长针线虫Longidorus pinus的形态和分子鉴定
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摘要
在传统形态学分类鉴定的基础上,结合18S和28S核糖体(r DNA)序列分析方法,对采自山西介休市枣树根际土样中的线虫进行了种类鉴定。结果表明,雌虫热杀死后呈J型或C型,体长3 480～4 366μm,唇区明显缢缩,齿尖针70～82μm,导环距头前端31～34μm,阴门位于体中部,尾短,圆锥形;雄虫未发现;1龄幼虫的替代口针插入齿托内部,尾短,圆锥形,齿尖针44μm,替代口针长度52μm。其鉴定代码为A23-B1-C23-D4-E1-F2-G23-H23-I1-J2-K6。系统发育研究表明,试验种群的18S和28S区序列与Longidorus pinus相应序列(18S:MF674014～MF674015;28S:MF674016～MF674019)在同一进化枝上,置信度分别达100%和96%。通过形态和分子特征,将该种群鉴定为Longidorus pinus Xu,Ye,Wang&Zhao,2018。Longidorus pinus在枣树上首次报道,枣树为该线虫的新寄主。
On the basis of classification and identification of traditional morphology,combining 18 S and 28 S ribosome(r DNA)sequence analysis,species of nematodes in jujube rhizosphere soil samples collected from Jiexiu city of Shanxi province were identified.The results showed that females were J-or C-shaped after heat killing,body length was 3 480-4 366 μm,the lip region was distinctly set off,odontostyle 70-82 μm long,guide ring situated at 31～34 μm from anterior end,vulva in form of a transverse slit,tail short,conoid,male was not found.The first-stage juvenile characterised by having replacement odontostyle tip inserted into odontophore base,tail short conoid,odontostyle length 44 μm long,replacement odontostyle length 52 μm long.The codes for identifying the species of Longidorus pinus were A23-B1-C23-D4-E1-F2-G23-H23-I1-J2-K6.The sequence alignments and phylogenetic relationships were analysed using data from 18 S and 28 S,respectively.The 18 S of the new population was grouped with Longidorus pinus(MF674014-MF674015)with 100%(pp)supported and 28 S of the new population was also clustered together with Longidorus pinus(MF674016-MF674019)with 96%(pp)supported clade.Based on their morphological and molecular studies,this population was identified as Longidorus pinus Xu,Ye,Wang & Zhao,2018.Longidorus pinus was first reported on Jujube trees,Jujube tree was a newhost for Longidorus pinus.
On the basis of classification and identification of traditional morphology,combining 18 S and 28 S ribosome(r DNA)sequence analysis,species of nematodes in jujube rhizosphere soil samples collected from Jiexiu city of Shanxi province were identified.The results showed that females were J-or C-shaped after heat killing,body length was 3 480-4 366 μm,the lip region was distinctly set off,odontostyle 70-82 μm long,guide ring situated at 31～34 μm from anterior end,vulva in form of a transverse slit,tail short,conoid,male was not found.The first-stage juvenile characterised by having replacement odontostyle tip inserted into odontophore base,tail short conoid,odontostyle length 44 μm long,replacement odontostyle length 52 μm long.The codes for identifying the species of Longidorus pinus were A23-B1-C23-D4-E1-F2-G23-H23-I1-J2-K6.The sequence alignments and phylogenetic relationships were analysed using data from 18 S and 28 S,respectively.The 18 S of the new population was grouped with Longidorus pinus(MF674014-MF674015)with 100%(pp)supported and 28 S of the new population was also clustered together with Longidorus pinus(MF674016-MF674019)with 96%(pp)supported clade.Based on their morphological and molecular studies,this population was identified as Longidorus pinus Xu,Ye,Wang & Zhao,2018.Longidorus pinus was first reported on Jujube trees,Jujube tree was a newhost for Longidorus pinus.
引文
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[15]徐玉梅,赵增旗,王建明,等.山西新记录Tripylina zhejiangensis记述[J].山西农业科学,2014,42(8):887-889.
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[18]XU Y M,ZHAO Z Q,WANG J M,et al.A newspecies of the genus Tripylina Brzeski,1963(Nematoda:Enoplida:Trischistomatidae)from Shanxi province,China[J].Zootaxa,2013,3630(3):561-570.
[19]CHEN Q,HOOPER D J,LOOF P A A,et al.A revised polytomous key for the identification of species of the genus Longidorus Micoletzky,1922(Nematoda:Dorylaimoidea)[J].Fundamental and applied Nematology,1997,20(1):15-28.
