CN114586584B - 一种基于全周期生物***的甜椒绿色轻简控害方法 - Google Patents
一种基于全周期生物***的甜椒绿色轻简控害方法 Download PDFInfo
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Abstract
本发明公开了一种基于全周期生物***的甜椒绿色轻简控害方法,具体针对甜椒种植全周期中以黑斑病和白粉病为代表的病害及以蓟马和粉虱为代表的虫害,构建“一灭、二防、三抗、四控、五治”的绿色轻简控害方法,操作简便、控害高效、经济环保,在保证甜椒高品质和产量的同时,显著减少了化学农药的使用,对甜椒的绿色生产及保护生态环境具有重要意义。
Description
技术领域
本发明涉及蔬菜病虫害绿色防控技术领域,更具体的说是涉及一种基于全周期生物***的甜椒绿色轻简控害方法。
背景技术
甜椒是世界各地广泛栽培的大宗、优良蔬菜品种之一,也是国内周年化供应的重要蔬菜之一。甜椒的维生素含量在果菜中最高,还富含胡萝卜素、辣椒红素、梓檬酸、苹果酸及多种糖类、蛋白质和矿物质等营养物质,具改善食欲、促进消化和防治坏血病等功效,深受人们的喜爱。
我国甜椒种植主要分布在东南、华北和东北等沿海区域,特别是在山东、海南和福建等地区已形成规模化生产基地。由于甜椒消费量不断增加,生产规模逐年扩大,种植面积达40万公顷以上,约占椒类种植总面积的1/4,年产值达100亿以上,不仅成为蔬菜发展高增长的带动产业,实现了农业增效和农民增收,也带动了种植区域周边地区的农资、加工和物流等第二、三产业的发展。
但随着甜椒产业的发展与壮大,生长栽培模式逐步向设施化和集约化转变。栽种方式的变革,为以黑斑病和白粉病为代表的病害及以蓟马和粉虱为代表的虫害提供了有利的生存环境,在甜椒种植区呈蔓延扩展态势,直接、间接影响了甜椒的产量和品质,急需解决。
目前,针对甜椒病虫害仍缺乏***防控思维和贯穿整个甜椒种植全程的“绿色、轻简”控害技术的有效集成,“头痛医头、脚痛医脚”,化学农药过量施用现象普遍存在,导致病虫抗药性上升、有益生物减少和药害残留等生态和食品安全问题日益突出,与我国农业的绿色引领导向冲突不断。
因此,如何提供一种绿色、轻简的控害方法用于保证甜椒高品质和产量的同时,显著减少化学农药的使用,是本领域技术人员亟需解决的技术问题。
发明内容
有鉴于此,本发明的目的在于提供一种基于全周期生物***的甜椒绿色轻简控害方法。本发明针对甜椒种植全程中以黑斑病和白粉病为代表的病害及以蓟马和粉虱为代表的虫害,构建“一灭、二防、三抗、四控、五治”的绿色轻简控害方法,操作简便、控害高效、经济环保,在保证甜椒高品质和产量的同时,显著减少了化学农药的使用,对甜椒的绿色生产及保护生态环境具有重要意义。
为了实现上述目的,本发明采用如下技术方案:
一种基于全周期生物***的甜椒绿色轻简控害方法,具体包括以下步骤:
(1)一灭
种植甜椒之前,每亩撒施由钩状木霉菌TH-01菌粉3-5kg、蜡蚧轮枝菌LL-01菌粉3-5kg、硅藻土10-15kg和生石灰20-30kg混拌而成的菌土进行翻耕,并用由钩状木霉菌TH-01菌粉和蜡蚧轮枝菌LL-01菌粉混合制得的菌剂进行浸种或浸苗;
(2)二防
种植甜椒之后,在温室、大棚风口和门口处增设银灰条膜和防虫网,定期维护棚膜、门窗和防虫网;畦面覆盖地膜;
(3)三抗
日常肥水增施乙酰水杨酸粉剂和钩状木霉菌TH-01菌粉;
(4)四控
在阴雨天气,于中午前后进行放风;出苗后在植株上方悬挂黄色诱虫板和蓝色诱虫板,且每片黄色诱虫板和蓝色诱虫板上各加装一个蓟马信息素诱芯和粉虱信息素诱芯;在设施温棚内部四周间作伴生植物;
(5)五治
当诱虫量≤3头/板时,释放微小花蝽或淡翅小花蝽,以及携菌加州新小绥螨或携菌智利小植绥螨;
当3<诱虫量≤5头/板时,在释放天敌之前,利用蜡蚧轮枝菌LL-01菌粉进行植株和地面喷雾;
当诱虫量>5头/板时,轮换施用苦参·印楝素乳油、硅藻土可湿性粉剂、乙基多杀菌素悬浮剂和噻虫嗪悬浮剂进行植株和地面喷雾;
当病株率≤3%时,利用钩状木霉菌TH-01菌粉进行植株和地面喷雾;
当病株率>3%时,轮换施用多抗霉素可湿性粉剂、中生菌素可湿性粉剂、氯溴异氰尿酸可溶性粉剂和苯甲·咪鲜胺水乳剂进行植株和地面喷雾;
中耕时,在畦间每亩撒施由钩状木霉菌TH-01菌粉2-3kg、蜡蚧轮枝菌LL-01菌粉2-3kg、麦麸3-5kg以及3-5瓶、净含量≥20000只/瓶的日本毛绥螨或地理厉螨混拌而成的基质。
