全 文 ：第 50 卷 第 7 期
2 0 1 4 年 7 月
林 业 科 学
SCIENTIA SILVAE SINICAE
Vol. 50，No. 7
Jul．，2 0 1 4
doi:10．11707 / j．1001-7488．20140712
Ｒeceived time: 2013 － 02 － 03; Ｒevised time: 2014 － 02 － 13．
Foundation project: This project was supported by the National Natural Science Foundation of China ( 31200487 ) and National Science and
Technology Support Program (2012BAK11B03) ．
* Corresponding author: Hu Jiafu． We thank Ｒothamsted Nematode Collection at Fera，UK and USDA Nematode Collection，USA for providing the
paratypes．
Devibursaphelenchus hunanensis的重新描述*
郭 恺1 顾建锋2 王江岭2 胡加付1
(1．浙江农林大学林业与生物技术学院森林保护重点实验室 临安 311300; 2．宁波出入境检验检疫局技术中心 宁波 315012)
摘 要: 对分离自宁波马尾松的 Devibursaphelenchus hunanensis 进行重新描述，其鉴定特征如下: 侧区有 3 条侧线，
较长的口针(雄虫为 15 ～ 20 μm，雌虫为 18 ～ 22 μm)，无基部球; 无阴门盖，非常短的后阴子宫囊; 雌虫直肠和肛门模
糊或者不存在; 交合刺直线长 16 ～ 19 μm，远端有盘状突; 从腹面观，存在清晰的交合伞; 具捕食性。并对其 rDNA 的
SSU 区、ITS1 /2 区、D2D3 LSU 区和部分 mtCOI 基因测序并提交 NCBI。结合形态和分子数据，将 D． hunanensis 和 D．
eproctatus 的模式标本进行比较，发现二者特征高度吻合，D． eproctatus 为 D． hunanensis 的同物异名。
关键词: 形态学;形态测量值;系统发育;分子;分类;捕食性
中图分类号: S763. 1 文献标识码: A 文章编号: 1001 － 7488(2014)07 － 0082 － 08
Ｒedescription of Devibursaphelenchus hunanensis (Yin et al．，1988)
(Nematoda: Ektaphelenchinae) from Pinus massoniana in China with the
Synonymy of D． eproctatus (Sriwati et al．，2008)
Guo Kai1 Gu Jianfeng2 Wang Jiangling2 Hu Jiafu1
(1. Key Laboratory of Forest Protection，College of Forestry and Biotechnology，Zhejiang Agricultural and Forestry University Lin’an 311300;
2. Technical Centre，Ningbo Entry-Exit Inspection and Quarantine Bureau Ningbo 315012)
Abstract: Devibursaphelenchus hunanensis isolated from Pinus massoniana is redescribed． The identification features are
as follows: three incisures in the lateral field; relatively long stylet ( about 15 － 20 μm and 18 － 22 μm for males and
females) with wide lumen and without basal knobs and vulva flap，and very short postuterine sac． Female rectum and anus
indistinct or disappear． The spicules are in medium size，16 － 19 μm long in chord，and the arcuate with a terminal
cucullus． There is a distinct bursa flap in ventral view． D． hananensis is predatory． The partial SSU region，the ITS1 /2
region，the D2D3 LSU region of rDNA and partial mtCOI gene were sequenced and deposited in the GenBank database．
With the morphological and molecular data，the isolate was compared with type specimens of D． hunanensis and D．
eproctatus． Their features are highly consistent，thus D． hunanensis and D． eproctatus is regarded as synonymous of D．
hunanensis．
Key words: morphology; morphometrics; phylogeny; molecular; taxonomy; predatory
Bursaphelenchus hunanensis ( Yin et al．，1988 )
was described from dead wood of Pinus massoniana in
Hunan Province， China． Braasch ( 2009 ) re-
established genus Devibursaphelenchus Kakuliya，
1967， and transferred B． hunanensis to genus
Devibursaphelenchus．
Bursaphelenchus eproctatus ( Sriwati et al．，2008
was first isolated from dead Japanese black pine in
Japan，and it was also replaced to Devibursaphelenchus
