CN115152773B - 防治小麦茎基腐病的丙硫菌唑与咯菌腈的组合物 - Google Patents
防治小麦茎基腐病的丙硫菌唑与咯菌腈的组合物 Download PDFInfo
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- 238000011160 research Methods 0.000 description 1
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- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- QGHREAKMXXNCOA-UHFFFAOYSA-N thiophanate-methyl Chemical group COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC QGHREAKMXXNCOA-UHFFFAOYSA-N 0.000 description 1
- QPDUQKTYZRXRBC-UHFFFAOYSA-N triazole-4-thione Chemical class S=C1C=NN=N1 QPDUQKTYZRXRBC-UHFFFAOYSA-N 0.000 description 1
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- 238000004383 yellowing Methods 0.000 description 1
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Abstract
本发明公开了一种防治小麦茎基腐病的丙硫菌唑与咯菌腈的组合物,主要由以下质量分数的原料组成:丙硫菌唑1~5%,咯菌腈2~5%、分散剂5~10%、增效剂1.2~2.5%、增稠剂1~3%、木霉菌‑改性壳聚糖‑聚乙二醇复合物1~2%、消泡剂0.5~1.5%,余量为水。本发明中所添加的木霉菌‑改性壳聚糖‑聚乙二醇复合物具备良好的抗菌性,渗透性及粘附性,能够很好地保护种子免受病菌的侵害,具有良好的防治小麦茎基腐病的功效,增加小麦的产量。
Description
技术领域
本发明涉及农药技术领域,尤其涉及一种防治小麦茎基腐病的丙硫菌唑与咯菌腈的组合物。
背景技术
茎基腐病(Crown rot)是由多种病原真菌引起的一种世界性土传病害,在世界主要麦类作物种植区均有发生,且近年来呈现出越来越严重的趋势。小麦茎基腐病主要是由镰孢菌(Fusarium spp.)引起的,从小麦分蘖到成熟期均可发生,可使茎基部叶鞘和茎秆变褐,茎节坏死,严重病株早期可致死苗或后期出现白穗,造成空瘪籽粒而影响小麦产量。
小麦茎基腐病在小麦各个生长发育阶段都能产生危害,在不同时期危害小麦植株的部位不同,病害的表现症状也存在差异。小麦种子在萌发前受到病菌侵染,幼芽鞘受害出现褐色斑痕,病害严重时造成种子腐烂死亡,影响小麦出苗率。小麦出苗至返青期病菌侵染麦苗,发病程度较轻植株,其地上部分感病症状表现不明显,在胚芽鞘、地中茎和叶鞘处出现褐色或黑色病斑,根部出现腐烂病症;发病较重植株,幼苗地面以上叶面发黄,生长缓慢、植株矮小;发病严重时植株枯萎死亡。