CN103999859B - α-倒捻子素在防治植物病害中的应用 - Google Patents
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Abstract
本发明公开了α-倒捻子素在防治植物病害中的应用。试验证明α-倒捻子素具有农用杀菌活性。该化合物在供试浓度为100mg/L时,对芒果炭疽病菌、芒果蒂腐病菌、番茄灰霉病菌和玉米大斑病菌也有很强的抑菌作用,EC50值介于18mg/L-35mg/L;对玉米大斑病菌和番茄灰霉病菌也有一定的抑制孢子萌发作用,EC50分别为66.20mg/L和50.43mg/L;同时,活体试验结果表明,该化合物对橡胶树白粉病菌还有很好的保护和治疗作用在供试浓度为500mg/L时,对橡胶树白粉病菌的保护和治疗作用分别为75.61%和70.12%,高于对照药剂咪鲜胺。上述结果说明α-倒捻子素对植物病菌真菌具有广谱的杀菌作用。这为进一步利用山竹果壳创制新型无公害植物源农药奠定了基础。
Description
技术领域
本发明涉及α-倒捻子素在防治植物病害中的应用,特别涉及α-倒捻子素及其作为杀菌剂在防治植物病害方面的应用。
背景技术
山竹(Garcinia mangostana L.),又称山竹子、莽吉柿、凤果,被誉为“果中皇后”,是藤黄科(Guttiferae)藤黄属(Garcinia)常绿乔木山竹的果实。原产于印度尼西亚和马来西亚,是一种典型的热带水果。主要分布于泰国、越南、马来西亚、印度尼西亚、菲律宾等东南亚国家,在中国广西、海南等地区广泛栽培。山竹果皮在东南亚国家一直作为传统医药用于治疗腹痛、腹泻、痢疾等疾病。近年许多研究人员对其进行了化学成分和生物活性的研究,口山酮、黄酮和多酚类等多种活性物质,具有抗氧化、抗肿瘤等多种活性。α-倒捻子素具有抗氧化、抗炎、抗癌等多种药理活性。近期有研究表明α-倒捻子素对紫外线屏蔽和酪氨酸酶抑制作用,可用作防晒和美白化妆品的开发。
经检索,目前国内尚无关于α-倒捻子素农用杀菌活性方面的报道。国内外对该化合物的研究主要集中在医用药理活性上,中国发明专利“α-倒捻子素在制备阿尔兹海默氏病药物中的应用(ZL201110040746.1)”和“α-,γ-倒捻子素衍生物及其在制备抗癌药物中的应用(201210402155.9)”公开了α-倒捻子素在制备医药中的应用,中国发明专利“一种从山竹皮中提取α-倒捻子素的方法(201310461749.1)”、“一种倒捻子素的制备方法(201010181155.1)”、“一种倒捻子素(mangostin)的全合成方法(201110302040.8)”、“一种纯化α-倒捻子素的方法(201110117537.2)”和“从山竹果皮中提取α-倒捻子素的方法(ZL200810017640.8)”公开了α-倒捻子素的全合成及制备方法,但未见α-倒捻子素对植物病原真菌抑制作用的报道。
发明内容
本发明的目的在于提供α-倒捻子素的一个新用途。α-倒捻子素是项目组采用生物活性追踪法从山竹果壳中分离获得具有良好杀菌活性的成分,α-倒捻子素的化学结构如式I所示。
本发明提供的α-倒捻子素的新用途是α-倒捻子素在防治植物病害中的应用。
本发明提供的α-倒捻子素在防治植物病害中的应用,所述植物病害为由下述病原真菌引起的植物病害:辣椒疫霜病菌(Phytophthora capsici Leonian)、番茄灰霉病菌(Botrytis cirerea)、水稻纹枯病菌(Thanatephorus cucumeris)、水稻稻瘟病菌(Pyricularia oryzae)、玉米大斑病菌(Exserohilum turcicum),芒果炭疽病菌(Colletotrichum gloeosporioides)、芒果蒂腐病菌(Botryodiplodia theobromae)、香蕉炭疽病菌(Gloeosporium musae)、香蕉枯萎病菌(Fusarium oxysporum f.sp cuben)、香蕉冠腐病菌(Fusarium semitectum)、橡胶树白粉病菌(Oidium heveae Steinm)、橡胶树棒孢霉落叶病菌(Corynespora cassiicola)、橡胶树褐根病菌(Phellinus noxius Corner)、橡胶树白根病菌(Rigidoporus lignosus)甘蔗赤腐病菌(Colletotrichum fuleatum Went.)