by Braasch ( 2009 ) ． Because a few specimens were
available then，the measurements were based only on 6
males and females，and molecular methods were not
applied．
During a survey of Bursaphelenchus spp． in dead
or declining Pinus massoniana in Ningbo，Zhejiang，
China since 2005， one nematode species were
repeatedly detected． With further morphological and
molecular study，and compared with type specimens of
第 7 期 郭 恺等: Devibursaphelenchus hunanensis 的重新描述
D． hunanensis and D． eproctatus，our specimens were
identified as D． hunanensis， and D． eproctatus is
regarded as synonymy of D． hunanensis．
1 Materials and methods
1. 1 Nematode culturing and morphological observations
Sawn samples taken from dead or declining pine
wood ( from Yuyao and Cicheng in Ningbo，China)
were cut into small pieces less than 1 cm wide．
Nematodes were extracted by a modified Baermann
funnel technique for 24 h． The feeding habit of the
collected nematodes was observed on a slide in water
and recorded． Multiplication on agar-fungi plates
( Botryotinia fuckeliana ) failed． Measurements were
made on specimens fixed in TAF and processed to
glycerin following the method of Seinhorst ( 1959 ) ．
Light micrographs were made using a Zeiss Imager Z1
microscope equipped with a Zeiss AxioCam MＲm CCD
camera． Paratypes of D． hunanensis ( slide 86c /8 /1 )
were obtained on loan from the Ｒothamsted Nematode
Collection at Fera，UK． Paratypes of D． eproctatus
( slides E-02 and E-03 ) were obtained on loan from
USDA Nematode Collection，USA．
1. 2 Molecular analyses
DNA samples of D． hunanensis were prepared
according to Li et al． (2008 ) ． Four sets of primers
( synthesised by Invitrogen，Shanghai，China ) were
used in the PCＲ analyses to amplify the partial SSU
region，the ITS1 /2 region，the D2D3 LSU region of
rDNA and partial mtCOI gene，respectively． Primers
for amplification of SSU were forward primer K4f (5-
ATG CAT GTC TAA GTG GAG TAT TA -3) and
reverse primer K1r (5-TTC ACC TAC GGC TAC CTT
GTT ACG ACT -3) (Penas et al．，2006) ． Primers for
amplification of ITS1 /2 were forward primer F194 (5-
CGT AAC AAG GTA GCT GTA G -3) (Ferris et al．，
1993) and reverse primer 5368r (5- TTT CAC TCG
CCG TTA CTA AGG -3) (Vrain，1993) ． Primers for
amplification of D2 /D3 LSU were forward primer D2A
(5-ACA AGT ACC GTG AGG GAA AGT TG-3) and
reverse primer D3Br (5-TCG GAA GGA ACC AGC
TAC TA-3) ( De Ley et al．，1999 ) ． Primers for
amplification of mtCOI were forward primer COI-F1
(5-CCT ACT ATG ATT GGT GGT TTT GGT AAT
TG-3) and COI-Ｒ2 ( 5-GTA GCA GCA GTA AAA
TAA GCA CG-3) ( Kanzaki et al．，2005 ) ． PCＲ
conditions were as described by Ye et al． (2007) and
Li et al． ( 2008 ) ． PCＲ products were separated on
1% agarose gels and visualised by staining with
ethidium bromide． PCＲ products of sufficiently high
quality were purified for cloning and sequencing by
Invitrogen，Shanghai，China． For ITS-ＲFLP profiles，
suitable aliquots of the amplied ITS rDNA were
digested for at least 3 h at 37 ℃ using 10 U of each of
the ve restriction endonucleases ＲsaI，HaeIII，MspI，