小麦拔节期病菌侵染主要危害小麦基部叶鞘和茎秆,发病植株茎基部的叶鞘颜色由绿色逐渐变为暗褐色;随着病菌侵染进一步发展蔓延,在小麦的根部、茎秆和第1~2茎节出现褐色病斑;危害严重时茎基部节间变深褐色、易折断,出现死苗现象,在降雨多、田间湿度大的情况下,在茎节处可以看见白色或红色粉状物,若剖开叶鞘,在其内部可见灰白色菌丝充满腔内。小麦孕穗期,在感病植株茎秆上出现病斑,发病植株结实率降低。在小麦灌浆期,植株茎基部部分分蘖节死亡,导致植株枯萎死亡;感染病害比较严重的植株,其叶片自下而上开始干枯,并且出现白穗现象;如果人工拔起病株,茎基部容易折断,在病株根部可见根毛、主根表皮脱落,根冠部变黑并粘附土粒,此时籽粒停止灌浆,形成的籽粒不饱满甚至无籽,严重影响小麦产量。
小麦茎基腐病是土传病害,使用药剂拌种或包衣可以在一定程度上减轻该病的发生。研究报道,甲基托布津、多菌灵、嘧菌酯、戊唑醇、氟硅唑、丙环唑、灭菌唑、苯醚甲环唑、咯菌腈、萎锈灵等杀菌剂对禾谷镰刀菌(F.graminearum)均有一定的抑制作用。播种时利用杀菌剂对小麦进行拌种对茎基腐病有一定的防治效果,但药效不能在小麦的整个生长期均有效,小麦生长后期防治效果会显著下降。
丙硫菌唑(Prothioconazole)是由拜耳公司开发和创制的脱甲基化抑制剂,其化学结构不同于以前的三唑类,属于另外一类新型的三唑硫酮类杀菌剂,丙硫菌唑的作用机理是抑制真菌中甾醇的前体——羊毛甾醇或2,4-亚甲基二氢羊毛甾醇l4位上的脱甲基化作用,即脱甲基化抑制剂(DMIs)。不仅具有很好的内吸活性,优异的保护治疗和铲除活性,且持效期长。通过大量的田间药效试验,结果表明丙硫菌唑对作物不仅具有良好的安全性,防病治病效果好,而且增产明显,同三唑类杀茵剂相比,丙硫菌唑具有更广谱的杀菌活性。丙硫菌唑主要用于防治谷类(如花生、油菜、水稻、小麦、大麦)、豆类、甜菜和大田蔬菜等作物上的众多病害,如对小麦和大麦上的白粉病、纹枯病、枯萎病、叶斑病、锈病、菌核病、网斑病、云纹病等具有很好的防治效果;此外还能防治油菜和花生的土传病害,如菌核病,以及主要叶面疾病,如灰霉病、黑斑病、褐斑病、黑胫病、菌核病和锈病等病害。由于丙硫菌唑杀菌谱广,所以其特点为应用范围广、使用时机灵活、植物吸收迅速、有良好的耐雨淋性,且持效期长,这些优点综合在一起成为防治植物病害的新标杆。
咯菌腈,也叫氟咯菌腈,是一种广谱、非内吸性吡咯类杀菌剂。咯菌腈的作用机理是通过抑制病菌内葡萄糖磷酰化有关酶的转移,并抑制真菌菌丝体的生长,最终导致病菌死亡,从而杀灭病菌的目的。当咯菌腈用作种子处理时,有效成分在搅拌及种子发芽时,大部分药剂吸附在种子表面成膜,形成一个稳定持久的保护圈,通过渗透作用,一部分进入种子内部,可以同时杀死种子表面及种皮内的病菌。对作物种子安全性极好,不影响种子出苗,并能促进种子提前出苗,出苗齐,长势壮。对人和动物非常安全,是化学合成的仿生制剂,其毒性比食盐还低,非常适合无公害蔬菜的生产。
专利CN112970760A公开了一种三元复配杀菌剂及其在小麦茎基腐病防治中的应用,所述的三元复配杀菌剂包括氟吡菌胺、霜霉威和腐植酸铜三种有效成分,且三种成份的质量份数比为6:(6~12):(4~9)。本发明的三元复配杀菌剂在小麦茎基腐病防治中的应用,不仅能提高小麦茎基腐病的防效;同时还能增加小麦营养、提高小麦的生长势,缓解小麦茎基腐病抗药性的发生,该三元复配杀菌剂在田间表现出一定的增产效果,小麦增产量达609.65kg/667m2,增产率达18.97%;对小麦茎基腐病的防效达到80.07%,且用药最佳时间在发病初期,对小麦茎基腐病的防控具有较高的参考作用,不影响小麦产量。
