CN113729074B - 一种用于防治梨炭疽病的保鲜剂及其应用 - Google Patents
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Abstract
本发明提供一种用于防治梨炭疽病的保鲜剂，保鲜剂中活性成分是由药剂A、B和竹醋液组成，药剂A和B组合为吡唑醚菌酯和戊唑醇组合、或者、吡唑醚菌酯和甲基硫菌灵组合。可有效控制炭疽病对梨储藏期的危害，有效减少药剂的使用量，延缓炭疽病对抗药剂的抗药性，提高保鲜效果，降低贮藏损失，具有显著的经济和社会效益。
Description
技术领域
本发明属于植物保鲜剂领域，具体涉及一种用于防治梨炭疽病的保鲜剂及其应用。
背景技术
梨炭疽病又称苦腐病，是由盘长孢状刺盘孢侵染所引起的、发生在梨上的病害。病原为盘长孢状刺盘孢(Colletrichum gloeosporioidesPenz.)。病原菌以菌丝体在僵果或病枝上越冬，第二年条件适宜时对梨树产生侵染。采摘后随梨子生活力降低而逐渐发病，往往在贮藏期间继续扩大甚至造成大量果实腐烂，造成一定的经济损失。梨炭疽病发生普遍，在世界各地梨产区均有发生。
目前，梨贮藏保鲜方法有化学保鲜剂、天然保鲜剂、生物保鲜剂、气调贮藏等。使用化学保鲜剂具有低成本、高防效等优势，目前仍是一种重要的保鲜防腐手段。化学保鲜剂虽然能起到防腐保鲜作用，但大多具有不同程度的毒性和残留，对人体具有潜在的危害。
竹醋液属于天然保鲜剂，是竹材碳化、热解过程中产生的副产品，由有机酸、酚类、酮类、醇类及脂类等多种组分组成，对多种细菌和真菌具有较强的抑制作用，是一种良好的天然源防腐保鲜剂的开发材料。因此通过竹醋液与化学保鲜剂的复配，开发研制防治梨炭疽病的保鲜剂，对化学保鲜剂使用量的降低，增强保鲜效果起到积极的作用。
发明内容
本发明提供一种可用于防治梨炭疽病的保鲜剂及其应用。本发明提供如下技术方案：
一种用于防治梨炭疽病的保鲜剂，保鲜剂中活性成分是由药剂A、B和竹醋液组成，药剂A和B组合为吡唑醚菌酯和戊唑醇组合、或者、吡唑醚菌酯和甲基硫菌灵组合。本发明所述的保鲜剂中药剂A和药剂B的质量配比为25∶1～1∶10，化学农药质量百分含量为1～5％，竹醋液质量百分含量在60～80％。优选的药剂A和药剂B的质量配比为10∶1-1∶10、5∶1-1∶5。
本发明优选的药剂A和药剂B的组合为吡唑醚菌酯和戊唑醇。
本发明所述的保鲜剂的剂型为水乳剂和水悬浮剂。本发明保鲜剂的使用方法为喷雾或浸果。可以有效的防治梨炭疽病产生的病害，起到减少化学保鲜的使用。
本发明所述剂型为水乳剂，各组分的重量百分含量为：
本药剂组合物水乳剂的加工步骤为：将原药和溶剂相混合加入高速均质釜中，经高速剪切混合均匀，搅拌0.1-0.5h，形成均一油相；将竹醋液调节PH值后(PH＜7)和乳化剂混合，搅拌器搅拌均匀，形成均一水相；打开高速均质釜中高速剪切机，将水相缓慢加入高速均质釜中，均质0.5-2h，制得本发明所述的组合物水乳剂。
本发明所述剂型为水悬浮剂，各组分的重量百分含量为：
本药剂组合物水悬浮剂的加工步骤为：先将助剂及水混合，经高速剪切混合均匀，加入药剂A和药剂B原药，在砂磨机中磨0.5～2小时，使微粒粒径全部在5μm以下，制得本发明所述的组合物水悬浮剂。
本发明所述的溶剂是环己烷、二氯乙烷、正己烷、乙酸乙酯、三氯甲烷、环己烷、丙酮、N，N-二甲基甲酰胺、甲苯、四氯化碳、乙二醇中的一种或几种。
本发明所述的增稠剂是硅酸镁铝、黄原胶、羟甲基纤维素、阿拉伯树胶、聚乙烯吡咯烷酮、黄原胶中的一种或是几种。
本发明所述的乳化剂是烷基酚聚氧乙烯醚磷酸盐、烷基酚聚氧乙烯醚甲醛缩合物、二苯基酚聚氧乙烯聚甲醛缩合物、二苯基酚聚氧乙烯聚氧丙烯醚、吐温-60、NNO、MOA-7、601、A-110、TX-10、宁乳700#、5202中的一种或几种。
本发明有益技术效果：
本发明制备得到的保鲜剂可有效控制炭疽病对梨储藏期的危害，有效减少药剂的使用量，延缓炭疽病对抗药剂的抗药性，提高保鲜效果，降低贮藏损失，具有显著的经济和社会效益。
附图说明
图1：复配组合1对梨炭疽病的保鲜效果图
图2：复配组合3对梨炭疽病的保鲜效果图
具体实施方式