HinfI and AluI ( Takara， Japan ) following the
manufacturer’s instructions． Fragments were resolved
by electrophoresis in a 2. 5% agarose gel and stained
with ethidium bromide．
For Cicheng isolate，sequences of the partial SSU
region，the ITS1 /2 region，the D2D3 LSU region of
rDNA and partial mtCOI gene were sequenced，and
ITS-ＲFLP profiles following the methods described by
Wang et al．(2012) ． For Yuyao isolate，only the ITS1 /
2 region was sequenced．
The ITS and partial LSUsequences were analysed
and aligned using the program ClustalW implemented
in MEGA version 4. 0 ( Tamura et al．， 2007 ) ．
Phylogenetic trees were generated with the Neighbor
Joining ( NJ) method using the Tajima-Nei distance
option． Bootstrapping analysis was performed with
1 000 repicates．
2 Ｒesults
2. 1 Measurements
Morphometric values of the original description
and two isolates from Ningbo are compared in Tab． 1．
2. 2 Description
1) Female Body slender，cylindrical，annulated，
ventrally curved when killed by gentle heat． Cuticle
marked by ne trans-verse striae． Lateral elds with
three incisures． Lip region offset， 6. 5 － 9. 7 μm
diam．，2. 9 － 4. 2 μm high( Fig． 1A) ． Stylet 18 － 22
μm long，lacking basal knobs． Median bulb elongate-
oval，median bulb valve well developed; plate located
slightly posterior to middle of bulb． Pharyngeal gland
lobe well developed， ca six body diam． long，
overlapping intestine dorsally． Nerve ring located
posterior to median bulb． Excretory pore ca 10 － 15
μm posterior to median bulb，hemizonid conspicuous，
located just posterior to excretory pore ( Fig． 1B，C) ．
Ovary single，outstretched，oocytes arranged in multiple
38
林 业 科 学 50 卷
rows from anterior to middle part of ovary． Spermatheca
and quadricolumella obscure． Uterine region roundish．
Vulva not protuberant， vulval flap absent， vagina
sllightly inclined anteriorly (Fig. 1I － J) ． Postuterine
sac short，less than one body diam． long． Ｒectum and
anus very difficult to discern and probably non-
functional． Intestine terminating as blind sac． Tail
tapering to conoid with bluntly rounded or pointed
terminus(Fig. 1L － O) ．
Fig． 1 D． hunanensis
A: Anterior body; B，C: Median bulb region ( show excretory pore position) ; D: Feeding on Aphelenchoides sp． ;
E － H: Male tail; I，J: Vulva region; K: Male tail ( ventral view) ; L － O:Female tail． Scale bars = 10 μm．
2) Male Anterior body region and cuticle similar
to female，body strongly curved ventrally when killed by
gentle heat． Testis outstretched， anterior half with
sperm cells，spermatocytes arranged in two rows． Sperm
spherical． Spicule mitten-shaped，condylus rounded，
slightly elongated，lamina smoothly and symmetrically
curved， rostrum conical with bluntly pointed tip，
junction of rostrum and calomus smoothly curved，
cucullus small． Tail strongly recurved with pointed tip，
bursa spade-shaped(Fig． 1E － H) ． Two pairs of caudal
papillae present: one pair located slightly precloacal
and the second subventral pair located just anterior to
