高山杜鹃枯梢病病原菌鉴定及品种抗病性调查
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摘要
在云南省昆明市晋宁区高山杜鹃盆花种植基地发现枯梢病,田间发病率达20%以上。发病植株顶梢枯死,茎杆溃疡,最终枯萎死亡。对病原菌进行组织分离培养和致病性测定,结果表明,病原菌菌落初期白色,后期呈灰黑色;分生孢子器球形,黑色;分生孢子单胞,近椭圆形至纺锤形,大小为(17.2～32.8)μm×(4.7～7.8)μm。以ITS通用引物对病原菌基因组DNA进行扩增并测序,同源性比对显示其与小新壳梭孢(Neofusicoccum parvum)的一致性达99%,结合形态学鉴定结果,确定该病原菌为小新壳梭孢。病原菌的最适生长温度为25℃;p H 5～7时菌丝生长加快,pH 7时最适于菌丝生长;以蔗糖为碳源、硫酸铵为氮源时菌丝生长最好。不同高山杜鹃品种对小新壳梭孢的抗性存在差异,18个品种中免疫品种2个,高抗品种2个,抗病品种3个,中感品种3个,感病品种4个,高感品种4个。
A new branch disease on Rhododendron was observed in a planting base in Jinning District,Kunming,Yunnan. The disease incidence was more than 20%. Disease symptoms included twig dieback and stem canker. Pathogens were isolated from the disease samples. The pathogenicity test,morphological and molecular identification were studied and cultivar resistance of Rhododendron were evaluated. The results showed that the mycelium was white initially and turned gray-black later. Conidia were one-celled,ellipsoidal to fusiform,with the length of 17.2 to 32.8 μm and the width of 4.7 to 7.8 μm.The internal transcribed spacer(ITS)region was amplified using the ITS1/ITS4 primer pair. BLAST analysis of the sequence showed 99% homology with the sequence of Neofusicoccum parvum. Based on the morphology and ITS sequence analysis,the pathogen was identified as N. parvum. The optimum temperature for mycelium growth was 25 ℃. The mycelial growth rate increased rapidly at pH 5–7,and the optimum pH value was 7. The utilization rates of sucrose and ammonium sulfate were the highest among all carbon and nitrogen sources,respectively. Different cultivars exhibited different disease resistances,including two immune cultivars,two highly resistant cultivars,three resistant cultivars,three moderately susceptible cultivars,four susceptible cultivars and four highly susceptible cultivars.
A new branch disease on Rhododendron was observed in a planting base in Jinning District,Kunming,Yunnan. The disease incidence was more than 20%. Disease symptoms included twig dieback and stem canker. Pathogens were isolated from the disease samples. The pathogenicity test,morphological and molecular identification were studied and cultivar resistance of Rhododendron were evaluated. The results showed that the mycelium was white initially and turned gray-black later. Conidia were one-celled,ellipsoidal to fusiform,with the length of 17.2 to 32.8 μm and the width of 4.7 to 7.8 μm.The internal transcribed spacer(ITS)region was amplified using the ITS1/ITS4 primer pair. BLAST analysis of the sequence showed 99% homology with the sequence of Neofusicoccum parvum. Based on the morphology and ITS sequence analysis,the pathogen was identified as N. parvum. The optimum temperature for mycelium growth was 25 ℃. The mycelial growth rate increased rapidly at pH 5–7,and the optimum pH value was 7. The utilization rates of sucrose and ammonium sulfate were the highest among all carbon and nitrogen sources,respectively. Different cultivars exhibited different disease resistances,including two immune cultivars,two highly resistant cultivars,three resistant cultivars,three moderately susceptible cultivars,four susceptible cultivars and four highly susceptible cultivars.
引文
Cheng Yan-lin,Liang Jun,LüQuan,Zhang Xing-yao.2011.Advances in Botryosphaeriaceae:identification,phylogeny and molecular ecology.Acta Ecologica Sinica,31(11):3197-3207.(in Chinese)程燕林,梁军,吕全,张星耀.2011.葡萄座腔菌科研究进展:鉴定,系统发育学和分子生态学.生态学报,31(11):3197-3207.
Espinoza J G,Brice?o E X,Chávez E R,úrbez-Torres J R,Latorre B A.2009.Neofusicoccum spp.associated with stem canker and dieback of blueberry in Chile.Plant disease,93:1187-1194.
Fang Zhong-da.1998.Methodology for plant pathology.Beijing:China Agricultural Press.(in Chinese)方中达.1998.植病研究方法.北京:中国农业出版社.
Li Xiu-yan,Zhang Ge,Chen Jun.1999.Study on the pathogen of rhododendron dieback.Forest Pest and Disease,(1):30-31.(in Chinese)李秀艳,张革,陈军.1999.杜鹃花枯梢病病原菌的研究.森林病虫通讯,(1):30-31.
Liu Xiao-qing,Su Jia-le,Li Chang,He Li-si,Chen Lu.2011.A new alpine rhododendron cultivar‘Fuli Jinling’.Acta Horticulturae Sinica,38(11):2237-2238.(in Chinese)刘晓青,苏家乐,李畅,何丽斯,陈璐.2011.高山杜鹃新品种‘富丽金陵’.园艺学报,38(11):2237-2238.
Pintos V C,Redondo F V,Mansilla V J P,Aguin C O.2011.First report of dieback on hybrid Rhododendrons caused by Neofusicoccum luteum and N.parvum in Spain.Plant Disease,95(2):221.
