本文主要研究内容
作者林连男（2019）在《中华散尾鬼笔活性物质的提取优化及作用机制研究》一文中研究指出：当今植物病虫害的防治仍然侧重于使用化学农药,随着单一农药品种使用时间的增长,林木害虫和病原菌都产生了一定的耐药性,这致使杀虫剂、杀菌剂的用量在近年来逐渐增大,大量使用化学农药已给人类社会造成非常严重环境污染与经济损失。在国家的倡导下,生物农药的使用比例逐年增加,寻求高效、安全、无污染的绿色新型生物农药显得尤为迫切。真菌中含有大量抗虫、抗菌、抗病毒等活性物质。本论文在前期试验中证明,中华散尾鬼笔具有抑菌活性,显示出中华散尾鬼笔在生物防治领域具有应用开发的潜力,作为生物源农药资源具有重要开发前景。本研究通过野外采集中华散尾鬼笔,采用不同提取方式提取杀虫抑菌活性物质,并进行提取条件优化,分离纯化中华散尾鬼笔粗取物,并对森林害虫和病原菌进行生物测定,研究了粗提取对害虫和病原菌酶活的影响及作用机制,获得如下研究成果:(1)用乙醇对中华散尾鬼笔的子实体进行活性物质提取,利用Plackett-Burman和Box-Behnken试验设计方法,对影响中华散尾鬼笔乙醇提取物的5个因素进行筛选,对关键因素进行优化,结果确定中华散尾鬼笔乙醇提取物的最佳工艺条件为:乙醇浓度60%、料液比1:80(g/mL)、超声时间80min,在此条件下中华散尾鬼笔杀虫物质的提取率为31.21%,与提取率的理论值31.36%相近。(2)应用扫描电子显微镜观察超声辅助提取与浸提法,对中华散尾鬼笔物料表面的不同影响。结果表明,对中华散尾鬼笔粉末进行超声辅助提取,提取率为31.23%,使用温水浸提的方法为24.10%;并且由温水浸提提取的表面组织是波浪状和光滑的,与其相比,超声处理的中华散尾鬼笔样品的表面形态松散、粗糙。(3)利用GC-MC分析中华散尾鬼笔乙醇提取物中的化学成分,得出样品中存在戊酸(Pentanoic acid)、十六烷酸,乙基酯(Pentanoic acid Hexadecanoic acid,ethyl ester)等7种化合物。(4)测定中华散尾鬼笔抑菌活性物质主要存在于发酵液中,之后对发酵液中的活性物质进行萃取及萃取条件优化。结果显示,中华散尾鬼笔的活性物质存在于发酵液中,发酵液中活性物质最佳的提取溶剂为正丁醇,最佳料液比为1:3,最佳提取次数为3次。(5)应用硅胶柱层析等分离纯化技术,并用薄层层析跟踪检测,对中华散尾鬼笔正丁醇提取物进行分离纯化。得到中华散尾鬼笔抑菌提取物L134。(6)通过研究中华散尾鬼笔抑菌物质L134对拟盘多毛孢孢子萌发、菌丝的生长过程、膜透性、细胞壁完整性和关键酶系的影响,初步阐明了提取物L134对拟盘多毛孢的生物防治机理。提取物L134能够抑制拟盘多毛孢孢子萌发,并且对菌丝生长有抑制作用EC50为0.305 mg/mL;影响拟盘多毛孢菌丝体SOD、POD、PPO、CAT、CL的活性,导致致病力下降,氧自由基清除系统出现障碍,MDA含量升高,膜质过氧化加重;L134能对拟盘多毛孢菌丝的细胞壁产生不良影响,同时对其菌丝生长的外部形态也产生了负面作用。(7)对中华散尾鬼笔提取物L134的稳定性测定的结果表明:提取物L134在不同pH值处理下比较稳定;处理温度在65℃以下比较稳定;应用不同金属离子处理中华散尾鬼笔提取物L134,其抑菌活性无显著差异;在氧化剂和还原剂处理的中华散尾鬼笔提取物L134的试验中,氧化剂会影响中华散尾鬼笔的抑菌活性物质的稳定性,但在紫外线照射下稳定性良好。(8)最后为了验证试验结果,分别测试不同提取物对舞毒蛾和多种病原菌的生物活性。结果表明:中华散尾鬼笔乙醇提取物对舞毒蛾幼虫有良好的杀虫活性,死亡率达到59.77%;中华散尾鬼笔正丁醇提取物对樟子松枯梢病(Sphaeropsis visci)、樟子松黑点针枯病(Pestalotiopsis sp.)、杨树烂皮病(Cytospora chrysosperma)、尖孢镰刀菌(Fusarium oxysporum)、灰葡萄孢菌(Botrytis cinerea)等真菌均具有抑制活性。
Abstract
dang jin zhi wu bing chong hai de fang zhi reng ran ce chong yu shi yong hua xue nong yao ,sui zhao chan yi nong yao pin chong shi yong shi jian de zeng chang ,lin mu hai chong he bing yuan jun dou chan sheng le yi ding de nai yao xing ,zhe zhi shi sha chong ji 、sha jun ji de yong liang zai jin nian lai zhu jian zeng da ,da liang shi yong hua xue nong yao yi gei ren lei she hui zao cheng fei chang yan chong huan jing wu ran yu jing ji sun shi 。zai guo jia de chang dao xia ,sheng wu nong yao de shi yong bi li zhu nian zeng jia ,xun qiu gao xiao 、an quan 、mo wu ran de lu se xin xing sheng wu nong yao xian de you wei pai qie 。zhen jun zhong han you da liang kang chong 、kang jun 、kang bing du deng huo xing wu zhi 。