CN112301087A - 一种通过真菌孢子萌发和生物活性测定相结合科学评价真菌杀虫剂产品质量的方法 - Google Patents
一种通过真菌孢子萌发和生物活性测定相结合科学评价真菌杀虫剂产品质量的方法 Download PDFInfo
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Abstract
本发明公开了一种通过真菌孢子萌发和生物活性测定科学评价真菌杀虫剂产品质量的方法。真菌杀虫剂是一类重要的生物杀虫剂，其主要活性成份为真菌孢子，对高温等较为敏感。现有对真菌杀虫剂产品质量评价的主要指标是真菌孢子萌发率，但经历了高温等的真菌孢子除了萌发率变化，其对害虫的致病力是否有变化,真菌孢子的萌发力能否直接代表真菌的致病力,这方面鲜有报道。本技术通过真菌孢子萌发和生物活性测定相结合科学评价真菌杀虫剂产品质量，先测定孢子萌发率，在萌发率低于90％时进一步测定产品对靶标害虫对象的致病力。本发明对于真菌杀虫剂产品质量的科学评价具有直接指导作用，同时对于真菌杀虫剂生产加工以及运输贮存过程中的质量控制也具有重要意义。
Description
一、技术领域
本发明属于病虫害防治技术领域，具体涉及一种通过真菌孢子萌发和生物活性测定相结合科学评价真菌杀虫剂产品质量的方法。
二、技术背景
病虫害一直是农作物安全高效生产的重要威胁，长期以来，其防控主要依赖化学农药。化学农药虽在病虫害有效防控中发挥了巨大作用，但其带来的抗药性、安全性等问题难以回避。随着人们生活水平的提高和环境保护意识的增强，对于农作物病虫害的控制，无论是从以安全为目标的绿色防控需要出发，还是就避开病虫害的抗药性来考虑，以生物防治为核心的病虫害绿色防控应是理想手段，是未来发展趋势。杀虫真菌是一类重要的病原微生物，在自然界中广泛存在，一些已被成功开发为真菌杀虫剂，如白僵菌、绿僵菌等，在玉米螟、松毛虫等农林害虫的生物防治中取得了明显成效。
农药产品质量评价是保证其田间控害效果发挥的重要环节。对于生物农药而言，因其生物源活性成份对环境条件敏感性更高，故产品质量的科学评价更为重要。真菌杀虫剂产品的主要活性成份为真菌孢子，故对高温等较为敏感。真菌杀虫剂产品在真菌发酵生产、剂型加工以及运输贮存时，可能会经历高温.造成对真菌孢子的损伤。因此，现有对真菌杀虫剂产品质量评价的主要指标是孢子萌发率。经历了高温的真菌孢子除了萌发率，其对害虫的致病力又会有怎么的变化？真菌孢子的萌发力能否直接代表真菌的致病力？这方面鲜有报道。而真菌致病力才是真菌杀虫剂最根本的质量评价标准，如果孢子萌发率与真菌致病力不一致，则当前以测定孢子萌发率评价真菌杀虫剂产品质量的方法则不全面，也不尽合理。本研究针对上述问题，将发明一种通过真菌孢子萌发和生物活性测定相结合科学评价真菌杀虫剂产品质量的方法。本发明对于客观合理评价真菌杀虫剂产品质量具有重要指导作用，同时对于真菌杀虫剂生产加工以及运输贮存过程中的质量控制也具有积极指导意义。
三、发明内容
1、发明目的
提供一种理想的真菌杀虫剂产品质量评价方法，具体涉及一种通过真菌孢子萌发和生物活性测定相结合科学评价真菌杀虫剂产品质量的方法，以更客观评价产品质量并合理指导产品生产和运输过程的质量把控。
2、技术方案
本发明具体涉及一种真菌杀虫剂产品质量评价方法，其特征在于利用真菌孢子萌发和生物活性测定相结合来科学评价真菌杀虫剂产品质量的方法。
3、有益效果
本发明的方法与现有技术相比，产生以下有益效果：(1)与常规的真菌杀虫剂质量评价仅以孢子萌发率作为评价指标，本发明通过孢子萌发和生物活性测定相结合来评价产品质量，对产品质量评价更为客观全面；(2)有助于提升产品生产、运输以及贮存过程中的质量控制水平。
四、具体实施方式
本发明的实施例是采用室内测定的方法。
测定方法如下：
1、爪哇棒束孢不同温度处理
将同一批加工的真菌杀虫剂50亿孢子/ml爪哇棒束孢油悬剂按每份20ml密封包装后，置于25、30、36、42、48、54℃烘箱中，每个温度处理重复4次即放置4份，分别在放置7和14天后取出，测定其中分生孢子萌发率及对烟粉虱的毒力。
2、爪哇棒束孢孢子萌发率测定
从各温度处理后的爪哇棒束孢油悬剂中取出100微升，将其稀释10000倍后在马铃薯葡萄糖琼脂培养基(PDA)上培养，26±1℃培养24h，显微镜下观察孢子的萌发，记录已萌发和未萌发孢子数量，计算萌发率。
3、爪哇棒束孢对烟粉虱致病力测定
采用喷雾法测定爪哇棒束孢对烟粉虱的毒力。用5×106个孢子/ml爪哇棒束孢孢子悬浮液喷雾处理带有烟粉虱4龄若虫的番茄叶片，每张叶片上有若虫20-30头，喷雾处理后晾干，放在一次性塑料培养皿中，同时以清水作为空白对照，每处理重复4次。处理后10天内每天检查烟粉虱的死亡情况，记录死亡虫和活虫数，计算死亡率。
