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日本松材线虫和南京松材线虫对雪松和马尾松的致病性研究 总被引：5，自引：0，他引：5
用来自日本的木质包装材料上截获的松材线虫和南京松材线虫进行对雪松Ｃｅｄｒｕｓｄｅｏｄａｒａ和马尾松Ｐｉｎｕｓｍａｓｏｎｉａｎａ的致病力试验，发现日本松材线虫能使４～５年生雪松苗２０％发病，１０％左右枯死，而南京松材线虫则没有致病力；１０年生雪松对两种松材线虫具有高度抗性。这两种松材线虫都能使马尾松中度感病和１０％左右的枯死 相似文献
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分离自对湖南郴州、耒阳、张家界地区的病死马尾松样品,采用形态鉴别和分子生物学鉴别相结合的方法对其进行鉴定.形态鉴定结果显示分离检出线虫更符合拟松材线虫特征.同时采用ITS-PCR-RELP方法对病死马尾松中分离线虫进行分子鉴定,ITS区经EcolI和HindⅢ酶切后得到110、160、242、404bp 4条特征带,与拟松材线虫相同,而与松材线虫有明显区别.而且ITS及CoI基因序列信息显示,湖南病死马尾松中分离线虫与松材线虫的同源性分别为92.9%、89.3%,而与拟松材线虫的同源性高达97.2%、95.2%.研究结果表明湖南病死马尾松中分离的线虫为拟松材线虫. 相似文献
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松材线虫种群遗传多样性AFLP标记的建立及其应用 总被引：2，自引：0，他引：2
松材线虫是极具危险性的外来入侵生物,其引起的松材线虫病,目前正在我国部分地区迅速扩展和蔓延,对我国林业生产造成了严重的经济损失。开展松材线虫的种群遗传学研究,是了解其成功入侵和爆发成灾内在机理的重要途径。但迄今为止,尚未找到很有效的分子标记方法来检测松材线虫入侵种群的遗传变异。本文采用AFLP分子标记技术,通过对各反应体系和反应条件的优化及对多态性引物组合的筛选,成功地建立了松材线虫的AFLP分子标记实验体系,并筛选出52对高效多态性的引物组合。应用4对引物对27个松材线虫种群样品进行遗传多样性检测,结果表明,AFLP是进行松材线虫种群遗传学研究的一种很灵敏和可靠的分子标记。此外,本文还对AFLP技术在松材线虫研究中的应用前景进行了讨论。 相似文献
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松材线虫rDNA的测序和PCR-SSCP分析 总被引：37，自引：3，他引：37
本文为克服形态鉴定的不足,用分子生物学的方法鉴别松材线虫。线虫核糖体DNA(r DNA)的内部转录间隔区(ITS1)区(约308bp)的测序结果显示:松材线虫种内区别很小,不超过1bp;拟松材线虫种内区别较大,最大达7bp;这2种线虫的种间区别为32~39bp。根据以上测序结果,本文结合单条线虫DNA的提取技术,对14个松材线虫和拟松材线虫样本进行了单链构象多态性(PCR-SSCP)分析,结果表明PCR-SSCP分析技术可明确区分这2种线虫,该技术可为单条松材线虫的鉴定提供一套灵敏而可靠的方法。 相似文献
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松材线虫(Bursaphelenchus xylophilus)是一种对我国林业危害极大的外来入侵种,具有繁殖力强,扩散范围广的特点。为探究松材线虫的发育调控机制,本研究通过转录组分析和实时荧光定量PCR技术,比较了不同虫态松材线虫发育差异表达基因及功能富集分析。利用LC/MS技术检测了松材线虫代谢物中的蛔甙成分及含量。结果显示:繁殖型和扩散型虫态的差异表达基因主要与滞育和发育调控相关。繁殖型虫态的daf-9基因表达水平显著高于扩散型松材线虫,扩散型LIV期线虫的daf-12、daf-2及daf-16基因表达量显著高于繁殖型松材线虫。Ln和LIII的蛔甙成分比较分析结果显示:繁殖型松材线虫短链蛔甙asc-C3、asc-ΔC7含量显著高于扩散型松材线虫,长链蛔甙asc-C9含量低于扩散型松材线虫。推测松材线虫DAFs基因与蛔甙可能参与松材线虫扩散型虫态的形成。 相似文献
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松材线虫是进境木质包装中的重要检疫对象之一。本研究以进境木质包装中截获的12种伞滑刃属线虫,包括4种不同来源的松材线虫、3种不同来源的拟松材线虫、豆伞滑刃线虫、大尖尾伞滑刃线虫、阿苏里伞滑刃线虫、伯氏伞滑刃线虫及拟小刺伞滑刃线虫为材料,进行实时荧光PCR检测。结果显示,仅松材线虫可观察到明显的荧光强度变化,而其他线虫的荧光强度没有变化。通过研磨样品的方法,提取单条松材线虫的DNA,进行实时荧光PCR试验,检测成功率为100%,且不论是雄虫、雌虫或幼虫均能检测出来。该研究对于我国口岸进境木质包装中松材线虫的检测具有一定的实际意义。 相似文献
