CN113812426B - 一种蒜秸还田农作物根腐防治剂的制作方法及产品 - Google Patents
一种蒜秸还田农作物根腐防治剂的制作方法及产品 Download PDFInfo
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Abstract
本发明公开了一种蒜秸还田农作物根腐防治剂的制作方法及产品,其原料基于农业废弃物蒜秸,然后经过抗根腐因子浸提、蒜秸碳化、炭质营养液浸提、碳基载菌等技术,最终将提取的营养组分、载菌碳基和蒜根腐解的粘土基质复配而成。本发明在实现蒜秸废弃物还田的同时,可以将根腐病的病发率下调90.23%,病发后植株死亡率下降77.27%。
Description
技术领域
本发明属于农作物病害防治技术领域,具体涉及到一种蒜秸还田农作物根腐防治剂的制作方法及产品。
背景技术
根腐病是农作物春苗时期盛行的一种造成农作物根部腐烂的疾病,其通常会造成农作物的区域性群体死亡,甚至大面积绝产,一旦病发将对农业造成严重的经济损失。
根腐病一方面源于酶菌、镰刀菌和疫酶等多重病害菌的累计叠加侵染;另一方面,农用田地的粘性、板结、透气不良将促进农作物根腐的发生。针对这种病害,一些列的化学杀菌剂农药被研发和使用,如甲霜恶霉灵、多菌灵、恶霉灵、福美双等产品,但是这些方法仅适用于针对单株发病农作物进行灌根处理,不适用于大范围根腐病爆发的抑制。并且化学试剂的使用反而会破坏土壤微生态和理化属性,从而导致病害的恶行循环。
因此,本领域亟需一种兼具调节土壤微生物群和土壤理化作用的天然农作物根腐防治剂。
发明内容
本部分的目的在于概述本发明的实施例的一些方面以及简要介绍一些较佳实施例。在本部分以及本申请的说明书摘要和发明名称中可能会做些简化或省略以避免使本部分、说明书摘要和发明名称的目的模糊,而这种简化或省略不能用于限制本发明的范围。
鉴于上述和/或现有技术中存在的问题,提出了本发明。
因此,本发明的目的是,克服现有技术中的不足,提供一种蒜秸还田农作物根腐防治剂的制作方法。
为解决上述技术问题,本发明提供了如下技术方案:一种蒜秸还田农作物根腐防治剂的制作方法,包括,
抗腐因子浸提:将蒜叶、蒜秸和磷酸盐缓冲液混匀,捣碎成糊状,再加入磷酸盐缓冲液,混匀后微波处理,抽滤得滤液和浸提滤渣,滤液为抗腐因子;
蒜秸生物炭的制备:将浸提滤渣烘干至恒重,磨粉过筛,将得到的蒜秸粉在氮气的填充下升温至300~700℃并保持2h,随后以10℃/min降温至250℃并保持1h,自然冷却至室温,并将产物再次研磨过筛,制得蒜秸生物炭;
碳质营养液的制备:将抗腐因子浸提液以磷酸调节至pH=1~3,将浸提液与蒜秸生物炭混匀,超声清洗,抽滤分离,得到碳质营养液;
碳基载菌体的制备:将复配菌液、碳质营养液、蒜秸生物炭混匀,震荡培养,将混合体系冷风吹干,制得碳基载菌体;
蒜根腐解基质制备:将蒜根与腐植土、凹凸棒土、腐熟污泥、麦饭石粉混匀,混合物含水量保持在60~80%,堆积日晒自然发酵30~60天,制得蒜根腐解基质;
按重量份数计,将抗腐因子浸提液1~3份、碳基载菌体7~9份、蒜根腐解基质3~5份复配,即得蒜秸还田农作物根腐防治剂。
作为本发明所述蒜秸还田农作物根腐防治剂的制作方法的一种优选方案,其中:所述抗腐因子浸提,包括,按重量份数计,蒜叶3~5份、蒜秸1~3份和pH=6的磷酸盐缓冲液6~8份混匀并捣碎成糊状,再次加入磷酸盐缓冲液30~50份,混匀后以540W功率微波处理30~60min,抽滤得滤液。
