松材线虫侵染对马尾松、黑松水分及其相关代谢的影响

 以盆栽马尾松(Pinus massoniana)幼苗和离体水培黑松(P.thunbergii)、马尾松为材料,研究了松材线虫(Bursaphe-lenchus xylophilus)侵染对寄主水分、游离脯氨酸及叶绿素含量的影响以及由此引起的幼苗萎蔫与干旱萎蔫间的差异。松苗和离体枝接种松材线虫后,茎部相对含水量逐渐下降,早期下降缓慢,中后期加快。叶片变化滞后于茎部,相对含水量的明显下降发生在叶片褐变萎蔫症状出现之后(病害发展后期)。游离脯氨酸含量、叶绿素含量变化完全随叶片相对含水量的变化而变化。水分状况是影响受侵植株叶片脯氨酸和叶绿素含量的决定因子。自然干旱枯萎和由松材线虫侵染引起枯萎的松幼苗,两者从枯萎方式到茎、叶相对含水量的变化均存在显著差异。     

黑松13个抗病家系对松材线虫的抗性反应及组织病理学观察

 为评估自日本引进的抗性黑松对松材线虫(Pine Wood Nematode,PWN)的抗病性,选用3个不同来源的PWN强毒和中强毒虫株采用皮接法接种,对1.5年生的黑松13个优良家系进行抗病性测定。结果表明,引进的黑松大部分家系对3个PWN虫株都表现出一定的抗病性,但针对不同虫株,抗病各家系间差异明显,其中有7个家系(3、4、5、6、9、11和12号)表现良好。同时,为探讨引进黑松家系感染PWN后的组织病理反应,对12号抗性家系进行组织病理学研究。观察表明,松材线虫在抗性黑松和普通黑松中的纵向扩散速度差别不明显,但其在抗性黑松内的横向扩散速度比在普通黑松内要慢,且抗性黑松体内木射线细胞和围绕树脂道泌脂细胞的薄壁细胞,出现病变要比普通黑松晚;在接种后同一时间取茎段的同一部位发现,抗性松苗组织解剖结构及细胞的病变和破坏程度要比普通松苗轻。     

松材线虫对马尾松的致病性及其早期诊断试验

本试验通过从枯死松树分离繁殖的松材线虫 Bursaphelenchus xylophilus 接种3～4年生马尾松苗,观察到接种株第45天开始出现外观症状,三个月内接种株相继枯死。在枯死株中分离出大量松材线虫,表明松材线虫对马尾松具有明显的致病性。试验还通过酸性品红染色追踪树液流,观察到松材线虫侵染早期,其松脂管的薄壁细胞受破坏,松脂从松脂管渗漏至周围的管胞中,使水溶性染剂不能着色而出现白斑,树液移动受阻。而对照株中染剂可在植株管胞中自由移动,不会产生白斑,松脂流动只限于松脂管内。而在这段时间接种株和对照株外观上生长正常,并无明显差异。因此,通过酸性品红染色追踪树液流,观察植株根茎横切面白斑的出现可作为早期诊断马尾松松材线虫病的一种方法。     

松材线虫伴生细菌与松树萎蔫病关系的初步研究

利用马尾松水培离体松枝作接种材料,接种消毒后的松材线虫、线虫虫体上分离到细菌分离物GD1以及两者的混合体。结果表明,单独接种消毒后的松材线虫蔌细菌分离物GD1均不能使松枝萎蔫,只有两者混合接种才能使松枝发病枯死。从而说明了松材线虫的伴生细菌对松树萎蔫病的发生起着重要的作用。     

马尾松(Pinus massoniana)感染松材线虫(Bursaphelenchus xylophilus)的研究

 1987-1988年在江苏南京-镇江松材线虫疫区内采(征)集143份自然枯死或濒死马尾松样本,在94份(65.7%)样本中查见松材线虫。有关林分中马尾松的病枯情况不一,在句容桥头一下蜀一线、江宁汤山和六合方山等处有过相当多的马尾松发病死亡。表明松材线虫是当地马尾松自然枯死的主要原因。人工测定了无黑松林区5年生浙江富阳马尾松苗及混栽黑松林区4-5年生江苏马尾松苗对两种松材线虫分离物的反应,结果指出,供试松苗都表现中度感染或高度感染,并有20%或70-80%的病枯株。接种林间马尾松,初步证实马尾松对松材线虫的抗性随树龄变化。1-2年生马尾松苗高感松材线虫,病枯株率高达85.7%;树龄25年以上的成材树病枯株率约30%;而10年左右的幼树则无一发病死亡。     

