摘要
本研究于2005～2007年间,对四川都江堰的川芎,雅安天全二郎山的川牛膝,德阳中江的丹参、芍药等重要药材的病害进行了调查,并对川牛膝黑根病进行了初步诊断,分离鉴定了川芎、川牛膝的内生、附生真菌。针对危害严重的川芎根腐病进行了病原鉴定、生物防治和化学防治等研究。
经调查川芎病害主要有根腐病、炭疽病、灰霉病、叶枯病、白粉病、病毒病和菌核病等7种,其中根腐病危害最大,发生最严重;川牛膝有5种病害,即白锈病、褐斑病、炭疽病、根结线虫病和黑根病;丹参有6种病害,即丹参根腐病、白绢病、叶枯病、根结线虫病、病毒病和丹参菟丝子等。还有3种芍药病害,芍药轮纹病、灰霉病和白粉病。
从30个川芎样品中共分离鉴定出4种内生真菌,9种附生真菌,内生真菌以镰孢属和木霉属为主,附生真菌中木霉、镰刀菌和毛霉为主,内生、附生真菌以木霉和镰刀菌为主要种群。
川牛膝根部内生真菌,1年生病根种类少,只有镰刀菌和黑孢霉,1年生健根分离到5种真菌,分别是木霉属、腐霉属、丝核菌、曲霉和黑孢霉,共有的真菌是黑孢霉;2年生病根和健根都分离得到了4个属的真菌,病根内生真菌中,镰刀菌和黑孢霉出现的频率比较高,镰刀菌为44.4%,黑孢霉为38.7%,还分离得到了少量的木霉、链格孢;健根中分离频率最高的是曲霉属,达到了83.4%;另外,枝孢霉属、黑孢霉和茎点霉真菌也偶见。在病根和健根中都出现的真菌种类是黑孢霉;3年生病根中分离得到12个属,而健根中仅分离到5个属,共同出现的真菌种类是镰刀菌、曲霉、青霉、枝孢霉和黑孢霉属真菌。
川牛膝根部附生真菌分离结果表明,1年生病根附生真菌分离得到10个类群,其中以镰刀菌和木霉为主,镰刀菌的分离频度最大为51.9%,1年生健根分离得到的真菌只有3个属,分别是曲霉、柱孢属和丝核菌。1年生病健根中都出现的真菌是柱孢属和丝核菌两类;2年生病根分离得到7种真菌,分别是镰刀菌、柱孢属、毛霉属、丝核菌、黑孢霉、茎点霉和链格孢,以镰刀菌最多。2年生健根共得到5种附生真菌,镰刀菌的分离频度最大为33.3%;3年生病根附生真菌的种类丰富,共有14种,镰刀菌最多,3年生健根分离得到5中附生真菌。病健根共有的真菌是镰刀菌、青霉和黑孢霉。
经分离鉴定,川芎根腐病的病原菌是茄腐皮镰刀菌[Fusarium solani(Mart)Sacc]。从川牛膝黑根病1、2、3年生病根中分离得到了3种共有的真菌种类,分别是黑孢霉、镰刀菌和柱孢属真菌,回接健康植株,无发病症状,通过综合分析,初步诊断黑根病不属于侵染性病害。
对峙培养结果表明,所试40个木霉菌株对茄腐皮镰刀菌都有不同程度的抑制作用,只有1株没有抑制作用,其中T131-21的抑制效果最好。供试的细菌对病原菌的抑制效果不明显,生防意义不大。木霉生长速度测定结果表明,多数菌株生长迅速,为空间和营养资源的强竞争者。非挥发性代谢产物的拮抗试验中,菌株T131-2,T150-17的抑制率(63.82%)极显著高于其他菌株。生防盆栽试验结果表明,每种预定处理方案对川芎根腐病都有一定的防治效果,处理E(先接生防菌后接病原菌与生防菌)的防治效果最好。
选择了6种杀菌剂进行药剂试验,采用含药平板法测定它们对茄腐皮镰刀菌菌丝生长的抑制作用,并测定了抑菌中量(EC_(50))。结果表明,6种杀菌剂对川芎根腐病的菌丝生长均有抑制作用,一品甲托最好,其次是百菌清、根腐灵、代森锰锌和多菌灵,治根神医最弱。盆栽试验结果也表明一品甲托的防治效果较好,与室内药剂筛选结果一致。
Disease investigation of some important medicinal plants were carried out in Dujiangyan,Er'lang Mountain of Tianquan Ya'an and Zhongjiangof Deyang from 2005 to 2007.Research as isolation and identification of pathogen of Ligusticum Chuanxiong root rot disease,studied on biological and chemical control of root rot of Ligusticum Chuanxiong.The causes of root rot of Chuanxiong and black root of Achyranthis were analyzed.To isolate and identify the Endophytic and Epiphytic fungus of chuanxiong and Achyranthis.