作为本发明所述蒜秸还田农作物根腐防治剂的制作方法的一种优选方案,其中:所述蒜秸生物炭的制备,包括,
将浸提滤渣烘干至恒重,并磨粉过60目筛,将蒜秸粉在氮气的填充下以10℃/min升温至400℃并保持2h,随后以10℃/min降温至250℃并保持1h,自然冷却至室温并将产物,研磨过60目筛。
作为本发明所述蒜秸还田农作物根腐防治剂的制作方法的一种优选方案,其中:所述碳质营养液的制备,包括,
将抗腐因子浸提液以10M的磷酸调节至pH=1,随后将浸提液15~30份与蒜秸生物炭1~3份混匀,并以40~60kHz超声清洗1h,抽滤分离碳质营养液和清洗后的蒜秸生物炭。
作为本发明所述蒜秸还田农作物根腐防治剂的制作方法的一种优选方案,其中:所述复配菌液,其制备方法,包括,
分别取毛壳菌、木霉、枯草芽孢杆菌、Geminicoccus roseus、Vicinamibactersilvestris,分别配置浓度均为109cfu·mL-1的菌悬液配制;
按重量份数计,取上述6种菌悬液等质量比复配即得复配菌液。
作为本发明所述蒜秸还田农作物根腐防治剂的制作方法的一种优选方案,其中:所述碳基载菌体的制备,其中,以质量份数计,复配菌液3~5份、碳质营养液3~5份、蒜秸生物炭6~10份。
作为本发明所述蒜秸还田农作物根腐防治剂的制作方法的一种优选方案,其中:所述碳基载菌体的制备,其中,以35℃、180r·min-1震荡培养3~5h,冷风吹干温度为10~15℃。
作为本发明所述蒜秸还田农作物根腐防治剂的制作方法的一种优选方案,其中:所述蒜根腐解基质制备,包括,
按重量份数计,将蒜根10~50份与腐植土20~30份、凹凸棒土20~30份、腐熟污泥20~30份、麦饭石粉20~30份混匀,混合物含水量保持在60~80%,堆积日晒自然发酵30~60天。
作为本发明所述蒜秸还田农作物根腐防治剂的制作方法的一种优选方案,其中:所述蒜秸还田农作物根腐防治剂,其中,按重量份数计,抗腐因子为2份、碳基载菌体为9份、蒜根腐解基质为4份。
本发明的目的是,克服现有技术中的不足,提供一种蒜秸还田农作物根腐防治剂的制作方法制得的农作物病害防治剂产品。
本发明有益效果:
(1)本发明的蒜秸含有丰富的大蒜素、蒜氨酸等活性抗菌因子,是根腐病菌的天然抵抗者,将这些抗菌抗腐因子进行浸提有利于蒜秸资源的充分且直接的利用,抗腐因子浸液的补充将根腐的发病率降低了61.94%,病发后死亡率降低了28.26%,根腐病发后植株的死亡率降低43.55%。
(2)碳质营养液的浸提一方面有利于蒜秸生物炭嵌合的营养组分可以直接参与营养供给,另一方面嵌合组分的洗脱有利于生物炭表面活性位点的暴露,从而初进土壤的改良,碳质营养液的制备可将根腐防治剂的抗根腐效果提高8.86%。
(3)碳基载菌体的制备是根腐防治剂的核心技术,生物炭负载的菌体包括腐植质降解菌和土壤益生菌,因此碳基载菌体可以兼具土壤修复、腐植质补充、根腐菌抑制的作用,载菌碳基可以将抗根腐能力提高70.23%,可以将发病植株的死亡率降低90.77%。
(4)蒜根腐解基质是本发明主要的营养成分源,蒜根分泌物的腐解能力搭配蒜根根际的微生物群,可以将矿土组分腐解为植株可直接利用的有机质,从而达到恶补根际营养的目的,并且这可以将根腐防治剂的抗根腐能力提升8.55%。
