油菜田日本看麦娘对高效氟吡甲禾灵抗药性的初步研究
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摘要
本文以我国长江中下游地区油菜田恶性杂草日本看麦娘(Alopecurus japonicus)为主要研究对象，初步研究了其种子休眠解除及萌发温度条件；通过种子生物测定法和整株测定法测定了不同地区不同用药年限油菜田日本看麦娘种群对乙酰辅酶A羧化酶(Acetyl coenzyme A carboxylase，简称ACCase)抑制剂类除草剂高效氟吡甲禾灵(haloxyfop-R-methyl)的抗药性及对其它药剂的交互抗性水平；并采用植物抗性生理生化测定技术，初步研究了日本看麦娘对高效氟吡甲禾灵产生抗性的机理。
对日本看麦娘种子休眠及萌发适宜温度的研究表明：日本看麦娘萌发适宜温度为15-20℃。通过地下沉积10天，滁州和合肥种子即可解除休眠，萌发率达87.5％和90％；而句容点种子不能被解除，萌发率仅为1.67％。采用低温(4℃)下清水浸泡对三地种子都可解除休眠，且萌发时间也较一致。说明日本看麦娘种子存在一定的休眠，但通过短暂的低温、较高的湿度条件即可解除休眠。
采用种子生物测定法和整株测定法对江苏、安徽11个点不同用药年限油菜田日本看麦娘对高效氟吡甲禾灵的抗药性测定结果基本相符。种子生物测定结果显示，江苏句容点日本看麦娘对此药剂的相对抗性倍数达61.13，整株测定结果为1331.68，说明句容点的日本看麦娘对此药剂已产生明显抗性。交互抗性检测结果显示，日本看麦娘对芳氧基苯氧基丙酸酯类(aryloxyphenoxypropionates，简称AOPPs)其它药剂已产生了不同程度的抗性，抗性水平依次为精吡氟禾草灵(fluazifop-P-butyl)＞精喹禾灵(quizalofop-P-ethyl)＞精噁唑禾草灵(fenoxaprop-P-ethyl)；对具相同作用靶标的环已烯酮类(cyclohexanediones，简称CHDs)除草剂烯禾啶(sethoxydim)的抗性水平相对较低。室内测定结果与田间反映的实际情况吻合。
以抗性(句容)和敏感(滁州)两种日本看麦娘种群为试验材料，研究了高效氟吡甲禾灵对这两地日本看麦娘抗氧化酶(antioxidase)活性、活性氧(active oxygen)、还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)、丙二醛(MDA)、脯氨酸(proline)、叶绿素(chlorophyll)和可溶性蛋白质(soluble protein)含量的影响。结果表明：药剂处理后，敏感种群超氧化物歧化酶(SOD)活性急剧下降，过氧化物酶(POD)活性在药后14天迅速上升，过氧化氢酶(CAT)活性先升高，后降低，而抗性种群SOD活性保持稳定，POD和CAT活性都先升高后降低，但CAT活性变化幅度比敏感种群小：敏感种群GSH含量随着时间的推移不断升高，超氧阴离子(O_2·~-)、过氧化氢(H_2O_2)、GSSG和MDA的含量也都逐渐提高，抗性种群则保持不变；敏感种群脯氨酸含量显著提高，叶绿素含量先升高后下降，而抗性种群则无显著变化；两种群可溶性蛋白质含量都先降低后升高，但抗性种群含量高于敏感种群。采用同位素标记法测定了抗性及敏感种群靶标酶ACCase活性，结果表明高效氟吡甲禾灵明显抑制了敏感种群ACCase活性。说明高效氟吡甲禾灵对敏感种群的生理生化指标及靶标酶活性影响较大。
In this paper, the Japanese Foxtail (Alopecurus japonicus Steud.), a kind of worstgrass weed in oilseed rape fields in the middle and lower reach of Yangtze valley in China,was chosen as a main research object. The methods of breaking dormancy of A.japonicusseeds and the optimal germinating temperature were studied, the resistance level of A.japonicus to haloxyfop-R-methyl, which is an ACCase inhibitor belonging toaryloxyphenoxypropionates (AOPPs), cross-resistance and the resistacne mechanism of A.japonicus was investigated.
It was found that the optimal germinating temperature range of A.japonicus was 15—20℃. The dormancy of seeds from Chuzhou and Hefei could be broken by being buriedunderground 7 cm for 10 d, whose germination rates reach to 87.5％and 90％,respectively. While the germinate rate of seeds from Jurong was only 1.67％. Dormancycould also be broken by being soaked in water at 4℃, and Jurong, Chuzhou and Hefeipopulations could germinate at the same time. It was indicated that the short dormancy of A.japonicus seeds could be broken by lower temperature and higer humidity.
