CN115500350B - 一种稻绿核菌生长抑制剂 - Google Patents
一种稻绿核菌生长抑制剂 Download PDFInfo
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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
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- A01N31/16—Oxygen or sulfur directly attached to an aromatic ring system with two or more oxygen or sulfur atoms directly attached to the same aromatic ring system
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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
- A01N31/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic oxygen or sulfur compounds
- A01N31/08—Oxygen or sulfur directly attached to an aromatic 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
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- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
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Abstract
本发明提供了一种稻绿核菌生长抑制剂，该稻绿核菌生长抑制剂含有丁香酚、香芹酚或2，4‑二叔丁基苯酚中的一种或一种以上。本发明提供的稻绿核菌生长抑制剂，其中丁香酚、香芹酚或2，4‑二叔丁基苯酚均为天然产物，毒性低无污染，低浓度情况下对水稻稻曲病病原菌稻绿核菌的菌丝生长和孢子萌发具有显著的抑制作用，为稻曲病的防治提供了一种高效低毒无污染的抑菌剂。
Description
技术领域
本发明涉及农产品质量安全领域，具体的说涉及一种稻绿核菌生长抑制剂。
背景技术
稻曲病是由稻绿核菌Ustilaginoidea virens(Cooke)Tak引起的一种水稻穗部谷粒病害，是世界上水稻三大主要病害之一，广泛分布于亚洲、非洲、南美洲、欧洲各地的水稻主产区。自20世纪80年代开始，随着灌溉与施肥水平的提高、高产型品种的普及推广以及气候环境的变化，稻曲病在我国发生范围不断扩大，危害不断加重。稻绿核菌不仅引起水稻稻曲病，显著增加水稻空秕粒率、降低粒重，造成20％～30％的产量损失，其厚垣孢子所产生的稻曲菌素(Ustiloxin)和黑粉菌素(Ustilaginoidins)等真菌毒素对植物和人畜均有较大的毒害作用，严重威胁稻米质量安全。
为防控稻曲病，在水稻孕穗期早期使用杀菌剂目前已被证明存在一定的成效，但常用的苯甲·丙环唑、己唑·嘧菌酯、丙环·咪鲜胺、多菌灵等化学试剂的长期施用会导致环境污染、病原菌耐药性增强、引入新的食品安全风险等一系列问题，因此亟待开发一种绿色、环保、高效的新型杀菌剂防治稻绿核菌引起的水稻稻曲病。
发明内容
本发明提供了一种稻绿核菌生长抑制剂，该稻绿核菌生长抑制剂含有丁香酚、香芹酚或2，4-二叔丁基苯酚中的一种或一种以上；
具体的说，本发明的一种稻绿核菌生长抑制剂，其中丁香酚、香芹酚或2，4-二叔丁基苯酚在使用时先由少量二甲基亚砜溶解，然后加入水，配制成水溶液；
其中丁香酚、香芹酚或2，4-二叔丁基苯酚的浓度分别为至少0.1mmol/L；
更为具体的说，丁香酚、香芹酚或2，4-二叔丁基苯酚的浓度分别为0.1-5mmol/L；在具体用于水稻中稻曲病治疗时，由于复杂的水稻生长环境，丁香酚、香芹酚或2，4-二叔丁基苯酚的浓度可能比实验室条件下的浓度更高。
本发明提供的稻绿核菌生长抑制剂，其具备以下优点：
(1)本发明首次发现了丁香酚、香芹酚和2，4-二叔丁基苯酚可以有效抑制稻绿核菌的菌落生长和孢子萌发，低浓度即可达抑制率100％；
(2)丁香酚、香芹酚和2，4-二叔丁基苯酚均是天然产物，安全、易降解、无污染、毒性低。丁香酚还是植物源天然产物，在中国、美国和欧盟等多个国家和地区作为香料和食品添加剂可以合法使用，且没有制定最大残留或使用限(GB 1886.129-2022食品安全国家标准-食品添加剂-丁香酚，GB 2760-2011食品安全国家标准-食品添加剂使用标准)；香芹酚也是一种植物源天然产物，呈辛香、凉香、药草香，毒性低，已被列入允许使用的食品用天然香料名单(GB 2760-2011食品安全国家标准-食品添加剂使用标准)；
(3)来源广泛。丁香酚可以采用自富含丁香酚的植物精油通过碱处理单离而得，也可以以俞创木酚和烯丙基氯等为原料化学合成而得，生产成本低；香芹酚天然存在于百里香等植物中，来源广泛、提取简单，也可对百里香酚经磺化后再经碱处理而得，生产成本低；2，4-二叔丁基苯酚是许多植物和微生物挥发代谢物的组成成分，来源广泛，提取简单，也可通过酯化反应等大规模合成，成本低
(4)本发明的抑菌剂使用简单，无需复杂操作，易于大规模应用。