附图说明
为了更清楚地说明本发明实施例的技术方案,下面将对实施例描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动性的前提下,还可以根据这些附图获得其它的附图。其中:
图1为本发明实施例中碳基载菌体对土壤根腐菌的干预图。
具体实施方式
为使本发明的上述目的、特征和优点能够更加明显易懂,下面结合说明书实施例对本发明的具体实施方式做详细的说明。
在下面的描述中阐述了很多具体细节以便于充分理解本发明,但是本发明还可以采用其他不同于在此描述的其它方式来实施,本领域技术人员可以在不违背本发明内涵的情况下做类似推广,因此本发明不受下面公开的具体实施例的限制。
其次,此处所称的“一个实施例”或“实施例”是指可包含于本发明至少一个实现方式中的特定特征、结构或特性。在本说明书中不同地方出现的“在一个实施例中”并非均指同一个实施例,也不是单独的或选择性的与其他实施例互相排斥的实施例。
本发明中所用材料:
Chaetomium sp.VIP(i)2518:莱耀标准品网;
Trichodermaharzianum VIP(i)13056:莱耀标准品网;
Bacillus subtilis CMCC(B)63501:莱耀标准品网;
Geminicoccus roseus BNCC316989:北京北纳创联生物技术研究院;
Vicinamibacter silvestris DSM 29464:宁波明舟生物科技有限公司;
菌种使用前进行活化和富集培养,以分光光度计法鉴定使菌群浓度大于1×109CFU/g;
新鲜蒜秸:山东省济宁市金乡县大蒜农田;
OTF-1200X管式炉:合肥科晶材料技术有限公司;
腐植土:石家庄万世石材有限公司;
腐熟污泥:山东鲁兴环保科技有限公司;
其他原料与设备无特殊说明,均为普通市售。
实施例1
本实施例提供一种蒜秸还田农作物根腐防治剂的制作方法,包括:
(1)原料预处理:新鲜的农业废弃物蒜秸、蒜叶和蒜根均为现制现用。选取新鲜蒜秸并去除霉变、枯黄的表皮和蒜叶。收集削除的新鲜蒜根,保留根际泥土;
(2)抗腐因子浸提:按重量份数计,将蒜叶4份、蒜秸2份和pH=6的磷酸盐缓冲液7份混匀并捣碎成糊状,再次加入磷酸盐缓冲液40份,混匀后以540W功率微波处理45min,抽滤得滤液用于碳质营养液制备,剩余滤渣用于蒜秸生物炭制备;
(3)蒜秸生物炭的制备:将浸提滤渣烘干至恒重,并磨粉过60目筛,将蒜秸粉在氮气的填充下以10℃/min升温至400℃碳化温度并保持2h,随后以10℃/min降温至250℃并保持1h,最终自然冷却至室温并将产物再次研磨过60目筛;
(4)碳质营养液的制备:将抗腐因子浸提液以10M的磷酸调节至pH=1,随后将浸提液20份与蒜秸生物炭3份混匀,并以60kHz超声清洗1h,抽滤分离碳质营养液和清洗后的蒜秸生物炭;
(5)碳基载菌体的制备:分别以腐植基质降解菌:毛壳菌、木霉;土壤益生菌:枯草芽孢杆菌、Geminicoccus roseus、Vicinamibacter silvestris;配制109cfu·mL-1的菌悬液。按重量份数计,取6种菌悬液等质量比复配的菌液4份、碳质营养液4份、蒜秸生物炭7份混匀,以35℃、180r·min-1震荡培养4h,随后将混合体系10℃冷风吹干。