beginning of bursal flap(Fig． 1K) ．
48
第 7 期 郭 恺等: Devibursaphelenchus hunanensis 的重新描述
2. 3 Comparison with original description
Fig． 2 Photograph of D． hunanensis paratypes
A: Anterior body of D． eproctatus; B: Female tail of D． hunanensis;
C: Male tail of D． eproctatus; D: Male tail of D． hunanensis． Scale bars = 10 μm．
The measurements of the two isolates from Ningbo
and the original measurements of D． hunanensis and
D． eproctatus are very similar． Only that in D．
hunanensis，the stylet length is slightly longer (19 － 21
μm and 20 － 26 μm vs 16 － 20 μm and 18 － 22 μm for
males and females，respectively) ．
Sriwati et al．(2008) stated that D． eproctatus is
distinguished from D． hunanensis by the presence of
three lateral lines vs four，rostrum squared vs rounded
and absence vs presence of a functional rectum and
anus． Yin et al． (1988) described that the lateral as
having“2 longitudinal，refractive lines on either side
on a narrow refractive band”． In our observation of the
paratypes，there’s only three lateral lines，and which
is accord with Sriwati et al． ( 2008 ) ． Also in our
observation of paratypes of D． hunanensis， only
indistinct anus remaining sometimes exist ( Fig． 2A －
B)，so no clear rectum and anus was shown in D．
hunanensis． And as to the rostrum，we didn’t see any
difference from paratypes of D． hunanensis and D．
eproctatus (Fig． 2C，D) ． Yin et al． (1988) described
that the distal end of spicules obtuse， without
cucullus，but we can see that the small culullus in
paratypes of D． hunanensis． So D． eproctatus is
regarded as being synonymous with D． hunanensis．
The samples of both isolates from Ningbo are very
similar to D． hunanensis． Sriwati et al．(2008) described
the female tail terminus as“pointed”，but Yin et al．
(1988 ) described it as “finely rounded”． While in
isolated from Ningbo，both types of female tail terminus
is found，so we conclude that this is due to interspecies
variation． The spicules shape are very similar． The
spicules length along arc is 18. 8 － 20. 8 μm in the
58
林 业 科 学 50 卷
original description ( Sriwati et al．，2008 )，which is
not accord with our measurements ( Tab． 1 )，so the
three paratype males loaned from USDA Nematode
Collection，USA were remesured． It’s spicule length
in chord is 17. 1，17. 3 and 17. 6 μm，in arc is 14. 3，
14. 3， 14. 4 μm． So it is now clear that when
measuring the spicule length in arc，Sriwati et al．
( 2008 ) measured it from the condylus ． According to
Tab． 1 Morphometrics of two isolates of D． hunanensis from Ningbo，China compared with original data(mean ± SD)
μm
Item
Female Male
Japanese isolate Yuyao isolate Cicheng isolate Japanese isolate Yuyao isolate Cicheng isolate
n 6 15 15 6 15 15
L
639 ± 56. 0
(580 － 714)
675 ± 38. 8
(613 － 745)
715 ± 35. 4
(670 － 759)
545 ± 42. 3
(550 － 595)
548 ± 30. 0
(500 － 613)
575 ± 44. 6
(502 － 657)
a
35. 9 ± 3. 5
(32. 6 － 41. 9)
37. 5 ± 2. 7
(34. 1 － 42. 6)
33. 0 ± 2. 5
(29. 5 － 38. 9)
33. 8 ± 1. 4
(31. 5 － 35. 1)
36. 9 ± 1. 7
(34. 5 － 39. 5)
33. 3 ± 3. 1
(25. 7 － 37. 4)
b
7. 4 ± 0. 1