五、实施效果：
1、不同温度处理7天后杀虫真菌爪哇棒束孢孢子的萌发率及对烟粉虱的致病力
比较了不同温度处理7天后爪哇棒束孢孢子的萌发率及对烟粉虱的致病力。25℃处理后，爪哇棒束孢孢子萌发率为91.5％，此温度处理下，5×106个孢子/ml爪哇棒束孢孢子对烟粉虱4龄若虫的致死率为100％；30℃处理后孢子的萌发率为90％，较25℃处理仅下降了1.6％，此处理对烟粉虱的致死率为95％，较25℃处理下降了5％，30℃处理孢子萌发率和致死率均与25℃处理差异不大；36℃处理后孢子萌发率降至83.5％，对烟粉虱的致死率则降为55％，与25℃处理相比分别下降了8.7％和45％，两者下降比例差异大；42℃处理后孢子萌发率和致死率分别为82％和42％，与25℃处理相比分别下降了10.4％和58％；48℃处理后孢子萌发率和致死率分别为78％和38％，与25℃处理相比分别下降了14.8％和62％；54℃处理后孢子萌发率和致死率分别降至62.5％和23％，与25℃处理相比分别下降了31.7％和77％。从各温度处理7d后爪哇棒束孢孢子萌发和致病力变化来看，在36℃及以上温度处理后，爪哇棒束孢孢子萌发和致病力影响程度不一致，其中对致病力的影响更大，孢子萌发不能真实反映其致病力。因此，要客观评价真菌杀虫剂产品质量，在真菌孢子萌发率低于90％时，需要进一步对致病力作测定。
2、不同温度处理14天后杀虫真菌爪哇棒束孢的孢子萌发率及对烟粉虱的致病力
比较了不同温度处理14天后爪哇棒束孢孢子的萌发率及对烟粉虱的致病力。25℃处理后，爪哇棒束孢孢子萌发率为90％，5×106个孢子/ml爪哇棒束孢孢子对烟粉虱4龄若虫的致死率为100％；30℃处理后孢子的萌发率为90％，较25℃处理未有下降，此处理对烟粉虱的致死率为89％，较25℃处理下降了11％；36℃处理后孢子萌发率降至81％，对烟粉虱的致死率则降为43％，与25℃处理相比分别下降了10％和57％，两者下降比例差异较大；42℃处理后孢子萌发率和致死率分别为78％和40％，与25℃处理相比分别下降了13.3％和60％；48℃处理后孢子萌发率和致死率分别为68.5％和26％，与25℃处理相比分别下降了23.9％和74％；54℃处理后孢子萌发率和致死率分别降至58％和15％，与25℃处理相比分别下降了35.6％和85％。从各温度处理14d后爪哇棒束孢孢子萌发和致病力变化来看，同样表现出了36℃及以上高温处理后，爪哇棒束孢孢子萌发和致病力影响程度不一致，其中对致病力的影响更大，孢子萌发不能真实反映其致病力。由此说明真菌杀虫剂产品质量的客观评价需要真菌孢子萌发率测定和致病力测定，在孢子萌发率低于90％时，致病力测定更为重要。
Claims (2)
1.一种真菌杀虫剂产品质量科学评价方法，其特征在于通过真菌孢子萌发和生物活性测定相结合来评价真菌杀虫剂产品质量。
2.根据权利要求1所述的通过真菌孢子萌发和生物活性测定相结合来评价真菌杀虫剂产品质量，其特征在于先测定真菌孢子萌发率，在萌发率低于90％时，再进一步测定生物活性。
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103756913A (zh) * | 2013-12-24 | 2014-04-30 | 华南农业大学 | 一种玫烟色棒束孢菌株 |
| CN107212022A (zh) * | 2017-05-24 | 2017-09-29 | 江苏省农业科学院 | 一种利用杀虫真菌降低烟粉虱体内植物病毒含量的方法 |
| CN108102929A (zh) * | 2017-12-26 | 2018-06-01 | 华南农业大学 | 一种抗吡蚜酮的爪哇棒束孢及其应用 |
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| CN103756913A (zh) * | 2013-12-24 | 2014-04-30 | 华南农业大学 | 一种玫烟色棒束孢菌株 |
| CN107212022A (zh) * | 2017-05-24 | 2017-09-29 | 江苏省农业科学院 | 一种利用杀虫真菌降低烟粉虱体内植物病毒含量的方法 |
| CN108102929A (zh) * | 2017-12-26 | 2018-06-01 | 华南农业大学 | 一种抗吡蚜酮的爪哇棒束孢及其应用 |
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