(6)蒜根腐解基质制备:按重量份数计,将蒜根30份与腐植土25份、凹凸棒土25份、腐熟污泥25份、麦饭石粉25份混匀,混合物含水量保持在70%,堆积日晒自然发酵45天;
(7)按重量份数计,抗腐因子2份、碳基载菌体9份、蒜根腐解基质4份复配,即得蒜秸还田农作物根腐防治剂。
实施例2
本实施例提供一种蒜秸还田农作物根腐防治剂的制作方法,包括:
(1)原料预处理:新鲜的农业废弃物蒜秸、蒜叶和蒜根均为现制现用。选取新鲜蒜秸并去除霉变、枯黄的表皮和蒜叶。收集削除的新鲜蒜根,保留根际泥土;
(2)蒜秸生物炭的制备:将蒜秸烘干至恒重,并磨粉过60目筛,将蒜秸粉在氮气的填充下以10℃/min升温至400℃碳化温度并保持2h,随后以10℃/min降温至250℃并保持1h,最终自然冷却至室温并将产物再次研磨过60目筛;
(3)碳质营养液的制备:以10M的磷酸配制pH=1的酸性溶液,随后将溶液20份与蒜秸生物炭3份混匀,并以60kHz超声清洗1h,抽滤分离碳质营养液和清洗后的蒜秸生物炭;
(4)碳基载菌体的制备:分别以,腐植基质降解菌:毛壳菌、木霉;土壤益生菌:枯草芽孢杆菌、Geminicoccus roseus、Vicinamibacter silvestris;配制109cfu·mL-1的菌悬液。按重量份数计,取6种菌悬液等质量比复配的菌液4份、碳质营养液4份、蒜秸生物炭7份混匀,以35℃、180r·min-1震荡培养4h,随后将混合体系10℃冷风吹干。
(6)蒜根腐解基质制备:按重量份数计,将蒜根30份与腐植土25份、凹凸棒土25份、腐熟污泥25份、麦饭石粉25份混匀,混合物含水量保持在70%,堆积日晒自然发酵45天;
(7)按重量份数计,碳基载菌体9份、蒜根腐解基质4份复配,即得一种蒜秸还田农作物根腐防治剂。
实施例3
本实施例与实施例1相比,蒜秸生物炭制备的碳化温度为300℃,其他制备工艺条件均与实施例1相同。
实施例4
本实施例与实施例1相比,蒜秸生物炭制备的碳化温度为500℃,其他制备工艺条件均与实施例1相同。
实施例5
本实施例与实施例1相比,蒜秸生物炭制备的碳化温度为600℃,其他制备工艺条件均与实施例1相同。
以原子吸收光谱法测定总碳和总氮;关于固定碳的测定的标准DB23/T2422—2019测定固定碳;以国标GB 5009.4-2010测定灰分;以马弗炉灼烧减重量计算挥发分含量;以碳酸氢钠法测定速效磷;以乙酸铵提取—火焰光度计法测定速效钾。
表1碳化温度对成品生物炭品质的影响
由表1可知,300℃碳化的蒜秸生物炭速效磷和速效钾含量最高,超过这一温度将严重削弱生物炭的速效肥效。并且300℃处理下的总碳和固定碳含量最低,这意味着蒜秸碳化程度最低,因此300℃制成的蒜秸生物炭表面将富含调理土壤的活性官能团,并且可以嵌合丰富的碳营养基质。而超过这一温度导致碳化完全化,官能团和活性成分将被剥夺。
实施例6
本实施例提供一种蒜秸还田农作物根腐防治剂的制作方法,包括:
(1)原料预处理:新鲜的农业废弃物蒜秸、蒜叶和蒜根均为现制现用。选取新鲜蒜秸并去除霉变、枯黄的表皮和蒜叶。收集削除的新鲜蒜根,保留根际泥土;
(2)抗腐因子浸提:按重量份数计,将蒜叶4份、蒜秸2份和pH=6的磷酸盐缓冲液7份混匀并捣碎成糊状,再次加入磷酸盐缓冲液40份,混匀后以540W功率微波处理45min,抽滤得滤液用于碳质营养液制备,剩余滤渣用于蒜秸生物炭制备;
