朝鲜孢囊线虫——浙江省孢囊线虫新记录种
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摘要
在2015—2016年间对浙江省孢囊线虫的种类及分布进行了调查,从浙江省临安市雷竹根围采集到一个孢囊线虫群体,采用形态学和分子生物学方法对其进行了鉴定。利用光学显微镜和扫描电子显微镜对孢囊和2龄幼虫(J2)的形态进行系统的观察和测量,主要形态特征为:孢囊近圆形,有明显的短颈和阴门锥,无下桥、泡囊突和膜孔,阴门裂长45.3μm;2龄幼虫体长528.5μm,头部缢缩不明显,具2个唇环数,口针长19.6μm;侧区具3条侧线;尾圆锥形,末端尖细,尾长75.4μm,透明尾占尾长的55%~75%。对该线虫群体rDNA的内转录间隔区(internal transcribed spacer,ITS)和28SD2D3区进行特异性扩增,分别得到1 006bp和786bp的片段,与朝鲜孢囊线虫(Heterodera koreana)的序列相似度分别高达99.7%和99.6%;综合形态学、分子鉴定及系统进化分析等,将分离自雷竹的孢囊线虫群体鉴定为朝鲜孢囊线虫,系浙江省孢囊线虫的新记录种。
Cyst-forming nematode is an important plant parasitic nematode group,in which over 80 species were reported and described.There are many economically important cyst-forming nematodes species in China,such as Heterodera glycines,Heterodera avenae,and Heterodera filipjevii.During 2015—2016,a survey for cystforming nematodes in Zhejiang,eastern China,revealed the distribution of a cyst-forming nematode population from Phyllostachys praecox in Lin'an,Zhejiang,eastern China.Morphological characteristics and morphometrics of cysts and second-stage juveniles(J2s)were observed and measured using light microscope and scanning electron microscope.Cysts were characterized by appearing light to dark brown in coloration,globose to subspherical shape of body contour,distinct neck and cone,lacking underbridge,bullae and fenestrae,body length ranging from 575.0to 1 230.0μm,body width ranging from 397.0to 872.0μm,and vulva slit length ranging from 36.7to 50.9μm.J2 was characterized by hemispherical labial region,nearly continuous with round body contour and stylet knobs,three incisures in lateral field,tail tapering and tail hyaline region accounting for 55%-75%,body length ranging from 426.3to 608.0μm;body width ranging from 15.9to 29.3μm;stylet length ranging from 17.7to21.9μm;dorsal gland orifice ranging from 4.3to 6.2μm;tail length ranging from 68.7to 84.5μm.Amplificationof the ITS and D2/D3 fragments of the cyst-forming nematodes from bamboo with universal primers AB28(ATATGCTTAAGTTCAGCGGGT)and TW81(GTTTCCGTAGGTGAACCTGC),D2A(ACAAGTACCGTGA GGGAAAGTTG)and D3B(TCGGAAGGAACCAGCTACTA)yielded PCR fragments with sizes of 1 006bp(GenBank:KX640828)and 786bp(GenBank:KX640827),respectively,the sequences of ITS and D2/D3 of cyst nematodes parasitizing on bamboo were 99.7% and 99.6% similar to that of Heterodera koreana deposited in GenBank,respectively.Combined with morphological,morphometrics and molecular phylogeny analysis,the results showed that the cyst nematodes parasitizing on bamboo from Lin'an,Zhejiang,China is H.koreana,is a new record in Zhejiang,Eastern China.