下面通过实施例来具体说明本发明的内容。在本发明中，以下实施例是为了更好地阐述本发明，并不是用来限制本发明的范围。下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。
实施例1竹醋液及杀菌剂对供试植物病原菌的毒力
供试作物病菌：梨炭疽病
测定方法：采用菌丝生长速率法测定竹醋液及杀菌剂对供试植物病原菌的毒力。
将供试竹醋液用无菌水稀释到适当浓度。取稀释后的竹醋液1mL，加入24mL PDA培养基中混合均匀，倒入培养皿中制成含竹醋的培养平板。以加入同量无菌水的PDA培养基平板作为空白对照(CK)。
供试杀菌剂以乙腈溶解并稀释至适当浓度，分别取杀菌剂溶液1mL，与24mLPDA培养基混合均匀，倒入培养皿中制成不同浓度的含药培养基。以加入同量乙腈的PDA培养基平板作为空白对照(CK)。
无菌环境下，用打孔器将固体PDA培养基培养的供试菌种制成直径为5.0mm的菌饼。接种供试病原菌菌饼(菌丝面朝下)。将接种后的PDA培养基平板倒置，放于28℃恒温箱中培养。培养至CK菌丝长至培养皿85％左右，用抗游标卡尺测量菌落直径。计算菌丝生长抑制率，求得各单剂毒力回归方程、相关系数和EC50。
表1各单剂与竹醋液对梨炭疽病的毒力
实施例2复配组合联合作用测试
根据各单剂和竹醋液的EC50，通过正交试验，设计各种配比组合，将各配比组合用无菌水稀释到适当浓度。取稀释后的竹醋液1mL，加入24mL PDA培养基中混合均匀，倒入培养皿中制成含竹醋的培养平板。以加入同量无菌水的PDA培养基平板作为空白对照(CK)。
无菌环境下，用打孔器将固体PDA培养基培养的供试菌种制成直径为5.0mm的菌饼。接种供试病原菌菌饼(菌丝面朝下)。将接种后的PDA培养基平板倒置，放于28℃恒温箱中培养。培养至CK菌丝长至培养皿85％左右，用抗游标卡尺测量菌落直径。计算菌丝生长抑制率，求得各单剂毒力回归方程、相关系数、EC 50和共毒系数。
以共毒系数CTC值评判复配剂的联合毒力作用。CTC值小于80为拮抗作用，80-120之间时为相加作用，120-200具有一定增效作用，大于200为明显增效。
表2正交实验设计表
注：列表中的1、2、3、4代表EC50的倍数，以1号实验组为例A1B1C1表示实验各组分浓度比＝1倍竹醋液EC50∶1倍药剂AEC50∶1倍药剂B EC50；A、B为甲基硫菌灵、戊唑醇或吡唑醚菌酯。
表3复配组合(吡唑醚菌酯·戊唑醇·竹醋液)对梨炭疽病的毒力
通过表3可以看出，复配组合(吡唑醚菌酯·戊唑醇·竹醋液)联合毒力指数CTC大多大于200，说明复配剂1具有明显的增效作用。
表4复配组合(吡唑醚菌酯·甲基硫菌灵·竹醋液)对梨炭疽病的毒力
通过表4可以看出，复配组合(吡唑醚菌酯·甲基硫菌灵·竹醋液)联合毒力指数CTC均大于120，说明复配剂具有明显的增效作用。
实施例3活体抑菌活性测定
测试方法：实施例中配方与不同药剂进行浸果处理，晾干后每个果子扎孔(3mm深)，每个果子扎孔3个。每个孔中贴上5mm菌饼，每组处理6个梨子。室温培养6天，测定病斑直径，验证了药剂对炭疽病的防治效果。
测试药剂：
表5复配剂成分组成(1L)及防治对象
| 复配组名称 | 竹醋液 | 甲基硫菌灵 | 戊唑醇 | 吡唑醚菌酯 | 助剂 | 剂型 | 防治对象 |
| (复配剂1)吡唑醚菌酯·戊唑醇·竹醋液 | 745.7ml | / | 11.4g | 17g | 225.9g | 水乳剂 | 炭疽病 |
| (复配剂3)吡唑醚菌酯·甲基硫菌灵·竹醋液 | 844.6ml | 43.9g | / | 43.9g | 67.6g | 悬浮剂 | 炭疽病 |
对比药剂：30％吡唑醚菌酯水乳剂、43％戊唑醇乳油、70％甲基硫菌灵可湿性粉剂
测试结果
表6：复配剂1及对照组活体实验防治效果分析表
通过表6可以看出，活体防治效果表明，配剂1按照800稀释时，防治效果略低于30％吡唑醚菌酯4000倍，与43％戊唑醇4000倍相当；按照1000倍稀释时，防治效果明显优于43％戊唑醇5000倍。
复配剂1的800倍稀释药液，每升中含0.014g吡唑醚菌酯和0.021g戊唑醇，单剂吡唑醚菌酯1200倍稀释药液，每升含0.250g吡唑醚菌酯，化学农药有效成分总量减少了85.6％；单剂戊唑醇4000倍稀释药液，每升含0.107g戊唑醇，化学农药有效成分总量减少了67.0％；