(3)蒜秸生物炭的制备:将浸提滤渣烘干至恒重,并磨粉过60目筛,将蒜秸粉在氮气的填充下以10℃/min升温至400℃碳化温度并保持2h,随后以10℃/min降温至250℃并保持1h,最终自然冷却至室温并将产物再次研磨过60目筛;
(4)碳基载菌体的制备:分别以,腐植基质降解菌:毛壳菌、木霉;土壤益生菌:枯草芽孢杆菌、Geminicoccus roseus、Vicinamibacter silvestris;配制109cfu·mL-1的菌悬液。按重量份数计,取6种菌悬液等质量比复配的菌液4份、抗腐因子浸提液体4份、蒜秸生物炭7份混匀,以35℃、180r·min-1震荡培养4h,随后将混合体系10℃冷风吹干。
(6)蒜根腐解基质制备:按重量份数计,将蒜根30份与腐植土25份、凹凸棒土25份、腐熟污泥25份、麦饭石粉25份混匀,混合物含水量保持在70%,堆积日晒自然发酵45天;
(7)按重量份数计,抗腐因子2份、碳基载菌体9份、蒜根腐解基质4份复配,即得一种蒜秸还田农作物根腐防治剂。
以液相色谱-质谱联用仪测定亲水性和疏水性有机物,并将检测结果分类。
表2碳质营养液中活性组分的含量(mg/L)
如表2所示,300℃的碳化温度(实施例1)所产生的活性营养组分优于更高的碳化温度。并且,这些活性营养组分从生物炭上洗脱,其可直接在农作物营养供给中直接发挥作用,另一方面,洗脱后的生物炭表面不再受其他组分干预,活性位点充分暴露,对土壤的修复效率大大提升。未洗脱制备碳质营养液的实施例6,其总可溶性有机物减少了300.32mg/L,而疏水性有机物和亲水性可溶有机物分别下降了64.45mg/L和235.87mg/L。并且因为活性组分的填充和干扰,实施例6的蒜秸生物炭并不能充分发挥其吸附和土壤调理功能。
实施例7
本实施例提供一种蒜秸还田农作物根腐防治剂的制作方法,包括:
(1)原料预处理:新鲜的农业废弃物蒜秸、蒜叶和蒜根均为现制现用。选取新鲜蒜秸并去除霉变、枯黄的表皮和蒜叶。收集削除的新鲜蒜根,保留根际泥土;
(2)抗腐因子浸提:按重量份数计,将蒜叶4份、蒜秸2份和pH=6的磷酸盐缓冲液7份混匀并捣碎成糊状,再次加入磷酸盐缓冲液40份,混匀后以540W功率微波处理45min,抽滤得滤液用于碳质营养液制备,剩余滤渣用于蒜秸生物炭制备;
(3)蒜秸生物炭的制备:将浸提滤渣烘干至恒重,并磨粉过60目筛,将蒜秸粉在氮气的填充下以10℃/min升温至400℃碳化温度并保持2h,随后以10℃/min降温至250℃并保持1h,最终自然冷却至室温并将产物再次研磨过60目筛;
(4)碳质营养液的制备:将抗腐因子浸提液以10M的磷酸调节至pH=1,随后将浸提液20份与蒜秸生物炭3份混匀,并以60kHz超声清洗1h,抽滤分离碳质营养液和清洗后的蒜秸生物炭;
(5)蒜根腐解基质制备:按重量份数计,将蒜根30份与腐植土25份、凹凸棒土25份、腐熟污泥25份、麦饭石粉25份混匀,混合物含水量保持在70%,堆积日晒自然发酵45天;
(6)按重量份数计,抗腐因子2份、蒜秸生物炭9份、蒜根腐解基质4份复配,即得一种蒜秸还田农作物根腐防治剂。