附图说明
图1丁香酚对稻绿核菌菌丝生长的抑制作用
图2丁香酚对稻绿核菌孢子萌发的抑制作用
图3香芹酚对稻绿核菌菌丝生长的抑制作用
图4 2，4-二叔丁基苯酚对稻绿核菌菌丝生长的抑制作用
具体实施方式
下列实施例中所用到的材料及其来源或制备方法：
(1)试剂和材料
丁香酚、香芹酚、2，4-二叔丁基苯酚、二甲基亚砜(国药集团化学试剂有限公司)
蔗糖、琼脂、Tween 20(上海源叶生物科技有限公司)；
马铃薯(普通市售产品)；
稻绿核菌菌株HWD-2(中国典型培养物保藏中心CCTCC NO:2011023)。
(2)仪器
Milli-Q超纯水仪(美国Millipore公司)；
AL104分析天平(瑞士梅特勒-托利多仪器有限公司)；
SX-500高压灭菌锅(日本TOMY公司)；
RI 250培养箱(美国Thermo Fisher scientific公司)；
Heraeus Multifuge X3高速离心机(美国Thermo Fisher scientific公司)；
BX-2显微镜(日本奥林巴斯公司)。
(3)菌株培养基的制备
PSA培养基：去皮马铃薯200g切成小块，煮沸30min后取滤液，加入蔗糖20g、琼脂16g，用蒸馏水定容至1000mL，115℃高压灭菌30min，降温到55℃左右倒平板，每平板20mL。
PSB液体培养基：去皮马铃薯200g切成小块，煮沸30min后取滤液，加入蔗糖20g，用蒸馏水定容至1000mL，115℃高压灭菌30min。
实施例1丁香酚对稻绿核菌菌丝生长的抑制作用
量取适量的丁香酚溶于的10mL含0.5％ Tween 20的无菌超纯水，过滤除菌后加入90mL灭菌后的PSA培养基，使得最终添加浓度分别达到0.1、0.2、0.5、1、2和5mmol/L，获得含丁香酚的PSA培养基。将稻绿核菌菌株HWD-2接种于PSA培养基，28℃黑暗活化培养14天后，切取外沿菌饼(直径约5mm)，置于含丁香酚的培养基平板中间，28℃黑暗培养14天后，用垂直十字法测量稻绿核菌菌落直径，以不含丁香酚的PSA培养基为对照组，计算生长抑制率：
生长抑制率/％＝[(对照菌落直径-处理菌落直径)/(对照菌落直径)]×100％
如图1所示，当浓度达到0.5mmol/L时，丁香酚对稻绿核菌的生长产生明显抑制，抑制率为30.38％；抑制效果随着丁香酚浓度的升高而增强，当丁香酚的浓度为1mmol/L时，抑制率达到76.59％；当丁香酚浓度达到2mmol/L时完全抑制稻绿核菌的生长。
实施例2丁香酚对稻绿核菌孢子萌发的抑制作用
将活化后的稻绿核菌菌饼加入PSB培养基，28℃，150rpm黑暗活化培养7天后，培养液变为绿色，过滤菌丝，去孢子液，显微镜观察孢子浓度，血球计数板计数，用无菌水调整至106个/mL。取30μL孢子悬浮液，均匀涂布于含不同浓度丁香酚(0.1、0.2、0.5、1、2和5mmol/L)的PSA培养基表面，28℃黑暗活化培养24小时，显微镜观察孢子萌发情况。孢子萌发标准为芽管长度超过孢子短测直径长度。以不含丁香酚的PSA培养基为对照组，计算孢子萌发抑制率：
孢子萌发抑制率/％＝[(对照孢子萌发率-处理孢子萌发率)/(对照孢子萌发率)]×100％
结果显示(图2)，当浓度达到0.1mmol/L时，丁香酚即抑制稻绿核菌的孢子萌发，抑制率为83.8％，0.5mmol/L时抑制率为98.3％，1mmol/L以上时抑制率为100％。
实施例3
(1)香芹酚对稻绿核菌菌丝生长的抑制作用
试验方法同实施例1，试验结果如下：
结果显示：如图3所示，当浓度达到0.2mmol/L时，香芹酚即对稻绿核菌的生长产生明显抑制(P<0.01)，抑制率为44.32％；抑制效果随着香芹酚浓度的升高而增强，当香芹酚为0.5mmol/L时，抑制率达到62.31％；当香芹酚浓度达到1mmol/L时完全抑制稻绿核菌的生长。
(2)香芹酚对稻绿核菌孢子萌发的抑制作用
试验方法同实施例2。
结果显示：当浓度达到0.1mmol/L和0.2mmol/L时，香芹酚即几乎完全抑制稻绿核菌的孢子萌发，抑制率分别为为99.0％和99.5％，0.2mmol/L以上时完全抑制孢子萌发(抑制率为100％)。
实施例4
(1)2，4-二叔丁基苯酚对稻绿核菌菌丝生长的抑制作用
试验方法同实施例1。
结果如图4所示，当浓度达到0.1mmol/L时，2，4-二叔丁基苯酚即对稻绿核菌的生长产生明显抑制(P<0.01)，抑制率为52.29％；抑制效果随着2，4-二叔丁基苯酚浓度的升高而增强，当2，4-二叔丁基苯酚的浓度为0.2mmol/L时，抑制率达到86.87％；当2，4-二叔丁基苯酚浓度达到0.5mmol/L时完全抑制稻绿核菌的生长.
(2)2，4-二叔丁基苯酚对稻绿核菌孢子萌发的抑制作用
试验方法同实施例2。
结果显示，当浓度达到0.1mmol/L及以上时，2，4-二叔丁基苯酚即完全抑制稻绿核菌的孢子萌发，抑制率为100％。
Claims (2)
1.香芹酚水溶液用于制备稻绿核菌生长抑制剂的用途，其中香芹酚水溶液中香芹酚的浓度为0.1-5 mmol/L。
2.香芹酚水溶液用于抑制稻绿核菌孢子萌发的用途，其中香芹酚水溶液中香芹酚的浓度为0.1-5 mmol/L。
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| CN110235905A (zh) * | 2018-03-09 | 2019-09-17 | 江苏莱科化学有限公司 | 一种杀菌组合物 |
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