、甘蔗梢腐病(Gibberella fujikuroi(Sawada)Wollenw)、甘蔗黑穗病(Ustilago scitamineaSydow.)、甘蔗眼斑病菌(Bipolaris sacchari(Butler.)Shoemaker)、荔枝霜霉病菌(Peronophythora litchii)中的一种或两种以上任意组合。所述病原真菌优选为橡胶树白粉病菌、芒果炭疽病菌、芒果蒂腐病菌、玉米大斑病菌和番茄灰霉病菌中的一种或两种以上任意组合。所述病原真菌最优选为芒果炭疽病菌和/或芒果蒂腐病菌。
所述防治植物病害为杀死引起植物病害的植物病原菌或抑制引起植物病害的植物病原菌生长。所述抑制引起植物病害的植物病原菌生长是抑制所述植物病原菌的菌丝生长和/或抑制所述植物病原菌的孢子萌发和生长。
同上述相同的道理,α-倒捻子素也可以制成植物病原真菌的抑制剂,因此,α-倒捻子素在制备植物病原菌的抑制剂中的应用也属于本发明的保护范围。所述的植物病原菌是上述引起植物病害的植物病原菌。
上述α-倒捻子素可以通过化学合成,也可以通过从山竹的果壳粉经溶剂提取,乙酸乙酯萃取,硅胶柱层析,最后经HPLC制备获得该化合物。
为评价α-倒捻子素的农用杀菌活性,本发明分别采用生长速率法和孢子萌发法来评价本发明的化合物对植物病原真菌的抑制菌丝生长作用和抑制孢子萌发作用;对活性较为敏感的植物病原真菌将采用机值分析法进一步测定其EC50,以此来评价本发明的化合物。
试验证明,本发明提供的α-倒捻子素在100mg/L的浓度下,对多种重要的热带作物病害尤其是几种热带水果和蔬菜病害有着显著的抑制菌丝生长作用,对芒果炭疽病菌、芒果蒂腐病菌具有很好的抑菌作用,抑制率分别为90.38%和91.23%;对番茄灰霉病菌和玉米大斑病菌也有很强的抑菌作用,其抑制率分别为85.71%和86.10%;对橡胶树棒孢霉落叶病菌、香蕉炭疽病菌、水稻纹枯病菌、水稻稻瘟病菌、辣椒疫霉病菌和荔枝霜疫霉病菌也具有较好的抑菌作用,其抑制率介于75-85%之间。
总之,本发明提供的化合物具有广谱的杀菌活性,对多种热带作物病害具有显著抑菌作用,为热带作物病害防治困难,药剂缺乏提供天然源杀菌活性物质;该化合物可用于防治芒果炭疽病菌等多种重要的热带作物病害;将其作为杀菌剂,其高效、低毒、安全等特点符合当前新农药创制的要求。
具体实施方式
下述实施例中,所述百分含量如无特殊说明,均为质量百分含量。
下述实施例中所述实验方法,如无特殊说明,均为常规方法。
为了更清楚的理解本发明以及本发明α-倒捻子素在防治植物病害中的应用,以下通过发明人给出的依本发明技术方案所完成的如下实施例对本发明作进一步的详细描述,但这些仅是本发明较好的实施例,该领域的技术熟练人员可以根据本发明的内容对本发明作出一些非本质的改进和调整。
实施例1、α-倒捻子素在防治植物病害中的应用
发明人在对山竹农用杀菌活性物质的分离纯化中,采用活性追踪法发现α-倒捻子素具有对植物病原菌的防治作用。以下述实验为例,验证α-倒捻子素在防治植物病害中的作用。
一、α-倒捻子素的制备
参考文献的方法(张成中,卜其涛,刘发贵,郑毅男.山竹果壳中α-倒捻子素和水溶性多糖的含量测定.药物分析杂志,2012,32(2):345-248)将干燥的山竹果壳粉碎过40目筛,以80%(体积百分浓度)乙醇溶液冷浸过夜后,经超声提取机(频率:1200Hz,每次超声20min,工作间歇:0.2s,全程时间:45min,温度:50℃,重复提取2次)提取,过滤后滤液经旋转蒸发仪减压浓缩得乙醇提取物,将其提取物加水悬浮后,用乙酸乙酯萃取,分别得乙酸乙酯萃取物和水相。乙酸乙酯萃取物与硅胶(80-100目)拌匀,于40℃烘箱烘干,石油醚湿法上硅胶(200-300目)柱,然后以氯仿为洗脱剂进行洗脱,得到其浓缩物,将该浓缩物经乙腈溶解,HPLC制备,乙腈-水(85:15)为流动相减压回收溶剂,冷冻干燥后得到其制备品。将制备品与α-倒捻子素经TLC检测、熔点测定及HPLC检测,发现其为同一物质,故确认为α-倒捻子素,经检测,其纯度>95%。