将各实施例产品以40kg/亩的用量施用在山东省济宁市金乡县20年重茬蒜龄的绝产大蒜土壤中,施用方式为跟随底肥施用。收集原始土壤和本发明处理6个月后的土壤进行土壤微生物总DNA提取。在质控和过滤之后以ITS引物进行土壤真菌ITS基因扩征,随后以NovaSeq6000高通量测序仪进行基因序列鉴定。将测定的序列进行质控、拼接、聚类和注释,以获得土壤环境中所有根腐菌信息。
如图1所示,原始根腐病害土壤的优势菌群被Gibberella等根腐菌所主导,施用实施例7,即未负载土壤益生菌农作物根腐防治剂(实施例7)可以分别将根腐菌Fusarium、Gibellulopsis和Plectosphaerella的丰度分别降低至原来的45.74%、67.86%和81.32,特别是根腐主导菌从原来的32.10%降低至18.47%,此时的根腐菌将不再是土壤的主导菌属。而负载了土壤益生菌的农作物根腐防治剂(实施例1)的根腐菌抑制效果进一步提升。本发明对Gibberella、Fusarium、Gibellulopsis和Plectosphaerella的抑制程度分别达到最高的79.89%、78.26%、90.86%和91.31%。
实施例8
本实施例提供一种蒜秸还田农作物根腐防治剂的制作方法,包括:
(1)原料预处理:新鲜的农业废弃物蒜秸、蒜叶和蒜根均为现制现用。选取新鲜蒜秸并去除霉变、枯黄的表皮和蒜叶。收集削除的新鲜蒜根,保留根际泥土;
(2)抗腐因子浸提:按重量份数计,将蒜叶4份、蒜秸2份和pH=6的磷酸盐缓冲液7份混匀并捣碎成糊状,再次加入磷酸盐缓冲液40份,混匀后以540W功率微波处理45min,抽滤得滤液用于碳质营养液制备,剩余滤渣用于蒜秸生物炭制备;
(3)蒜秸生物炭的制备:将浸提滤渣烘干至恒重,并磨粉过60目筛,将蒜秸粉在氮气的填充下以10℃/min升温至400℃碳化温度并保持2h,随后以10℃/min降温至250℃并保持1h,最终自然冷却至室温并将产物再次研磨过60目筛;
(4)碳质营养液的制备:将抗腐因子浸提液以10M的磷酸调节至pH=1,随后将浸提液20份与蒜秸生物炭3份混匀,并以60kHz超声清洗1h,抽滤分离碳质营养液和清洗后的蒜秸生物炭;
(5)碳基载菌体的制备:分别以,腐植基质降解菌:毛壳菌、木霉;土壤益生菌:枯草芽孢杆菌、Geminicoccus roseus、Vicinamibacter silvestris;配制109cfu·mL-1的菌悬液。按重量份数计,取6种菌悬液等质量比复配的菌液4份、碳质营养液4份、蒜秸生物炭7份混匀,以35℃、180r·min-1震荡培养4h,随后将混合体系10℃冷风吹干。
(7)按重量份数计,抗腐因子2份、碳基载菌体9份份复配,即得一种蒜秸还田农作物根腐防治剂。
实施例9
本实施例与实施例1相比,蒜根腐解基质由蒜根10份、腐植土30份、凹凸棒土30份、腐熟污泥30份、麦饭石粉30份复配发酵,其他制备工艺条件均与实施例1相同。
实施例10
本实施例与实施例1相比,蒜根腐解基质由蒜根20份、腐植土27.5份、凹凸棒土27.5份、腐熟污泥27.5份、麦饭石粉27.5份复配发酵,其他制备工艺条件均与实施例1相同。
实施例11
本实施例与实施例1相比,蒜根腐解基质由蒜根30份、腐植土25份、凹凸棒土25份、腐熟污泥25份、麦饭石粉25份复配发酵,其他制备工艺条件均与实施例1相同。
实施例12
本实施例与实施例1相比,蒜根腐解基质由蒜根40份、腐植土22.5份、凹凸棒土22.5份、腐熟污泥22.5份、麦饭石粉22.5份复配发酵,其他制备工艺条件均与实施例1相同。