进一步,上述钩状木霉菌TH-01的分类命名为钩状木霉(Trichoderma hamatum),保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址为北京市朝阳区北辰西路1号院3号,保藏编号为CGMCCNo.40078,保藏日期为2022年02月15日;蜡蚧轮枝菌LL-01的分类命名为蜡蚧轮枝菌(Lecanicillium lecanii),保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址为北京市朝阳区北辰西路1号院3号,保藏编号为CGMCCNo.21940,保藏日期为2021年3月31日;钩状木霉菌TH-01菌粉的孢子含量为3亿/g,蜡蚧轮枝菌LL-01菌粉的孢子含量为5亿/g。
进一步,上述步骤(1)中,所述菌剂稀释至100-200倍液。
采用上述进一步技术方案的有益效果在于,通过菌土翻耕和菌剂浸种或浸苗,能够杀灭土壤中及种子、种苗表面的病原菌和虫蛹,减少初代病、虫基数。通过增设银灰条膜和防虫网,能够防止外部病虫进入温室;通过畦面覆盖地膜,既能够防止植株病虫入土及地下病虫出土,减少初代和次代病虫基数,又能降低透光率,抑制杂草生长,防止病虫滋生。
进一步,上述步骤(3)中,所述乙酰水杨酸粉剂的质量浓度为98%,用量为20-30g/亩;所述钩状木霉菌TH-01菌粉的用量为40-60g/亩。
采用上述进一步技术方案的有益效果在于,通过日常肥水增施乙酰水杨酸粉剂和钩状木霉菌TH-01菌粉,能够促进植株光合作用和新陈代谢,加快营养和生殖生长,诱导激活植株防御反应,增强植株免疫能力,提高植株抗逆性。此外,钩状木霉菌TH-01还能改善植物生长的微环境,与病原菌形成竞争作用,在根围和叶围迅速生长,抢占植物体表面的位点,形成类似靴子作用的保护罩,阻止病虫直接接触到根系和叶片表面,还对病虫进行重寄生,分泌胞外酶溶解、穿透寄主细胞壁,吸取寄主营养,进而杀死寄主,以此减轻病虫为害程度,促根壮苗护花保果。
进一步,上述步骤(4)中,所述放风的时间为2-3h;所述黄色诱虫板和蓝色诱虫板的用量为1:1,各20-40片/亩,悬挂位置为植株上方10-15cm处,更换频率为20-40天/次;伴生植物为芹菜和茼蒿,种植宽度为20-30cm。
采用上述进一步技术方案的有益效果在于,通过悬挂黄色诱虫板和蓝色诱虫板,能够诱集、监控害虫种群;通过在设施温棚内部四周间作伴生植物,能够驱避、调控蓟马、粉虱种群。
进一步,上述步骤(5)中,当诱虫量≤3头/板时,所述微小花蝽或淡翅小花蝽的释放量为300-500头/亩,释放频率为30-40天/次;所述携菌加州新小绥螨或携菌智利小植绥螨所携的菌为蜡蚧轮枝菌LL-01菌粉,成螨量≥200只/袋,释放量为70-100袋/亩,释放频率为30-40天/次;
当3<诱虫量≤5头/板时,所述蜡蚧轮枝菌LL-01菌粉的用量为60-100g/亩;所述喷雾的次数为1-2次,频率为8-12天/次;
当诱虫量>5头/板时,所述苦参·印楝素乳油的质量浓度为1%,用量为70-100mL/亩;所述硅藻土可湿性粉剂的质量浓度为88%,用量为1200-1400g/亩;所述乙基多杀菌素悬浮剂的质量浓度为6%,用量为12-16g/亩;所述噻虫嗪悬浮剂的质量浓度为21%,用量为14-18mL/亩;所述喷雾的次数为1-2次,频率为8-12天/次;
当病株率≤3%时,所述钩状木霉菌TH-01菌粉的用量为70-120g/亩;所述喷雾的次数为1-3次,频率为8-12天/次;
当病株率>3%时,所述多抗霉素可湿性粉剂的质量浓度为10%,用量为100-120g/亩;所述中生菌素可湿性粉剂的质量浓度为3%,用量为50-80g/亩;所述氯溴异氰尿酸可溶性粉剂的质量浓度为50%,用量为40-50g/亩;所述苯甲·咪鲜胺水乳剂的质量浓度为35%,用量为60-80mL/亩;所述喷雾的次数为1-3次,频率为8-12天/次。
采用上述进一步技术方案的有益效果在于,以虫治虫,以螨携菌治虫,以菌治虫,以药治虫(作为应急预案),以菌、螨协同共治土壤中的病原菌和虫蛹,减少后续病虫发生。
其中,苦参·印楝素乳油为植物源药剂,源于天然,对植物不易产生药害,对害虫具有强烈的拒食、驱避和抑制生长发育的作用,在植株体内有良好的内引传导特性,在环境中易降解,对人、畜、鸟类和蜜蜂安全,对靶标害虫不易产生抗药性;
