022-7.
The previously described poplar chitinase, WIN6, is induced during infestation by gypsy moth (Lymantria dispar L.) larvae, thus suggesting a role in defense against insect pests. To test this hypothesis, we produced tomato seedlings infected with a recombinant potato virus X (PVX), which produces WIN6, and tested its insecticidal properties on Colorado potato beetle [CPB; Leptinotarsa decemlineata (Say)], which is a serious pest of tomatoes and other crops. The advantage of PVX is that plant material is ready for insect bioassay within 3-4 weeks of constructing the recombinant virus. Considering that production of transgenic tomato seedlings using Agrobacterium takes at least 6 months, this hastens the rate at which genes can be examined. Upon insect bioassay, only 47% CPB neonates feeding on leaves containing >0.3% w/w WIN6 developed to 2nd instar while 93% of controls reached 2nd instar. To our knowledge this is the first plant chitinase that retards development of an insect pest.
之前描述的杨树几丁质酶WIN6,在舞毒蛾(Lymantria dispar L.)幼虫侵染期间被诱导产生,因此表明其在抵御害虫方面发挥作用。为了验证这一假设,我们培育了感染重组马铃薯X病毒(PVX)的番茄幼苗,该病毒可产生WIN6,并测试了其对科罗拉多马铃薯甲虫[CPB;马铃薯叶甲(Leptinotarsa decemlineata (Say))]的杀虫特性,这种甲虫是番茄和其他作物的严重害虫。PVX的优势在于,在构建重组病毒后的3 - 4周内,植物材料即可用于昆虫生物测定。考虑到利用农杆菌生产转基因番茄幼苗至少需要6个月,这加快了基因检测的速度。经过昆虫生物测定,取食含>0.3% w/w WIN6叶片的CPB幼虫中,只有47%发育到二龄,而对照组有93%发育到二龄。据我们所知,这是第一种能延缓害虫发育的植物几丁质酶。