实施例13
本实施例与实施例1相比,蒜根腐解基质由蒜根50份、腐植土20份、凹凸棒土20份、腐熟污泥20份、麦饭石粉20份复配发酵,其他制备工艺条件均与实施例1相同。
实施例14
本实施例与实施例1相比,蒜秸还田农作物根腐防治剂由抗腐因子1份、碳基载菌体9份、蒜根腐解基质5复配而成,其他制备工艺条件均与实施例1相同。
实施例15
本实施例与实施例1相比,蒜秸还田农作物根腐防治剂由抗腐因子3份、碳基载菌体9份、蒜根腐解基质3复配而成,其他制备工艺条件均与实施例1相同。
实施例16
本实施例与实施例1相比,蒜秸还田农作物根腐防治剂由抗腐因子2份、碳基载菌体8份、蒜根腐解基质5复配而成,其他制备工艺条件均与实施例1相同。
实施例17
本实施例与实施例1相比,蒜秸还田农作物根腐防治剂由抗腐因子3份、碳基载菌体7份、蒜根腐解基质5复配而成,其他制备工艺条件均与实施例1相同。
将各实施例产品以40kg/亩的用量施用在山东省济宁市金乡县20年重茬蒜龄的绝产大蒜土壤中,施用方式为跟随底肥施用。在施用次年的返春生长期(3月、4月、5月)对大蒜根腐病发病率进行统计。
表3各实施例处理下的根腐病发病情况
如表3所示,实施例1为最优实施例,其施用后根腐病的病发率下降了90.23%,病发后植株死亡率下降77.27,并且病死率为0。而任一制备方法的改变或组分比例的改变都将破坏这一优越效果。
实施例2与实施例1相比,抗腐因子的缺失将根腐的发病率提高了61.94%,病发后死亡率提高了28.26%,这意味着一旦根腐病发,植株的死亡率在43.55%。因此,蒜秸提取的抗腐因子是蒜秸还田农作物根腐防治剂的核心功能组分。实施例3-5与实施例1相比,高温碳化的蒜秸生物炭不利于根腐病的防治,其防治效果将降低9.08%-11.00%。实施例6和8与实施例1相比,碳质营养液和蒜根腐解基质的缺失使本发明根腐防治剂的抗根腐能力分别下降了8.86%和13.32%,这是营养活性因子和腐质的缺失引起的。实施例7与实施例1相比,缺少腐殖质降解菌和土壤益生菌的负载将导致抗根腐能力下降70.23%,并且一旦病发植株死亡率高达90.77%。因此,碳基载菌体的制备是本发明的核心技术。
由实施例9-13与实施例1对比可知,蒜根腐解基质由蒜根30份、腐植土25份、凹凸棒土25份、腐熟污泥25份、麦饭石粉25份复配发酵而成的根腐防治剂效果最佳,而作为腐解的主要贡献者的蒜根比例发生改变时,最佳效果将被破坏,其根腐防治效果下降约5.56%-8.55%。由实施例14-17与实施例1对比可知,根腐防治剂的最佳复配比例是抗腐因子2份、碳基载菌体9份、蒜根腐解基质4份复配。这一比例被破坏将导致抗根腐效果下降约1.63-2.69%。
应说明的是,以上实施例仅用以说明本发明的技术方案而非限制,尽管参照较佳实施例对本发明进行了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的精神和范围,其均应涵盖在本发明的权利要求范围当中。
Claims (9)
1.一种蒜秸还田农作物根腐防治剂的制作方法,其特征在于:包括,
抗腐因子浸提:将蒜叶、蒜秸和磷酸盐缓冲液混匀,捣碎成糊状,再加入磷酸盐缓冲液,混匀后微波处理,抽滤得滤液和浸提滤渣,滤液为抗腐因子;
