木霉菌对杨树树皮溃疡病菌拮抗作用的研究

应用哈茨木霉（Trichodema harziaunm)T88菌株和深绿木霉（T.atroviride)T95菌株分别与杨树烂皮病菌（Cytospora chrysosperma)和杨树水泡溃疡病菌（Dothiorella gregaria)进行了平皿对峙培养和载片对峙培养；并研究了两种木霉菌产生的挥发性代谢物对病原菌菌落的影响和其非挥发性代谢物对菌丝干重的影响；分别在光学显微镜和扫描电镜下观察了两种木霉菌的重寄生现象。结果表明：对峙培养中木霉菌T88和T95对两种病原菌均有明显的抑制作用。两种木霉菌均能产生挥发性代谢物质并不同程度地抑制病菌菌落的生长，在密闭条件下T95的抑制率最高达到80．30％。两种木霉菌产生的非挥发性代谢物质可以强烈抑制病原菌的生长、明显降低其菌丝干重，并具有热稳定性。在光学显微镜和扫描电镜下，可以观察到木霉菌的菌丝在病原菌的菌丝上平行或波浪式生长，并在其上产生钩状分枝、吸器或附着胞吸附于病原菌的菌丝上，或穿透病原菌的菌丝生长，或使病原菌的菌丝细胞生质浓缩和菌丝断裂。     

毛白杨内生菌优势种毛壳ND35室内拮抗作用的研究

对毛白杨内生真菌区系种群分析 ,筛选出优势种毛壳ND35 ,并测试了它对 6种常见林果病害病原真菌的拮抗作用。结果表明 ,毛壳ND35是一种广谱性拮抗菌 ,对供试的 6种病菌存在不同程度的抑制作用。可降低病菌菌丝的生长率和孢子的萌发率 ,并对小核菌的菌核进行重寄生 ,从而抑制其萌发。毛壳ND35的菌丝还产生吸器、附着枝等结构对立枯丝核菌 (Rhizoctoniasolani)等病菌的菌丝进行重寄生 ,在显微镜下可观察到病菌的菌丝被缠绕、穿透、断裂等现象。抑菌带和抑菌圈的产生证明抗生活动是毛壳ND35的主要拮抗作用机制之一。对活体毛白杨组培苗进行人工接种试验 ,结果用毛壳ND35预先接种的苗木发病率为零 ,明显低于只接烂皮病菌的对照发病率 5 3 33%,进一步证实了毛壳ND35对植物病害的生物防治潜能。     

樟子松赤枯病拮抗真菌的筛选

在健康的樟子松针叶上分离得到25种真菌,从中选出具有生防潜能的11种用于控制樟子松赤枯病的病原菌拮抗菌株的筛选。拮抗真菌与病原菌的平板对峙试验结果表明:长枝木霉Trichoderma longibrachiatum、血红毛壳Chaetomium cruentum和印度毛壳C.indicum能有效抑制该病原菌,其中长枝木霉对病原菌菌丝的抑制效果最强,抑制率为72.81%,后2种的抑制率分别为49.16%和42.36%。     

绿木霉菌株T43对松烂皮病菌的抑制效果及其抑菌机理

通过对峙培养和非挥发性代谢产物对病原菌生长的抑制]作用,对引进绿木霉(Trichoderma virene)菌株T43及其抑菌活性物质对松树烂皮病菌(Cenangium ferruginosum)的抑制效果进行了研究;从菌株T43发酵液乙酸乙酯提取物对松树烂皮病菌的3种保护酶(SOD、CAT和POD)活力影响的角度,初步探讨了提取物对病原菌的抑菌机理.研究结果表明:对峙培养48 h后,菌株T43对松烂皮病菌相对抑制效果为1.00,72 h后菌株T43与病原菌形成明显的拮抗线,并抑制病原菌的生长.菌株T43乙酸乙酯提取物对病原菌株的室内生长抑制率为60.50％.菌株T43发酵液乙酸乙酯提取物对病原菌的3种保护酶具有明显的抑制作用.     