感染松材线虫病松树滑刃目线虫调查

 本研究首次在全国范围内调查感染松材线虫病松树体内滑刃目线虫的种群组成,包括种类记述和数量组成分析。结果表明:在300份样品中共分离得到10种滑刃目线虫,隶属于3科、4属,其中有2个国内新记录种;它们是松材线虫、拟松材线虫、霍夫曼尼伞滑刃线虫、薄荷滑刃线虫、大核滑刃线虫、小麦长尾线虫(国内新记录种)、爱尔密那长尾线虫(国内新记录种)、李氏长尾线虫、吴氏长尾线虫和外滑刃科线虫一种;描述并且图示比较它们的形态特征。种群数量组成分析表明:松材线虫在感病松树体内占据绝对的优势,削弱了松树线虫群落组成的多样性,即线虫群落结构趋向单一化。研究还表明,利用长尾属线虫的捕食习性来控制中国松材线虫病的流行与危害值得进一步探索与研究。     

不同地理种群的松材线虫对马尾松的致病力差异

本文对我国主要病区及日本的４个松材线虫地理种群,对４～６年生马尾松的致病力进行了研究。结果表明,江苏和广东种群对马尾松的致病力较强,感病指数皆为０．９４,而安徽和日本种群的致病力较弱,感病指数分别０．７５和０．６９。     

松材线虫对黑松一些生理生化指标的影响（英文）

 松材线虫病是松树的一种毁灭性病害。本文对接种松材线虫后3~4年生黑松茎部的一些生理生化指标的变化进行了研究。结果显示,接种线虫后,松苗的可溶性糖含量降低,在病情发展中期和后期比早期降低更显著。松苗的可溶性蛋白质和抗坏血酸含量逐渐降低,游离氨基酸含量也降低。松树茎部的可溶性蛋白质含量可用于病害的早期诊断。此外,对这4个生理生化指标在病害致病机理中的作用进行了讨论。     

松材线虫自然侵染后松树感病进程中光合作用及抗氧化酶活性的变化

根据松树针叶变色程度及流脂状况,野外分别选取不同感病阶段的黑松、马尾松为对象,以研究松材线虫自然侵染黑松和马尾松后,两种松树的光合作用及其相关生理生化变化过程。结果表明,黑松、马尾松在感病初期,针叶净光合速率(Pn)大于健康阶段数值,随后逐渐下降,感病重期为负值,即松材线虫的侵入破坏了黑松、马尾松的光合作用。由叶绿素荧光参数分析可看出,光合作用的下降是由于光合系统Ⅱ反应中心实际进行光化学反应的效率(ФPSⅡ)下降引起的,而ФPSⅡ的下降是由于PSⅡ反应中心捕获光能效率Fv'/Fm'下降和开放的PSⅡ中心数目qP下降共同导致。两种松树在感病进程中,针叶叶绿素含量和类胡萝卜素含量整体呈下降趋势,但在感病初期含量上升,这也是引起光合作用速率下降的因素之一。此外,针叶含水量变化也一直呈下降趋势,是引起松树萎蔫,叶绿素含量下降,光合作用速率下降的主要原因之一。实验表明,随着松材线虫侵染的加重,植株体内与抗性相关的抗氧化酶系统活性也会发生变化。黑松和马尾松过氧化氢酶(CAT)活性随着感病进程逐渐下降,过氧化物酶(POD)活性和超氧化物歧化酶(SOD)活性略有上升,但不显著,说明松材线虫的侵入破坏了黑松、马尾松的抗氧化酶系统,导致松树自我保护作用减弱,同时由于两种松树热耗散能力(NPQ)的下降,产生的多余电子进一步破坏了抗氧化酶系统,导致光合速率下降。研究表明,松材线虫的侵染,对寄主光合作用产生了破坏,同时寄主也作出相应防御反应,但随着松材线虫的大量繁殖,寄主自身保护作用逐渐丧失。     

松材线虫病组织病理学研究

 利用马尾松水培离体松枝作接种试验材料,对松材线虫侵染早期寄主细胞和组织所发生的一系列病理变化作了观察。切片结果表明,接种枝树脂道的细胞最先受到线虫侵袭的破坏。由核变形、胞质凝聚标志的轴向和射线薄壁细胞的死亡,是寄主对线虫侵染的第一个明显的组织病理学反应。这一反应随线虫侵入时间而日益显著,死亡薄壁细胞数量和程度上的逐步增加,反映了病害的发展进程。     