硅藻土可湿性粉剂为矿物源药剂,由硅藻镶嵌的外壳沉积形成,具有似针一样尖锐的外壳,其粉末的每一细微颗粒都带有非常锐利的边缘和尖锐的突刺,当害虫爬过时若粘附于其体表,它即可由害虫之运动刺穿其外壳或柔软蜡壳结构,可以入侵到昆虫的表皮,甚至进入害虫体内,不仅能引起害虫呼吸、消化、生殖、运动等***紊乱,而且能吸收3-4倍于自身重量的水分,致使害虫体液锐减,使昆虫维持生命的体液外漏,在失去10%以上的体液后死亡;硅藻土也会吸收昆虫身体的外层蜡质,使害虫脱水死亡;还能改良土壤,起到保湿、疏松土质、延长药效肥效时间、助长农作物生长的功效;
乙基多杀菌素悬浮剂为微生物代谢产物药剂,能够作用于烟碱型乙酰胆碱受体(nAChr)和γ-氨基丁酸(GABA)受体,但又与吡虫啉等烟碱类杀虫剂和已知的阿维菌素在作用位点是不同的,独一无二的作用方式使其对害虫具有快速触杀和摄食毒性;迄今为止,尚未发现某类杀虫剂产品能以相同的作用方式影响昆虫的神经***,因而它的使用对已有的杀虫剂产生交互抗性的风险很小;同时对鸟类、鱼类、蚯蚓和水生植物低毒;在实际应用中,对蜜蜂几乎无毒;适用于有害生物综合治理;
噻虫嗪悬浮剂为高效低风险化学药剂,对害虫具有胃毒、触杀及内吸活性,用于叶面喷雾及土壤灌根处理,其施药后迅速被内吸,并传导到植株各部位,对刺吸式或锉吸式害虫如蓟马、粉虱等有良好的防效;
多抗霉素可湿性粉剂为微生物代谢产物药剂,属于广谱性抗生素类杀菌剂,具有较好的内吸传导作用,其作用机理是干扰病菌细胞壁几丁质的生物合成,使菌体细胞壁不能进行生物合成导致病菌死亡;芽管和菌丝接触药剂后,局部膨大、破裂、溢出细胞内含物,而不能正常发育,导致死亡;因此还具有抑制病菌产孢和病斑扩大的作用;
中生菌素可湿性粉剂为微生物代谢产物药剂,其抗菌谱广,具有触杀、渗透作用;能通过抑制病原细菌蛋白质的肽键生成,最终导致细菌死亡;对真菌可抑制菌丝的生长、抑制孢子的萌发,起到防治真菌性病害的作用;可刺激植物体内植保素及木质素的前体物质的生成,从而提高植物的抗病能力;同时具有一定的增产作用;
氯溴异氰尿酸可溶性粉剂为高效低风险化学药剂,其作用机理是喷施到作物表面能慢慢地释放Cl和Br,形成次氯酸(HCIO)和次溴酸(HBrO)分子,高效、广谱,具内吸性,可杀灭各种细菌、藻类、真菌和病菌,使用安全、简便、用量少、杀菌效果良好;还具一定的促进作物营养生长作用;
苯甲·咪鲜胺水乳剂为高效低风险化学药剂,通过抑制病原菌甾醇的生物合成起作用,高效、广谱、低毒,具有内吸、传导预防、保护、治疗、铲除等多重作用,对多种病原菌均具有良好的防效;
经由上述的技术方案可知,与现有技术相比,本发明的有益效果如下:
1、高效:对甜椒病虫的控害效果达82%-96%,提升控害效果11%-25%;
2、环保:减少化学农药使用量25%-33%;
3、经济:降低控害成本33%-46%;
4、增益:提高甜椒产量11%-13%和商品合格率12%-14%。
附图说明
图1为菌株TH-01的菌落特征;
图2为菌株TH-01的显微特征;
图3为接种菌株TH-01处理后的甜椒黑斑病菌;
图4为空白对照的甜椒黑斑病菌;
图5为菌株LL-01对花蓟马的侵染;
图6为菌株LL-01对花蓟马的致死率。
具体实施方式
下面对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1
基于全周期生物***的甜椒绿色轻简控害方法,具体包括以下步骤:
(1)一灭
种植甜椒之前,每亩撒施由3亿孢子/g的钩状木霉菌TH-01菌粉3kg、5亿孢子/g的蜡蚧轮枝菌LL-01菌粉3kg、硅藻土10kg和生石灰20kg混拌而成的菌土进行翻耕,并用由3亿孢子/g的钩状木霉菌TH-01和5亿孢子/g的蜡蚧轮枝菌LL-01菌粉混合制得的菌剂200倍液进行浸种或浸苗;
(2)二防
种植甜椒之后,在温室、大棚风口和门口处增设银灰条膜和防虫网,定期维护棚膜、门窗和防虫网;畦面覆盖地膜;
(3)三抗
日常肥水增施98%乙酰水杨酸粉剂25g/亩和3亿孢子/g的钩状木霉菌TH-01菌粉50g/亩;
(4)四控
在阴雨天气,于中午前后放风3h;出苗后在植株上方10cm处以1:1组合悬挂黄色诱虫板和蓝色诱虫板各30片/亩,且每片黄色诱虫板和蓝色诱虫板上各加装一个蓟马信息素诱芯和粉虱信息素诱芯,隔30天更换诱虫板和诱芯1次;在设施温棚内部四周间作30cm宽的伴生植物芹菜和茼蒿;
(5)五治