Qi Hui-xia,Li Shuang-min,Yu Jin-yong,Liu Rui,Yang Wen-lan.2008.Pathogen identification and biological characteristics of anthracnose in Hydrangea macrophylla.Nonwood Forest Research,26(3):84-87.(in Chinese)齐慧霞,李双民,余金咏,刘锐,杨文兰.2008.八仙花炭疽病病原菌鉴定及其生物学特性.经济林研究,26(3):84-87.
Tang Shi-jie,Ma Jian-xia,Guo Zhong-ren,Tang Geng-guo.2005.Occurence of common diseases and insect pests of Rhododendron L.and their controlling methods.Journal of Jiangsu Forestry Science&Technology,32(3):24-25.(in Chinese)汤诗杰,马建霞,郭忠仁,汤庚国.2005.杜鹃花常见病虫害的发生与防治.江苏林业科技,32(3):24-25.
úrbez J R,Leavitt G M,Voegel T M,Gubler W D.2006.Identification and distribution of Botryosphaeria spp.associated with grapevine cankers in California.Plant Disease,90(12):1490-1503.
Wills W H,Lambe R C.1986.Studies on Rhododendron dieback caused by Botryosphaeria dothidea.Journal American Rhododendron Society,40(4):22.
Xu Cheng-nan.2016.Etiology,molecular detection and control of blueberry stem blight in China[Ph.D.Dissertation].Baoding:Agricultural University of Hebei.(in Chinese)徐成楠.2016.中国蓝莓枝枯病的病原学、分子检测及防治技术研究[博士论文].保定:河北农业大学.
Yu Lei,Zhao Jian-rong,Rarisara I,Xu Sheng-guang,Wu Xu.2013.Identification of the pathogen causing twigs and stem dieback in blueberry.Acta Phytopathologica Sinica,43(4):421-425.(in Chinese)余磊,赵建荣,Rarisara Impaprasert,徐胜光,吴旭.2013.蓝莓枝枯病病原菌鉴定.植物病理学报,43(4):421-425.
Espinoza J G,Brice?o E X,Chávez E R,úrbez-Torres J R,Latorre B A.2009.Neofusicoccum spp.associated with stem canker and dieback of blueberry in Chile.Plant disease,93:1187-1194.
Fang Zhong-da.1998.Methodology for plant pathology.Beijing:China Agricultural Press.(in Chinese)方中达.1998.植病研究方法.北京:中国农业出版社.
Li Xiu-yan,Zhang Ge,Chen Jun.1999.Study on the pathogen of rhododendron dieback.Forest Pest and Disease,(1):30-31.(in Chinese)李秀艳,张革,陈军.1999.杜鹃花枯梢病病原菌的研究.森林病虫通讯,(1):30-31.
Liu Xiao-qing,Su Jia-le,Li Chang,He Li-si,Chen Lu.2011.A new alpine rhododendron cultivar‘Fuli Jinling’.Acta Horticulturae Sinica,38(11):2237-2238.(in Chinese)刘晓青,苏家乐,李畅,何丽斯,陈璐.2011.高山杜鹃新品种‘富丽金陵’.园艺学报,38(11):2237-2238.
Pintos V C,Redondo F V,Mansilla V J P,Aguin C O.2011.First report of dieback on hybrid Rhododendrons caused by Neofusicoccum luteum and N.parvum in Spain.Plant Disease,95(2):221.
Qi Hui-xia,Li Shuang-min,Yu Jin-yong,Liu Rui,Yang Wen-lan.2008.Pathogen identification and biological characteristics of anthracnose in Hydrangea macrophylla.Nonwood Forest Research,26(3):84-87.(in Chinese)齐慧霞,李双民,余金咏,刘锐,杨文兰.2008.八仙花炭疽病病原菌鉴定及其生物学特性.经济林研究,26(3):84-87.
Tang Shi-jie,Ma Jian-xia,Guo Zhong-ren,Tang Geng-guo.2005.Occurence of common diseases and insect pests of Rhododendron L.and their controlling methods.Journal of Jiangsu Forestry Science&Technology,32(3):24-25.(in Chinese)汤诗杰,马建霞,郭忠仁,汤庚国.2005.杜鹃花常见病虫害的发生与防治.江苏林业科技,32(3):24-25.
úrbez J R,Leavitt G M,Voegel T M,Gubler W D.2006.Identification and distribution of Botryosphaeria spp.associated with grapevine cankers in California.Plant Disease,90(12):1490-1503.
Wills W H,Lambe R C.1986.Studies on Rhododendron dieback caused by Botryosphaeria dothidea.Journal American Rhododendron Society,40(4):22.
Xu Cheng-nan.2016.Etiology,molecular detection and control of blueberry stem blight in China[Ph.D.Dissertation].Baoding:Agricultural University of Hebei.(in Chinese)徐成楠.2016.中国蓝莓枝枯病的病原学、分子检测及防治技术研究[博士论文].保定:河北农业大学.
Yu Lei,Zhao Jian-rong,Rarisara I,Xu Sheng-guang,Wu Xu.2013.Identification of the pathogen causing twigs and stem dieback in blueberry.Acta Phytopathologica Sinica,43(4):421-425.(in Chinese)余磊,赵建荣,Rarisara Impaprasert,徐胜光,吴旭.2013.蓝莓枝枯病病原菌鉴定.植物病理学报,43(4):421-425.