全 文 ：第 3 期 国际会议学术论文摘要选登
tions，which thus verified the precision of our resistant and re-
silience system．
References:
［1］Bergdahl D R． Impact of pinewood nematode in North America:
present and future［J］． J Nematol，1988，20，260 － 265．
［2］Chai X M，Jiang P． Occurrence and Control Of Pine Wilt Disease
［M］． Beijing:China Agric Press，2003．
［3］Cheng H R，Lin M S，Li W Q． Pine wilt disease found on the Pi-
nus thunbergii in Nanjing［J］． For Pest Dis，1983 (4) :1 － 5．
［4］Dwinell L D． The pinewood nematode:regulation and mitigation
［J］． Annu Rev Phytopathol，1997，35:153 － 166．
［5］Dropkin V H，Foudin A，Kondo E，et al． Pinewood nematode:a
threat to U． S． forests? ［J］Plant Dis，1981，65:1022 － 1027．
［6］Enda，N． The damage of pine wilt disease and control in Asia
［J］． For Pests，1997，46:182 － 188．
［7］Holling C S． Resilience and stability of ecological systems［J］．
Annu Rev Ecol Syst，1973(4) :1 － 24．
［8］Shi Juan，Luo Youqing，Song Jiying，et al． Traits of masson pine
affecting attack of pine wood nematode［J］． Journal of Integrative
Plant Biology，2007，49 (12) :1 － 9
［9］Shi Juan，Luo Youqing，Wu Haiwei，et al． Impact of invasion of
pine wood nematode on the growth of dominant shrub Pleioblastus
amarus in Pinus massoniana communities［J］． For Stud China，
2009，11(1) :61 － 63．
［10］Shi Juan，Luo Youqing，Wu Haiwei，et al． Evaluation Criteria
and Indicator System of Resistance and Resilience of Pine Forest
Ecosystem Against Bursaphelenchus xylophilus Invasion:Forest
Regeneration:Ecology，Management and Economics［M］． New
York:Nova Science Publishers，
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2009．
http:/ /www． nldxb． com［doi:10． 3969 / j． issn． 1000 － 2006． 2011． 03． 033
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IUFRO2009 国际松萎蔫病研讨会学术论文摘要选登
感染松萎蔫病的葡萄牙海岸松中内生细菌的分离
Vieira dos Santos M C 1，2，Proena D A N3，Fonseca L1，2，
Abrantes I M de O1，2，Morais P V 1，3
(1． IMAR-CMA，Departamento de Zoologia，Faculdade de Ci ncias e Tecnologia，Universidade de Coimbra (FCTUC) ，
3004-517 Coimbra，Portugal;2． Departamento de Zoologia，FCTUC，3004-517 Coimbra，Portugal;3． Departamento de
Bioqu mica，FCTUC，3001-401 Coimbra，Portugal)
摘要:葡萄牙海岸松被松材线虫感染后的迅速死亡表明有其他生物的介入。内生细菌是一类普遍
存在的生物，它在大部分植物中能形成种群。松材线虫携带的细菌也有可能导致海岸松的迅速死
亡。笔者通过培养分离得到的细菌分离物，利用扩增性核糖体 DNA 限制性酶切片段分析
(ARDRA)和变性梯度凝胶电泳分析(DGGE) ，了解被感染的和未被感染的海岸松内生细菌的种群
结构，以分析鉴定与松材线虫相关的细菌。从样树和松材线虫中分离的细菌 Alphaproteobacteria、
Bacteroidetes 属仅存在于内生菌中，而 Actinobacteria、Firmicutes 被发现仅与松材线虫相关。ARDRA
和 DGGE 分析证明内生菌种群结构多样性的差异可能与松材线虫的入侵有关。这种细菌种群的多
样性差异似乎与 Burkholderia、Pseudomonas 和 Luteibacter 属中的某些菌株有关。而从树中分离出
来的 Janthinobacterium agaricidamnosum、P． lutea 和 Dyella yeojuensis 与松材线虫相关。
关键词:松材线虫;松萎蔫病;海岸松;种群结构
中图分类号:S763 文献标志码:D 文章编号:1000 － 2006(2011)03 － 0149 － 02
941
南 京 林 业 大 学 学 报 (自 然 科 学 版 ) 第 35 卷
Endophytic bacteria isolated from pine wilt-affected Pinus pinaster trees
Vieira dos Santos M C1，2，Proena D A N3，Fonseca L1，2，Abrantes I M de O1，2，Morais P V1，3
(1． IMAR-CMA，Departamento de Zoologia，Faculdade de Ci ncias e Tecnologia，Universidade de Coimbra (FCTUC) ，
3004-517 Coimbra，Portugal;2． Departamento de Zoologia，FCTUC，3004-517 Coimbra，Portugal;3． Departamento de
Bioqu mica，FCTUC，3001-401 Coimbra，Portugal)
The characteristic rapid death of maritime pine，Pinus pi-
naster，after infection by the pinewood nematode (PWN) ，Bur-
saphelenchus xylophilus，suggests the involvement of other or-
ganisms in pine wilt disease (PWD)． Endophytic bacteria are