当诱虫量≤3头/板时,每亩释放微400头小花蝽或淡翅小花蝽,以及80袋、成螨量≥200只/袋、携带5亿孢子/g蜡蚧轮枝菌LL-01菌粉的携菌加州新小绥螨或携菌智利小植绥螨,每隔35天释放一次;
当3<诱虫量≤5头/板时,在释放天敌之前,利用5亿孢子/g的蜡蚧轮枝菌LL-01菌粉80g/亩进行植株和地面喷雾1,每次间隔12天;
当诱虫量>5头/板时,轮换施用植物源药剂1%苦参·印楝素乳油80mL/亩、和微生物代谢产物药剂6%乙基多杀菌素悬浮剂15g/亩进行植株和地面喷雾2次,每次间隔12天;
当病株率≤3%时,利用3亿孢子/g的钩状木霉菌TH-01菌粉80g/亩进行植株和地面喷雾2次,每次间隔8天;
当病株率>3%时,轮换施用微生物代谢产物药剂10%多抗霉素可湿性粉剂110g/亩和高效低风险化学药剂35%苯甲·咪鲜胺水乳剂65mL/亩进行植株和地面喷雾2次,每次间隔10天;
中耕时,在畦间每亩撒施由3亿孢子/g的钩状木霉菌TH-01菌粉2kg、5亿孢子/g的蜡蚧轮枝菌LL-01菌粉2kg、麦麸4kg以及4瓶、净含量≥20000只/瓶的日本毛绥螨或地理厉螨混拌而成的基质。
实施例2
基于全周期生物***的甜椒绿色轻简控害方法,具体包括以下步骤:
(1)一灭
种植甜椒之前,每亩撒施由3亿孢子/g的钩状木霉菌TH-01菌粉4kg、5亿孢子/g的蜡蚧轮枝菌LL-01菌粉4kg、硅藻土12kg和生石灰25kg混拌而成的菌土进行翻耕,并用由3亿孢子/g的钩状木霉菌TH-01和5亿孢子/g的蜡蚧轮枝菌LL-01菌粉混合制得的菌剂100倍液进行浸种或浸苗;
(2)二防
种植甜椒之后,在温室、大棚风口和门口处增设银灰条膜和防虫网,定期维护棚膜、门窗和防虫网;畦面覆盖地膜;
(3)三抗
日常肥水增施98%乙酰水杨酸粉剂20g/亩和3亿孢子/g的钩状木霉菌TH-01菌粉40g/亩;
(4)四控
在阴雨天气,于中午前后放风2h;出苗后在植株上方12cm处以1:1组合悬挂黄色诱虫板和蓝色诱虫板各20片/亩,且每片黄色诱虫板和蓝色诱虫板上各加装一个蓟马信息素诱芯和粉虱信息素诱芯,隔20天更换诱虫板和诱芯1次;在设施温棚内部四周间作25cm宽的伴生植物芹菜和茼蒿;
(5)五治
当诱虫量≤3头/板时,每亩释放微300头小花蝽或淡翅小花蝽,以及70袋、成螨量≥200只/袋、携带5亿孢子/g蜡蚧轮枝菌LL-01菌粉的携菌加州新小绥螨或携菌智利小植绥螨,每隔30天释放一次;
当3<诱虫量≤5头/板时,在释放天敌之前,利用5亿孢子/g的蜡蚧轮枝菌LL-01菌粉60g/亩进行植株和地面喷雾1次,每次间隔8天;
当诱虫量>5头/板时,轮换施用植物源药剂1%苦参·印楝素乳油100mL/亩和微生物代谢产物药剂6%乙基多杀菌素悬浮剂16g/亩进行植株和地面喷雾1次,每次间隔8天;
当病株率≤3%时,利用3亿孢子/g的钩状木霉菌TH-01菌粉70g/亩进行植株和地面喷雾1次,每次间隔10天;
当病株率>3%时,轮换施用微生物代谢产物药剂10%多抗霉素可湿性粉剂120g/亩和高效低风险化学药剂35%苯甲·咪鲜胺水乳剂80mL/亩进行植株和地面喷雾1次,每次间隔8天;
中耕时,在畦间每亩撒施由3亿孢子/g的钩状木霉菌TH-01菌粉2kg、5亿孢子/g的蜡蚧轮枝菌LL-01菌粉2kg、麦麸3kg以及3瓶、净含量≥20000只/瓶的日本毛绥螨或地理厉螨混拌而成的基质。
实施例3
基于全周期生物***的甜椒绿色轻简控害方法,具体包括以下步骤:
(1)一灭
种植甜椒之前,每亩撒施由3亿孢子/g的钩状木霉菌TH-01菌粉5kg、5亿孢子/g的蜡蚧轮枝菌LL-01菌粉5kg、硅藻土15kg和生石灰30kg混拌而成的菌土进行翻耕,并用由3亿孢子/g的钩状木霉菌TH-01和5亿孢子/g的蜡蚧轮枝菌LL-01菌粉混合制得的菌剂200倍液进行浸种或浸苗;
(2)二防
种植甜椒之后,在温室、大棚风口和门口处增设银灰条膜和防虫网,定期维护棚膜、门窗和防虫网;畦面覆盖地膜;
(3)三抗
日常肥水增施98%乙酰水杨酸粉剂30g/亩和3亿孢子/g的钩状木霉菌TH-01菌粉60g/亩;
(4)四控
在阴雨天气,于中午前后放风3h;出苗后在植株上方15cm处以1:1组合悬挂黄色诱虫板和蓝色诱虫板各40片/亩,且每片黄色诱虫板和蓝色诱虫板上各加装一个蓟马信息素诱芯和粉虱信息素诱芯,隔40天更换诱虫板和诱芯1次;在设施温棚内部四周间作30cm宽的伴生植物芹菜和茼蒿;
(5)五治