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宁波口岸在对一批来自美国的火炬松原木线虫检疫时,发现少量口针退化的幼虫,疑似松材线虫滞育型4龄幼虫,但未发现繁殖型幼虫或成虫。由于这种现象极为少见,实验室进一步对这些幼虫进行观察测计和人工培养,并挑取单条线虫作核糖体28S基因扩增测序和序列分析。结果表明,该种线虫为松材线虫。由于松材线虫滞育型4龄幼虫头部口针退化,且尾部有尾尖突,不但难以判断为松材线虫,甚至容易被误判为腐生线虫,从而导致检测差错。因此,本文对松材线虫滞育型4龄幼虫形态进行描述,可供其他实验室检测鉴定时参考。 相似文献
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云南松毛虫是福建省沿海地区湿地松的重要害虫之一。该虫一年发生1代,以卵越冬,卵于翌年2月下旬孵化,幼虫7龄,历期158d～195d,以取食嫩针叶为主,幼虫老熟时下树,藏于草丛里或树干基部,在枯枝落叶中结茧化蛹,预蛹期50d～70d,成虫寿命5d～10d.其发生与林分的林木组成、气候、天敌等因子密切相关.试验表明：用25%灭幼脲3号,白僵菌与0.4%氰戊菊酯粉剂按4︰1混合制成的粉炮防治幼虫,有良好效果。 相似文献
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松材线虫自然侵染后对不同松树组织结构的影响 总被引：2，自引：0，他引：2
利用滑走切片技术从显微水平研究松材线虫（Bursaphelenchus xylophilus）在自然状态下入侵,对马尾松（Pinus massoniana）和黑松（Pinus thunbergii）显微结构的影响。结果表明,马尾松和黑松的组织结构在感病初期即表现出病变特征,并随感病时间推移病变加剧。马尾松在感病初期皮层薄壁细胞开始木质化,伴有细胞破裂并融合成空腔;至感病中期周皮的栓内层细胞、皮层薄壁细胞以及韧皮部薄壁细胞均发生木质化,细胞内含物增多,甚至形成层细胞也发生一定程度的木质化现象,树脂道因周围细胞破裂而变形;至感病晚期,木材以外所有部分完全被木质化、大量细胞破裂。黑松发病症状与马尾松大致相同但发病时间稍晚,发病程度也较轻,这可能是因为两者对松材线虫病的抗性强弱有差异。同一时期的组织病理学特征在空间上也存有差异,这与松材线虫入侵的位置及其在树体内的分布有关。 相似文献
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An interaction between the fungal pathogenSphaeropsis sapinea (Fr.) Dyko & Sutton, causal organism of Sphaeropsis canker, and the Israeli pine bast scaleMatsucoccus josephi (Homoptera: Margarodidae), causal agent of pine decline, was studied, since both were prevalent on Aleppo pine (Pinus halepensis) stands.Sphaeropsis sapinea was isolated from larvae (stage II) and molts of the Israeli pine bast scale collected in four Aleppo pine stands. When Aleppo
pine saplings were infested with pine bast scale and subsequently inoculated withS. sapinea most plants died, whereas mortality of seedlings inoculated withS. sapinea alone without previous scale infestation was negligible. Scale infestation of saplings without subsequent fungal inoculation
caused the death of one third of the plants. Wounding of pine shoot apices by removal of needle fascicles enhanced infection
byS. sapinea. The results support the assumption thatM. josephi can play an important role in transmitting the fungal inoculum to the host tree as well as enabling its penetration to the
shoots through wounds in the bark.