Through the investigation of chuanxiong disease,seven types of diseases had been found,including root rot disease,anthrax disease,grey-mildewy disease,leaf blight disease,powdery mildew disease,virus disease and sclerotium disease.Root rot harms the most among them.Five diseases had been found in Achyranthis,which were white-rust disease,brown-spot disease,anthrax disease,meloidogyne disease and black root disease.Six diseases had been found in salvia,which were root rot disease, southern blight disease,leaf blight disease,meloidogyne disease,virus disease and dodder disease.three disease have been found in chinese herbaceous peony,which were Peony ring spot disease,grey mildewy disease and powdery mildew disease.
Four types of Endophytic fungus and nine types of Epiphytic fungus had been isolated and identified among thirty samples of chuanxiong.Endophytic fungus mainly consists of Fusarium spp.and Trichoderma spp.Epiphytic fungus mainly consists of Trichoderma spp.,Fusarium spp.and Mucor spp..Trichoderma spp.and Fusarium spp. were main populations of Endophytic fungus and Epiphytic fungus.To Endophytic fungus of root of Achyranthis,one-year kind had few of Pathogensexcept for Fusarium spp and Nigrospora sp.Five types of fungus could been isolated from one-year healthy root,which were Trichoderma spp.、Pythium spp.、Rhizoctonia sp.、Aspergillus spp.and Nigrospora sp.,and Nigrospora sp.were their commonage.Four genuses of fungus had been separated in both two-year sick-root and healthy-root.Among the Endophytic fungus of sick-root,Fusarium spp.and Nigrospora sp.occur more frequently,Fusarium spp.44.4%,Nigrospora sp 38.7%and little Trichoderma spp.、Alternaria spp.were also been found.To healthy root,the highest separation frequency was Aspergillus spp., which attains to 83.4%,besides,Cladosporium spp.、Nigrospora sp.and Phoma sp.can been seen occasionly.Nigrospora was found both in sick-root and healthy-root.Twelve genuses have been obtained in the separation of three-year sick-root while five genuses have been obtained in the separation of healthy-root.Fusarium spp,Aspergillus spp.、Penicillium spp.、Cladosporium spp.and Nigrospora sp.appear in every case.
The separation of Epiphytic fungus shows that ten genuses had been obtained in the separation in one-year sick root,mainly being Fusarium spp.and Trichoderma spp.. The highest separation frequency was 51.9%.Only three types of genus had been isolated in one-year healthy root,which were Aspergillus spp.、Cylindrocarpon spp.and Rhizoctonia sp.respectively.Cylindrocarpon spp.and Rhizoctonia sp.appeared in both one-year sick root and healthy root.Seven types of fungus had been isolated in the separation of two-year sick root,which are Fusarium spp,Cylindrocarpon spp., Penicillium spp.,Rhizoctonia sp.,Nigrospora sp.,Phoma sp.and Alternaria spp.,and Fusarium spp appears most often.Five attached fungus had been isolated in two-year healthy root and the highest separation frequency of Fusarium spp is 33.3%.fourteen types of attached fungus have been isolated in three-year sick root,among which,most of which was Fusarium spp.Five types of attached fungus had been isolated in the separation of three-year healthy root.Fusarium spp,Penicillium spp.and Nigrospora sp.appear in both sick root and healthy root.
Through the separation and identification,the pathogeny of root rot of chuanxiong was Fusarium solani.Three common types of fungus which were Nigrospora sp.、Fusarium spp.and Cylindrocarpon spp.,were obtained in the separation process from one-year sick root,two-year sick root and three-year sick root of black root disease of Achyranthis,beening transplanted to healthy plant,no disease was found.Through analysis,initial Diagnosis could been drawn that black root disease done not belong to infectious diseases.
The result of dural culture result showed that forty Trichoderma spp.strains had inhibition ability to Fusarium solani,except one and T131-21 has the greatest effect.The bacteria studied make unobviously inhibition ability to pathogeny and it was of little significance to take biological control.The testing of growing speed of Trichoderma spp. revealed that most microbial strain grow fast,and were strong competitors striving for space and nutrition resources.In the inhibition experiment of non-volatilization,T131-2, T150-17,which reaches 63.82%,has more serious inhibition ability than other microbial strains.Planting-in-pot experiment in biological way showed that every prearranged project can prevent and cure root-rot of chuangxiong to some extent but method E has the best effect.
Six fungicides were selected to test their EC_(50) and their Inhibition effect on Fusarium solani(Mart)Sacc.The result showed that six medicaments can restrain hypha from growing,thiopanate-meththiy/thiram does the best while hymexazol does the worst. chlorothalonil、Tetramethyl thiuram disulfide、suifer.mancozeb and carbendazim have some effect.The result that thiopanate-meththiy/thiram done the best verifies the in-door experiment.
引文
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