当诱虫量≤3头/板时,每亩释放微500头小花蝽或淡翅小花蝽,以及100袋、成螨量≥200只/袋、携带5亿孢子/g蜡蚧轮枝菌LL-01菌粉的携菌加州新小绥螨或携菌智利小植绥螨,每隔40天释放一次;
当3<诱虫量≤5头/板时,在释放天敌之前,利用5亿孢子/g的蜡蚧轮枝菌LL-01菌粉100g/亩进行植株和地面喷雾2次,每次间隔12天;
当诱虫量>5头/板时,轮换施用矿物源药剂88%硅藻土可湿性粉剂1400g/亩和高效低风险化学药剂21%噻虫嗪悬浮剂18mL/亩进行植株和地面喷雾2次,每次间隔12天;
当病株率≤3%时,利用3亿孢子/g的钩状木霉菌TH-01菌粉120g/亩进行植株和地面喷雾3次,每次间隔12天;
当病株率>3%时,轮换施用微生物代谢产物药剂3%中生菌素可湿性粉剂80g/亩和高效低风险化学药剂50%氯溴异氰尿酸可溶性粉剂50g/亩进行植株和地面喷雾3次,每次间隔12天;
中耕时,在畦间每亩撒施由3亿孢子/g的钩状木霉菌TH-01菌粉3kg、5亿孢子/g的蜡蚧轮枝菌LL-01菌粉3kg、麦麸5kg以及5瓶、净含量≥20000只/瓶的日本毛绥螨或地理厉螨混拌而成的基质。
对比例
常规化学措施,具体包括以下步骤:
(1)分别在种植30d、60d和75d后,用25%吡虫啉可湿性粉剂6-8g/亩和25%的咪鲜胺乳油40-50mL/亩进行植株和地面喷雾1次;
(2)在种植90d后,用20%啶虫脒可溶液剂5-7g/亩和10%的苯醚甲环唑水分散粒剂30-50g/100亩进行植株和地面喷雾3次,每次间隔15天;
(3)在种植135d后,用21%噻虫嗪悬浮剂15-20mL/亩和40%腈菌唑可湿性粉剂10-15g/亩进行植株和地面喷雾3次,每次间隔15天;
(4)在种植180d后,用22.4%螺虫乙酯悬浮剂20-30mL/亩和20%***酮乳油40-45mL/亩进行植株和地面喷雾3次,每次间隔15天。
性能测试
一、一株钩状木霉菌TH-01的鉴定及其对甜椒黑斑病菌的抑菌活性
1、材料与方法
(1)供试材料:供试木霉菌采自福建省漳州市诏安县甜椒种植基地中甜椒植株基部的凋落物,编号为TH-01;供试黑斑病菌Cladosporium cladosporioides也采自该基地甜椒植株的病叶,均采用PDA培养基分离、纯化和保存。
(2)菌株鉴定:挑取菌株TH-01单孢接种于察氏培养基平板,置于28±1℃、相对温度90±5%、光周期12L﹕12D的光照培养箱内培养4d,先用单反相机拍摄菌落形态特征,再用生物显微镜拍摄菌丝、分生孢子梗和分生孢子的形态特征,初步鉴定菌株TH-01的分类地位。用无菌手术刀在培养基上刮取约0.2g的菌丝和孢子,使用真菌基因组DNA提取试剂盒提取所分离菌株的总DNA,采用rDNA-ITS基因通用引物(ITS1:5’-TCCGTAGGTGAACCTGCGG-3’和ITS4:5’-TCCTCCGCTTATTGATATGC-3’)进行PCR扩增,后测定序列;将所得的基因序列进行BLAST比对,把相关种属序列导入MEGA7软件,利用Neighbor joining法构建序列的***发育树,根据序列间同源性的高低最终判定所分离菌株的种类。
(3)抑菌测定:采用平板对峙法,测定菌株TH-01对甜椒黑斑病菌的抑菌活性。先用5mm打孔器在纯化好的甜椒黑斑病菌边缘打孔,将菌饼接入直径9mm的PDA平板培养基中心,再用无菌接菌环在距菌饼25mm的四个方向分别接入菌株TH-01单孢,以四周未接菌株TH-01的处理为空白对照,置于光照培养箱中培养4d,测定病原菌的菌落直径,并根据公式(对照组病原菌落直径-处理组病原菌落直径)/(对照组病原菌落直径-菌饼直径)×100%计算菌株TH-01的抑菌率。
2、结果与分析
(1)菌株鉴定:菌株TH-01在PDA培养基平板上生长良好;菌落初始白色、近圆形,逐渐向四周扩展,并从中央产生绿色孢子;菌落周围有白色菌丝的生长带,中央区域的孢子层逐渐变成绿色或黄绿色;菌丝具分隔、多分枝,宽2.9-3.7μm;分生孢子梗对生或互生、梗瓶状,顶部长有近圆形或椭圆形、大小为(2.8-3.6)μm×(3.2-3.9)μm的分生孢子,据此初步鉴定菌株TH-01属木霉属Trichoderma。
对菌株TH-01的总DNA进行ITS扩增,获得长度为582bp的序列片段,与NCBI/BLAST中的相关种属进行同源性比对,发现该基因序列分别与登录号为KC403936、KC403948和MG827166的Trichoderma hamatum序列的相似性高达99.83%、98.80%和98.80%;结合形态特征,最终确定菌株为钩状木霉菌(菌落特征如图1所示,显微特征如图2所示)。