上述α-倒捻子素也可以通过化学合成获得。
二、α-倒捻子素对植物病原真菌的抑制活性测定
1.供试菌种
辣椒疫霉病菌(Phytophthora capsici Leonian,购买于中国农业微生物菌株保藏管理中心,保藏编号ACCC No.36073)、番茄灰霉病菌(Botrytis cirerea,购买于中国农业微生物菌株保藏管理中心,保藏编号ACCC No.30387)、水稻稻瘟病菌(Pyriculariaoryzae,购买于中国农业微生物菌株保藏管理中心,保藏编号ACCC No.30320)、玉米大斑病菌(Exserohilum turcicum,购买于中国农业微生物菌株保藏管理中心,保藏编号ACCC No.36780)、芒果炭疽病菌(Colletotrichum gloeosporioides,购买于中国农业微生物菌株保藏管理中心,保藏编号ACCC No.31219)、香蕉炭疽病菌(Gloeosporiummusae,购买于中国农业微生物菌株保藏管理中心,保藏编号ACCC NO.31245)、香蕉冠腐病菌(Fusarium semitectum,购买于中国农业微生物菌株保藏管理中心,保藏编号ACCC No.31945)、橡胶树棒孢霉落叶病菌(Corynespora cassiicola,购买于中国农业微生物菌株保藏管理中心,保藏编号ACCC No.37597)。
橡胶树白粉病菌(Oidium heveae Steinm)(杨意伯,谭志琼,郑服丛,张荣意.橡胶树白粉病杀菌剂生物测定接种方法研究.植物保护,2011,27(2):152-155)、橡胶树褐根病菌(Phellinus noxius Corner)(贺春萍,李锐,吴伟怀,郑肖兰,吴川颖,梁艳琼.12种杀菌剂对橡胶树褐根病菌的毒力测定.热带作物学报,2013,34(10):1987-1990)、橡胶树白根病菌(Rigidoporus lignosus)(贺春萍,吴海理,李锐,邱晓聪,郑肖兰,吴伟怀,郑服丛.橡胶树白根病菌生物学研究.热带作物学报,2010,31(11):1981-1985)、甘蔗赤腐病菌(Colletotrichum fuleatum Went.)(梁艳琼,雷照鸣,贺春萍,郑肖兰,李锐,吴伟怀.甘蔗赤腐病菌生物学特性研究.热带作物学报,2013,34(5):967-972)、甘蔗梢腐病(Gibberella fujikuroi(Sawada)Wollenw)(吴伟怀,李锐,郑服丛.甘蔗梢腐病原菌的鉴定及其室内毒力测定.甘蔗糖业,2006,(2):10-14)、甘蔗黑穗病(Ustilago scitaminea Sydow.)(吴伟怀,谢祖建,贺春萍,郑肖兰,李锐,郑服丛.甘蔗黑穗病菌冬孢子生物学特性及杀菌剂对其萌发的影响.热带作物学报,2009,30(1):1674-1678)、甘蔗眼斑病菌(Bipolarissacchari(Butler.)Shoemaker)(吴伟怀,郑肖兰,李锐,黄振良,贺春萍.甘蔗眼斑病菌部分生物学特性及其室内毒力测定.广东农业科学,2011,23:77-79)、水稻纹枯病菌(Thanatephorus cucumeris)(张静,冯岗,袁旭超,陈从珍,林勇.百里香酚抑菌活性初探.中国农学通报.2009,25(21):277-280)、芒果蒂腐病(Botryodiplodia theobromae)(高兆银,胡美姣,李敏,杨凤珍.壳聚糖对热带水果采后病原菌的抑菌活性研究.中国南方果树,2008,37(4):51~53)、荔枝霜疫霉菌(Peronophythora litchii)(吴光旭,刘爱媛,陈维信.