球毛壳抗生物质对板栗疫病菌的抑制作用

通过液体发酵培养球毛壳ND35菌株,有机溶剂萃取,旋转蒸发仪浓缩,获得球毛壳抗生物质的粗提物,并对该粗提物进行了平板抑菌活性检测和板栗水培枝条抑菌活性检测。结果表明：球毛壳ND35抗生物质能有效地抑制板栗疫病菌的菌丝生长和分生孢子的萌发;板栗离体水培枝条抑菌效果达66.29%,与对照相比,在一定程度上降低了发病率,抑制了板栗疫病菌在枝条上的扩展和定殖。这为将来球毛壳抗生物质在田间应用打下了基础。     

用螺旋毛壳(Chaetomium spirale) ND35菌防治果树腐烂病的试验研究

报道了在离体和田间条件下螺旋毛壳(C. spirale) ND35菌株对苹果树腐烂病的初步抑菌试验结果.PDA平板对峙培养的结果显示在毛壳ND35和苹果树腐烂病菌的两菌落间出现明显的抑菌带;毛壳ND35培养液粗提液比麦芽浸膏培养液的粗提液有更强的抑制病菌菌丝生长的作用.温室和田间生防试验表明用接种毛壳ND35的处理,其苹果树腐烂病发病率明显低于其它处理,包括直接种苹果树腐烂病菌和烫伤处理,及健康对照.再分离试验表明毛壳ND35可成功地在苹果树枝干部位定殖,而且固体麦麸菌剂的定殖率高于玉米粉液体菌剂的定殖率.但是玉米粉液体菌剂在田间的生防效率较好.     

木霉菌株T88对7种病原真菌的拮抗作用

将木霉菌株Ｔ８８与７种常见的林果病原真菌进行平板对峙培养及显微观察,结果表明木霉菌株Ｔ８８对立枯丝核菌Ｒｈｉｚｏｃｔｏｎｉａｓｏｌａｎｉ等供试的７种病原真菌都有明显的拮抗作用,可观察到Ｔ８８对病原菌菌丝的缠绕、使细胞质变稀薄、菌丝消解或穿透等现象,以及对齐整小核菌Ｓｃｌｅｒｏｔｉｕｍｒｏｌｆｓｉ的菌核的重寄生作用,使其发生腐烂,并抑制了白绢病菌菌核的萌发。     

利用三种梢栖真菌生物防治落叶松枯梢病的研究

从落叶松梢栖真菌筛选出对落叶松枯梢病病原菌有抑制作用的菌株,对落叶松枯梢病进行生物防治。将自落叶松梢部分离到的11种梢栖真菌与落叶松枯梢病病原菌进行对峙培养,根据被覆盖程度、抑菌率、拮抗系数和防治预试验的结果,综合评价后发现:粪生粪壳、深绿木霉和球毛壳3种梢栖真菌对落叶松枯梢病有良好的拮抗作用。利用这3个菌株在黑龙江省勃利县通天一林场国富沟进行林间防治试验,结果表明:防治效果为粪生粪壳>代森锰锌400倍液>深绿木霉>球毛壳;深绿木霉和粪生粪壳喷洒浓度50%和球毛壳喷洒浓度75%时,真菌多样性最多,梢栖真菌系统较其他浓度更为稳定,与化学防治相比,既达到了防治目的又具有不污染环境的优势。     

杨树内生真菌的分离和拮抗生防菌的筛选

从健康杨树植株的叶(芽)、皮、枝中分离获得154株内生真菌,经形态观察分类鉴定归属于3个目、5个科、13个属中.利用两点对峙培养拮抗试验、孢子萌发试验和人工接种试验,从中筛选杨树烂皮病病原菌的拮抗菌.试验结果表明所获内生真菌(4属)菌株对病原菌菌丝生长和孢子萌发均存在不同程度的抑制作用,其中,青霉属真菌(Penicillium sp.)的菌株拮抗效果表现最明显.青霉菌可迅速对病原菌菌落包围、覆盖,从而使植物病原菌生长受到抑制,孢子萌发试验中,3 d的培养滤液就可对病原菌孢子萌发产生抑制作用,表现出一定的生防潜能.     