(7. 2 － 7. 4)
7. 5 ± 0. 4
(6. 6 － 8. 2)
8. 1 ± 0. 7
(7. 2 － 9. 6)
6. 3 ± 0. 4
(5. 7 － 6. 9)
6. 6 ± 0. 4
(5. 9 － 7. 4)
6. 8 ± 0. 3
(6. 2 － 7. 3)
b —
2. 9 ± 0. 1
(2. 7 － 3. 1)
3. 2 ± 0. 2
(3. 0 － 3. 5)
—
2. 7 ± 0. 2
(2. 4 － 3. 1)
3. 0 ± 0. 2
(2. 7 － 3. 4)
c — — —
14. 2 ± 0. 7
(13. 3 － 15. 0)
15. 2 ± 0. 8
(14. 0 － 16. 3)
14. 8 ± 0. 5
(14. 3 － 16. 1)
c — — —
2. 7 ± 0. 2
(2. 3 － 3. 0)
2. 9 ± 0. 2
(2. 5 － 3. 2)
2. 8 ± 0. 2
(2. 2 － 3. 2)
V or T
78. 5 ± 1. 4
(78. 7 － 80. 8)
77. 4 ± 0. 7
(76. 5 － 79. 2)
78. 1 ± 0. 8
(76. 4 － 78. 8)
25. 2 ± 1. 9
(22. 9 － 27. 6)
32. 2 ± 4. 3
(25. 3 － 40. 1)
32. 7 ± 3. 9
(27. 9 － 40. 8)
Max body diam
17. 8 ± 0. 9
(17 － 19)
18. 1 ± 1. 5
(15. 5 － 20. 2)
21. 7 ± 1. 9
(18. 8 － 24. 7)
16. 2 ± 1. 5
(15 － 18)
14. 9 ± 0. 9
(13. 7 － 16. 8)
17. 4 ± 1. 9
(14. 6 － 22. 0)
Lip diam
8. 7 ± 0. 4
(8 － 9)
7. 8 ± 0. 5
(6. 5 － 8. 5)
8. 7 ± 0. 5
(7. 9 － 9. 7)
8. 3 ± 0. 8
(7 － 9)
6. 7 ± 0. 3
(6. 2 － 7. 0)
8. 0 ± 0. 9
(6. 4 － 9. 3)
Lip height
4. 3 ± 0. 5
(4 － 5)
3. 2 ± 0. 2
(2. 9 － 3. 5)
3. 5 ± 0. 3
(3. 2 － 4. 2)
4. 1 ± 0. 4
(4 － 5)
2. 9 ± 0. 3
(2. 5 － 3. 5)
3. 6 ± 0. 2
(3. 3 － 3. 8)
Stylet length
20. 5 ± 1. 2
(19 － 22)
20. 3 ± 0. 9
(18. 8 － 22. 4)
20. 3 ± 1. 0
(18. 4 － 21. 9)
17. 7 ± 2. 3
(15 － 20)
17. 5 ± 1. 2
(15. 7 － 19. 3)
18. 6 ± 0. 7
(17. 4 － 20. 0)
Median bulb length
20. 3 ± 1. 6
(18 － 23)
21. 4 ± 0. 9
(20. 0 － 22. 5)
21. 5 ± 1. 7
(20. 0 － 25. 5)
19. 0 ± 0. 7
(18 － 20)
19. 3 ± 0. 9
(17. 5 － 20. 6)
19. 7 ± 2. 2
(14. 7 － 22. 1)
Median bulb diam
10. 9 ± 0. 9
(10 － 11)
11. 6 ± 1. 0
(10. 0 － 13. 9)
13. 9 ± 2. 1
(12. 0 － 17. 6)
10. 3 ± ． 07
(10 － 11)
10. 1 ± 0. 5
(9. 5 － 11. 0)
11. 3 ± 1. 0
(9. 6 － 12. 7)
Excretory pore position —
103. 4 ± 6. 3
(93. 0 － 113. 8)
97. 6 ± 14. 6
(77. 0 － 113. 6)
—
89. 5 ± 4. 6
(78. 5 － 98. 6)
91. 3 ± 8. 1
(73. 0 － 99. 5)
Spicule( chord) — — — —
17. 8 ± 0. 5
(16. 6 － 18. 5)
17. 5 ± 0. 7
(16. 5 － 19. 4)
Spicule( curved median line)① — — —
19. 8 ± 0. 6
(18. 8 － 20. 8)
16. 0 ± 0. 5
(15. 2 － 17. 0)
14. 5 ± 0. 6
(13. 1 － 15. 4)
Spicule( dorsally line) — — — —
21. 4 ± 1. 0
(19. 7 － 23. 0)
21. 2 ± 1. 1
(19. 1 － 22. 5)
Spicule( arc from the middle of
the condylus to the end)
— — — —
20. 0 ± 0. 5
(19. 4 － 20. 7)
19. 1 ± 0. 8
(17. 5 － 20. 3)
Ovary or testis length
185. 6 ± 31. 6
(152 － 243)
252. 7 ± 37. 8
(198. 0 － 325. 2)
295. 9 ± 46. 3
(202. 0 － 356. 0)
136. 7 ± 4. 9
(129 － 143)
175. 8 ± 21. 7
(145. 9 － 223. 0)
194. 9 ± 13. 4
(179. 0 － 222. 0)
Post-uterine sac length
10. 1 ± 1. 9
(7 － 13)
8. 5 ± 0. 6
(7. 6 － 9. 4)
13. 4 ± 3. 2
(10. 6 － 19. 5)
— — —
Tail length — — —
38. 4 ± 3. 0
(35 － 42)
36. 1 ± 2. 1
(32. 8 － 40. 0)
39. 3 ± 3. 5
(34. 2 － 45. 8)
Anal body diam — — — —
12. 2 ± 0. 5
(11. 5 － 13. 0)
14. 1 ± 1. 0
(12. 7 － 16. 5)
①Spicules length along arc could be measured from the condylus by the scale bar of the original light micrographs and drawings．
68
第 7 期 郭 恺等: Devibursaphelenchus hunanensis 的重新描述
Ｒyss et al． (2005)，the spicule length along arc should
start from the middle of the capitulum．
So both isolates from Ningbo is identified as D．