The resistance level of A. japonicus to haloxyfop-R-methyl were evaluated byseed-bioassay and whole-plant test, which were collected from different oilseed rape fieldswhere haloxyfop-R-methyl had been continuously applied for different years, and similarresults were obtained by both of methods. In these two methods, the resistance index of A.japonicus to haloxyfop-R-methyl, which were collected from Jurong in Jiangsu province,were 61.13 and 1331.68, respectively, which demonstrated the emergence of A. japonicuspopulation resistant to haloxyfop-R-methyl in fields. The results of cross-resistance revealed that this resistant population was also resistant to some other AOPPs, in which theresistance level was: fluazifop-P-butyl>quizalofop-P-ethyl>fenoxaprop-P-ethyl.However, a relative low resistance level of this resistant population to sethoxydim wasobserved, which was a cyclohexanedione (CHD) herbicide sharing the same target site withAOPPs. And the results obtained in laboratory were agreed with the real fact in fields.
To characterize the biochemical differences in haloxyfop-R-methyl-resistant and-sensitive populations of A. japonicus collected from Jurong and Chuzhou, the activities ofantioxidant oxygens, content of reduced/oxydized glutathione (GSH/GSSG),malondialdehyde (MDA), proline, chlorophyll and soluable protein in shoots and leaves ofindividuals treated by haloxyfop-R-methyl were studied. The results showed that theactivities of superoxide dismutase (SOD) decreased significantly in sensitive population,but kept stable in resistant population. The activity of peroxidase (POD) in sensitivepopulation rose obviously in 14 days after treatment, while the eatalase (CAT) increased inearlier period and then decreased, the trends of activities of POD and CAT in resistantpopulation were the same as that of CAT in sensitive population. The content of GSH keptincreasing in experiments, which could also be observed in the contents of superoxideanion radical (O_2·~-), hydrogen peroxide (H_2O_2), GSSG, MDA and proline, but not changingin the resistant population. The trends of soluable protein in both populations were oppositeto that of Chlorophyll content in sensitive population. In addition, the activities of aeetylcoenzyme A carboxylase (ACCase) in both of the population were determined. The resultdemonstrated that ACCase activity in sensitive population was inhibited obviously byhaloxyfop-R-methyl. It may be concluded that this herbicide greatly affected some of thephysiological and biochemical pathways in sensitive population, except for ACCase.
对日本看麦娘种子休眠及萌发适宜温度的研究表明：日本看麦娘萌发适宜温度为15-20℃。通过地下沉积10天，滁州和合肥种子即可解除休眠，萌发率达87.5％和90％；而句容点种子不能被解除，萌发率仅为1.67％。采用低温(4℃)下清水浸泡对三地种子都可解除休眠，且萌发时间也较一致。说明日本看麦娘种子存在一定的休眠，但通过短暂的低温、较高的湿度条件即可解除休眠。
采用种子生物测定法和整株测定法对江苏、安徽11个点不同用药年限油菜田日本看麦娘对高效氟吡甲禾灵的抗药性测定结果基本相符。种子生物测定结果显示，江苏句容点日本看麦娘对此药剂的相对抗性倍数达61.13，整株测定结果为1331.68，说明句容点的日本看麦娘对此药剂已产生明显抗性。交互抗性检测结果显示，日本看麦娘对芳氧基苯氧基丙酸酯类(aryloxyphenoxypropionates，简称AOPPs)其它药剂已产生了不同程度的抗性，抗性水平依次为精吡氟禾草灵(fluazifop-P-butyl)＞精喹禾灵(quizalofop-P-ethyl)＞精噁唑禾草灵(fenoxaprop-P-ethyl)；对具相同作用靶标的环已烯酮类(cyclohexanediones，简称CHDs)除草剂烯禾啶(sethoxydim)的抗性水平相对较低。室内测定结果与田间反映的实际情况吻合。
以抗性(句容)和敏感(滁州)两种日本看麦娘种群为试验材料，研究了高效氟吡甲禾灵对这两地日本看麦娘抗氧化酶(antioxidase)活性、活性氧(active oxygen)、还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)、丙二醛(MDA)、脯氨酸(proline)、叶绿素(chlorophyll)和可溶性蛋白质(soluble protein)含量的影响。结果表明：药剂处理后，敏感种群超氧化物歧化酶(SOD)活性急剧下降，过氧化物酶(POD)活性在药后14天迅速上升，过氧化氢酶(CAT)活性先升高，后降低，而抗性种群SOD活性保持稳定，POD和CAT活性都先升高后降低，但CAT活性变化幅度比敏感种群小：敏感种群GSH含量随着时间的推移不断升高，超氧阴离子(O_2·~-)、过氧化氢(H_2O_2)、GSSG和MDA的含量也都逐渐提高，抗性种群则保持不变；敏感种群脯氨酸含量显著提高，叶绿素含量先升高后下降，而抗性种群则无显著变化；两种群可溶性蛋白质含量都先降低后升高，但抗性种群含量高于敏感种群。采用同位素标记法测定了抗性及敏感种群靶标酶ACCase活性，结果表明高效氟吡甲禾灵明显抑制了敏感种群ACCase活性。说明高效氟吡甲禾灵对敏感种群的生理生化指标及靶标酶活性影响较大。
