CN117426380A - 防治青花菜黑腐病的植物源杀菌剂组合 - Google Patents
防治青花菜黑腐病的植物源杀菌剂组合 Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N35/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical
- A01N35/04—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical containing aldehyde or keto groups, or thio analogues thereof, directly attached to an aromatic ring system, e.g. acetophenone; Derivatives thereof, e.g. acetals
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/90—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P1/00—Disinfectants; Antimicrobial compounds or mixtures thereof
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
本发明涉及生物农药领域,具体公开一种防治青花菜黑腐病的植物源杀菌剂组合,其有效成分由0.2%苯丙烯菌酮和0.5%苦参碱复配而成,本发明的技术方案下,室内杀菌剂毒力测定结果表明,0.2%苯丙烯菌酮和0.5%苦参碱2种植物源杀菌剂对青花菜黑腐病均具有较强的抑制作用;当0.2%苯丙烯菌酮和0.5%苦参碱的质量比为3:7~6:4时,对青花菜黑腐病病原菌Xcc的共毒系数均大于120,表现为增效作用;室内盆栽试验表明,0.2%苯丙烯菌酮和0.5%苦参碱的质量比为3:7~6:4时,防治效果均远高于200 mg/L 15%农用链霉素。
Description
技术领域
本发明涉及生物农药领域,具体涉及一种防治青花菜黑腐病的植物源杀菌剂组合。
背景技术
青花菜(Brassica oleracea) , 又称西兰花、绿菜花、花菜薹等, 属十字花科芸薹属甘蓝变种, 可食用部分花球由多个肉质花梗和发育不全的花蕾组成。近年来,随着集约化种植程度的提高以及种植年限的增加,青花菜黑腐病逐渐加重,已成为制约青花菜产业发展的重要因素。
青花菜黑腐病是一种由野油菜黄单胞菌野油菜致病变种(XanthomonasCampestris Pv. Campestris,简称Xcc)引起的一种细菌性病害。其主要传播途径包括种子、土壤和病残体。病菌通过水孔、气孔或伤口入侵叶片,导致叶片发病,进而蔓延到茎部和球茎,引起植株枯萎和球茎的腐烂。此外,黑腐病也会诱发软腐病、菌核病等其他病害的发生,造成植物组织的严重腐烂,严重影响青花菜的产量和品质,对种植业造成重要的经济损失。
多雨和湿热的环境更是病害暴发的重灾区。加上连作、过高氮肥、植株徒长和虫害等都会造成青花菜黑腐病加重。目前对该种疾病的预控和控制主要集中在严格执行对种子和育苗期培养基质的杀菌消毒工作,加强在栽培管理和实施轮作等方面,此外,高效的防治黑腐病主要以化学防治为主,如使用氢氧化铜、噻菌铜、甲霜灵、链霉素等,但长期的农药使用,会导致病菌的抗药性增强,为加强防治效果,通常会增加使用次数以及使用量,这导致青花菜的农药残留量的增加,为环境污染、食品安全埋下隐患。植物源杀菌剂是从植物中提取出的有效成分,有抑制植物病原物的作用,具有高效、低毒、在自然中易降解等特点。市场上有多种植物源杀菌剂,如苦参碱、丁子香酚、乙蒜素、春雷霉素等,均在多种作物病害防治上得到了推广应用并取得了显著的效果,目前在青花菜黑腐病防治未见相关研究报道。
发明内容
本发明提供了一种防治青花菜黑腐病的植物源杀菌剂组合,为防治青花菜黑腐病,提高产量提供了效果更好、毒害副作用更小的方法。
为实现上述目的,本发明提供如下技术方案:
一种防治青花菜黑腐病的植物源杀菌剂组合,其有效成分由0.2%苯丙烯菌酮和0.5%苦参碱复配而成。
作为优选,所述0.2%苯丙烯菌酮和0.5%苦参碱的质量比为3:7~6:4。
进一步的,作为最优选,所述0.2% 苯丙烯菌酮和0.5%苦参碱的质量比为3:7。