hunanensis．
2. 4 Molecular profiles and phylogenetic status
The partial SSU region ( JN122012，924 bp)，the
ITS1 /2 region ( JN122010 for Cicheng isolate and
JN122011 for Yuyao isolate，both 918 bp)，the D2D3
LSU region of rDNA ( JN122009，807 bp) and partial
mtCOI gene sequences ( JN122013， 675 bp ) are
deposited in the GenBank database． The ITS region
sequences of the two isolates of D． hunanensis ( from
Cicheng and Yuyao in Ningbo) only differs at 2 base
positions，but they are shorter than another isolate
“NB” also from Ningbo ( EU400449 )， a 29 bp
sequence is lost (Tab． 2) ． The sequence of the D2D3
LSU region of Cicheng isolate and isolate“NB”differs
only at 1 base pair．
The molecular phylogenetic status of D．
hunanensis has already done in Gu et al． (2010) ． The
ITS-ＲFLP pattern of Cicheng isolate is shown in
Fig． 3，which is identical with the pattern shown in
Burgermeister et al． ( 2009 ) as B． hunanensis
(Fig． 3) ．
Tab． 2 Sizes of PCＲ products and DNA restriction fragments obtained in ITS-ＲFLP analysis and calculated on
sequencing results of the ITS1 /2 regions bp
Devibursaphelenchus species PCＲ product
Ｒestriction fragments①
Ｒsa Ⅰ Hae Ⅲ Msp Ⅰ Hinf Ⅰ Alu Ⅰ
D． hunanensis
( isolates from Cicheng and Yuyao)
918 374
305
167
72
580
338
736
182
468
182
163
63
42
612
293
13
D． hunanensis②
( isolate“NB”from Ningbo，
EU400449)
947 374
305
196
72
580
367
765
182
497
181
164
63
42
641
293
13
D． wangi
966 403
335
155
73
717
249
769
197
488
433
45
798
73
65
15
15
①Fragment sizes ( bp) were calculated with a computer program DNASTAＲ MapDraw 5. 01．②According to Burgermeister et al．，2009．
In Fig． 4 and 5，Devibursaphelenchus，Ektaphelenchoides
and Ektaphelenchus forms a well-supported clade，D．
wangi and D． hunanensis are very close，though D． lini
is more close to Ektaphelenchoides pini．
2. 5 Diagnosis and relationships
D． hunanensis is characterized by three incisures
in the lateral field; relatively long stylet ( about 15 － 20
μm and 18 － 22 μm for males and females) with wide
lumen and lacking basal knobs; vulva flap absent，
very short postuterine sac; female rectum and anus
indistinct or disappear; the spicules are of medium
size，16 － 19 μm long in chord，and arcuate with a
terminal cucullus; presence of a distinct bursa flap in
ventral view; and a predatory lifestyle．
Braasch ( 2009 ) re-established the genus
Devibursaphelenchus Kakuliya， 1967 belonging to
Ektaphelenchinae which contains six species: D．
typographi Kakuliya，1967; D． eproctatus (Sriwati et al．，
2008) Braasch，2009; D． hunanensis (Yin et al．，1988)
Braasch，2009; D． lini (Braasch，2004) Braasch，2009;
D． teratospicularis (Kakuliya et al．，1965) Braasch，2009
and D． wangi Gu et al．，2010． D． eproctatus is now
regarded as synonymous with D． hunanensis．
Fig． 3 ITS-ＲFLP pattern of D． hunanensis
M:Molecular size marker ( 100 bp ladder ) ; Lane 1: rDNA
amplification product; Lanes 2 － 6: Digestion products obtained with
ＲsaⅠ，HaeⅢ，MspⅠ，HinfⅠ and AluⅠ ． Sizes of PCＲ product