ben lun wen zai qian ji shi yan zhong zheng ming ,zhong hua san wei gui bi ju you yi jun huo xing ,xian shi chu zhong hua san wei gui bi zai sheng wu fang zhi ling yu ju you ying yong kai fa de qian li ,zuo wei sheng wu yuan nong yao zi yuan ju you chong yao kai fa qian jing 。ben yan jiu tong guo ye wai cai ji zhong hua san wei gui bi ,cai yong bu tong di qu fang shi di qu sha chong yi jun huo xing wu zhi ,bing jin hang di qu tiao jian you hua ,fen li chun hua zhong hua san wei gui bi cu qu wu ,bing dui sen lin hai chong he bing yuan jun jin hang sheng wu ce ding ,yan jiu le cu di qu dui hai chong he bing yuan jun mei huo de ying xiang ji zuo yong ji zhi ,huo de ru xia yan jiu cheng guo :(1)yong yi chun dui zhong hua san wei gui bi de zi shi ti jin hang huo xing wu zhi di qu ,li yong Plackett-Burmanhe Box-Behnkenshi yan she ji fang fa ,dui ying xiang zhong hua san wei gui bi yi chun di qu wu de 5ge yin su jin hang shai shua ,dui guan jian yin su jin hang you hua ,jie guo que ding zhong hua san wei gui bi yi chun di qu wu de zui jia gong yi tiao jian wei :yi chun nong du 60%、liao ye bi 1:80(g/mL)、chao sheng shi jian 80min,zai ci tiao jian xia zhong hua san wei gui bi sha chong wu zhi de di qu lv wei 31.21%,yu di qu lv de li lun zhi 31.36%xiang jin 。(2)ying yong sao miao dian zi xian wei jing guan cha chao sheng fu zhu di qu yu jin di fa ,dui zhong hua san wei gui bi wu liao biao mian de bu tong ying xiang 。jie guo biao ming ,dui zhong hua san wei gui bi fen mo jin hang chao sheng fu zhu di qu ,di qu lv wei 31.23%,shi yong wen shui jin di de fang fa wei 24.10%;bing ju you wen shui jin di di qu de biao mian zu zhi shi bo lang zhuang he guang hua de ,yu ji xiang bi ,chao sheng chu li de zhong hua san wei gui bi yang pin de biao mian xing tai song san 、cu cao 。(3)li yong GC-MCfen xi zhong hua san wei gui bi yi chun di qu wu zhong de hua xue cheng fen ,de chu yang pin zhong cun zai wu suan (Pentanoic acid)、shi liu wan suan ,yi ji zhi (Pentanoic acid Hexadecanoic acid,ethyl ester)deng 7chong hua ge wu 。(4)ce ding zhong hua san wei gui bi yi jun huo xing wu zhi zhu yao cun zai yu fa jiao ye zhong ,zhi hou dui fa jiao ye zhong de huo xing wu zhi jin hang cui qu ji cui qu tiao jian you hua 。jie guo xian shi ,zhong hua san wei gui bi de huo xing wu zhi cun zai yu fa jiao ye zhong ,fa jiao ye zhong huo xing wu zhi zui jia de di qu rong ji wei zheng ding chun ,zui jia liao ye bi wei 1:3,zui jia di qu ci shu wei 3ci 。