Cyst-forming nematode is an important plant parasitic nematode group,in which over 80 species were reported and described.There are many economically important cyst-forming nematodes species in China,such as Heterodera glycines,Heterodera avenae,and Heterodera filipjevii.During 2015—2016,a survey for cystforming nematodes in Zhejiang,eastern China,revealed the distribution of a cyst-forming nematode population from Phyllostachys praecox in Lin'an,Zhejiang,eastern China.Morphological characteristics and morphometrics of cysts and second-stage juveniles(J2s)were observed and measured using light microscope and scanning electron microscope.Cysts were characterized by appearing light to dark brown in coloration,globose to subspherical shape of body contour,distinct neck and cone,lacking underbridge,bullae and fenestrae,body length ranging from 575.0to 1 230.0μm,body width ranging from 397.0to 872.0μm,and vulva slit length ranging from 36.7to 50.9μm.J2 was characterized by hemispherical labial region,nearly continuous with round body contour and stylet knobs,three incisures in lateral field,tail tapering and tail hyaline region accounting for 55%-75%,body length ranging from 426.3to 608.0μm;body width ranging from 15.9to 29.3μm;stylet length ranging from 17.7to21.9μm;dorsal gland orifice ranging from 4.3to 6.2μm;tail length ranging from 68.7to 84.5μm.Amplificationof the ITS and D2/D3 fragments of the cyst-forming nematodes from bamboo with universal primers AB28(ATATGCTTAAGTTCAGCGGGT)and TW81(GTTTCCGTAGGTGAACCTGC),D2A(ACAAGTACCGTGA GGGAAAGTTG)and D3B(TCGGAAGGAACCAGCTACTA)yielded PCR fragments with sizes of 1 006bp(GenBank:KX640828)and 786bp(GenBank:KX640827),respectively,the sequences of ITS and D2/D3 of cyst nematodes parasitizing on bamboo were 99.7% and 99.6% similar to that of Heterodera koreana deposited in GenBank,respectively.Combined with morphological,morphometrics and molecular phylogeny analysis,the results showed that the cyst nematodes parasitizing on bamboo from Lin'an,Zhejiang,China is H.koreana,is a new record in Zhejiang,Eastern China.
引文
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[2]SUBBOTIN S A,MUNDO-OCAMPO M,BALDWIN J G,et al.Systematics of cyst nematodes(Nematoda:Heteroderinae)(8B).BRILL,2010:249-253.
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[9]ZHUO K,WANG H H,ZHANG H L,et al.Heterodera guangdongensis n.sp.(Nematoda:Heteroderinae)from bamboo in Guangdong Province,China:a new cyst nematode in the Cyperi group.Zootaxa,2014,3881(5):488-500.
[10]DING Z,NAMPHUENG J,HE X F,et al.First report of the cyst nematode(Heterodera elachista)on rice in Hunan Province,China.Plant Disease,2012,96(1):151.
[11]王宏洪,卓侃,张洪玲,等.孢囊线虫中国新记录种—朝鲜孢囊线虫(Heterodera koreana).植物病理学报,2012,42(5):551-555.WANG H H,ZHUO K,ZHANG H L,et al.Heterodera koreana,a new record species from China.Acta Phytopathologica Sinica,2012,42(5):551-555.(in Chinese with English abstract)
[12]陈品三,郑经武,马承铸.艾球孢囊线虫(Globodera artemisiae)在我国的发现初报.中国农业科学,1994,27(3):89.CHEN P S,ZHENG J W,MA C Z.A new record of the genus Globodera(G.artemisiae)from Beijing,China.Scientia Agricultura Sinica,1994,27(3):89.(in Chinese)
[13]ZHENG J W,ZHANG Y,LI X Q,et al.First report of the soybean cyst nematode,Heterodera glycines,on soybean in Zhejiang,Eastern China.Plant Disease,2009,93(3):319.
[14]谢辉.植物线虫分类学.安徽:科学技术出版社,2000:21-22,32-34.XIE H.Taxonomy of Plant Nematodes.Anhui:Science and Technology Press,2000:21-22,32-34.(in Chinese)
[15]宋志强,王暄,林宇,等.土壤中南方根结线虫2龄幼虫和卵的实时荧光PCR检测和定量.植物保护学报,2013,40(3):255-260.SONG Z Q,WANG X,LIN Y,et al.Detection and quantification of Meloidogyne incognita in soil sample using real-time PCR.Acta Phytophylacica Sinica,2013,40(3):255-260.(in Chinese with English abstract)
[16]SUBBOTIN S A,STURHAN D,RUMPENHORST H J,et al.Morphological characterization of the Heterodera avenae species complex(Tylenchida:Heteroderidae).Nematology,2003,5(4):515-538.