http://www.phytoparasitica.org posting Dec. 20, 2004. 相似文献
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马尾松松材线虫病与松材蓝变相关性研究 总被引：1，自引：0，他引：1
研究表明:松材线虫引起的马尾松(Pinus massoniana)病树死亡后期,必然有蓝变(Blue-stain)现象。松材蓝变程度与松树树干部位相关性不大。但是,对于同一棵树,若蓝变程度高,则每一部位的蓝变程度都比较高;若蓝变程度低,则每部位的蓝变程度可能都比较低。蓝变程度与松材线虫(Bursaphelenchus xylophilus)的含量也没有正相关关系。在松材线虫病木的检疫过程中,松木蓝变仅可以作为松材线虫病的参考指标。 相似文献
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N. Kim H. W. Jeon M. Mannaa S.-I. Jeong J. Kim J. Kim C. Lee A. R. Park J.-C. Kim Y.-S. Seo 《Plant pathology》2019,68(3):434-444
Pine wilt disease (PWD) is the most destructive disease threatening pine worldwide. The disease is mainly caused by the pine wood nematode, Bursaphelenchus xylophilus, which is vectored by pine sawyer longhorn-beetles, Monochamus spp. This study aimed to select resistance-inducing pine endophytic bacteria for management of PWD. To set up a defence-related genes expression pattern for screening, four chemical inducers (salicylic acid, γ-aminobutyric acid (GABA), β-aminobutyric acid and α-aminobutyric acid) were tested in vitro on pine calli and in vivo on pine seedlings. Treatment with GABA had the greatest reduction in PWD severity on pine seedlings. The pattern of defence-related gene expression in calli treated with GABA was used to select potential resistance-inducing bacterial strains. In addition, 92 bacterial strains were isolated from pine tree needles and stems and were tested for expression of defence-related genes in pine calli in vitro. Among the tested strains, 13 showed a similar pattern to GABA treatment in at least four tested defence-related genes and were selected for the seedling assay. From the seedling assay, three bacterial strains (16YSM-E48, 16YSM-P180 and 16YSM-P39) showed significant reduction in PWD severity compared to the untreated control. Moreover, among the selected strains, cell-free culture supernatant of strain 16YSM-P180 significantly reduced PWD severity in inoculated pine seedlings. The selected strains were identified based on the 16S rRNA sequence as Pseudomonas putida 16YSM-E48, Curtobacterium pusillum 16YSM-P180 and Stenotrophomonas rhizophila 16YSM-P39. These selected strains are suggested as potential alternatives for management of PWD by induction of systemic resistance. 相似文献
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Yasuhiro Mori Fumihiko Miyahara Yuji Tsutsumi Ryuichiro Kondo 《European journal of plant pathology / European Foundation for Plant Pathology》2008,122(4):529-538
To study the relationship between resistance to pine wilt disease and the migration or proliferation of pine wood nematodes
(PWN) (Bursaphelenchus xylophilus), we conducted experiments using clonally-propagated Japanese black pines (Pinus thunbergii) with pre-evaluated individual resistance levels. Bark including the cortical resin canals—one of the main migration pathways
of PWN—was removed by girdling, but neither the migration of PWN nor symptom development of pine wilt disease were inhibited
by this treatment in non-resistant clones. Histological observations showed no significant differences in the lumen area or
the number of cortical- and xylem- axial resin canals between resistant and susceptible clone groups from a half-sib family.