专利CN111996125 B一种球毛壳菌在防治小麦茎基腐病中的应用,所述球毛壳菌的保藏名称为12XP1-2-3,保藏编号为CGMCC No.17183,保藏单位为中国微生物菌种保藏管理委员会普通微生物中心,保藏日期为2019年1月23日。田间试验结果表明,用球毛壳菌12XP1-2-3进行小麦种子包衣处理,可以使病茎率、病级和病情指数分别降低19.0%-41.3%、43.7%和4.7%-45.4%。
现有技术中所公开的防治小麦茎基腐病的制剂通常是选用两种或两种以上的杀菌剂进行复配或者生物防治剂对小麦种子进行拌种或包衣,然而,由于杀菌剂通常为有机物,其在水中的溶解性很低,简单的复配很难使得杀菌剂发挥出最佳的功效,而单独的生物制剂的防治效果则有限。因此,研发出一种抗菌能力佳,渗透性强,防治效果好的组合物用于防治小麦茎基腐病十分必要。
发明内容
有鉴于现有技术中的上述缺陷,本发明所要解决的技术问题制备是一种防治效果好,渗透性强的防治小麦茎基腐病的组合物。
本发明的技术方案:
一种防治小麦茎基腐病的丙硫菌唑与咯菌腈的组合物,包括以下质量分数的组分:丙硫菌唑1~5%,咯菌腈2~5%、分散剂5~10%、增效剂1.2~2.5%、增稠剂1~3%、木霉菌-改性壳聚糖-聚乙二醇复合物1~3%、消泡剂0.5~1.5%,余量为水;
优选的,所述分散剂是由木质素磺酸钠、苯乙基酚聚氧乙烯醚、吐温80混合而成。
进一步的,所述分散剂中木质素磺酸钠、苯乙基酚聚氧乙烯醚、吐温80的质量比为1:1~3:2~5。
优选的,所述增效剂为氮素增效剂、有机硅聚氧乙烯醚化合物、氮酮类增效剂中的一种或几种。
优选的,所述增稠剂为黄原胶、羧甲基纤维素、聚氧乙烯的一种或几种。
优选的,所述消泡剂为有机硅消泡剂。
所述木霉菌-改性壳聚糖-聚乙二醇复合物的制备方法包括如下步骤:
S1将哈茨木霉菌按照1~2wt%的接种量接种于PDA培养基上,于25~30℃有氧条件下培养3~6天,得到活化菌株;
S2将活化的哈茨木霉菌用5mm打孔器取3~4个菌饼接枝到PDA液体培养基上,在25~30℃,密闭发酵培养,得到发酵液;
S3将步骤S2中的20~30重量份发酵液接种到固体发酵培养基中,在25~30℃的有氧条件下固体发酵72~96h,30~40℃烘干后,研磨,密封,得到分生孢子粉;
S4称取5~6重量份的壳聚糖加入到40~45重量份的水中,搅拌30~40min,再加入1~2mol/L的盐酸调节pH至5,得到溶液A,称取5~6重量份的咖啡酸加入到40~45重量份的四氢呋喃中,得到溶液B,将溶液A、溶液B混合,加入1~2重量份的脂肪酶,在60~65℃下搅拌20~24h,反应结束后,于45~50℃,0.07~0.09MPa下浓缩除去四氢呋喃,再加入50~60重量份的水,于4500~5000rpm下离心5~10min取下清液,下清液中加入50~60重量份的乙醇,有沉淀出现,过滤,沉淀于﹣30~﹣40℃,7~10Pa下冷冻干燥得到改性壳聚糖;
S5称取1.5~2重量份步骤S4中的改性壳聚糖、0.5~1.0重量份的聚乙二醇分散于80~90重量份水中,加入0.1~0.2重量份的冰醋酸,再加入1.0~1.5重量份的甘油,在60~80下搅拌20~24h,得到混合物;
S6称取1.0~1.5重量份步骤S3中的分生孢子粉加入到步骤S5的混合物中,在摇床上以150~200rpm混合15~20min,得到均质的木霉菌-改性壳聚糖-聚乙二醇复合物。
优选的,所述PDA培养基的组分包括:马铃薯汁20~25g,葡萄糖2~3g、琼脂1~2g,加水定容至100mL,pH=6.8~7.0。