该钩状木霉菌TH-01的分类命名为钩状木霉(Trichoderma hamatum),保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址为北京市朝阳区北辰西路1号院3号,保藏编号为CGMCCNo.40078,保藏日期为2022年02月15日。
该钩状木霉菌TH-01的ITS序列为:
AGGGATCATTACGGAGCTTCACTCCCAACCCAATGTGAACGTTACCAAACTGTTGCCTCGGCGGGGTCACGCCCCGGGTGCGTCGCAGCCCCGGAACCAGGCGCCCGCCGGAGGAACCAACCAAACTCTTTCTGTAGTCCCCTCGCGGACGTATTTCTTACAGCTCTGAGCAAAAATTCAAAATGAATCAAAACTTTCAACAACGGATCTCTTGGTTCTGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGAATCATCGAATCTTTGAACGCACATTGCGCCCGCCAGTATTCTGGCGGGCATGCCTGTCCGAGCGTCATTTCAACCCTCGAACCCCTCCGGGGGATCGGCGTTGGGGATCGGGACCCCTCACACGGGTGCCGGCCCCGAAATACAGTGGCGGTCTCGCCGCAGCCTCTCCTGCGCAGTAGTTTGCACAACTCGCACCGGGAGCGCGGCGCGTCCACGTCCGTAAAACACCCAACTTTCTGAAATGTTGACCTCGGATCAGGTAGGAATACCCGCTGAACTTAAGCATATCAAAAACCGGGAGGAA,如SEQ ID NO.1所示。
(2)抑菌活性:菌株TH-01可有效抑制甜椒黑斑病菌生长,在平板对峙试验中,仅接菌4d处理组的菌株TH-01生长就完全覆盖黑斑病菌,黑斑病菌落直径仅为13.77±1.22mm(如图3所示),而对照处理的黑斑病菌落直径达38.00±2.43mm(如图4所示),抑菌率平均高达73.42%。
由此可见,菌株TH-01对甜椒黑斑病菌具有良好的抑菌活性和重寄生作用,生防潜力良好。
二、蜡蚧轮枝菌LL-01对甜椒蓟马的致病力
1、材料与方法
供试甜椒蓟马为优势种花蓟马Frankliniella intonsa,采自福建省漳州市诏安县甜椒种植基地,取交代羽化3d的成虫备用。
供试蜡蚧轮枝菌Lecanicillium lecanii LL-01现保存于福建省农业科学院植物保护研究所,也保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址为北京市朝阳区北辰西路1号院3号,保藏编号为CGMCC No.21940,保藏日期为2021年3月31日,记载在申请日为2021年05月17日、申请号为202110534601.0、名称为“一株蜡蚧轮枝菌LL-01及其生防应用”的专利申请文件中,现可通过正常渠道购买获得。
采用察氏平板培养基活化,取培养7d的菌株LL-01,往培养皿中加入10mL含0.05%吐温-80的无菌水,过滤得分生孢子原液,用无菌水稀释成104、105、106、107和108孢子/mL处理浓度,以0.05%吐温-80无菌水为对照,3次重复。分别往每个处理菌液中挑取30头甜椒蓟马成虫,浸泡10s并于滤纸上晾干2min后挑到依次铺有花泥、滤纸和甜椒花的培养皿中,花梗用湿棉花包裹并与滤纸紧贴,培养皿用保鲜膜封口,并用0.27mm的昆虫针在膜上密扎细孔通气;将培养皿置于温度26±1℃、相对湿度90±5%、光周期12L∶12D的人工气候箱内,每天定时观察直至所有处理中试虫的死亡数停止发生变化为止,统计菌株LL-01对甜椒蓟马的致死率。
2、结果与分析
在菌株LL-01侵染7d后,花蓟马的死亡数停止发生变化(如图5所示);104和105孢子/mL处理的致死率相对较低,最高仍仅44.25%;106孢子/mL处理的致死率虽达57.47%,但与105孢子/mL处理差异不显著;107和108孢子/mL处理的致死率相对较高,分别达78.16%和87.36%,两者均显著高于其他3个处理(如图6所示)。
由此可见,菌株LL-01能有效侵染花蓟马,生防潜力良好。