本发明达到的技术效果为:室内杀菌剂毒力测定结果表明,0.2%苯丙烯菌酮和0.5%苦参碱2种植物源杀菌剂对青花菜黑腐病均具有较强的抑制作用;当0.2%苯丙烯菌酮和0.5%的质量比为3:7~6:4时,对青花菜黑腐病病原菌Xcc的共毒系数均大于120,表现为增效作用,其中,当苯丙烯菌酮和苦参碱的质量比为3:7时,表现为显著增效作用;室内盆栽试验表明,0.2%苯丙烯菌酮和0.5%苦参碱的质量比为3:7~6:4时,防治效果均远高于200 mg/L15%农用链霉素。
具体实施方式
下面结合实施例进一步说明本发明,这些实施例仅作为说明本发明的例子,不限制本发明的范围。下述实施例中的实验方法,如无特殊说明,均为常规方法。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 不同植物源杀菌剂的抑菌活性
1试验方法
1.1供试菌株
青花菜黑腐病病原菌Xcc由本课题组分离获得。
1.2供试植物源杀菌剂
供试药剂及相关信息见表1。
供试药剂用0.1%吐温-80水溶液稀释,配置成单剂母液。
表1 供试药剂、生产厂家及浓度
1.3Xcc活化及菌落计数
将Xcc划线接种于马铃薯-葡萄糖琼脂培养基(PDA)中,于30℃恒温培养箱中培养1-2 d,待菌落长好备用。取灭菌试管,加入2ml LB培养基,挑取单菌落放入LB培养基中,摇床110 rpm,30℃恒温培养10 h。采用梯度稀释法稀释菌悬液至10-8。取10-6-10-8对应浓度试管中的菌液10 μl分别涂布于LB琼脂培养基中,放入30 ℃恒温培养箱培养16 ~ 18 h后计算菌落数。
1.4单剂对Xcc生长的抑制作用测定
采用牛津杯抑菌圈法进行抑菌测定。将灭菌的PDA培养基加热融化,冷却至55~50℃左右,倾倒入培养皿,凝固冷却后加入50 μl浓度为1.2×107cfu/ml的病原菌。取牛津杯***培养基中,等距放置4个,每个牛津杯中加入250 μl植物源杀菌剂,另设无菌水作为空白对照(Control)。每种杀菌剂的每个浓度设置3个重复,30 ℃恒温培养箱避光培养至对照菌落直径生长至培养皿直径的 2/3~3/4 时,用直尺测量各菌落直径,采用十字交叉法垂直测量,取其平均值,计算Xcc生长抑制率。将菌丝生长抑制率换算为生物统计机率值(y),药剂浓度换算成对数(x),根据浓度对数与机率值回归法,得到线性回归方程,并由此计算供试药剂处理下Xcc的相关性系数以及EC50。Xcc生长抑制率的计算公式为:Xcc生长抑制率=[(对照组病原菌菌落直径-处理组病原菌菌落直径)/对照组病原菌菌落直径]×100%。
2试验结果
5种杀菌剂对Xcc生长的抑制效果结果如表2所示,其中,苯丙烯菌酮对Xcc生长抑制作用最强,当苯丙烯菌酮的质量浓度为10 mg/L时,其对Xcc的抑制作用最低为40.02%,当苯丙烯菌酮的质量浓度为80 mg/L时,抑制作用最高达68.54%;苦参碱效果较次,对Xcc最高抑制作用为67.42%;春雷霉素的抑制效果最差,当其质量浓度为80 mg/L时,对Xcc的生长抑制率仅为39.20%。
通过标准曲线计算5种杀菌剂对Xcc的毒力回归方程、相关性系数和EC50,结果如表3所示。5种杀菌剂的毒力回归方程的相关性系数均高于0.96,说明杀菌剂浓度与Xcc的抑制效果呈正相关,其中,苯丙烯菌酮的EC50为22.18 mg/L,相对于其它杀菌剂,数值最小,表明苯丙烯菌酮对Xcc生长的抑制效果最好;苦参碱的抑菌效果次之,其 EC50为 39.71 mg/L。参考5种杀菌剂对Xcc的 EC50,可知5种杀菌剂的有效成分对 Xcc的抑制作用由强到弱依次为苯丙烯菌酮>苦参碱>蛇床子素>丁子香酚>春雷霉素。
表2 5种植物源杀菌剂对Xcc生长的抑制效果
注:同一列数据中的不同小写字母表示不同处理组在0.05水平上差异的显著性。下同。
表3 5种植物源杀菌剂对Xcc的毒力分析
实施例2 植物源杀菌剂复配组合配方筛选
1试验方法
在预实验中,根据上述5种杀菌剂的毒力测定试验结果,参考不同单剂的作用机理,配置好0.2%苯丙烯菌酮和0.5%苦参碱两单剂的EC50溶液,以其体积之比为 0:10、1:9、2:8、3:7、4:6、5:5、6:4、7:3、8:2、9:1、10:0 等不同比例混合,观察Xcc在含上述不同复配制剂的平板中的生长情况,计算平均抑制率(结果见表4)。基于预实验结果,将0.2%苯丙烯菌酮和0.5% 苦参碱按质量比为3:7、4:6、5:5、6:4、7:3复配。试验混合药剂浓度为10、20、40、60和80 mg/L。采用牛津杯抑菌圈法进行抑菌测定,具体方法见实施例1。测量抑菌圈直径,绘制标准曲线,计算不同处理组复配药剂对Xcc的毒力回归方程、相关系数和 EC50,所得结果根据孙云沛等提出的共毒系数(CTC)法来评判两种药剂的联合毒力作用。 CTC 小于80为拮抗作用,大于120为增效作用,大于200为显著增效作用,80~120之间为相加作用。共毒系数按以下公式计算:
