松墨天牛携带的松材线虫PCR检测技术

利用本研究筛选出的一对松材线虫特异性PCR引物(上游引物:5'-CTACGTGCTGTTGTTGAGTTGGC-3',下游引物:5'-TGGTGCCTAACATTGCGCGA-3'),从松材线虫DNA中,扩增出一条长度403 bp(基因库登陆号:DQ855275)的特异性DNA片段.该引物可以将松材线虫DNA与松墨天牛组织以及拟松材线虫、畸刺伞滑刃线虫、变异伞滑刃线虫、小角伞滑刃线虫、利昂伞滑刃线虫、湖南伞滑刃线虫、红松滑刃线虫、滑刃属线虫、斯坦纳长尾线虫、小茎线虫、剑尾齿杆双胃线虫和寄生小杆属线虫等12种线虫的DNA区分开来.在此基础上,建立一套利用PCR技术直接从受感染的松墨天牛组织中检测出松材线虫的技术,并在实际运用中得到检验.     

松材线虫伴生细菌的分离鉴定及其致病性

从松材线虫虫体上分离到两个细菌菌株:GD1和R,经鉴定,菌株GD1为坚强芽孢杆菌.利用水培马尾松离体松枝作接种材料,接种消毒后的松材线虫(Bx)、GD1、R、Bx GD1和Bx R.结果表明,松材线虫和坚强芽孢杆菌混合接种,松枝发病,松枝流脂减少至停止,蒸腾作用降低,单独接种松材线虫或坚强芽孢杆菌,松枝不发病.菌株R对松材线虫病的发生没有影响.提出松材线虫及其伴生细菌均是松材线虫病不可缺少的致病因素.     

Monitoring of <Emphasis Type="Italic">Pinus massoniana</Emphasis> spatial pattern changes based on RS and GIS techniques

Our research focused on Pinus massoniana information extracted from remote sensing images based on the knowledge detection and decision tree algorithm and established a spatial pattern model, combining quantitative theoretical ecology with remote sensing （RS） and geometric information system （GIS） techniques. Applying information extraction methods and a spatial pattern model, we studied P. massoniana spatial patterns changes before and after the invasion by pine wood nematode （Bursaphelenchus xylophilus） in Fuyang and Zhoushan counties, Zhejiang Province, east China. The P. massoniana spatial patterns are clustering, whether the invasion happened or not. But the degree of clustering is different. Our results show good agreement with field data. Applying the results, we analyzed the relationship between spatial patterns and the invasion level. Then we drew the elementary conclusion that there are two kinds of patterns for pine wood nematode to spread： continuous and discontinuous diffusion. This approach can help monitor and evaluate the changes in ecological systems.     

松材线虫检疫样品前处理机的研制

本文简要介绍了自行研制的松材线虫检疫样品前处理机的构造及其应用。该设备可以将日常检疫松材线虫的样品切割成符合检疫要求的木条,大大提高了松材线虫的检疫效率。适合在各口岸一线检疫部门广泛应用。     

松材线虫病发病过程特征变化的研究

为提高松树病死木松材线虫检出率,及时、准确诊断疫木和疫点,根据病害的外部症状,把松材线虫病的发病过程分为四个阶段,对各阶段的松材线虫分布、松褐天牛发生情况、含水率的变化、木材蓝变情况等特征进行研究。结果表明：病死木上松材线虫的分布由多到少为：在发病前期,上部〉中部〉下部;发病后期,上部〈中部〈下部。松材线虫的数量与松褐天牛老熟幼虫的数量呈正相关,最佳的取样阶段为第Ⅱ和第Ⅲ阶段靠上部梢段取样,第Ⅳ阶段偏下部取样。     

环氧乙烷对松木片中松材线虫的熏蒸效果

在18℃、60%-70%RH的室内条件下,研究了不同剂量环氧乙烷(EO)对松木片中松材线虫的熏蒸效果.结果表明:EO对松材线虫具有一定的熏杀作用,松材线虫的死亡率(y)与EO剂量(x)呈线性相关[y=-44.998+1.838x(p=0.00948)],EO对松材线虫的LD50为51.686 g.m-3,LD99.9为78.835 g.m-3;松木片对EO的吸附作用很大,当EO剂量为56 g.m-3时,24 h浓度值为1.157 g.m-3,吸附量达到89.974 mg.L-1,且吸附量随剂量的增大而增加,吸附主要发生在熏蒸开始后4 h内,4 h吸附率可达94%以上.因此,不推荐将EO用于集装箱内松木片松材线虫的处理.     

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。     

Interspecific hybridization between Bursaphelenchus xylophilus and Bursaphelenchus mucronatus

Bursaphelenchus xylophilus is the pathogen that causes pine wilt disease, which has greatly damaged forests and ecosystems in countries of East Asia and Europe.Bursaphelenchus mucronatus is closely related to B. xylophilus in morphology and host plant specificity. A longrunning debate has existed regarding whether these two species can successfully produce hybrid offspring. In the present study, we performed in the laboratory, hybridization of two B. xylophilus nematode isolates from China and Japan and three B. mucronatus isolates from China, Japan and France. Nematode isolates of B. xylophilus were successfully crossed with B. mucronatus isolates, and the rate of hybridization was relatively high; however, some hybrid offspring died. Successful hybridization occurred between B. xylophilus and B. mucronatus isolates from China, and 22 generations of hybrids were produced. All F1 hybrids could be backcrossed with their parents and produce offspring. Variation in mucro length among the hybrid offspring and their parents was observed. The hybrid offspring and their parents were inoculated into 3-months-old black pine(Pinus tunbergii) seedlings. Weaker pathogenicity of hybrid offspring was observed compared with that of their parents, and significantly fewer offspring nematodes than parents were reisolated from pines. Therefore, the offspring of B. xylophilus and B. mucronatus may exist in the forest and could influence disease epidemics.     

生防细菌对松材线虫灭杀效果初步研究

为了寻找高效低毒且对环境友好的松材线虫病生防菌株,本研究测定了10株对根结线虫有较好防治效果的生防菌对松材线虫的室内灭杀作用,并且选取杀虫活性最好的菌株A4做了其发酵乙醇抽提物对松材线虫LC50值的测定和林间防治效果的测定。结果表明:10株生防菌株的发酵原液对松材线虫都有较好的杀虫效果,其中A4更是在将发酵液稀释10倍之后仍然具有大于50%的杀虫活性,表明该菌株具有较大的生防潜力;室内毒力测定显示菌株A4的发酵抽提物对松材线虫处理24 h的LC50值为0.291 mg/mL;林间防效实验表明菌株A4的发酵抽提物能够显著降低松树枯死率。