三、效果验证
在福建省漳州市诏安县的甜椒种植基地,在种植前选取4个大棚(长×宽=40m×8m)。其中三个大棚分别按照实施例1-3进行基于全周期生物***的绿色轻简控害处理,作为处理组;另一个大棚按照对比例基地常规的化学措施进行控害,作为对照组。处理组和对照组的其他肥水管理等农事措施均一致。至甜椒采收结束后,对比实施例1-3和对照组的控害效果、化学农药使用量、控害成本及甜椒产量和商品合格率。
对比结果如表1所示。
表1实施例1-3和对照组对比结果
由表1可知,按照实施例1-3对甜椒进行基于全周期生物***的绿色轻简控害,对甜椒病虫的控害效果达82%-96%。与常规的化学措施相比,提升控害效果11%-25%,减少化学农药使用量25%-33%,降低控害成本33%-46%,提高甜椒产量11%-13%和商品合格率12%-14%,经济效益显著。
对所公开的实施例的上述说明,使本领域专业技术人员能够实现或使用本发明。对这些实施例的多种修改对本领域的专业技术人员来说将是显而易见的,本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下,在其它实施例中实现。因此,本发明将不会被限制于本文所示的这些实施例,而是要符合与本文所公开的原理和新颖特点相一致的最宽的范围。
序列表
<110> 福建省农业科学院植物保护研究所
<120> 一种基于全周期生物***的甜椒绿色轻简控害方法
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 582
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
agggatcatt acggagcttc actcccaacc caatgtgaac gttaccaaac tgttgcctcg 60
gcggggtcac gccccgggtg cgtcgcagcc ccggaaccag gcgcccgccg gaggaaccaa 120
ccaaactctt tctgtagtcc cctcgcggac gtatttctta cagctctgag caaaaattca 180
aaatgaatca aaactttcaa caacggatct cttggttctg gcatcgatga agaacgcagc 240
gaaatgcgat aagtaatgtg aattgcagaa ttcagtgaat catcgaatct ttgaacgcac 300
attgcgcccg ccagtattct ggcgggcatg cctgtccgag cgtcatttca accctcgaac 360
ccctccgggg gatcggcgtt ggggatcggg acccctcaca cgggtgccgg ccccgaaata 420
cagtggcggt ctcgccgcag cctctcctgc gcagtagttt gcacaactcg caccgggagc 480
gcggcgcgtc cacgtccgta aaacacccaa ctttctgaaa tgttgacctc ggatcaggta 540
ggaatacccg ctgaacttaa gcatatcaaa aaccgggagg aa 582
Claims (9)
1.一种基于全周期生物***的甜椒绿色轻简控害方法,其特征在于,具体包括以下步骤:
(1)一灭
种植甜椒之前,每亩撒施由钩状木霉菌TH-01菌粉3-5kg、蜡蚧轮枝菌LL-01菌粉3-5kg、硅藻土10-15kg和生石灰20-30kg混拌而成的菌土进行翻耕,并用由钩状木霉菌TH-01菌粉和蜡蚧轮枝菌LL-01菌粉混合制得的菌剂进行浸种或浸苗;
所述钩状木霉菌TH-01的分类命名为钩状木霉(Trichoderma hamatum),保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址为北京市朝阳区北辰西路1号院3号,保藏编号为CGMCC No.40078,保藏日期为2022年02月15日;
所述蜡蚧轮枝菌LL-01的分类命名为蜡蚧轮枝菌(Lecanicillium lecanii),保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址为北京市朝阳区北辰西路1号院3号,保藏编号为CGMCC No.21940,保藏日期为2021年3月31日;