开口箭提取物对荔枝霜疫霉菌的抑制作用及其对荔枝果实的贮藏效果.中国农业科学,2006,39(8):1703-1708)。以上10种病菌可以用文献((1)杨意伯,谭志琼,郑服丛,张荣意.橡胶树白粉病杀菌剂生物测定接种方法研究.植物保护,2011,27(2):152-155;(2)贺春萍,李锐,吴伟怀,郑肖兰,吴川颖,梁艳琼.12种杀菌剂对橡胶树褐根病菌的毒力测定.热带作物学报,2013,34(10):1987-1990;(3)贺春萍,吴海理,李锐,邱晓聪,郑肖兰,吴伟怀,郑服丛.橡胶树白根病菌生物学研究.热带作物学报,2010,31(11):1981-1985;(4)梁艳琼,雷照鸣,贺春萍,郑肖兰,李锐,吴伟怀.甘蔗赤腐病菌生物学特性研究.热带作物学报,2013,34(5):967-972;(5)吴伟怀,李锐,郑服丛.甘蔗梢腐病原菌的鉴定及其室内毒力测定.甘蔗糖业,2006,(2):10-14;(6)吴伟怀,谢祖建,贺春萍,郑肖兰,李锐,郑服丛.甘蔗黑穗病菌冬孢子生物学特性及杀菌剂对其萌发的影响.热带作物学报,2009,30(1):1674-1678;(7)吴伟怀,郑肖兰,李锐,黄振良,贺春萍.甘蔗眼斑病菌部分生物学特性及其室内毒力测定.广东农业科学,2011,23:77-79;(8)张静,冯岗,袁旭超,陈从珍,林勇.百里香酚抑菌活性初探.中国农学通报.2009,25(21):277-280;(9)高兆银,胡美姣,李敏,杨凤珍.壳聚糖对热带水果采后病原菌的抑菌活性研究.中国南方果树,2008,37(4):51~53;(10)吴光旭,刘爱媛,陈维信.开口箭提取物对荔枝霜疫霉菌的抑制作用及其对荔枝果实的贮藏效果.中国农业科学,2006,39(8):1703-1708)所述的方法分离获得,中国热带农业科学院环境与植物保护研究所已保存。
2.试验方法
2.1离体活性检测
为评价α-倒捻子素的农用杀菌活性,本发明采用生长速率法和孢子萌发法来评价本发明的化合物对植物病原真菌的抑制菌丝生长作用和抑制孢子萌发作用;采用机值分析法拟合本发明的化合物对部分活性较高的植物病原真菌的EC50,以此来评价本发明的化合物。采用的抑制菌丝生长速率法和抑制孢子萌发法测定α-倒捻子素对上述植物病原真菌的抑制活性,参照方中达(方中达.植病研究方法.北京:中国农业出版社,1998)的方法。并在倒置显微镜下(400×)对孢子萌发和菌丝生长情况作细致的观察和记录。α-倒捻子素对菌丝生长抑制作用的测定以50mg/L作为筛选浓度,对表现抑制活性较高的病原菌进行抑制菌丝生长和孢子萌发的毒力测定,设置5-7个浓度处理。以上每处理均重复3次(3个培养皿或3个载玻片),以空白培养基为对照。
菌丝生长抑制率(%)=(对照菌落直径-处理菌落直径)/对照菌落直径×100%
孢子萌发抑制率(%)=(对照萌发率-处理萌发率)/对照萌发率×100%
2.2数据分析
实验结果用SPSS 13.0进行统计分析,将抑制率转化为几率值,浓度转换成对数后进行线性回归,EC50和回归方程均使用机值分析法计算。采用单因素方差分析DMRT法多重比较,P<0.05为具有差异显著性。
采用生长速率法测定了α-倒捻子素对供试20种植物病原真菌的抑菌活性(表1),结果表明,在供试浓度为100mg/L时,α-倒捻子素对植物病菌真菌的敏感性不同,其中,对芒果炭疽病菌和芒果蒂腐病菌的抑菌效果最高,其抑制率分别为90.38%和91.23%;对番茄灰霉病菌和玉米大斑病菌也有很强的抑菌作用,其抑制率分别为85.71%和86.10%;对橡胶树棒孢霉落叶病菌、香蕉炭疽病菌、水稻纹枯病菌、水稻稻瘟病菌、辣椒疫霉病菌和荔枝霜疫霉病菌也具有较好的抑菌作用,其抑制率介于75-85%之间;对橡胶树褐根病菌、橡胶树白根病菌、甘蔗赤腐病菌、甘蔗梢腐病菌、甘蔗黑穗病菌也有一定的抑菌作用,其抑制率介于50-70%;而对甘蔗眼斑病菌、香蕉枯萎病菌、香蕉冠腐病菌、油菜菌核病菌和番茄早疫病菌的抑菌活性较差,其抑制率低于50%。
