ITS鉴定真菌病原:一例司法鉴定研究实例

本研究选择一例疑似空气污染物伤害松树的司法鉴定案作为研究实例,在现场勘察和初步形态鉴定的基础上,应用ITS序列分析鉴定了对疑似空气污染物伤害松树的病原做了分子鉴定。我们对委托鉴定的疑似空气污染物伤害松树的地及周边地区进行了勘查,现场勘查结果并不支持空气污染物伤害松树的看法。为明确松树枯死的原因,对取样病枝上的疑似病原物进行了显微镜观察和培养性状观察,根据显微镜下孢子的形态和病原菌的培养性状,初步判断导致马尾松枯死的病原可能为拟盘多毛孢属(Pestalotipsis)真菌。为进一步确定病原种类,我们应用分子生物学技术对危害马尾松的病原物进行了ITS分子鉴定。测定了疑似真菌的ITS序列,并进行了比对和系统发育分析。研究结果表明,导致松树枯死的病原菌系韦司梅拟盘多毛孢(Pestalotipsis vismiae)。在本研究案例中,利用真菌ITS序列的系统发育分析,快速、准确地鉴定了真正导致松树枯死的病原,为今后类似司法鉴定案件审理提供了证据鉴定的新手段。     

高度重视虫传危险性森林病害——松材线虫病

松材线虫病 ,又名松树萎蔫病 ,是松树的一种毁灭性病害 ,具有发病致死速度快、传播蔓延迅速、防治难度大等特点 ,因此被称为“松树癌症” ,在国内外均被列为重要的植物检疫对象。松树一旦感染此病 ,最快的4 0多天即可枯死 ;病害流行很快 ,一般从发病到松林毁灭只需 3～ 5年。该病目前主要分布在北美和东北亚 ,在美国、加拿大、墨西哥、日本、韩国、中国 ,葡萄牙也有分布 ,但危害程度不一 ,以日本受害最重。在北美三国 ,该病虽严重影响了木材出口 ,但并未对松林造成严重危害。而在东北亚 ,该病导致松树大量死亡 ,尤其在日本 ,该病分布全国 ,…     

黏质沙雷氏菌AHPC29对松树萎蔫病的防治效果及其增菌条件

【目的】从松材线虫的媒介天牛蛹室及其气管中获得的黏质沙雷氏菌Serratia marcescens AHPC29对松材线虫具有致病能力,本研究旨在探究黏质沙雷氏菌AHPC29对松材线虫引起的松树萎蔫病的防治效果以及该菌株在实验室的增菌条件。【方法】通过对温室内人工感染松材线虫的松树灌溉菌剂,分析黏质沙雷氏菌AHPC29对松树萎蔫病的作用效果;通过单组分筛选和正交实验确定培养基组分和培养条件对其生长的影响,探究黏质沙雷氏菌AHPC29的最佳增菌条件。【结果】对于感染松材线虫的松树,灌溉黏质沙雷氏菌AHPC29菌液的处理组生长状态优于对照组,并且树内松材线虫含量显著降低;黏质沙雷氏菌AHPC29增菌的最佳培养基配比为0.1%乳糖、0.5%复合氨基酸、0.5% KNO₃、1.5% MgSO₄,其中影响最大的组分为氮源;培养时其菌液最佳接种量为7%,最佳装液量为40%,最佳转速为180 r·min^-1,最佳温度为30 ℃,最适培养时长为36 h。【结论】本研究获得了黏质沙雷氏菌AHPC29的最佳增菌条件,并证实该菌具有良好的松树萎蔫病防治应用潜力,为其作为生防菌的应用提供了理论基础。     

松树的菌根及其造林意义

松树是北半球分布非常广泛的树种,南半球现在也广泛引种栽培.全世界大约有80-100多种.由于松树生长迅速、材质优良、适应性强、用途广泛,所以在世界各国林木引种和绿化造林中都占特殊地位.然而,大部分松树的根毛较少,菌根则是它们吸收水分和养分的一个主要器官.因此,有无菌根是松树成活与良     

甘蔗宿根矮化病病原菌Leifsonia xyli subsp.xyli 基因组学研究进展

综述甘蔗宿根矮化病病原细菌Leifsonia xyli subsp.xyli(Lxx)基因组组成特征、基因组进化、基因组中致病相关基因与寄主适应性等方面的研究进展,并对该病原茵、引起番茄细菌性萎蔫和溃疡的病原细菌、马铃薯环腐病痛原细菌的基因组和致病相关基因进行比较,发现它们存在同源致病基因,如celA、pat1基因,其致病机理可能有相似性.     