优选的,所述PDA液体培养基的组分包括:马铃薯12~15g,葡萄糖3~5g,磷酸氢二钾0.8~1.5g,氯化钠1~3g,加水定容至1000mL,pH=4.5~5。
优选的,所述固体培养基的组分包括:麦麸30~50g,豆粕20~40g,稻壳10~30g,矿物质元素0.5~2g,加水至混合物密度范围为3~7g/mL,pH=4.5~5。
本发明还提供了所述组合物的使用方法,包括以下步骤:
S1用所述复配杀菌组合物与小麦种子按照1:500~600的重量比拌种,充分搅拌10~30min;
S2在阴凉处晾干种子,种皮见干后可播种。
优选的,所述拌种温度为10~20℃。
优选的,所述晾干温度为30~40℃。
壳聚糖是农业中使用的生物聚合物之一,应用广泛。壳聚糖作为种子处理剂使用时,表现为抗性诱导剂,可诱导幼苗生理上的增强幼苗和植物的防御能力,增强植物的先天防御能力。将像木霉菌这样的生物控制剂与其他天然种子包衣物质(如壳聚糖)结合起来,可以提高种子的活力和在水分胁迫下的发芽率,并对病原体的入侵产生协同作用。然而,壳聚糖结构表面存在大量的活性基团如氨基、羧基等,导致其很容易被降解,因此常常对其改性进行修饰以提高水溶性及稳定性。
本发明人发现,通过将木霉菌-改性壳聚糖-聚乙二醇复合物添加到杀菌剂中对种子进行处理,具有良好的抗菌效果。由于壳聚糖具有良好的粘附性和抗菌性,在本发明中被用作种子处理的木霉菌传递载体。在此基础上,本发明还利用脂肪酶与咖啡酸制备了改性壳聚糖,脂肪酶作为一种良好的生物催化剂成功提高了接枝率,而咖啡酸中的羧基能够与壳聚糖中的氨基和羟基反应,形成羟基和酯基,使得壳聚糖的晶体结构被破坏从而水溶性增强,而活性基团由于接枝了咖啡酸从而得到了保护。同时咖啡酸作为一种酚酸具有良好的抑菌性,这导致接枝后的改性壳聚糖具备了更强的抑菌活性。对聚乙二醇作为一种可生物降解的、水溶性的聚合物,能够很好地保护像木霉菌这样的生物控制剂,并帮助其进行渗透。为了提高组合物在种子表面的润湿性和复合物的粘附性,在组合物中加入甘油作为表面活性剂,降低看表面张力,并保持极性和非极性基团之间的平衡,加强了渗透效果。所制备的木霉菌-改性壳聚糖-聚乙二醇复合物可以在种子周围创造一个保护性的微环境,保证微生物与种子的粘附,改善植物的生长,对于防治小麦的茎基腐病具有良好的效果。
与现有技术相比,本发明的有益效果:
(1)所制备的木霉菌-改性壳聚糖-聚乙二醇复合物中壳聚糖在添加脂肪酶的条件下与咖啡酸进行反应,得到改性壳聚糖,脂肪酶作为一种良好的生物催化剂成功提高了接枝率,改性后的壳聚糖水溶性及稳定性增强,同时咖啡酸作为一种酚酸具有良好的抑菌性,这导致接枝后的改性壳聚糖具备了更强的抑菌活性;
(2)聚乙二醇作为一种可生物降解的、水溶性的聚合物,能够很好地保护像木霉菌这样的生物控制剂,并帮助其进行渗透。为了提高组合物在种子表面的润湿性和复合物的粘附性,在组合物中加入甘油作为表面活性剂,降低看表面张力,并保持极性和非极性基团之间的平衡,渗透效果进一步加强;
(3)过将木霉菌-改性壳聚糖-聚乙二醇复合物添加到杀菌剂中对种子进行处理,具有良好的抗菌效果,对于小麦茎基腐病具有优异的防治效果,并且不会影响小麦的产量。
具体实施方式
下面,通过具体实施例对本发明的技术方案进行详细说明,但是应该明确提出这些实施例用于举例说明,但是不解释为限制本发明的范围。
本发明实施例中部分原料的参数如下:
哈茨木霉菌(Trichoderma harzianum),中国普通微生物菌种保藏管理中心,CGMCC编号:5.1213。