(5)ying yong gui jiao zhu ceng xi deng fen li chun hua ji shu ,bing yong bao ceng ceng xi gen zong jian ce ,dui zhong hua san wei gui bi zheng ding chun di qu wu jin hang fen li chun hua 。de dao zhong hua san wei gui bi yi jun di qu wu L134。(6)tong guo yan jiu zhong hua san wei gui bi yi jun wu zhi L134dui ni pan duo mao bao bao zi meng fa 、jun si de sheng chang guo cheng 、mo tou xing 、xi bao bi wan zheng xing he guan jian mei ji de ying xiang ,chu bu chan ming le di qu wu L134dui ni pan duo mao bao de sheng wu fang zhi ji li 。di qu wu L134neng gou yi zhi ni pan duo mao bao bao zi meng fa ,bing ju dui jun si sheng chang you yi zhi zuo yong EC50wei 0.305 mg/mL;ying xiang ni pan duo mao bao jun si ti SOD、POD、PPO、CAT、CLde huo xing ,dao zhi zhi bing li xia jiang ,yang zi you ji qing chu ji tong chu xian zhang ai ,MDAhan liang sheng gao ,mo zhi guo yang hua jia chong ;L134neng dui ni pan duo mao bao jun si de xi bao bi chan sheng bu liang ying xiang ,tong shi dui ji jun si sheng chang de wai bu xing tai ye chan sheng le fu mian zuo yong 。(7)dui zhong hua san wei gui bi di qu wu L134de wen ding xing ce ding de jie guo biao ming :di qu wu L134zai bu tong pHzhi chu li xia bi jiao wen ding ;chu li wen du zai 65℃yi xia bi jiao wen ding ;ying yong bu tong jin shu li zi chu li zhong hua san wei gui bi di qu wu L134,ji yi jun huo xing mo xian zhe cha yi ;zai yang hua ji he hai yuan ji chu li de zhong hua san wei gui bi di qu wu L134de shi yan zhong ,yang hua ji hui ying xiang zhong hua san wei gui bi de yi jun huo xing wu zhi de wen ding xing ,dan zai zi wai xian zhao she xia wen ding xing liang hao 。(8)zui hou wei le yan zheng shi yan jie guo ,fen bie ce shi bu tong di qu wu dui wu du e he duo chong bing yuan jun de sheng wu huo xing 。jie guo biao ming :zhong hua san wei gui bi yi chun di qu wu dui wu du e you chong you liang hao de sha chong huo xing ,si wang lv da dao 59.77%;zhong hua san wei gui bi zheng ding chun di qu wu dui zhang zi song ku sao bing (Sphaeropsis visci)、zhang zi song hei dian zhen ku bing (Pestalotiopsis sp.)、yang shu lan pi bing (Cytospora chrysosperma)、jian bao lian dao jun (Fusarium oxysporum)、hui pu tao bao jun (Botrytis cinerea)deng zhen jun jun ju you yi zhi huo xing 。
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论文作者分别是来自东北林业大学的林连男，发表于刊物东北林业大学2019-11-18论文，是一篇关于中华散尾鬼笔论文,活性物质论文,次生代谢产物论文,提取工艺优化论文,作用机理论文,东北林业大学2019-11-18论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自东北林业大学2019-11-18论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：中华散尾鬼笔论文; 活性物质论文; 次生代谢产物论文; 提取工艺优化论文; 作用机理论文; 东北林业大学2019-11-18论文;