ubiquitous in nature and colonize a broad spectrum of plant spe-
cies． They colonize plants internally without harming them and
although they have been reported to promote plant growth and
stimulate plant defence mechanisms． Although such bacteria
may be involved in improving plant health，they have also been
seen as a potential cause of the low regeneration capacity of ma-
ture trees． The bacteria carried out by the PWN throughout pine
trees may also contribute to the rapid wilting of the maritime
pine trees． This study was made to assess the endophytic micro-
bial community structure of pine wilt affected and non-affected
P． pinaster，based on culture isolates，amplified ribosomal
DNA restriction analysis (ARDRA)and denaturing gradient
gel electrophoresis (DGGE)profiling，and to relate the results
to a possible association between the characteristics and bacteri-
a associated with the PWN． The microbial community from
trees and PWNs included strains from Beta and Gammapro-
teobacteria，but Alphaproteobacteria and Bacteroidetes were only
found in the endophytic community whereas Actinobacteria and
Firmicutes were only found to be associated with the nematode．
ARDRA and DGGE profiles showed differences in the diversity
of the endophytic microbial community that may be related to
the presence of the PWN． Those differences seem to be associ-
ated with the number of strains of the genera Burkholderia，
Pseudomonas and Luteibacter． Isolates belonging to the species
Janthinobacterium agaricidamnosum，P． lutea and Dyella yeo-
juensis were isolated from trees and were associated with the
PWN． This research documents the presence of bacteria associ-
ated with the PWN by molecular methods and the impact of the
nematode in the plant endophytic microbial community struc-
ture．
Reference:
［1］Davison J． Plant beneficial bacteria［J］． Biotechnology，1988，
6:282 － 286．
［2］Guo D S，Zhao B G，Gao R． Observation of the site of pine wood
nematode where bacteria are carried with SEM and TEM［J］． J
Nanjing For Univ，2000，24(4) :69 － 71．
［3］Han Z M，Hong Y D，Zhao B G． A study on pathogenicity of bac-
teria carried by pine wood nematodes［J］． J Phytopathology，
2003，151:683 － 689．
［4］Hoflich G，Wiehe W，Kuhn G． Plant growth stimulation by inoc-
ulation with symbiontic and associative rhizosphere mechanisms
［J］． Experientia，1994，50:897 － 905．
［5］Kloepper J W，Tipping B，Lifshitz R． Plant growth promotion me-
diated by bacterial rhizosphere colonizers［G］/ / Cregan D L K a P
B． The Rhizosphere and Plant Growth． The Netherlands:Kluwer
Academic Publishers，1991．
［6］Kloepper J W，Scher F M，Laliberte M，et al． Emergence-promo-
ting rhizobacteria:description，implications for agriculture［G］/ /
Swinburne T R． Iron，Siderophores，and Plant Disease． New
York:Plenum Press，1986．
［7］Lambert B，Joos H． Fundamental aspects of rhizobacterial plant
growth promotion research［J］． Trends Biotechnol，1989，7:215
－ 219．
［8］Oku H，Shiraishi T，Kurozumi S． Pine wilt toxin，the metabolite
of bacteria associated with a nematode［J］． J For Res，1980，67:
198 － 199．
［9］Zhao B G，Lin F． Mutualistic symbiosis between Bursaphelenchus
xylophilus and bacteria of the genus Pseudomonas［J］． For
Pathol，2005，29:339 － 345．
［10］Zhao B G，Wang H L，Han S F，et al． Distribution and pathoge-
nicity of bacteria species carried by Bursaphelenchus xylophilus in
China［J］． Nematology，2003，6:899 － 906．
051