壳聚糖,脱乙酰度>95%,粘度:100~200mPa·s。
脂肪酶,CNB20025,50000U/g,阿拉丁。
3,4-二甲基吡唑磷酸盐(DMPP),含量:98%,水分含量0.1%。
聚乙二醇,分子量5500~7000,水分≤1.0%。
禾谷镰刀菌(Fusarium graminearum),ATCC编号:Bio-83590。
对照例1
一种防治小麦茎基腐病的丙硫菌唑与咯菌腈的组合物的制备方法,包括以下步骤:
S1将哈茨木霉菌按照1wt%的接种量接种于PDA培养基上,于25℃有氧条件下培养3天,得到活化菌株,PDA培养基的组分包括:马铃薯汁20g,葡萄糖2g、琼脂1.5g,加水定容至100mL,pH=7.0;
S2将活化的哈茨木霉菌用5mm打孔器取3个菌饼接枝到PDA液体培养基上,液体培养基的组分包括:马铃薯12g,葡萄糖3g,磷酸氢二钾0.8g,氯化钠1g,加水定容至1000mL,pH=5,在25℃下密封发酵培养,得到发酵液;
S3将20g步骤S2中的发酵液接种到固体发酵培养基中,固体培养基的组分包括:麦麸30g,豆粕20g,稻壳10g,矿物质元素0.5g,加水至混合物密度范围为4g/mL,pH=5,在25℃有氧条件下固体发酵72h,30℃烘干后,研磨,密封,得到分生孢子粉;
S4称取丙硫菌唑10g,咯菌腈20g,木质素磺酸钠10g,苯乙基酚聚氧乙烯醚20g,吐温8020g,DMPP 12g,黄原胶10g,分生孢子粉10g,有机硅消泡剂5g,1L水,混合均匀,即得丙硫菌唑与咯菌腈的组合物。
实施例1
一种防治小麦茎基腐病的丙硫菌唑与咯菌腈的组合物的制备方法,包括以下步骤:
称取丙硫菌唑10g,咯菌腈20g,木质素磺酸钠10g,苯乙基酚聚氧乙烯醚20g,吐温8020g,DMPP 12g,黄原胶10g,木霉菌-改性壳聚糖-聚乙二醇复合物10g,有机硅消泡剂5g,1L水,混合均匀,即得丙硫菌唑与咯菌腈的组合物。
所述木霉菌-改性壳聚糖-聚乙二醇复合物的制备方法,包括以下步骤:
S1将哈茨木霉菌按照1wt%的接种量接种于PDA培养基上,于25℃有氧条件下培养3天,得到活化菌株,PDA培养基的组分包括:马铃薯汁20g,葡萄糖2g、琼脂1.5g,加水定容至100mL,pH=7.0;
S2将活化的哈茨木霉菌用5mm的打孔器取3个菌饼接枝到PDA液体培养基上,液体培养基的组分包括:马铃薯12g,葡萄糖3g,磷酸氢二钾0.8g,氯化钠1g,加水定容至1000mL,pH=5,在25℃密封发酵培养,得到发酵液;
S3将20g步骤S2中的发酵液接种到固体发酵培养基中,固体培养基采用如下方法制备:麦麸30g,豆粕20g,稻壳10g,矿物质元素0.5g,加水至混合物密度范围为4g/mL,pH=5;在25℃有氧条件下固体发酵72h,30℃烘干后,研磨,密封,得到分生孢子粉;
S4称取50g的壳聚糖加入到400mL的水中,搅拌30min,再加入2mol/L的盐酸调节pH至5,得到溶液A;称取50g的咖啡酸加入到400mL的四氢呋喃中,得到溶液B;将溶液A、溶液B混合,加入10g的脂肪酶,在60℃下搅拌24h,反应结束后,于45℃,0.09MPa下浓缩除去四氢呋喃,再加入500mL的水,于4500rpm下离心5min取下清液,下清液中加入500mL的乙醇,有沉淀出现,过滤,沉淀于﹣30℃,10Pa下冷冻干燥24h得到改性壳聚糖;
S5称取15g步骤S4中的改性壳聚糖、10g聚乙二醇分散于800mL水中,再加入10g甘油,在60℃下搅拌24h,得到混合物;
