新型抗烟草花叶病毒的药物筛选及作用机理初步研究

【作者】 严凯；

【导师】 宋宝安； 黄荣茂；

【作者基本信息】 贵州大学 ， 农药学， 2008， 硕士

【摘要】 本论文采用半叶枯斑法对本中心合成的200多个抗病毒新化合物进行室内生物活性测试,筛选发现了具有较高抗TMV活性的氰基丙烯酸酯类化合物2005196和2006172,即3-苄胺基-3-[1-（R）-4-氟苯基乙基胺基]-2-氰基丙烯酸酯和3-甲硫基-3-（二乙氧基乙基）膦酰基-2-氰基丙烯酸酯,在500μg/mL剂量下的活体治疗抑制率分别为55.6%和53.3%,活体治疗效果与对照药剂宁南霉素相当（活体治疗500μg/mL,51.7%）,在活体钝化条件下的抑制率分别达52.5%和80.8%,在活体保护条件下的抑制率分别达60.4%和44.7%;化合物2005196和2006172对TMV活体治疗的EC₅₀值分别为218.7μg/mL和275.4μg/mL。本文进行了抗TMV病盆栽药效试验研究,2005196和2006172药剂处理在250μg/mL剂量下的病情指数均明显低于空白对照区（P＜0.05）,2005196处理组第45天的防治效果达60.6%。同时观察发现,喷施2005196后,烟草叶片的颜色明显变绿。以上结果表明半叶法摩擦接种的室内活体治疗效果与盆栽试验药效具有一定可比性,这可能意味着2005196在室内生测心叶烟与盆栽试验普通烟K_-326叶片中的作用机制具有类似性。因此,本文进行了2005196和2006172抗TMV生化机理初步研究,证明了2005196和2006172处理接种TMV后的烟草植株中,PAL活性、POD活性、SOD活性、PPO活性及叶绿素含量等相关酶和调控物质在一定的时间内都具有相关性。2005196明显诱导了抗性相关酶活性的升高;同时也诱导了叶绿素含量的增加。研究了药剂对烟草病程相关蛋白（PR）的诱导作用,结果显示:药剂2005196诱导烟叶胞间蛋白的种类和TMV刺激烟叶产生的胞间蛋白的种类基本一致。更多还原

【Abstract】 Bioassay experiment by half-leaf method revealed that the compounds 2005196 （3-benzylamino-3-[1-（R）-4-fluorophenyl ethyl amidocyanogen]-2-cyanoacrylate） and 2006172 （3-methylthio-3-（diethoxy ethyl）phosphonyl-2-cyanoacrylate） had comparable curative activities of 55.6% and 53.3% with the standard Ningnanmycin（51.7%） against TMV at 500μg/mL in vivo. The median effective concenteations（EC5o） values were 218.7μg/mL and 275.4μg/mL, respectively. In addition, 2005196 and 2006172 had inactivation activities of 52.5% and 80.8%, protection activities of 60.4% and 44.7% respectively in vivo.On the basis of experimental results, a potted plant survey in lab on the bioactivities of the compounds against TMV on Nicotiana glutinosa was conducted. Result showed that the compound 2005196 led to the lysis of TMV symptom, with a the curative inhibition rate of 60.6% at 250μg/mL after 45 days （P<0.05）. In addition, tobacco leaves followed by treatment with the compound 2005196 were found to become green as compared to that of the control in lab. It implied that the curative effect in vivo by half-leaf method was comparable to the protective effect in lab. Furthermore, it also indicated that the compound 2005196 acted on both leaves of Nicotiana glutinosa indoor and that of K-₃26 in lab by the similar mechanism of action. The enzyme activities of PAL, POD and SOD, Chlorophyll content and Pathogenesis Related Proteins （PR） in tobacco inoculated with the compound 2005196 and 2006172 were measured. The result showed the compound could stimulate tobacco to enhance enzyme activity so as to develop resistance. Regarding PR, the compound 2005196 can stimulate tobacco to produce homogeneous proteins with TMV.更多还原