Sublethal infection of different pine species by the pinewood nematode

Bursaphelenchus xylophilus, the pinewood nematode and causal agent of pine wilt disease (PWD), is a globally destructive pine pest. PWD is favoured by high temperatures and drought, but B. xylophilus can also persist in symptomless hosts in areas not prone to PWD. We compared seedlings of different Pinus species that were inoculated with B. xylophilus when well-watered and under moderate temperatures, conditions that favour sublethal infections. We compared isolates of B. xylophilus in south-west Europe (Portugal) and south-east USA (Louisiana). P. pinea, P. pinaster, and P. radiata were challenged in both areas with inoculations of local B. xylophilus. P. sylvestris in Portugal and P. palustris in Louisiana were also tested. Seedling mortality was low, as expected, but nematode establishment was common. There were higher nematode densities and more damage in seedlings of P. sylvestris and P. radiata, followed by P. pinaster, with P. pinea and P. palustris proving to be more tolerant. Life history of hosts was a better predictor of resistance to B. xylophilus than geographical origin, even under sublethal conditions. Fast-growing species (P. radiata and P. pinaster) appeared most likely to be symptomless carriers in areas not prone to the development of PWD. Cold-adapted species (P. sylvestris) may still be suitable hosts for the pathogen, even at the sublethal conditions of their distribution areas. There were more nematodes per gram of host using nematode isolates from Portugal. Nonetheless, our results opposed the hypothesis that North American pinewood nematodes have such low virulence that they are obligatorily saprophytic.     

马尾松松材线虫病与松材蓝变相关性研究

 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。     

Induction of resistance against pine wilt disease caused by Bursaphelenchus xylophilus using selected pine endophytic bacteria

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.     

溴甲烷对松木片中松材线虫的熏蒸作用

采用二次正交旋转组合设计,在室内条件下研究了溴甲烷剂量、熏蒸处理时间和处理温度对松木片中松材线虫熏蒸效果的影响。结果表明,溴甲烷剂量是影响熏蒸效果的最主要因子;14、18 ℃和22 ℃条件下处理24 h,松材线虫死亡率达99.9%时的溴甲烷浓度分别为17.92、16.07、14.87 g/m3; 14、18、22 ℃条件下,松材线虫死亡率达99.9%时,溴甲烷浓度和时间乘积,即CT（99.9）分别为192.52、174.705、166.229 g•h/m3。溴甲烷可用于口岸松木片中松材线虫的除害处理。     

进境木包装松材线虫检验检疫制样方法比较

对来自日本、韩国的进境木包装 ,采用不同直径的电钻钻取木屑各 1 0 g和采用薄木片 1 0 g ,进行 6次重复制样进行线虫分离 ,结果表明差异显著。同时采用薄木片 1 0 g、30 g、6 0 g进行线虫分离 ,同样差异显著。进境木包装用电钻取样 ,获得线虫量较小 ,不能在实际工作中运用。薄木片制样 ,能较多的获得线虫 ,符合进境木包装检疫工作的需要。     

光照对松材线虫病潜育期的影响

应用相关回归方法，分析了黑松接种松材线虫后，病害的潜育期与环境中的日平均温度、日平均日照时数的关系。结果显示，病害潜育期与日平均温度呈显著负相关，与日平均日照时数呈显著负相关。     

Suppression of pine wilt disease by an antibacterial agent,oxolinic acid

BACKGROUND: Pine wilt disease (PWD) is very complex and has been reported to be caused by pine wood nematode, Bursaphelenchus xylophilus (Steiner & Buhrer) Nickle, and its accompanying bacteria. However, there is no report on the control of PWD by antibacterial agent. The present study was performed to investigate disease control efficacy of antibacterial agents against PWD. RESULTS: Among six antibacterial antibiotics tested, oxolinic acid (OA) showed the strongest antibacterial activity against five bacteria isolated from three strains of pine wood nematode. In in vivo assay, it effectively suppressed the development of PWD in three‐year‐old seedlings of Pinus densiflora Sieb. & Zucc.; it showed 71% control when injected at 3 mg per seedling. A mixture of OA and the nematicidal agent abamectin (Ab) showed higher disease control efficacy against PWD than either OA or Ab alone. In addition, OA alone and a mixture of OA and Ab also controlled PWD in approximately 20‐year‐old pine trees under field conditions. CONCLUSION: This is the first report on the suppression of PWD by OA. The result strongly indicates that PWD could be controlled by antibacterial antibiotic alone and a combination of antibacterial and nematicidal agents. Copyright © 2010 Society of Chemical Industry