S6称取10g步骤S3中的分生孢子粉加入到步骤S5的混合物中,在摇床上以150rpm混合15min,得到木霉菌-改性壳聚糖-聚乙二醇复合物。
实施例2
一种防治小麦茎基腐病的丙硫菌唑与咯菌腈的组合物的制备方法,包括以下步骤:
称取丙硫菌唑10g,咯菌腈20g,木质素磺酸钠10g,苯乙基酚聚氧乙烯醚20g,吐温8020g,DMPP 12g,黄原胶10g,改性壳聚糖-聚乙二醇复合物10g,有机硅消泡剂5g,1L水,混合均匀,即得丙硫菌唑与咯菌腈的组合物。
所述改性壳聚糖-聚乙二醇复合物的制备方法,包括以下步骤:
S1称取50g的壳聚糖加入到400mL的水中,搅拌30min,再加入2mol/L的盐酸调节pH至5,得到溶液A,称取50g的咖啡酸加入到400mL的四氢呋喃中,得到溶液B,将溶液A、溶液B混合,加入10g的脂肪酶,在60℃下搅拌24h,反应结束后,于45℃,0.09MPa下浓缩除去四氢呋喃,再加入500mL的水,于4500rpm下离心5min取下清液,下清液中加入500mL的乙醇,有沉淀出现,过滤,沉淀于﹣30℃,10Pa下冷冻干燥24h得到改性壳聚糖;
S2称取15g改性壳聚糖、10g聚乙二醇分散于800mL水中,再加入10g甘油,在60℃下搅拌24h,得到改性壳聚糖-聚乙二醇复合物。
实施例3
一种防治小麦茎基腐病的丙硫菌唑与咯菌腈的组合物的制备方法,包括以下步骤:
称取丙硫菌唑10g,咯菌腈20g,木质素磺酸钠10g,苯乙基酚聚氧乙烯醚20g,吐温8020g,DMPP 12g,黄原胶10g,有机硅消泡剂5g,1L水,混合均匀,即得丙硫菌唑与咯菌腈的组合物。
测试例1
对对照例及实施例所制备的丙硫菌唑与咯菌腈的组合物进行室内毒力测定,供试菌种采用小麦茎基腐病菌禾谷镰刀菌(Fusarium graminearum)。首先将200g马铃薯去皮切块,放入锅中加水煮沸20min,用四层纱布过滤,将滤液倒入锅中继续加热,依次加入琼脂20g,葡萄糖20g,搅拌混匀溶解后,补充水分至1000mL,冷却,分装,加塞,包扎。将装好PDA的三角瓶至于湿热灭菌锅中,压强0.1Mpa,温度121℃条件下,灭菌20min后取出,冷却后备用。用无菌水将不同丙硫菌唑与咯菌腈的组合物稀释为5mg/kg,然后使用移液管吸取2mL的药液放入18mL的PDA培养基中混匀后倒入培养皿,每个处理4次重复,以正常PDA培养基为对照。用打孔器打取直径为6mm的菌饼,在每个冷却凝固的平板中央放入打好的菌饼,用保鲜膜密封,做好标记,在25℃培养箱中培养5d,测量菌落直径(单位:mm)。用直尺以十字交叉法测量菌落直径,取其去除菌饼直径(菌饼直径:6mm)的平均值代表菌落大小。根据以下公式计算菌丝生长抑制率:菌丝生长抑制率=(对照菌落的直径-处理菌落的直径)/对照菌落的直径*100%,具体结果见表1。
表1抑菌结果表
实验方案 | 抑菌率/% |
对照例1 | 63.24 |
实施例1 | 70.16 |
实施例2 | 54.37 |
实施例3 | 50.11 |
通过抑菌实验可以得知,实施例1所制备的组合物抑菌率最高,对照例1中添加木霉菌孢子粉作为生物防治剂,木霉菌自身具备一定的抗菌性,对于禾谷镰刀菌具有一定的防治效果,然而单独添加生物防治剂的抑菌效果有限,然而实施例1中所添加的木霉菌-改性壳聚糖-聚乙二醇复合物由于壳聚糖具有良好的粘附性和抗菌性,对于禾谷镰刀菌具有一定的粘附效果,壳聚糖作为木霉菌传递载体能够帮助其更好地渗透,咖啡酸作为一种酚酸具有良好的抑菌性,接枝在壳聚糖表面使得制剂具备了更佳的抑菌活性。