[20]PENEVA V K,LAZAROVA S S,DE LUCA F,et al.Description of Longidorus cholevae sp.n.(Nematoda,Dorylaimida)from a riparian habitat in the Rila Mountains,Bulgaria[J].Zoo Keys,2013,330:1-26.
[2]BARSALOTE E M,PHAMH T,LAZAROVA S,et al.Description of Longidorus cheni sp.n.(Nematoda,Longidoridae)from China[J].Zookeys,2018,744:1-18.
[3]XU Y,YE W,WANG J,et al.Morphological and molecular characterisation of Longidorus pinus sp.n.(Nematoda:Longidoridae)from China and a key to known species of Longidorus in China[J].Nematology,2018,20(7):617-639.
[4]MARIE B E,TIAN Z,CAI R,et al.Description of two newrecords of genus Longidorus(Nematoda:Dorylaimida)in China[J].Journal of Zhejiang University,2018,44(1):31-40.
[5]ARCHIDONA-YUSTE A,JUAN A,NAVAS-CORTS,et al.Unravelling the biodiversity and molecular phylogeny of needle nematodes of the Genus Longidorus(Nematoda:Longidoridae)in olive and a description of six new species[J].PLo S One,2016,11(1):e0147689.
[6]CHEN Q W,LOOF P.A revised polytomous key for the identification of species of the genus Longidorus Micoletzky,1922(Nematoda:Dorylaimoidea).Supplement 1[J].Nematology,1999,20(1):15-28.
[7]DECRAEMER W,COOMANS A.Revision of some species of the genus Paralongidorus sensu Siddiqi et al.(1993),with a discussion on the relationships within the family Longidoridae(Nematoda:Dorylaimida)[J].Nematology,2007,9(5):643-662.
[8]LUCA F D,REYES A,GRUNDER J,et al.Characterization and Sequence Variation in the r DNA Region of Six Nematode Species of the Genus Longidorus(Nematoda)[J].Journal of Nematology,2004,36(2):147-52.
[9]EDWARD J C,MISRA S L,SINGH G R.Longidorus pisi n.sp.(Nematoda,Dorylaimoidea)associated with the rhizosphere of pisum sativum,from Uttar Pradesh,India[J].Japanese Journal of Applied Entomology and Zoology,1964,8(4):310-312.
[10]HETNS J.Further studies on South African Longidoridae(Nematoda)[J].South African Journal of Agricultural Science,1966,9:927-944.
[11]EVELEIGH E S,ALLEN W R.Description of,Longidorus diadecturus,n.sp.(Nematoda:Longidoridae),a vector of the peach rosette mosaic virus in peach orchards in southwestern Ontario,Canada[J].Canadian Journal of Zoology,1982,60(2):112-115.
[12]WHITEHEAD A G,HEMMING J R.A comparison of some quantitative methods for extracting small vermiform nematodes from soil[J].Annals of Applied Biology,1965,55(1):25-38.
[13]SEINHORST W.A rapid method for the transfer of nematodes from fixative to anhydrous glycerine[J].Nematologica,1959,4(1):67-69.
[14]HOOPER D J.Handling,fixing,staining and mounting nematodes[M]//SOUTHEY,J F.Laboratory methods for work with plant and soil nematodes.London,U K:Ministry of Agriculture,Fisheries and Food,1986:59-80.
[15]徐玉梅,赵增旗,王建明,等.山西新记录Tripylina zhejiangensis记述[J].山西农业科学,2014,42(8):887-889.
[16]徐玉梅,赵增旗,王建明,等.山西省新记录helicotylenchus indicus记述[J].山西农业大学学报(自然科学版),2013,33(6):517-519.
[17]ZHENG J W,SUBBOTIN S A,HE S S,et al.Molecular characterisationof some Asian isolates of Bursaphenlenchus xylophilus and Bursaphenlenchus mucronatususing PCR-RFLPs and sequences of ribosomal DNA[J].Russian Journal of Nematology,2003,11(1):17-22.
[18]XU Y M,ZHAO Z Q,WANG J M,et al.A newspecies of the genus Tripylina Brzeski,1963(Nematoda:Enoplida:Trischistomatidae)from Shanxi province,China[J].Zootaxa,2013,3630(3):561-570.
[19]CHEN Q,HOOPER D J,LOOF P A A,et al.A revised polytomous key for the identification of species of the genus Longidorus Micoletzky,1922(Nematoda:Dorylaimoidea)[J].Fundamental and applied Nematology,1997,20(1):15-28.
[20]PENEVA V K,LAZAROVA S S,DE LUCA F,et al.Description of Longidorus cholevae sp.n.(Nematoda,Dorylaimida)from a riparian habitat in the Rila Mountains,Bulgaria[J].Zoo Keys,2013,330:1-26.