表1 α-倒捻子素对16种植物病原真菌的抑菌活性
供试菌种 | 抑制率(%) | 供试菌种 | 抑制率(%) |
橡胶树棒孢霉落叶病菌 | 83.78 | 香蕉炭疽病菌 | 81.13 |
橡胶树褐根病菌 | 67.01 | 香蕉枯萎病菌 | 16.67 |
橡胶树白根病菌 | 63.41 | 香蕉冠腐病菌 | 82.76 |
甘蔗赤腐病菌 | 73.33 | 水稻纹枯病菌 | 81.25 |
甘蔗梢腐病菌 | 58.82 | 水稻稻瘟病菌 | 78.57 |
甘蔗黑穗病菌 | 53.85 | 番茄灰霉病菌 | 85.71 |
甘蔗眼斑病菌 | 36.00 | 荔枝霜疫霉病菌 | 75.21 |
芒果炭疽病菌 | 90.38 | 玉米大斑病菌 | 86.10 |
芒果蒂腐病菌 | 91.23 | 油菜菌核病菌 | 41.18 |
辣椒疫霉病菌 | 75.51 | 番茄早疫病菌 | 39.76 |
根据前述筛选结果,进一步测定了α-倒捻子素对前期筛选较为敏感的芒果蒂腐病菌等6种植物病原真菌的毒力,结果表明,α-倒捻子素对芒果蒂腐病菌的毒力最高,其EC50值为18.97mg/L;对芒果炭疽病菌、番茄灰霉病菌和玉米大斑病菌的抑菌作用也很强,其EC50值分别为24.52、31.88和35.04mg/L。
表2 α-倒捻子素对芒果蒂腐病菌等5种病菌的抑制菌丝生长毒力
供试病菌 | 毒力曲线 | 相关系数 | EC50(mg/L) | 95%置信限(mg/mL) |
芒果蒂腐病菌 | 1.8603x+2.6223 | 0.9788 | 18.97 | 17.64-20.41 |
芒果炭疽病菌 | 1.7781x+2.5293 | 0.9857 | 24.52 | 22.41-26.82 |
番茄灰霉病菌 | 1.8675x+2.1922 | 0.9926 | 31.88 | 28.15-34.86 |
玉米大斑病菌 | 2.3667x+1.3445 | 0.9736 | 35.04 | 32.89-37.33 |
α-倒捻子素对番茄灰霉病菌和玉米大斑病菌也有一定的抑制孢子萌发作用,对两种病菌的EC50分别为50.43mg/L和66.20mg/L。
表3 α-倒捻子素对玉米大斑病菌和番茄灰霉病菌的抑制孢子萌发毒力
供试病菌 | 毒力曲线 | 相关系数 | EC50(mg/L) | 95%置信限(mg/mL) |
番茄灰霉病菌 | 1.4887x+2.4652 | 0.9806 | 50.43 | 45.32-56.12 |
玉米大斑病菌 | 1.7799x+1.7591 | 0.9723 | 66.20 | 59.46-73.70 |
2.2盆栽试验法
橡胶树白粉病菌采自中国热带农业科学院试验场九队,将野外自然发病的橡胶树叶带回室内,采用单斑分离法得到纯菌株。轻轻弹去病叶表面的分生孢子后培养1d,使病原菌长出新鲜分生孢子。供试植物为产年生橡胶树芽接幼苗,品系为“热研7-33-97”,塑料棚隔离种植。等幼苗长出宽度为2-3cm的健康古铜色嫩叶后,将生长状态较一致的幼苗移入室内备用。将配置好的药液中加入0.2%Tw-20,用自来水稀释成500和100mg/L的供试液。咪鲜胺原药(97.5%)由湖北康宝泰精细化工有限公司生产。设咪鲜胺(100mg/L)为药剂对照,以清水为空白对照。
保护作用:将种有橡胶苗的花盆置于培养架上,喷施供试液,24h后以杨意伯等(杨意伯,谭志琼,郑服丛,张荣意.橡胶树白粉病杀菌剂生物测定接种方法研究.植物保护,2011,27(2):152-155)描述的喷雾法进行接菌,每处理3盆胶苗,每盆留橡胶苗1株。等清水对照充分发病后调查病情。根据公式(1)和(2)计算病情指数和相对防效,以DMRT法分析各处理的相对防效。橡胶树白粉病病情分级标准参照《橡胶树白粉病测报技术规程》(NY/T 1089-2006,橡胶树白粉病测报技术规程)。
治疗作用测定:试验方法同保护作用测定。区别是先在保湿条件下接种橡胶树白粉病,24h后再喷施供试药液。