测试例2
利用小米培养基进行病原菌扩繁并在室内进行盆栽试验,将小米洗净放入锅中加水煮沸2min,晾至表面无水后分装,封口,将小米的三角瓶于0.1Mpa,121℃下灭菌30min后取出,冷却后备用。在灭好菌的小米培养基中接入禾谷镰刀菌,放入25℃培养箱培养7d,定期摇晃确保菌落均匀分布在小米表面。小米培养基和无菌土以0.75:100的比例混匀,将本发明所制备的丙硫菌唑与咯菌腈的组合物对矮抗58小麦种子进行拌种,将菌土转入营养钵中,每钵播种10粒种子,并设置不拌种种植于带菌土壤中为对照,各处理设置3个重复,于25℃,湿度70%条件下培养1月后对出苗率、发病情况以及植株的发育情况进行调查。按照小麦茎基腐病的病情分级计算病情指数和防效:0级:整株无变褐症状;1级:植株仅第一叶鞘变褐,且变褐面积不超过1/2;2级:植株仅第一叶鞘变褐,且变褐面积超过1/2;3级:植株仅第一叶鞘变褐,变褐面积超过1/2,且褐色加深;4级:植株第二叶鞘变褐;5级:植株第三叶鞘变褐或植株死亡。病情指数=∑(各级病株数*代表数值)/(调查总株数*发病最重级的代表数值)*100%,防治效果=(对照平均病情指数-处理平均病情指数)/对照病情指数*100%。
表2室内盆栽试验防治效果
实验方案 | 病情指数/% | 防效/% |
对照例1 | 23.15a | 55.36 |
实施例1 | 18.24b | 64.82 |
实施例2 | 27.35a | 47.26 |
实施例3 | 42.89c | 17.29 |
对照组 | 51.86 | / |
通过室内盆栽防治试验结果可以看出,添加了木霉菌-改性壳聚糖-聚乙二醇复合物的丙硫菌唑与咯菌腈的组合物对于小麦茎基腐病具有良好的室内防治效果,这可能是由于壳聚糖作为木霉菌的传递载体能够帮助其在对小麦种子进行包衣时更好地进行渗透,咖啡酸的接枝也进一步提高了复合物的抗菌效果。聚乙二醇作为一种渗透保护剂,能够提高复合物在种子表面的粘附性,在组合物中加入甘油作为表面活性剂,可以降低表面张力,并保持极性和非极性基团之间的平衡,因而所制备的组合物可以在种子周围创造一个保护性的微环境,对于小麦茎基腐病的防治具有良好的效果。
测试例3
对对照例及实施例的组合物对小麦种子进行包衣处理,制剂用量均为10g/10kg种子,拌种后选择进行选择小麦茎基腐病常年发生的小麦地为供试点进行播种,土壤pH为6~7、肥力均匀、地块平整,测试组合物的田间防治效果。麦田栽培管理与常规管理相同。田间设置五个处理区,每个供试区面积为20m2,每处理重复3次,按随机区组排列,每个处理区边缘设保护行。作物生长期调查3次。第1次在冬前苗期,第2次在返青拔节期,第3次在小麦灌浆期。每个供试区采用5点取样法,每个点取25株苗。将植株连根挖出,保持根系完整。苗期调查时测量株高(土面以上至每株最高叶尖的平均高度为株高)、根长,以及地上部分鲜重和根重,与对照比较,计算不同处理对小麦苗期生长发育的影响。在收获期进行测产,每个供试区5点取样,每点取1m2小麦植株,实收测产,并折合每公顷产量,并计算增产率。增产率=(处理区产量-对照区产量)/对照区产量*100%。具体结果见表3。
表3田间防治及出苗效果结果表
