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1、2011,37(2):1562158PlantProtection玉米灰斑病研究方法:Ⅰ.玉米灰斑病菌孢子的培养1121,23赵正龙,吴兴兴,毛自朝,何月秋(1.云南农业大学农业生物多样性应用技术国家工程研究中心,昆明650201;2.云南农业大学农学与生物技术学院,昆明650201)摘要为了明确玉米灰斑病菌的产孢条件,将玉米灰斑病菌接种到马铃薯蔗糖(PSA)、燕麦番茄碳酸钙(OTCA)、玉米叶粉碳酸钙(MLPCA)、燕麦粉(OA)、粗面粉(CWPA)等5种培养基上,于25℃下培养,测定该菌生长速度、产孢时间
2、和产孢量。结果表明,菌丝在上述培养基上生长速度均较慢,每天平均生长0.33mm左右,其中以在OA和PSA上生长速度相对较快。基于上述结果,构建了玉米灰斑病菌孢子培养的二级分段培养法,即将菌丝块接种于加有10～15粒直径为5mm玻璃珠的马铃薯蔗糖(PS)液体培养基三角瓶里,在25℃,180r/min条件下振荡培养15d,形成菌丝悬液,再取菌丝悬浮液涂布在MLPCA培养基上,避光培养13d左右,可获得大量供接种的分生孢子。长期光照促进菌丝生长,不利于孢子的形成,而长期黑暗不利于菌丝生长,但有利于孢子的形成。关键词
3、玉米灰斑病菌;振荡培养;产孢;二级分段培养法中图分类号:S435.131.4文献标识码:ADOI:10.3969/j.issn.052921542.2011.02.035Researchmethodologyformaizegrayleafspot:Ⅰ.sporulationofCercosporazeae2maydis1121,2ZhaoZhenglong,WuXingxing,MaoZichao,HeYueqiu(1.TheNationalCenterforAgriculturalBiodiversity
4、Exploitation,YunnanAgriculturalUniversity,Kunming650201,China;2.FacultyofAgronomyandBiotechnology,YunnanAgriculturalUniversity,Kunming650201,China)AbstractThepathogenofmaizegrayleafspot,Cercosporazeae2maydis,wasinoculatedonpotatosucroseagar(PSA),oattomatopl
5、usCaCO3agar(OTCA),maizeleafpowderplusCaCO3agar(MLPCA),oatmealagar(OA)andcrudewheatpowderagar(CWPA),andincubatedat25℃formeasuringthemycelialgrowth,sporulationtimeandnumber.Theresultsshowedthatthemyceliagrewveryslowonthemedia,withanaverageof0.33mmperday.Theyg
6、rewfastestonOAandPSA,sloweronCWPAandmostslowlyonMLPCA.Basedontheresults,asporulationmethodwassetup,namelytwo2stepculturingmethod.ThemyceliawereinoculatedintoflaskwithPSand10-15glassballsof5mmdiameter,andshakenat25℃and180r/minfor15days,toculturethemycelialfr
7、agmentsuspension.ThenthesuspensionwassmearedonMLPCAmediumandincubatedforabout13daysat25℃withoutlight.Bythisway,alargenumberofsporescouldbeproducedforinoculation.Longilluminationpromotedthemycelialgrowth,butonthecontrary,longdarknesswasconducivetosporulation
8、.KeywordsCercosporazeae2maydis;shakingculture;sporulation;two2stepculturingmethod玉米灰斑病(maizegrayleafspot)也称玉米尾产量和品质。玉米发病后一般田块损失5%～[223]孢叶斑病,是由Cercosporazeae2maydisTehon&30%,严重田块达60%以上,甚至绝收。该病主[1]Daniel