毛壳菌和木霉菌防治山杨根腐病的试验

通过人工诱导试验证明山杨根腐病病原菌为锐顶镰孢菌,室内用毛壳菌和木霉菌,与锐顶 菌做对峙试验和室外防治试验,发现二者对锐顶镰菌都有较好的拮抗作用,是山杨根腐病理想的生防药剂。     

木霉菌株T88对7种病原真菌的拮抗作用

将木霉菌株T     

Temporal allocation of (14)C to extramatrical hyphae of ectomycorrhizal ponderosa pine seedlings

Ponderosa pine seedlings were inoculated with Hebeloma crustuliniforme either in growth pouches before they were transplanted to root-mycocosms (P seedlings), or at the time of transfer to root-mycocosms (V seedlings). Uninoculated seedlings served as controls (U seedlings). The use of root-mycocosms allowed examination of portions of hyphae separate from roots and rooting substrate but still in symbiosis with the host. The results thus provided a quantitative basis for estimating hyphal mass and carbon allocation to extramatrical hyphae. The amount of (14)CO(2) fixed after a 2-h exposure was greatest for P seedlings and least for uninoculated seedlings. Four and nine days after exposure, (14)C content was greatest in uninoculated seedlings and least in inoculated seedlings. In isotope distribution and dry mass accumulation, V seedlings were more similar to U than to P seedlings. Calculated on a dry weight basis, the allocation of isotope to mycelium suggested that extramatrical hyphae of P seedlings were a stronger sink for carbon than extramatrical hyphae of V seedlings. Differences in inoculation methods resulted in differences in carbon allocation and physiology of extramatrical hyphae that could affect seedling establishment and survival. Seedlings inoculated by one method cannot serve as surrogates for mycorrhizal seedlings produced by other inoculation techniques.     

An in vitro method for screening Colletotrichum gloeosporioides as a biological control agent for western hemlock dwarf mistletoe

An in vitro system using detached western hemlock branches infected with dwarf mistletoe was developed to screen the virulence of five isolates of Colletotrichum gloeosporioides, a hyperparasite of dwarf mistletoe shoots and berries. Detached branches infected with dwarf mistletoe were placed in nutrient‐saturated rock‐wool blocks and mistletoe shoots were inoculated with a conidial suspension of C. gloeosporioides. One month after inoculation, lesions on mistletoe stems and berries were well developed. Infection levels for individual isolates varied from 40% to 60% of shoots and 60% to 80% of berries. Significant differences were found between the isolates and control (p = 0.02 and 0.001, respectively) while no differences were noted between the isolates for both the shoots and berries. Parallel inoculation of mistletoe shoots detached from hemlock branches on moist filter paper and in rock‐wool blocks failed because these shoots deteriorated rapidly, fragmenting into segments within a week. This in vitro test may provide an alternative method for rapid screening of potentially virulent C. gloeosporioides isolates.     

Histology of current shoots of Japanese cedar (Cryptomeria japonica D. don) inoculated withCercospora sequoiae Ellis et Everhart

The histological reaction of current shoots of the Japanese cedar,Cryptomeria japonica, inoculated withCercospora sequoiae was investigated. Wound periderm was formed on current shoots of the cedar on and after the 4th day after inoculation. In the cells of the cork layer, lignin and suberin were deposited, and a phenolic substance was accumulated. In other inoculated samples, plasmolysis, contraction of the cells and necrosis of the tissues were also observed. On the 32nd day, wound perioderm and phellogen were observed in samples of the tissue inside the necrotic tissues in response to hyphal invasion. Occurrence of wound periderm and phellogen in tissues of the needles was confirmed in the inoculated samples although no hyphae were observed in and around the tissues. These results suggest that this fungus causes necrosis of host tissues and induces defense responses. A part of this paper was presented at the 106th Annual Meeting of the Japanese Forestry Society (1995).     

黄栌与大丽轮枝菌互作的组织病理学变化

采用石蜡切片和光学显微摄像的方法,对黄栌接种大丽轮枝菌后的组织病理学变化进行系统观察.接种大丽轮枝菌后黄栌不同组织部位受侵染的程度不同,叶部最高(18.83%),而枝条最低(6.72%).菌丝首先由根的伸长区及成熟区侵入,而后出现在距根尖1～3 cm处的皮层薄壁细胞的间隙,接着进入根部的导管内,最终经茎部及枝条的导管到达叶部.大耐轮枝菌主要以菌丝形态存在于木质部导管内,且在导管中不连续出现.在病菌侵染过程中,受侵染黄栌的导管壁、薄壁细胞、皮层细胞增厚并且木质化,髓部、树脂道及其附近的分泌细胞内含物增加,薄壁细胞及木射线内有胶状物出现,导管内部产生胶状物、侵填体等物质,以此缩小导管内径或将导管堵塞,从而阻止病菌的扩展,但大部分健康的导管仍然可以维持植株正常的水分运输.虽然导管的阻塞具有致萎作用,但不足以影响到正常的生命活动.