In this paper, the Japanese Foxtail (Alopecurus japonicus Steud.), a kind of worstgrass weed in oilseed rape fields in the middle and lower reach of Yangtze valley in China,was chosen as a main research object. The methods of breaking dormancy of A.japonicusseeds and the optimal germinating temperature were studied, the resistance level of A.japonicus to haloxyfop-R-methyl, which is an ACCase inhibitor belonging toaryloxyphenoxypropionates (AOPPs), cross-resistance and the resistacne mechanism of A.japonicus was investigated.
It was found that the optimal germinating temperature range of A.japonicus was 15—20℃. The dormancy of seeds from Chuzhou and Hefei could be broken by being buriedunderground 7 cm for 10 d, whose germination rates reach to 87.5％and 90％,respectively. While the germinate rate of seeds from Jurong was only 1.67％. Dormancycould also be broken by being soaked in water at 4℃, and Jurong, Chuzhou and Hefeipopulations could germinate at the same time. It was indicated that the short dormancy of A.japonicus seeds could be broken by lower temperature and higer humidity.
The resistance level of A. japonicus to haloxyfop-R-methyl were evaluated byseed-bioassay and whole-plant test, which were collected from different oilseed rape fieldswhere haloxyfop-R-methyl had been continuously applied for different years, and similarresults were obtained by both of methods. In these two methods, the resistance index of A.japonicus to haloxyfop-R-methyl, which were collected from Jurong in Jiangsu province,were 61.13 and 1331.68, respectively, which demonstrated the emergence of A. japonicuspopulation resistant to haloxyfop-R-methyl in fields. The results of cross-resistance revealed that this resistant population was also resistant to some other AOPPs, in which theresistance level was: fluazifop-P-butyl>quizalofop-P-ethyl>fenoxaprop-P-ethyl.However, a relative low resistance level of this resistant population to sethoxydim wasobserved, which was a cyclohexanedione (CHD) herbicide sharing the same target site withAOPPs. And the results obtained in laboratory were agreed with the real fact in fields.
To characterize the biochemical differences in haloxyfop-R-methyl-resistant and-sensitive populations of A. japonicus collected from Jurong and Chuzhou, the activities ofantioxidant oxygens, content of reduced/oxydized glutathione (GSH/GSSG),malondialdehyde (MDA), proline, chlorophyll and soluable protein in shoots and leaves ofindividuals treated by haloxyfop-R-methyl were studied. The results showed that theactivities of superoxide dismutase (SOD) decreased significantly in sensitive population,but kept stable in resistant population. The activity of peroxidase (POD) in sensitivepopulation rose obviously in 14 days after treatment, while the eatalase (CAT) increased inearlier period and then decreased, the trends of activities of POD and CAT in resistantpopulation were the same as that of CAT in sensitive population. The content of GSH keptincreasing in experiments, which could also be observed in the contents of superoxideanion radical (O_2·~-), hydrogen peroxide (H_2O_2), GSSG, MDA and proline, but not changingin the resistant population. The trends of soluable protein in both populations were oppositeto that of Chlorophyll content in sensitive population. In addition, the activities of aeetylcoenzyme A carboxylase (ACCase) in both of the population were determined. The resultdemonstrated that ACCase activity in sensitive population was inhibited obviously byhaloxyfop-R-methyl. It may be concluded that this herbicide greatly affected some of thephysiological and biochemical pathways in sensitive population, except for ACCase.
引文
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