天牛幼虫的外部形态及亚科检索图

<正> 天牛科是鞘翅目昆虫中的一个大科,全世界已知4万余种,国内记录已超过2600种。该科种类是农、林业重要害虫。松墨天牛(松天牛)Monochamus alternatus Hope因从国外传带松材线虫Bursaphelenchus xylophilus引起松树萎蔫病,致使南京中山陵等地的松林被毁。我国植检部门已将该种列入对外植物检疫     

中国的松树

松树是我们最常见的一种树,每个人可能都会熟悉它。但是,我们平常所讲“松树”这一名称,意义比较笼统,那是对松科(Pinaceae)松属(Pinus)若干种松树的统称。其实,我国松树有很多种,性状各有不同,分布地区也不一样,这里,仅就我国原产的松树作一个简单介绍,外国引种来的从略。松树的用途松树是很有用的树种,在我国现有的森林面积中,松林占了最大的比重,估计在60%左右,而今后造林事业中,松树仍然是最普遍最广泛的树种。松树的木材可做建筑用材、矿柱、枕木、樁木、或用来制家具,又可以制膠合板、火柴     

松材线虫(Bursaphelenchus xylophilus)入侵对马尾松(Pinus massoniana)林分生长的影响及相关生长模型

松材线虫（Bursaphelenchus xylophilus）是一种松树上发生严重的有害生物,它不仅改变了生态系统的结构和功能,而且改变了系统内生物的原有特性和地理分布。松材线虫及其引起的松树萎蔫病已对中国马尾松林（Pinus massoniana）的树木成长产生了巨大影响。基于此,使用＂每木调查法＂和＂样方法＂,对松材线虫入侵后的马尾松林内松树的各项生长指标因子进行了调查分析,其结果表明：自松材线虫1996年入侵所调查地区的松林后,对于受害松树不管是伐倒木（被伐倒）还是倒木（自然倒地）,其对周围马尾松胸径生长的影响是显著的,而对树高生长的影响不显著。最后建立了一系列的灰色和灰色-马尔可夫链数学模型,其预测结果精度高,可用于今后受害和未受害区马尾松林分因子的生长预测。     

植物病原棒形细菌的RAPD分析

采用RAPD分析技术对萎蔫短小杆菌落葵致病变种（Curtobacterium flaccumfaciens pv. basellae pv. Nov.）和萎蔫短小杆菌糖甜菜致病变种（Curtobacterium flaccumfaciens pv. beticola pv. Nov.）及植物病原棒形细菌4个属的15个菌株进行分类研究：从50条随机引物中筛选出20条,共产生225条带,多态性条带占804%。遗传相似矩阵及UPGMA聚类分析,表明这两个新致病变种与萎蔫短小杆菌属亲缘关系近,最小相似系数为0.6511;与其他属细菌亲缘关系较远;结合前人研究结果对植物病原棒形细菌新近提出的分类地位的改变进行了探讨。     

四川主要松树内生拟盘多毛孢

从5种松树健康的枝、叶、干中共分离了2,400个组织块,得到199株内生拟盘多毛孢Pestalotiopsis,形态学和分子生物学鉴定分属于11个种,平均分离率为8.29%。所得11种内生拟盘多毛孢都是首次在四川的松属植物中分离得到,其中葡萄生拟盘多毛孢P.uvicola和山桃草拟盘多毛孢P.gaurae为首次在松树上分离得到。基于ITS序列的分子系统发育分析显示松树内生拟盘多毛孢种类之间存在一定的遗传差异,部分内生拟盘多毛孢与病原拟盘多毛孢的ITS序列具有很高的同源性。     

东亚地区夏季干旱、强台风事件与松树枯萎病的关系

松树枯萎病的发生和流行给东亚的日本、中国和韩国带来了不小的损失,引起许多国家的重视. 本文应用实地观测和气象数据分析等方法,研究了东亚地区松树枯萎病的发生和流行特点,以及与灾害气象事件之间的关系.结果表明: 在日本、中国和韩国,持续的夏季干旱少雨和强台风等极端气象事件能够诱发松树枯萎.在极端干热的环境中,松树常出现能量代谢失调, 以至于受胁迫的松树整株枯萎;而在低温多雨的年份,松树枯萎少,甚至没有发生枯死现象.在松材线虫及其媒介昆虫侵染之前,松树的活力业已下降.松树枯萎病似乎应该局限在台风频发和持续干热的地区.在自然环境优越、少有台风和干热事件出现且没有不当扩大松树栽培范围的地区,松树枯萎病大面积发生的可能性不大.     

