中国生物防治学报
中國生物防治學報
중국생물방치학보
CHINESE JOURNAL OF BIOLOGICAL CONTROL
2014年
6期
780-786
,共7页
麦麦提艾力·热合曼%海利力·库尔班%郭立华%邱德文
麥麥提艾力·熱閤曼%海利力·庫爾班%郭立華%邱德文
맥맥제애력·열합만%해리력·고이반%곽립화%구덕문
激活蛋白%诱导抗病性%苯丙氨酸解氨酶%过氧化物酶%多酚氧化酶
激活蛋白%誘導抗病性%苯丙氨痠解氨酶%過氧化物酶%多酚氧化酶
격활단백%유도항병성%분병안산해안매%과양화물매%다분양화매
activator protein%induced resistance%phenylalanine ammonia lyase%peroxidase%polyphenol oxidase
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。
灰黴菌中提取純化16 kD的激活蛋白。番茄種子以20 mmol/L的HEPES緩遲液(pH 8.0)為對照,用2.5、5、10和20μg/mL的灰黴菌激活蛋白處理36 h,播種第45 d接種灰黴病,接種第7、14和21 d觀察髮病率;併用10μg/mL的激活蛋白溶液噴灑番茄幼苗第6、12、24、48、72和120 h取樣測定苯丙氨痠解氨酶(PAL)、過氧化物酶(POD)和多酚氧化酶(PPO)活性。結果顯示,10μg/mL激活蛋白處理番茄種子時,番茄抗病性最彊,誘抗效果為48%~54%;番茄幼苗噴灑激活蛋白處理第24 h,PAL活性達峰值、比對照組提高54%,處理第72 h,POD和PPO活性達峰值、分彆比對照組提高106%、122%。灰黴菌激活蛋白通過誘導抗病性相關酶的活性,提高番茄的抗病性,併且在最適濃度下誘導效果最佳。
회매균중제취순화16 kD적격활단백。번가충자이20 mmol/L적HEPES완충액(pH 8.0)위대조,용2.5、5、10화20μg/mL적회매균격활단백처리36 h,파충제45 d접충회매병,접충제7、14화21 d관찰발병솔;병용10μg/mL적격활단백용액분쇄번가유묘제6、12、24、48、72화120 h취양측정분병안산해안매(PAL)、과양화물매(POD)화다분양화매(PPO)활성。결과현시,10μg/mL격활단백처리번가충자시,번가항병성최강,유항효과위48%~54%;번가유묘분쇄격활단백처리제24 h,PAL활성체봉치、비대조조제고54%,처리제72 h,POD화PPO활성체봉치、분별비대조조제고106%、122%。회매균격활단백통과유도항병성상관매적활성,제고번가적항병성,병차재최괄농도하유도효과최가。
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.