蒜秸生物炭的制备:将浸提滤渣烘干至恒重,磨粉过筛,将得到的蒜秸粉在氮气的填充下升温至300~700℃并保持2h,随后以10℃/min降温至250℃并保持1h,自然冷却至室温,并将产物再次研磨过筛,制得蒜秸生物炭;
碳质营养液的制备:将抗腐因子浸提液以磷酸调节至pH=1~3,将浸提液与蒜秸生物炭混匀,超声清洗,抽滤分离,得到碳质营养液;
碳基载菌体的制备:将复配菌液、碳质营养液、蒜秸生物炭混匀,震荡培养,将混合体系冷风吹干,制得碳基载菌体;其中,所述复配菌液,其制备方法,包括,分别取毛壳菌、木霉、枯草芽孢杆菌、Geminicoccus roseus、Vicinamibacter silvestris,分别配置浓度均为109 cfu·mL-1的菌悬液配制;按重量份数计,取上述5种菌悬液等质量比复配即得复配菌液;
蒜根腐解基质制备:将蒜根与腐植土、凹凸棒土、腐熟污泥、麦饭石粉混匀,混合物含水量保持在60~80%,堆积日晒自然发酵30~60天,制得蒜根腐解基质;
按重量份数计,将抗腐因子浸提液1~3份、碳基载菌体7~9份、蒜根腐解基质3~5份复配,即得蒜秸还田农作物根腐防治剂。
2.如权利要求1所述蒜秸还田农作物根腐防治剂的制作方法,其特征在于:所述抗腐因子浸提,包括,按重量份数计,蒜叶3~5份、蒜秸1~3份和pH=6的磷酸盐缓冲液6~8份混匀并捣碎成糊状,再次加入磷酸盐缓冲液30~50份,混匀后以540W功率微波处理30~60min,抽滤得滤液。
3.如权利要求1所述蒜秸还田农作物根腐防治剂的制作方法,其特征在于:所述蒜秸生物炭的制备,包括,
将浸提滤渣烘干至恒重,并磨粉过60目筛,将蒜秸粉在氮气的填充下以10℃/min升温至400℃并保持2h,随后以10℃/min降温至250℃并保持1h,自然冷却至室温并将产物,研磨过60目筛。
4.如权利要求1所述蒜秸还田农作物根腐防治剂的制作方法,其特征在于:所述碳质营养液的制备,包括,
将抗腐因子浸提液以10M的磷酸调节至pH=1,随后将浸提液15~30份与蒜秸生物炭1~3份混匀,并以40~60kHz超声清洗1h,抽滤分离碳质营养液和清洗后的蒜秸生物炭。
5.如权利要求1所述蒜秸还田农作物根腐防治剂的制作方法,其特征在于:所述碳基载菌体的制备,其中,以质量份数计,复配菌液3~5份、碳质营养液3~5份、蒜秸生物炭6~10份。
6.如权利要求5所述蒜秸还田农作物根腐防治剂的制作方法,其特征在于:所述碳基载菌体的制备,其中,以35℃、180 r·min-1震荡培养3~5 h,冷风吹干温度为10~15℃。
7.如权利要求1所述蒜秸还田农作物根腐防治剂的制作方法,其特征在于:所述蒜根腐解基质制备,包括,
按重量份数计,将蒜根10~50份与腐植土20~30份、凹凸棒土20~30份、腐熟污泥20~30份、麦饭石粉20~30份混匀,混合物含水量保持在60~80%,堆积日晒自然发酵30~60天。
8.如权利要求1所述蒜秸还田农作物根腐防治剂的制作方法,其特征在于:所述蒜秸还田农作物根腐防治剂,其中,按重量份数计,抗腐因子为2份、碳基载菌体为9份、蒜根腐解基质为4份。
9.权利要求1~8中任一所述蒜秸还田农作物根腐防治剂的制作方法制得的农作物病害防治剂产品。
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