【作者】 黄世霞；

【导师】 王庆亚；

【作者基本信息】 南京农业大学 ， 植物学， 2004， 硕士

【摘要】 看麦娘(Alopecurus aequalis)为我国农田十大害草之一，危害大而又难以防除。尤其在我国长江中下游尤为严重。主要危害麦、油菜等夏收作物。本文以稻茬油菜田恶性禾本科杂草看麦娘为研究对象，研究了其种子休眠及萌发特性，开花生物学特性；并进行了看麦娘对高效盖草能、精禾草克及拿捕净产生抗药性的鉴定。结合形态解剖学、生理生化及分子生物学水平初步探讨了抗药性产生的机理。结果如下： 打破休眠不仅与处理物质有关也与处理时间有很大的关系。看麦娘种子于4℃条件下800ppm浓度的GA，为浸种液浸种5天，发芽率能大大提高至达80％以上。在不同种子储藏方式中，室温下土壤中浸水保存有利于其越夏，提高萌发率，也是稻油(麦)田看麦娘易于发生的原因之一。 采用种子测定法、整株测定法、花粉粒测定法进行了看麦娘对高效盖草能、精禾草克AOPP类除草剂产生的抗药性鉴定。表明采自溧阳(LYR)对高效盖草能产生高抗性，采自合肥的HFRⅠ，HFRⅡ，和江宁的JXRⅡ，JXRⅠ表现为中等水平的抗药性，抗性程度LYR＞HFRⅠ＞HFRⅡ＞JXRⅠ＞JXRⅡ。未使用除草剂处理过的油菜田或校园的看麦娘JJSⅠ，JJSⅡ，NAU，HFS均表现为敏感性。对精禾草克抗药性的研究表明JJSⅡ，NAU，HFS，JJSⅠ，仍为敏感性。LYR高抗；JXRⅡ＞HFRⅡ＞JXRⅠ＞HFRⅠ表现为抗性。 同时也进行了对拿捕净是否产生交互抗性的监测，发现南京江宁长期使用精禾草克、高效盖草能等乙酰辅酶A羧化酶抑制剂类除草剂约8年的油菜田里的看麦娘(JXRⅡ，JXRⅠ)对拿捕净产生较高水平的抗药性，HFRⅠ和HFRⅡ表现为中等水平的抗药性，LYR，JJSⅠ，JJSⅡ，NAU，HFS表现为敏感性。不同居群的看麦娘对拿捕净反应也有很大的区别，抗性程度JXRⅡ＞JXRⅡ＞HFRⅠ＞HFRⅠ＞LYR 选取抗(JXRⅡ)，感(JJSⅡ)两种看麦娘居群经高效盖草能茎叶处理(不用除草剂处理为对照)，处理后第6天左右，JJSⅡ的叶片电解质泄漏率极显著上升，比对照提高了160％；MDA则比对照上升了4倍，膜脂过氧化加剧；JJSⅡ的叶片的叶绿素含量比对照下降了约1.6倍，类胡萝卜素含量下降了约2倍；JJSⅡ的GSH含量在处理后2天就开始下降，均比对照低，而JXRⅡ的GSH含量均比对照高；抗、感居群经除草剂处理后，二者可溶性糖含量均增加，但敏感性居群JJSⅡ可溶性糖含量增加的幅度远远大于杭性居群JX租I;二者的POD表现也不同，处理后JXRJI的POD酶活却没有JJSll上升幅度大。说明这两种群体经除草剂胁迫后，除草剂对敏感性居群的生理变化影响大。 采用双向等位特异性聚合酶链式反应(B 1 d 1 reetional Axl ele一Specific PeR)，检测对拿捕净产生的抗性分子生物学基础，JXRll、JxR工能增出约SO0bp的特异片断，而其它只能扩.嘈出约780bp。推测该杭拿捕净生物型JXR工工、JXRI可脆是由于靶标酶ACCa se位点发.生突变，在研究其对高效盖草能产生杭药性分子基础时发现LYR能增出约49obp的片断，其它只能扩增出约1100bP，进一步证实了靶标酶AcCase可能存在多个突变位点，而产生不同模式的抗药性，同时也表明AOPP类和CHD类除草剂的作用位点是有差异的，更多还原

【Abstract】 Alopecurus Aequalis (S.) is one of the noxious weed in the field, which widely distributed in china, Japanese, kerora, and Turkey in Asian. Now it becomes a troublesome weed in our country, especially in the middle and lower region of Yangzi River. It mainly distributed in the field of wheat and oil rape. The Alopecurus Aequalis (S.) In the rape field was investigated in this paper, the dormancy, germination of the seed and biology of the anthesis of the species were studied, and the resistance to ACCase-inhibitor in Alopecurus Aequalis populations were identified. The modal anatomy, physiology and biochemistry, molecular biology was employed to discuss the mechanism of resistance. The results are as follows:There is exists 1-3 month’s dormancy for seeds of Alopecurus Aequalis, so it is necessary to find a rapid method to break the seed dormancy to make it germinate consistently. The result showed that it is related to the substance and the time of the treatment. The seed need to be immersed in GA3 with 800ppm concentration about 5 days, and the germination percentage will be enhanced to 80%. Treated with different preservation ways, storage in soil with immersed water is benefit to get through the summer that is one of the reasons of its occurrence easily in the field of rice-rape or rice-wheat.The bioassay to whole individual, to seeds and to pollen were used to detect the resistance of Alopecurus aequalis populations to haloxyfop-R, quizolofop-R , the results indicated that Alopecurus aequalis from the rape field without herbicide treatment is susceptible, while that population from the rape field in LiYang county where it has being used with the haloxyfop-R about 10 years has high resistance. The resistance level is: LYR>HFRI> HFRII> JXRI > JXRII.The cross- resistance in Alopecurus aequalis populations was also identified. The results showed that population of Alopecurus aequalis from the rape field without herbicide treatment is still susceptible to sethoxydim, and those populations of JXRII, JXRIfrom Jiangning whose resistance have been detected resistance to haloxyfop-R are high resistant to sethoxydim, while populations from LYR whose resistance are high to haloxyfop, but is susceptible to sethoxydim. This result suggested that different herbicide-resistant modes exist mAlopecurus aequalis populations.There was remarkable difference to Electrical conductivity, MDA, GSH, POD, chK car, soluble protein and sugar between the resistant and susceptible populations (JXRII, JJSII) after treated by haloxyfop-R ,This suggested the herbicide strongly affected the physiological and biochemical aspects of the susceptible biotype.After treated with herbicide for 6d, the Electrical conductivity of leaf in JJSII increased rapidly. The content of MDA rose four times in contrasted to the control, this implied membrane-lipid peroxidation was intensified. The content of chal and car decreased 1.6and2 times respectively; Two days later after treatment, GSH become lower, while the GSH from JXRII was higher contrasted to the control .the soluble sugar were increased both the susceptible and resistant biotype. But the former increased higher than the later, the activity of POD from JXRII increased slowly than that population from JJSII.The Bi-directional Allele-specific PCR was used to detect the base in molecular level of the resistance to sethoxydim. Only JXRII, JXRI can be amplified the SOObp specific-fragment, while others amplified the 780bp fragment. It is concluded that there probably exist the ACCase mutation within JXRII, JXRI. The base in molecular fragement level of the resistance to haloxyfop-R was studied, only LYR can be amplified the 490bp specific-fragment, while others amplified the 1100bp fragment which further verify there are many mutation points in the ACCase. At the same time, It also illuminates the target sites are different between AOPP and CHD.更多还原