and its restriction fragments are shown in Tab． 2．
78
林 业 科 学 50 卷
Fig． 4 Molecular phylogenetic status of D． hunanensis based on ITS sequences
Aphelenchus avenae served as the outgroup species． Numbers at branching points are bootstrap values obtained using 1 000 repetitions． Scale bar: Substitutions / site．
Fig． 5 Molecular phylogenetic status of D． hunanensis based on partial LSU sequences
A． avenae served as the outgroup species． Numbers at branching points are bootstrap values obtained using 1 000 repetitions． Scale bar: substitutions / site．
88
第 7 期 郭 恺等: Devibursaphelenchus hunanensis 的重新描述
D． hunanensis is distinguished from D． wangi Gu
et al．，2010 by the size of spicules (16. 5 － 19. 4 μm vs
14. 2 － 15. 6 μm long measured along chord );
different size of stylet (15 － 20 μm and 18 － 22 μm for
males and females，respectively vs 12. 4 － 16. 6 μm
and 16. 7 － 17. 4 μm); different c ratio of males ( c =
13. 3 － 16. 3 vs c = 16. 2 － 18. 8) ． Their ＲFLP pattern
are different ( Fig． 3 and Tab． 2 ) ． The range of ITS
sequence divergence between different isolates of D．
hunanensis is 0. 2%， but it’s 17. 9% － 18. 2%
between D． hunanensis and D． wangi． Also the range
of 28S sequence divergence between different isolates
of D． hunanensis is only 0. 1%，but it’s 9. 0% －
9. 2% between D． hunanensis and D． wangi．
2. 6 Feeding habitat
Tens of D． hunanensis，including males，females
and juveniles were found feeding on B． mucronatus，
B． rainulfi and Aphelenchoides sp． (Fig． 1D) ．
3 Discussion
Despite the clear rectum and anus of females，
Braasch (2009) temporaty transferred B． hunanensis to
Devibursaphelenchus． Now with the re-study of the
paratypes，we have come to a conclusion that really
those rectum and anus don’t exist，only sometimes
indistinct anus remainings in several samples． Also a
predatory behaviour supports the placement within a
genus of Ektaphelenchinae．
With the re-study of paratypes of D． epcrotatus，
we also decided that it is as synonymous with D．
hunanensis．
The wide-lumened stylet， indistinct or
nonfunctional rectum and anus suggests a predatory or
ecto-parasitic lifestyle， it’s possible that all
Devibursaphelenchus species are predatory on other
nematodes． It will be very interesting and of practical
interest to investgate the potential efficiency for
controling B． xylophilus in the future．
Ｒeferences
Braasch H． 2009． Ｒe-establishment of Devibursaphelenchus Kakuliya，
1967 ( Nematoda， Aphelenchoididae ) and proposal for a new
combination of several Bursaphelenchus species． Journal of Nematode
Morphology and Systematics，12(1) :1 － 5．
Braasch H． 2004． A new Bursaphelenchus species ( Nematoda:
Parasitaphelenchidae ) sharing characters with Ektaphelenchidae
from the People’s Ｒepublic of China． Zootaxa，624:1 － 10．
Burgermeiser W，Braasch H，Metge K，et al． 2009． ITS-ＲFLP analysis，
an effcient tool for differentiation of Bursaphelenchus species．
Nematology，11(5) : 649 － 668．
De Ley P， Félix M A， Frisse L M， et al． 1999． Molecular and
morphological characterisation of two reproductively isolated species