A bioassay using methanol extracts from resistant and susceptible clones showed that extracts from both clones showed similar
attractant effects to PWN, but neither had repellent effects. The resistant clones were multi-inoculated with PWN into three
split points to mimic migration in the stem. The proportion of damaged plants was not significantly different from that in
single-inoculated plants (control). In this experiment, the number of PWN detected from partially-damaged plants was much
higher than that from non-damaged plants. An inoculation test using stem cuttings showed that the population of PWN increased
in susceptible cuttings at 1–20 days after inoculation (dai), while it remained unchanged or gradually decreased in resistant
cuttings. These findings suggested that the factors contributing to resistance were associated with inhibiting the proliferation
of PWN, rather than inhibiting their migration. 相似文献
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两种分子技术检测松木中松材线虫的效果评价 总被引：1，自引：0，他引：1
为更好地解决当前松材线虫Bursaphelenchus xylophilus检测工作中频现的假阴性问题,通过不同分子技术检测效果的比较评价,摸索出了一套可提高松材线虫检测准确性的技术体系。针对早期感病松木的低线虫量,建立了线虫的最大量分离方法和基于rDNA ITS序列的分子检测体系,该体系可高效检测出单条松材线虫,准确率达93.75%。以来自浙江省不同疫区的96份松木样品为检测材料,比较了SCAR标记和ITS序列2种分子检测技术的阳性率。结果显示,通过首次PCR,ITS_Ⅰ和ITS_Ⅱ序列的检测阳性率分别为52.08%和55.21%,SCAR标记的检测阳性率为30.21%;通过第二次或巢式PCR,ITS_Ⅰ和ITS_Ⅱ序列的检测阳性率提高到了97.92%和100.00%,SCAR标记的检测阳性率提高到了59.38%,表明基于ITS序列的检测阳性率明显高于SCAR标记,且通过第二次或巢式PCR方法可进一步提高检测灵敏度,降低假阴性率。因此,通过线虫的最大量分离并基于rDNA ITS序列的分子检测可明显提高检测准确性,更适用于松材线虫的常规检测。 相似文献
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The relative importance of beetle species associated with Fusarium circinatum‐infected Monterey pines was investigated in three Monterey pine forests along the coast of central California, USA from April to November in 2004 and 2005. Fusarium circinatum was frequently isolated from Ips mexicanus and I. plastographus. The mean percentage isolation based upon numbers of I. mexicanus and I. plastographus carrying propagules of F. circinatum was 17·7 and 10·9% in 2004 and 16·7 and 17·3% in 2005, respectively. The mean percentage isolation was high in the spring and early summer and low in late summer and autumn in all three locations for both species. Isolation was higher from beetles emerging from harvested F. circinatum‐infected pine‐stems than for trapped beetles, 42·4% for I. mexicanus and 45·9% for I. plastographus. The mean (± SE) propagule load of trapped I. mexicanus was 269·5 (± 14·1) in 2004 and 281·7 (± 35·7) in 2005 and was 216·1 (± 28·9) in 2004 and 251·9 (± 28·4) in 2005 for I. plastographus. Mean propagule loads decreased from May to November in all locations for both species. Propagule loads of beetles emerged from infected stems were lower than that of trapped beetles, with means of 89·4 (± 23·2) and 93·0 (± 23·2) for I. mexicanus and I. plastographus, respectively. Thus beetles must acquire fungal propagules from more than one infected host. These results also suggest that higher contamination rates and propagule loads in spring and early summer may indicate a higher risk of pitch canker transmission, relative to late summer or autumn. 相似文献