混剂理论毒力指数(TTI) = A剂毒力指数 ×A剂在混剂中的比例(%) + B剂毒力指数 ×B剂在混剂中的比例(%)
在本试验中,设苦参碱为标准剂。
2试验结果
上述5种杀菌剂的毒力测定试验结果表明,苯丙烯菌酮和苦参碱对Xcc具有较好的抑制作用。苯丙烯菌酮通过破坏植物病菌的壁膜***,干扰细胞代谢过程而达到杀菌的目的;苦参碱对细菌和真菌均具有较好的抑制作用。基于以上原因,本试验进一步开展苯丙烯菌酮和苦参碱的复配试验,以明确二者复配是否存在增效作用。不同复配比例对Xcc生长的抑制效果结果如表5所示,0.2%苯丙烯菌酮和0.5%苦参碱按质量比3:7~7:3复配时,对Xcc的抑制作用明显。此外,0.2%苯丙烯菌酮和0.5%苦参碱的质量比为3:7或6:4时,对Xcc的毒力较强,其 EC50分别为10.71和10.79 mg/L(表6)。进一步计算2种杀菌剂不同复配组合的共毒系数(CTC),由表6可知,当0.2%苯丙烯菌酮和0.5%苦参碱的质量比为3:7~6:4时,均具有增效作用,其中配比为3:7时,对Xcc的毒力增效作用最强。
表4 预实验中不同比例苯丙烯菌EC50剂量和苦参碱EC50剂量对Xcc的平均抑制率
表5 复配药剂对Xcc生长的抑制效果
表6 复配药剂对Xcc的毒力测定
实施例3 青花菜黑腐病盆栽试验
1试验方法
盆栽保护效果试验在温室进行,采用盆栽方法测定药剂对青花菜黑腐病的防效。将Xcc接种于LB液体培养基中,30℃下恒温振荡培养3 h,菌液以生理盐水稀释至浓度为3.9×107cfu/mL,选择二叶一心时期长势一致的盆栽青花菜苗,吸取20 μL菌悬液注射于青花菜幼苗茎基部。使用常量喷雾器将不同浓度的苯丙烯菌酮和苦参碱复配剂喷洒于青花菜幼苗根部和叶面,药量以叶片两面湿润、药液不下滴为准,每株喷药约300 mL,对照组使用无菌生理盐水处理,并以防治青花菜黑腐病的杀菌剂15%农用链霉素可湿性粉剂作为商品杀菌剂对照。每个处理设 3 次重复,处理后的青花菜幼苗置于温室中培养(室温在24±1℃,相对湿度 90%,L/D=12 h/12 h,培养 7 d 后调查试验结果。
病症分级标准:
0 级:无病斑;
1 级:病斑面积占整个叶面积的 5%以下;
3 级:病斑面积占整个叶面积的 6%-10%;
5 级:病斑面积占整个叶面积的 11%-25%;
7 级:病斑面积占整个叶面积的 26%-50%;
9 级:病斑面积占整个叶面积的 50%以上。
根据分级标准计算病情指数和防效。计算公式如下:
2试验结果
由表7可知,在800 mg/L剂量下,0.2%苯丙烯菌酮和0.5%苦参碱的质量比为3:7~6:4时,防治效果均远高于200 mg/L 15%农用链霉素(70.25%),特别的,当苯丙烯菌酮和苦参碱的质量比为3:7时防治效果最好,防效为90.31%。
表7 供试药剂对青花菜黑腐病的防治效果
综上所述,室内杀菌剂毒力测定结果表明,0.2%苯丙烯菌酮和0.5%苦参碱2种植物源杀菌剂对青花菜黑腐病均具有较强的抑制作用;当0.2%苯丙烯菌酮和0.5%苦参碱质量比为3:7~6:4时,对青花菜黑腐病病原菌Xcc的共毒系数均大于120,表现为增效作用,其中,当苯丙烯菌酮和苦参碱的质量比为3:7时,表现为显著增效作用;室内盆栽试验表明,苯丙烯菌酮和苦参碱的质量比为3:7~6:4时,防治效果均远高于200 mg/L 15%农用链霉素(70.25%)。
以上所述仅为本发明的优选实施例,并非因此限制本发明的专利范围,凡是在本发明的构思下,利用本发明说明书内容所作的等效变换,或直接/间接运用在其他相关的技术领域均包括在本发明的专利保护范围内。
Claims (3)
1.一种防治青花菜黑腐病的植物源杀菌剂组合,其特征在于,其有效成分由0.2% 苯丙烯菌酮和0.5%苦参碱复配而成。
2.根据权利要求1所述的植物源杀菌剂组合,其特征在于,所述0.2%苯丙烯菌酮和0.5%苦参碱的质量比为3:7~6:4。
3.根据权利要求2所述的植物源杀菌剂组合,其特征在于,所述0.2% 苯丙烯菌酮和0.5%苦参碱的质量比为3:7。
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