[17]DELEY P,FELIX M A,FRISSE L M,et al.Molecular and morphological characterisation of two reproductively isolated species with mirror-image anatomy(Nematoda:Cephalobidae).Nematology,1999,1(6):591-612.
[18]VOVLAS N,LAMBERTI F,CHOO H Y.Description of Afenestrata koreana n.sp.(Nematoda:Heteroderinae),a parasite,of bamboo in Korea.Journal of Nematology,1992,24(4):553-559.
[19]MUNDO-OCAMPO M,TROCCOLI A,SUBBOTIN S A,et al.Synonymy of Afenestrata with Heterodera supported by phylogenetics with molecular and morphological characterisation of H.koreana comb.n.and H.orientalis comb.n.(Tylenchida:Heteroderidae).Nematology,2008,10(5):611-632.
[20]INSERRA R N,VOVLAS N,LEHMAN P S.Parasitism of fishpole bamboo roots by Afenestrata koreana.Nematropica,1999,29(1):105-111.
[21]MAAFI Z T,TAHERI Z M.First report of Korean cyst nematode,Heterodera koreana,parasitic on bamboo,Phyllostachys nigra,from Iran.Nematol,2015,47(3):167-168.
[22]徐祖祥,陈丁红,李良华,等.临安雷竹种植条件下土壤养分的变化.中国农学通报,2010,26(13):247-250.XU Z X,CHEN D H,LI L H,et al.A study on the change in soft nutrients under the condition of bamboo shoot planting in Lin'an.Chinese Agricultural Science Bulletin,2010,26(13):247-250.(in Chinese with English abstract)
[2]SUBBOTIN S A,MUNDO-OCAMPO M,BALDWIN J G,et al.Systematics of cyst nematodes(Nematoda:Heteroderinae)(8B).BRILL,2010:249-253.
[3]李惠霞,柳永娥,魏庄,等.新疆和西藏发现禾谷孢囊线虫.中国线虫学研究:第四卷,北京,中国农业科学技术出版社,2012:164-165.LI H X,LIU Y E,WEI Z,et al.The detection of Hetebodertl avenae from the cereal field in autonomous regions of Tibet and Xinjiang.Nematology Research in China:Vol.4,Beijing:China's Agricultural Science and Technology Press,2012:164-165.(in Chinese)
[4]陈品三,王明祖,彭德良.我国小麦禾谷孢囊线虫(Heterodera avenae Wollenweber)鉴定研究.植物病理学报,1992,22(4):339-342.CHEN P S,WANG M Z,PENG D L.Identification on cereal cyst nematode of wheat in China.Acta Phytopathologica Sinica,1992,22(4):339-342.(in Chinese with English abstract)
[5]PENG D L,YE W X,PENG H,et al.First report of the cyst nematode(Heterodera filipjevi)on wheat in Henan Province,China.Plant Disease,2010,94(10):1262.
[6]陈品三,郑经武,彭德良.中国异皮属线虫一新种(线虫纲:垫刃目:异皮科).动物分类学报,1996,21(1):18-24.CHEN P S,ZHENG J W,PENG D L.A new species of the genus Heterodera from China(Nematoda:Tylenchida:Heteroderoderidae).Acta Zootaxonomica Sinica,1996,21(1):18-24.(in Chinese with English abstract)
[7]WANG H H,ZHUO K,YE W M,et al.Heterodera fengi n.sp.(Nematoda:Heteroderinae)from bamboo in Guangdong Province,China:a new cyst nematode in the Cyperi group.Zootaxa,2013,3652(1):179-192.
[8]ZHUO K,WANG H H,YE W M,et al.Heterodera hainanensis n.sp.(Nematoda:Heteroderinae)from bamboo in Hainan Province,China:a new cyst nematode in the Afenestrata group.Nematology,2013,15(3):303-314.