所述钩状木霉菌TH-01菌粉的孢子含量为3亿/g,所述蜡蚧轮枝菌LL-01菌粉的孢子含量为5亿/g;
(2)二防
种植甜椒之后,在温室、大棚风口和门口处增设银灰条膜和防虫网,定期维护棚膜、门窗和防虫网;畦面覆盖地膜;
(3)三抗
日常肥水增施乙酰水杨酸粉剂和钩状木霉菌TH-01菌粉;
(4)四控
在阴雨天气,于中午前后进行放风;出苗后在植株上方悬挂黄色诱虫板和蓝色诱虫板,且每片黄色诱虫板上加装一个蓟马信息素诱芯和粉虱信息素诱芯,每片蓝色诱虫板上加装一个蓟马信息素诱芯和粉虱信息素诱芯;在设施温棚内部四周间作伴生植物;
(5)五治
当诱虫量≤3头/板时,释放微小花蝽或淡翅小花蝽,以及携菌加州新小绥螨或携菌智利小植绥螨;所述携菌加州新小绥螨或携菌智利小植绥螨所携的菌为蜡蚧轮枝菌LL-01菌粉;
当3<诱虫量≤5头/板时,在释放天敌之前,利用蜡蚧轮枝菌LL-01菌粉进行植株和地面喷雾;
当诱虫量>5头/板时,轮换施用苦参·印楝素乳油、硅藻土可湿性粉剂、乙基多杀菌素悬浮剂和噻虫嗪悬浮剂进行植株和地面喷雾;
当病株率≤3%时,利用钩状木霉菌TH-01菌粉进行植株和地面喷雾;
当病株率>3%时,轮换施用多抗霉素可湿性粉剂、中生菌素可湿性粉剂、氯溴异氰尿酸可溶性粉剂和苯甲·咪鲜胺水乳剂进行植株和地面喷雾;
中耕时,在畦间每亩撒施由钩状木霉菌TH-01菌粉2-3kg、蜡蚧轮枝菌LL-01菌粉2-3kg、麦麸3-5kg以及3-5瓶、净含量≥20000只/瓶的日本毛绥螨或地理厉螨混拌而成的基质。
2.根据权利要求1所述的一种基于全周期生物***的甜椒绿色轻简控害方法,其特征在于,步骤(1)中,所述菌剂稀释至100-200倍液。
3.根据权利要求1所述的一种基于全周期生物***的甜椒绿色轻简控害方法,其特征在于,步骤(3)中,所述乙酰水杨酸粉剂的质量浓度为98%,用量为20-30g/亩;所述钩状木霉菌TH-01菌粉的用量为40-60g/亩。
4.根据权利要求1所述的一种基于全周期生物***的甜椒绿色轻简控害方法,其特征在于,步骤(4)中,所述放风的时间为2-3h;所述黄色诱虫板和蓝色诱虫板的用量为1:1,各20-40片/亩,悬挂位置为植株上方10-15cm处,更换频率为20-40天/次。
5.根据权利要求1所述的一种基于全周期生物***的甜椒绿色轻简控害方法,其特征在于,步骤(4)中,所述伴生植物为芹菜和茼蒿,种植宽度为20-30cm。
6.根据权利要求1所述的一种基于全周期生物***的甜椒绿色轻简控害方法,其特征在于,步骤(5)中,当诱虫量≤3头/板时,所述微小花蝽或淡翅小花蝽的释放量为300-500头/亩,释放频率为30-40天/次;所述携菌加州新小绥螨或携菌智利小植绥螨的成螨量≥200只/袋,释放量为70-100袋/亩,释放频率为30-40天/次;
当3<诱虫量≤5头/板时,所述蜡蚧轮枝菌LL-01菌粉的用量为60-100g/亩;所述喷雾的次数为1-2次,频率为8-12天/次。
7.根据权利要求1所述的一种基于全周期生物***的甜椒绿色轻简控害方法,其特征在于,步骤(5)中,当诱虫量>5头/板时,所述苦参·印楝素乳油的质量浓度为1%,用量为70-100mL/亩;所述硅藻土可湿性粉剂的质量浓度为88%,用量为1200-1400g/亩;所述乙基多杀菌素悬浮剂的质量浓度为6%,用量为12-16g/亩;所述噻虫嗪悬浮剂的质量浓度为21%,用量为14-18mL/亩;所述喷雾的次数为1-2次,频率为8-12天/次。
8.根据权利要求1所述的一种基于全周期生物***的甜椒绿色轻简控害方法,其特征在于,步骤(5)中,当病株率≤3%时,所述钩状木霉菌TH-01菌粉的用量为70-120g/亩;所述喷雾的次数为1-3次,频率为8-12天/次。
9.根据权利要求1所述的一种基于全周期生物***的甜椒绿色轻简控害方法,其特征在于,步骤(5)中,当病株率>3%时,所述多抗霉素可湿性粉剂的质量浓度为10%,用量为100-120g/亩;所述中生菌素可湿性粉剂的质量浓度为3%,用量为50-80g/亩;所述氯溴异氰尿酸可溶性粉剂的质量浓度为50%,用量为40-50g/亩;所述苯甲·咪鲜胺水乳剂的质量浓度为35%,用量为60-80mL/亩;所述喷雾的次数为1-3次,频率为8-12天/次。
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