公式(1)
病情指数=[∑(病叶数*各级病叶)/(调查总叶数*最高代表级别值)]*100
公式(2)
防治效果%=[(对照病情指数-处理病情指数)/对照病情指数]*100
盆栽法结果显示,α-倒捻子素在500mg/L和100mg/L的处理浓度下对橡胶树白粉病菌表现出较高的保护和治疗作用,在供试浓度为500mg/mL时,对橡胶树白粉病菌的保护和治疗作用分别为75.61%和70.12%,显著高于对照药剂咪鲜胺。
表4 α-倒捻子素对橡胶树白粉病菌的保护和治疗作用
Claims (8)
1.α-倒捻子素在防治植物病害中的应用,其特征在于:所述植物病害为由下述病原真菌引起的植物病害:辣椒疫霉病菌(Phytophthora capsici Leonian)、番茄灰霉病菌(Botrytis cirerea)、水稻纹枯病菌(Thanatephorus cucumeris)、水稻稻瘟病菌(Pyricularia oryzae)、玉米大斑病菌(Exserohilum turcicum),芒果炭疽病菌(Colletotrichum gloeosporioides)、芒果蒂腐病菌(Botryodiplodia theobromae)、香蕉炭疽病菌(Gloeosporium musae)、香蕉冠腐病菌(Fusarium semitectum)、橡胶树棒孢霉落叶病菌(Corynespora cassiicola)、橡胶树褐根病菌(Phellinus noxius Corner)、橡胶树白根病菌(Rigidoporus lignosus)、甘蔗赤腐病菌(Colletotrichum fuleatum Went.)、荔枝霜霉病菌(Peronophythora litchii)中的一种或两种以上任意组合。
2.根据权利要求1所述的应用,其特征在于:所述病原真菌为芒果炭疽病菌、芒果蒂腐病菌、玉米大斑病菌和番茄灰霉病菌中的一种或两种以上任意组合。
3.根据权利要求2所述的应用,其特征在于:所述病原真菌为芒果炭疽病菌和/或芒果蒂腐病菌。
4.根据权利要求1-3中任意一项所述的应用,其特征在于:所述防治植物病害为杀死引起植物病害的植物病原菌或抑制引起植物病害的植物病原菌生长。
5.根据权利要求4所述的应用,其特征在于:所述抑制引起植物病害的植物病原菌生长是抑制所述植物病原菌的菌丝生长和/或抑制所述植物病原菌的孢子萌发和生长。
6.α-倒捻子素在制备植物病原真菌的抑制剂中的应用,其特征在于:所述植物病害为由下述病原真菌引起的植物病害:辣椒疫霉病菌(Phytophthora capsici Leonian)、番茄灰霉病菌(Botrytis cirerea)、水稻纹枯病菌(Thanatephorus cucumeris)、水稻稻瘟病菌(Pyricularia oryzae)、玉米大斑病菌(Exserohilum turcicum),芒果炭疽病菌(Colletotrichum gloeosporioides)、芒果蒂腐病菌(Botryodiplodia theobromae)、香蕉炭疽病菌(Gloeosporium musae)、香蕉冠腐病菌(Fusarium semitectum)、橡胶树棒孢霉落叶病菌(Corynespora cassiicola)、橡胶树褐根病菌(Phellinus noxius Corner)、橡胶树白根病菌(Rigidoporus lignosus)甘蔗赤腐病菌(Colletotrichum fuleatum Went.)、荔枝霜霉病菌(Peronophythora litchii)中的一种或两种以上任意组合。
7.根据权利要求6所述的应用,其特征在于:所述病原真菌为芒果炭疽病菌、芒果蒂腐病菌、玉米大斑病菌和番茄灰霉病菌中的一种或两种以上任意组合。
8.根据权利要求7所述的应用,其特征在于:所述病原真菌为芒果炭疽病菌和/或芒果蒂腐病菌。
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