[9]ZHUO K,WANG H H,ZHANG H L,et al.Heterodera guangdongensis n.sp.(Nematoda:Heteroderinae)from bamboo in Guangdong Province,China:a new cyst nematode in the Cyperi group.Zootaxa,2014,3881(5):488-500.
[10]DING Z,NAMPHUENG J,HE X F,et al.First report of the cyst nematode(Heterodera elachista)on rice in Hunan Province,China.Plant Disease,2012,96(1):151.
[11]王宏洪,卓侃,张洪玲,等.孢囊线虫中国新记录种—朝鲜孢囊线虫(Heterodera koreana).植物病理学报,2012,42(5):551-555.WANG H H,ZHUO K,ZHANG H L,et al.Heterodera koreana,a new record species from China.Acta Phytopathologica Sinica,2012,42(5):551-555.(in Chinese with English abstract)
[12]陈品三,郑经武,马承铸.艾球孢囊线虫(Globodera artemisiae)在我国的发现初报.中国农业科学,1994,27(3):89.CHEN P S,ZHENG J W,MA C Z.A new record of the genus Globodera(G.artemisiae)from Beijing,China.Scientia Agricultura Sinica,1994,27(3):89.(in Chinese)
[13]ZHENG J W,ZHANG Y,LI X Q,et al.First report of the soybean cyst nematode,Heterodera glycines,on soybean in Zhejiang,Eastern China.Plant Disease,2009,93(3):319.
[14]谢辉.植物线虫分类学.安徽:科学技术出版社,2000:21-22,32-34.XIE H.Taxonomy of Plant Nematodes.Anhui:Science and Technology Press,2000:21-22,32-34.(in Chinese)
[15]宋志强,王暄,林宇,等.土壤中南方根结线虫2龄幼虫和卵的实时荧光PCR检测和定量.植物保护学报,2013,40(3):255-260.SONG Z Q,WANG X,LIN Y,et al.Detection and quantification of Meloidogyne incognita in soil sample using real-time PCR.Acta Phytophylacica Sinica,2013,40(3):255-260.(in Chinese with English abstract)
[16]SUBBOTIN S A,STURHAN D,RUMPENHORST H J,et al.Morphological characterization of the Heterodera avenae species complex(Tylenchida:Heteroderidae).Nematology,2003,5(4):515-538.
[17]DELEY P,FELIX M A,FRISSE L M,et al.Molecular and morphological characterisation of two reproductively isolated species with mirror-image anatomy(Nematoda:Cephalobidae).Nematology,1999,1(6):591-612.
[18]VOVLAS N,LAMBERTI F,CHOO H Y.Description of Afenestrata koreana n.sp.(Nematoda:Heteroderinae),a parasite,of bamboo in Korea.Journal of Nematology,1992,24(4):553-559.
[19]MUNDO-OCAMPO M,TROCCOLI A,SUBBOTIN S A,et al.Synonymy of Afenestrata with Heterodera supported by phylogenetics with molecular and morphological characterisation of H.koreana comb.n.and H.orientalis comb.n.(Tylenchida:Heteroderidae).Nematology,2008,10(5):611-632.
[20]INSERRA R N,VOVLAS N,LEHMAN P S.Parasitism of fishpole bamboo roots by Afenestrata koreana.Nematropica,1999,29(1):105-111.
[21]MAAFI Z T,TAHERI Z M.First report of Korean cyst nematode,Heterodera koreana,parasitic on bamboo,Phyllostachys nigra,from Iran.Nematol,2015,47(3):167-168.
[22]徐祖祥,陈丁红,李良华,等.临安雷竹种植条件下土壤养分的变化.中国农学通报,2010,26(13):247-250.XU Z X,CHEN D H,LI L H,et al.A study on the change in soft nutrients under the condition of bamboo shoot planting in Lin'an.Chinese Agricultural Science Bulletin,2010,26(13):247-250.(in Chinese with English abstract)