植物抗病激活蛋白harpinXooc由水稻条斑病细菌hrp基因簇中hpa1基因编码。用构建于表达载体pET21a(+)上的hpa1诱导表达产物harpinXooc处理烟草,可激发烟草产生过敏反应,以及激活与烟草抗病信号途径相关的基因PR-1a、hin1和hsr203J的表达;处理水稻后,NPR1、OsPR1a、OsPR1b和PAL被激活。表明harpinXooc蛋白与植物互作后,通过水杨酸信号传导途径激活病程相关蛋白等防卫反应基因的转录表达,从而使植物产生系统获得抗病性。harpinXooc经加工后制成含量达1%的可溶性微颗粒制剂在水稻上进行应用,试验结果显示,harpinXooc蛋白可诱导水稻产生抗病性,防治水稻稻瘟病效果与杀菌剂稻瘟必克(三环唑)相当,防治水稻纹枯病和稻曲病效果与井冈霉素效果相当。对水稻增产的效果主要表现在增加粒实重上,增产达6%以上。
Harpin is a group of proteins produced by gram-negative plant pathogenic bacteria which triggers hypersensitive response in nonhost plants. Protein harpinXooc is encoded by the hpal gene in the hrp cluster of Xanthomonas oryzae pv. oryzicola. The gene was constructed into an expression vector pET21a ( + ) and the reconstruct was transformed into Escherichia coli BL21 (DE3). The engineered bacterium was induced to produce harpinXooc protein which triggers a hypersensitive response in tobacco. Reversetranscriptional polymerase chain reaction (RT-PCR) demonstrated that PR-1a, hin1 and hsr203J genes in tobacco, and NPR1, OsPR-1a, OsPR-1b, and PAL genes in rice were transcriptionally expressed after 8 h post spray with harpinXooc in induced through the salicylic acid si plants. This indicated that a systemic acquired resistance (SAR) was gnaling pathway marked with activation of PR-1a, hinl and hsr203J genes in tobacco and NPR1, OsPR-1a, and PAL genes in rice at 8 h point after harpinXooc interacted with tobacco and rice, respectively. The extracted protein was industrially developed into soluble granule, commercially termed Yilite as a bio-fungicide, containing 1% of harpinXooc. The biofungicide was sprayed twice in rice growing stage. Application of harpinXooc in rice showed that harpinXooc induced SAR in rice with control efficiencies equivalent to those by application of Tricyclazole and Jingangmycin, respectively, against Magnaporthe grisea, Rhizoctonia solani and Claviceps orysae-sativae. The rice yield was increased over 6% compared with that in the plots sprayed chemical fungicides. The factor for the yield increase was mainly due to the grain weight increased. This is the first report that hpal gene product induced systemic acquired resistance in plants and controlled rice diseases efficiently in the field.