Relationship between soil properties and incidence of pine wilt disease at stand level

This study was conducted to evaluate and compare soil properties between pine stands either damaged or undamaged by pine wilt disease in Jinju and Sacheon Cities, which are areas in Korea severely affected by the disease. Soil physical and chemical properties were generally similar between damaged and undamaged pine stands. There was no significant difference in soil factors related to soil nutrient fertility between damaged and undamaged pine stands, although the incidence of pine wilt disease was affected by soil clay content, which was significantly (P < 0.05) higher in the damaged (24.8%) than in the undamaged (20.1%) pine stands. However, this relationship should be treated with caution because small scale topographical variation may simply reflect the incidence of pine wilt disease. The results suggest that incidence of pine wilt disease was not influenced by soil nutrient properties at a small stand scale.     

Response of soil microbial communities to changes in a forest ecosystem brought about by pine wilt disease

Japan has suffered a lot from forestry losses due to pine wilt disease caused by pinewood nematode infestations. Studies were conducted regarding its causative agent and the effects of natural vegetation succession after pine wilt disease, but its effects on microorganisms were not given equal attention. This study determined the effects of pine wilt disease on light conditions, soil microbial biomass, litter decomposition, microbial abundance and the physical and chemical properties of the soil. Results showed that in a forest currently affected by pine wilt disease, there was higher light penetration, greater microbial biomass carbon, and a faster rate of litter decomposition. Microbial abundance was shown to be reduced in pine wilt affected areas. There were close correlations between the biological and physicochemical properties of the soil, but the reason for the decrease in microbial abundance is not yet well understood, and thus requires further study.     

Succession of secondary forests affected by pine wilt disease in western Japan followed on vegetation maps

Abstract. Succession, changes in the distribution pattern of forest vegetation, and Pinus forest survival following pine wilt disease were clarified based on phytosociological analysis and vegetation maps. Survival of Pinus forests was restricted to the early successional stages, which were located on ridges and the upper part of slopes. Subsequent to pine wilt disease, the succession progressed from early to late substages of Pinus forest, mixed deciduous and evergreen Quercus, to evergreen Quercus forest. Succession occurs in abandoned pine forests which apparently are in a bad state and are vulnerable to attacks by pine wilt disease.     

Development of secondary pine forests after pine wilt disease in western Japan

Abstract. The development of secondary Pinus densiflora (Japanese red pine) forests after pine wilt disease was studied through phytosociological analysis, estimation of forest structure before disease and size-structure, tree ring and stem analyses. Following the end of the disease, the growth of previously suppressed small oak trees was accelerated. This is quite different from the development of forests following fire, which starts with the establishment of pine seedlings. Pine wilt disease shifted the dominance of secondary forests from Pinus densiflora to Quercus serrata oak forest. In pine forests, disturbance by fire is important for forest maintenance. In contrast, disturbance by pine wilt disease leads to an acceleration of succession from pine forest to oak forest.     

松材线虫病发生点格局及影响因素

松材线虫病在中国大陆造成了巨大的生态与经济价值损失,南方地区尤为严重,分析松材线虫病空间分布、量化环境因素对其发生的影响对于松材线虫病的防控整治具有重要意义。本研究以江西省赣州市南康区松材线虫病为研究对象,采用核平滑密度、Ripley’s K函数、点过程模拟等空间点格局分析方法,探讨了区域松材线虫病发生的空间格局及其对环境变量的响应。结果表明: 研究区松材线虫病的发生不是随机分布,而是存在显著的空间聚集区。地形因子、植被因子和人类活动因子是影响松材线虫病空间异质性分布的主要因素。空间点格局分析表明,海拔、坡度、距最近道路距离、道路密度、距最近居民点距离、郁闭度和植被类型对松材线虫病的发生具有重要影响。在森林病害管理中,除了加强因人类活动引起病害传播源的管控外,还应该考虑地形、植被类型等特征进行松材线虫病害的综合预警监测。     

Pine wilt disease alters soil properties and root-associated fungal communities in Pinus tabulaeformis forest

Plant and Soil - The objective was to elucidate how pine wilt disease impacts soil properties and root-associated fungi in Pinus tabulaeformis forest. Soil responses to pine wilt disease were...     

