灰霉菌中提取纯化16 kD的激活蛋白。番茄种子以20 mmol/L的HEPES缓冲液（pH 8.0）为对照，用2.5、5、10和20μg/mL的灰霉菌激活蛋白处理36 h，播种第45 d接种灰霉病，接种第7、14和21 d观察发病率；并用10μg/mL的激活蛋白溶液喷洒番茄幼苗第6、12、24、48、72和120 h取样测定苯丙氨酸解氨酶（PAL）、过氧化物酶（POD）和多酚氧化酶（PPO）活性。结果显示，10μg/mL激活蛋白处理番茄种子时，番茄抗病性最强，诱抗效果为48%～54%；番茄幼苗喷洒激活蛋白处理第24 h，PAL活性达峰值、比对照组提高54%，处理第72 h，POD和PPO活性达峰值、分别比对照组提高106%、122%。灰霉菌激活蛋白通过诱导抗病性相关酶的活性，提高番茄的抗病性，并且在最适浓度下诱导效果最佳。%The Botrytis cinerea hypha has been sonication with HEPES buffer (pH 8.0), and the crude protein solution was heated at 100 ℃ for 30 min, the protein solution filtrate was loaded on RESOURCETMQ 5 mL column at flow rate of 2 mL/min. Two peaks were collected, and the both peaks were analyzed by SDS-PAGE, and then the target protein (Botrytis cinerea activator protein) was detected hypersensitive response (HR) on tobacco leaves. Tomato seeds were subjected for a period of 45 days to 2.5, 5, 10 and 20μg/mL of Botrytis cinerea activator protein (the molecular weight is16 kD) solution treatments, and 20 mmol/L HEPES buffer (pH 8.0) was used as control. Then inoculated by spray of 105 cfu/mL the gray mold spore suspension and moisture 48 hours in incubator, and observed the morbidity at the 7, 14 and 21th days of inoculation;And sprayed 10μg/mL protein solutions on tomato seedlings, then measured the phenylalanine ammonia-lyase (PAL), peroxidase (POD) and polyphenol oxidase (PPO) activities at 6, 12, 24, 48, 72 and 120 hours of treatment. The results showed that, the linear elution peak showing a single band with Coomassie Brilliant Blue R-250 staining, and this protein could induce the HR in plants. The disease resistance increased by 48%, 55% and 54% in seed treatment tomato respectively, and it showed most resistant at 10μg/mL of the protein concentration. The PAL activity reached the peak value after 24 h of treatment, and it increased by 54%than that of the control. The activities of POD and PPO reached the peak value at 72 h of spray activator protein on tomato seedlings, and it increased by 106%and 122%than that of the control, respectively. Conclusion that, the activator protein involves the improvement disease-resistance by enhancement of the resistance-related enzyme activities in tomato, and the resistant effect was most efficient at the optimal concentration of the protein.
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