with mirror-image anatomy ( Nematoda: Cephalobidae ) ．
Nematology，1(6) : 591 － 612．
Ferris V Ｒ，Ferris J M，Faghihi J． 1993． Variation in spacer ribosomal
DNA in some cyst-forming species of plant parasitic nematodes．
Fundamental and Applied Nematology，16(2) : 177 － 184．
Gu J，Wang J， Zheng J． 2010． Devibursaphelenchus wangi sp． n．
( Nematoda: Ektaphelenchinae ) feeding on Aphelenchoides sp．
Ｒussian Journal of Nematology，18(1) : 49 － 57．
Kakuliya G A． 1967． New nematode genus Devibursaphelenchus gen． n．
( Nematoda: Aphelenchoididae ) ． Bulletin of the Academy of
Sciences of the Georgian SSＲ，47: 439 － 443．
Kakuliya G A， Devdariani T G． 1965． A new nematode species
Bursaphelenchus teratospicularis Kakuliya et Devdariani，sp． nov．
( Nematoda， Aphelenchoididae ) ． Bulletin of the Academy of
Sciences of the Georgian SSＲ，38: 187 － 191．
Kanzaki N，Futai K． 2005． Description of Bursaphelenchus parvispicularis
n． sp． (Nematoda: Parasitaphelenchidae) isolated from a dead oak
tree，Quercus mongolica var． grosseserrata． Nematology，7 ( 5 ) :
751 － 759．
Li H，Trinh P Q，Waeyenberge L，et al． 2008． Bursaphelenchus chengi
sp． n． ( Nematoda: Parasitaphelenchidae ) isolated at Nanjing，
China，in packaging wood from Taiwan． Nematology，10 ( 3 ) :
335 － 346．
Penas A C，Metge K，Mota M，et al． 2006． Bursaphelenchus antoniae
sp． n． (Nematoda: Parasitaphelenchidae) associated with Hylobius
sp． from Pinus pinaster in Portugal． Nematology，8(5) : 659 － 669．
Ｒyss A，Vieira P，Mota M， et al． 2005． A synopsis of the genus
Bursaphelenchus Fuchs，1937 ( Aphelenchida: Parasitaphelenchidae )
with keys to species． Nematology，7:394 －458．
Seinhorst J W． 1959． A rapid method for the transfer of nematodes from
xative to anhydrous glycerin． Nematologica，4(1) : 67 － 69．
Sriwati Ｒ， Kantaki N， Phan L K， et al． 2008． Bursaphelenchus
eproctatus n． sp． ( Nematoda: Parasitaphelenchidae) isolated from
dead Japanese black pine，Pinus thunbergii Pars． Nematology，10
(1) : 1 － 7．
Tamura K， Dudley J， Nei M， et al． 2007． MEGA4: Molecular
evolutionary genetics analysis ( MEGA ) software version 4. 0．
Molecular Biology and Evolution，24(8) :1596 － 1599．
Vrain T C． 1993． Ｒestriction fragment length polymorphism separates
species of the Xiphinema americanum group． Journal of Nematology，
25(3) : 361 － 364．
Wang J，Gu J． 2012． Bursaphelenchus paraburgeri sp． n． (Nematoda:
Parasitaphelenchidae ) in packaging wood from Malaysia．
Nematology，14(1) :39 － 50．
Ye W， Giblin-Davis Ｒ M， Braasch H， et al． 2007． Phylogenetic
relationships among Bursaphelenchus species ( Nematoda:
Parasitaphelenchidae ) inferred from nuclear ribosomal and
mitochondrial DNA sequence data． Molecular Phylogenetics and
Evolution，43(3) : 1185 － 1197．
Yin K，Fang Y，Tarjan A C． 1988． A key to species in the genus
Bursaphelenchus with a description of Bursaphelenchus hunanensis
sp． n． ( Nematoda: Aphelenchoididae ) found in pine wood in
Hunan Province，China． Proceedings of Helmithological Society of
Washington，55(1) : 1 － 11．
(责任编辑 朱乾坤)
98