Using an evapo-transpiration model (ETpN) to predict the risk and expression of symptoms of pine wilt disease (PWD) across Europe

The pine wood nematode (PWN) Bursaphelenchus xylophilus is the causal agent of pine wilt disease (PWD), a xylem restricting disease of pine trees. PWN, a native of North America where it very rarely kills native pine trees, has spread internationally killing host trees in China, Japan, Korea, Taiwan and Portugal, with isolated incursions into Spain. Based on the locations where tree mortality has been recorded, it appears that pine trees growing in hot, dry conditions are more susceptible to pine wilt disease. This paper describes the ETpN model, an evapo-transpiration model (previously developed by Forest Research), which has been modified to incorporate the presence of PWN inside a tree and which predicts the regions of Europe that are likely to succumb to PWD. ETpN acts independently of the vector beetle (Monochamus spp.), predicting the likelihood of PWD on the assumption that a tree in a particular region has already been infested by the pine wood nematode. Different regions across Europe are included to investigate and demonstrate how different climates affect PWD incidence significantly. Simplified, “lite” and latency models have been developed to allow a non-specialist user to determine respectively the risk of PWD at a particular location and the likelihood of delays (latency) in expression of wilt symptoms.     

Pathogenicity and repulsion for toxin‐producing bacteria of dominant bacteria on the surface of American pine wood nematodes

Bacteria were isolated from the surface of two samples of American pine wood nematodes to identify methods of controlling pine wilt disease. The dominant bacterial strains were identified, and their toxicity and pathogenicity, in addition to their competitiveness with other pathogenic bacteria, were measured to ascertain how bacteria on the surface of American pine wood nematodes might be used to prevent and control pine wilt disease. The bacterial isolates show that the dominant bacteria carried by the two samples of pine wood nematodes are US4, US5, Smal‐007 and Rrad‐006. Based on routine staining, morphological observation and 16S rDNA sequence analysis, the four strains were identified as Delftia lacustris, Pseudomonas putida, Stenotrophomonas maltophilia and Rhizobium nepotum. The incubation of four dominant bacterial strains and Chinese dominant bacterial strains on the surface of aseptic nematodes and in nutrient broth showed that Smal‐007 and Rrad‐006 have strong competitiveness on the surface of pine wood nematodes. Using a bacterial culture medium to measure the propensity of pine seedlings to wilt, all the American dominant bacterial strains were shown to be less toxic than the Chinese dominant strains. If pine seedlings are inoculated with both bacterial and aseptic pine wood nematodes, American dominant bacterial strains present less pathogenicity than the Chinese dominant bacterial strains. In particular, Smal‐007 and Rrad‐006 show the lowest pathogenicity. If pine seedlings are inoculated with both bacterial and wild pine wood nematodes, American dominant bacterial strains significantly reduce the pathogenicity of wild pine wood nematodes isolated from Zhejiang Province, China. The effects of Smal‐007 and Rrad‐006 are confirmed as the most prominent. The American dominant strains Smal‐007 and Rrad‐006 satisfy two main requirements: excellent repulsion performance and low pathogenicity. Therefore, they can be used as candidate strains for biocontrol bacteria.     

化学通讯在松材线虫侵染和扩散中的作用

松材线虫为外来入侵种 ,由其引起的松材线虫病正在我国迅速扩散蔓延 ,造成我国部分地区松林资源的毁灭性破坏。松材线虫病的发生和流行与媒介天牛、寄主植物、共生真菌和细菌密切相关 ,松材线虫 -墨天牛 -松树 -共生微生物之间存在着广泛的化学联系 ,它们通过化学互作 ,调控松材线虫的行为 ,影响松材线虫的侵染和扩散