通过田间防治试验可以看出,经实施例1的组合物处理后的小麦种子不仅长势好,产量也高,这可能是由于添加的木霉菌-改性壳聚糖-聚乙二醇复合物具备良好的抗菌性,而聚乙二醇作为一种渗透保护剂,能够提高复合物在种子表面的粘附性,在组合物中加入甘油作为表面活性剂,可以降低表面张力,并保持极性和非极性基团之间的平衡,在包衣处理时能够在种子周围创造一个保护性的微环境,导致小麦在生长过程中不会受到病菌的侵害,从而表现出良好的长势,产量也有所增加。
Claims (6)
1.一种防治小麦茎基腐病的丙硫菌唑与咯菌腈的组合物,其特征在于,包括以下质量分数的原料:丙硫菌唑1~5%,咯菌腈2~5%、分散剂5~10%、增效剂1.2~2.5%、增稠剂1~3%、木霉菌-改性壳聚糖-聚乙二醇复合物1~2%、消泡剂0.5~1.5%,余量为水;
所述木霉菌-壳聚糖-聚乙二醇复合物的制备方法,包括如下步骤:
S1将哈茨木霉菌按照1~2wt%的接种量接种于PDA培养基上,于25~30℃有氧条件下培养3~6天,得到活化菌种;
S2将活化的哈茨木霉菌用5mm打孔器取3~4个菌饼接枝到PDA液体培养基上,在25~30℃,密闭发酵培养,得到发酵液;
S3取步骤S2中的发酵液20~30重量份接种到固体发酵培养基中,在25~30℃的有氧条件下固体发酵72~96h,30~40℃烘干后,研磨,密封,得到分生孢子粉;
S4称取5~6重量份的壳聚糖加入到40~45重量份的水中,搅拌30~40min,再加入1~2mol/L的盐酸调节pH至4~5,得到溶液A,称取5~6重量份的咖啡酸加入到40~45重量份的四氢呋喃中,得到溶液B,将溶液A、溶液B混合,加入1~2重量份的脂肪酶,在60~65℃下搅拌20~24h,反应结束后,浓缩除去四氢呋喃,再加入50~60重量份的水,于4500~5000rpm下离心5~10min取下清液,下清液中加入50~60重量份的乙醇,有沉淀出现,过滤取沉淀再冷冻干燥得到改性壳聚糖;
S5取1.5~2重量份步骤S4中的改性壳聚糖、0.5~1.0重量份的聚乙二醇、80~90重量份水、0.1~0.2重量份的冰醋酸,1.0~1.5重量份的甘油配制溶液,在60~80℃下搅拌20~24h,得到混合物;
S6称取1.0~1.5重量份步骤S3中的分生孢子粉加入到步骤S5的混合物中,在摇床上以150~200rpm混合15~20min,得到均质的木霉菌-改性壳聚糖-聚乙二醇复合物。
2.如权利要求1所述的丙硫菌唑与咯菌腈的组合物,其特征在于:所述分散剂中木质素磺酸钠、苯乙基酚聚氧乙烯醚、吐温80的质量比为1:1~3:2~5。
3.如权利要求1所述的丙硫菌唑与咯菌腈的组合物,其特征在于:所述增效剂为氮素增效剂、有机硅聚氧乙烯醚化合物、氮酮类增效剂中的一种或几种。
4.如权利要求1所述的丙硫菌唑与咯菌腈的组合物,其特征在于:所述增稠剂为黄原胶、羧甲基纤维素、聚氧乙烯的一种或几种,所述消泡剂为有机硅消泡剂。
5.如权利要求1所述的丙硫菌唑与咯菌腈的组合物,其特征在于,所述PDA培养基的组分包括:马铃薯汁20~25g,葡萄糖2~3g、琼脂1~2g,加水定容至100mL,pH=6.8~7.0;所述PDA液体培养基的组分包括:马铃薯12~15g,葡萄糖3~5g,磷酸氢二钾0.8~1.5g,氯化钠1~3g,加水定容至1000mL,pH=4.5~5。
6.如权利要求1所述的丙硫菌唑与咯菌腈的组合物,其特征在于,所述固体培养基的组分包括:麦麸30~50g,豆粕20~40g,稻壳10~30g,矿物质元素0.5~2g,加水至混合物密度范围为3~7g/mL,pH=4.5~5。
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