复配剂1的1000倍稀释药液，每升中含0.011g吡唑醚菌酯和0.017g戊唑醇，相对于单剂吡唑醚菌酯2400倍稀释药液，每升含0.125g吡唑醚菌酯，化学农药有效成分总量减少了77.3％；相对于单剂戊唑醇5000倍稀释药液，每升含0.086g戊唑醇，化学农药有效成分总量减少了67.0％；说明复配剂1对梨炭疽病具有良好的防治效果的同时达到农药减量化的目的。
表7：复配剂3及对照组活体实验防治效果分析表
通过表7可以看出，复配3按照800倍稀释时，防治效果略低于80％甲基硫菌灵800倍液，与30％吡唑嘧菌酯1200倍相当；按照1200倍稀释时，防治效果明显优于30％吡唑醚菌酯2400倍。
复配剂3的800倍稀释药液，每升中含0.055g甲基硫菌灵和0.055g吡唑醚菌酯，相对于单剂甲基硫菌灵800倍稀释药液，每升含0.875g甲基硫菌灵，化学农药有效成分总量减少了87.5％；相对于单剂吡唑醚菌酯1200倍稀释药液，每升含0.250g吡唑醚菌酯，化学农药有效成分总量减少了56.1％；
复配剂3的1200倍稀释药液，每升中含0.044g甲基硫菌灵和0.010g戊唑醇，相对于单剂甲基硫菌灵1200倍稀释药液，每升含0.583g甲基硫菌灵，化学农药有效成分总量减少了87.5％；相对于单剂吡唑醚菊酯2400倍稀释药液，每升含0.125g吡唑醚菌酯，化学农药有效成分总量减少了41.5％；
表明复配剂3对梨炭疽病具有较好的防治效果的同时达到农药减量化的目的。
实施例4竹醋液对杀菌组合联合作用测试
表8：复配1中加竹醋液与不加竹醋液对梨炭疽病的毒力对比
| 实验组 | 回归方程 | 相关系数 | 共毒系数 |
| 加竹醋液 | Y＝1.6745x-0.7154 | 0.9991 | 251.58 |
| 不加竹醋液 | Y＝1.3985X+0.0403 | 0.97944 | 185.11 |
竹醋液EC50∶吡唑醚菌EC50∶戊唑醇EC50＝4∶1∶2
本研究发明首次以竹醋液为主体配置的农药，通过表8可以看出，在复配1中竹醋液的使用明显提高了农药的共毒系数。
表9：复配3中加竹醋液与不加竹醋液对梨炭疽病的毒力对比
| 实验组 | 回归方程 | 相关系数 | 共毒系数 |
| 加竹醋液 | Y＝2.8501X-4.387 | 0.9547 | 289.96 |
| 不加竹醋液 | Y＝4.4942X-10.437 | 0.965 | 209.43 |
竹醋液EC50∶吡唑醚菌酯EC50∶甲基硫菌灵＝EC50＝2∶1∶1
本研究发明首次以竹醋液为主体配置的农药，通过表9可以看出，在复配3中竹醋液的使用明显提高了农药的共毒系数。
Claims (3)
1.一种用于防治梨炭疽病的保鲜剂，其特征在于，保鲜剂中活性成分是由药剂A、B和竹醋液组成，药剂A和B组合为吡唑醚菌酯和戊唑醇组合；
竹醋液EC50∶药剂A EC50∶药剂B EC50=1:1:1、1:2:3、1:3:4、1:4:2、2:1:2、2:2:4、2:3:3、2:4:1、3:1:3、3:2:1、3:3:2、3:4:4、4:1:4、4:2:2、4:3:1、4:4:3；
其中竹醋液EC50为11437mg/L,吡唑醚菌酯EC50为0.666mg/L,戊唑醇EC50为0.497mg/L；
保鲜剂的剂型为水乳剂和水悬浮剂。
2.一种用于防治梨炭疽病的保鲜剂，其特征在于，保鲜剂中活性成分是由药剂A、B和竹醋液组成，药剂A和B组合为吡唑醚菌酯和甲基硫菌灵组合；
竹醋液EC50∶药剂A EC50∶药剂B EC50=1:1:1、1:2:3、1:3:4、1:4:2、2:1:2、2:2:4、2:3:3、2:4:1、3:1:3、3:2:1、3:3:2、3:4:4、4:1:4、4:2:2、4:3:1、4:4:3；
其中竹醋液EC50为11437mg/L,吡唑醚菌酯EC50为0.666mg/L，甲基硫菌灵为EC50为0.747mg/L；
保鲜剂的剂型为水乳剂和水悬浮剂。
3.根据权利要求1或2所述的防治梨炭疽病的保鲜剂，其特征在于，使用方法为喷雾或浸果。
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