2种杨树可培养内生细菌的多样性和季节动态变化

为了解三倍体毛白杨和健杨94树干皮层组织中内生细菌的群落多样性和季节动态变化,利用传统的细菌分离培养,结合16S rDNA序列分析等方法对其进行分析.从2种杨树干部皮层中共计分离纯化细菌菌株5 32株,分属于26属47种,2种杨树内生细菌组成存在较大差异.其中,从三倍体毛白杨中分离得到1 8属30个种,以根瘤菌属和芽孢杆菌属为优势菌群;健杨94中分离得到16属22个种,以根瘤菌属、微杆菌属为优势菌群.杨树干部皮层植物内生细菌的多样性丰富.2种杨树内生细菌数量季节变化规律总体表现为夏秋2季均高于冬春2季.2种杨树在4个季节的优势菌群各不相同,随季节变化而变化.     

盘龙参内生真菌分布特征的研究

从盘龙参[Spiranthes sinensis(Pers)Ames]的根、茎、叶中共分离得到49株内生真菌,经形态观察,鉴定为3目、4科、9属。其中曲霉属、镰刀孢属、丝核菌属为优势种群,分别占已分离菌株数的16.3%、14.3%和14.3%。不同组织部位所分离得到的内生真菌在种群及数量上都存在差异:根中的优势属为镰刀孢属,占到了根中分离菌种数的23.5%;叶中的优势属是链格孢属,占到叶中分离菌种数的26.3%;茎中的优势属为长蠕孢属,占到叶中分离菌种数的23.1%。表明盘龙参内生真菌的分布具有一定的组织专一性。     

不同生态环境下银中杨内生菌群落结构及生态位变异

【目的】研究生长在3个不同地点的银中杨根和茎中内生细菌和真菌的多样性,为植物和微生物互作研究提供参考。【方法】分别在黑龙江省大庆市林源镇常围子村、齐齐哈尔市错海林场和北京市房山区韩村河东营苗圃,取银中杨根和茎,表面消毒后,提取微生物DNA,通过16S r RNA和内部转录间隔区(ITS)扩增子Illumina MiSeq测序以确定内生细菌和真菌群落的多样性。【结果】测序结果根据97%的序列相似性水平,将细菌和真菌的reads分别归类为1 541和240个OTU。与数据库比对后确定银中杨内生细菌群落主要以放线菌纲、β-变形菌纲、α-变形菌纲、γ-变形菌纲和拟杆菌纲为主,内生真菌群落主要以座囊菌纲、伞菌纲、子囊菌纲、和银耳纲为主。α多样性和β多样性结果表明,北京、大庆和齐齐哈尔3个地点银中杨的茎内生菌群落明显聚集;而根内生菌群落表现出依赖于植物器官和生长环境的现象。Mantel检验结果表明,p H值、土壤有机质(SOM)含量和钾(K)含量与杨树根内生菌群落显著相关(P0. 05);而氮(N)、磷(P)含量并不是解释银中杨根内生菌群落差异的重要因素。由此确定不同生态环境下生长的银中杨不同器官中的核心微生物,共获得23个核心细菌OTU,归属于6个纲; 22个核心真菌OTU,归属于7个纲。还可确定7个根内生细菌指示OTU:Actinophytocola、游动放线菌属、假诺卡氏菌属、红微菌属、链霉菌属、贪噬菌属和慢生根瘤菌属; 5个茎内生细菌指示OTU:双歧杆菌属、红球菌属、小杆菌属、粪杆菌属和微球菌属。2个根内生真菌指示OTU:小球腔菌属和Ilyonectria; 3个茎内生真菌指示OTU:格孢腔目、链格孢属和Endosporium。Up Set R结果表明:内生细菌中有51(3. 30%)个OTU被6组样本共有,6个组单独特有的OTU占总OTU数的4. 54%～15. 44%;内生真菌中有1(0. 42%)个OTU被6组样本共有,6个组单独特有的OTU占了总OTU数的2. 92%～29. 17%。【结论】银中杨根内生细菌和真菌群落结构取决于栽植环境中土壤的p H值、有机质含量和钾含量。不同植物器官可代表内生菌群落的独特生态位。本研究可确定与银中杨不同器官和不同生长环境条件相关的指示OTU和核心微生物。     

内蒙古赤峰地区主要树种根际固氮菌的分离和鉴定

用阿须贝培养基对内蒙古赤峰市4个地点的5种树木根际土壤样品中的固氮菌进行分离、提纯,共获得116株固氮菌株,基于16S r DNA序列最大相似性进行了分析鉴定。结果表明:红山区落叶松根际土壤中的固氮菌共有6个属,以芽孢杆菌属(Bacillus)和类芽孢杆菌属(Paenibacillus elgii)为主;松山区樟子松根际土壤中的固氮菌共有8个属,以节细菌属(Arthrobacter)和链霉菌属(Streptomyces)为主。旺业甸落叶松根际土壤中的固氮菌共有6个属,以芽孢杆菌属(Bacillus)和根癌土壤杆菌属(Agrobacterium tumefaciens)为主;旺业甸白桦根际土壤中的固氮菌共有4个属,假单胞菌属(Pseudomonas)为主。红山区白皮松根际土壤中的固氮菌共有5个属,以假单胞菌属(Pseudomonas)和芽孢杆菌属(Bacillus)为主。新城区白皮松根际土壤中的固氮菌共有10个属,以假单胞菌属(Pseudomonas)、芽孢杆菌属(Bacillus)和类芽孢杆菌属(Paenibacillus elgii)为主。从固氮菌种群分布看,新城区白皮松根际土壤中固氮菌种群分布最为广泛,从中分离的19株固氮菌分布在10个属内;节细菌属(Arthrobacter)固氮菌分布最为广泛,存在5种树种的根际土壤中。     

外源诱导尾叶桉非培养内生细菌的激活

【目的】探究桉树非培养内生细菌的激活,为应用内生细菌防治桉树青枯病提供参考。【方法】选用恶臭假单胞杆菌WCS358r、荧光假单胞杆菌WCS374r及其嗜铁素缺失突变体JM218和Mut2,荧光假单胞杆菌WCS417r及其脂多糖缺失突变体WCS417OA-(B4)作为外源激活菌株,在限菌系统中采用尾叶桉无菌实生苗与外源菌共培养方法,外源菌作为激活因子对桉树非培养内生细菌进行激活,使植物体内非培养内生细菌恢复可培养状态,并依次利用抗利福平检测、革兰氏染色反应和分子鉴定方法对分离到的内生细菌进行鉴定。【结果】3种外源菌WCS358r、WCS374r和WCS417r分别与同源同代桉树苗共培养后,WCS358r和WCS417r处理的桉树苗中均获得被激活的细菌。WCS358r可定殖于桉树体内,且在不同部位激活出2株解淀粉芽孢杆菌;WCS417r可在桉树根、茎和叶内定殖,且在根和茎内的定殖量显著高于叶片,但不能激活桉树体内的非培养内生细菌。不同种子桉树苗经WCS358r激活后,分离到的菌株种类存在差异,分别激活出链霉菌、苏云金杆菌、解淀粉芽孢杆菌和其他芽孢杆菌,且WCS358r在根、茎内的定殖量表现...     

遮阳网和肥料种类对铁皮石斛生长的影响

主要研究了遮阳网和肥料种类对铁皮石斛（Dendrobium candidum）生长的影响,选取了株高、茎长、茎粗、叶片数、顶叶叶长、叶宽等6个方面作为铁皮石斛的生长指标。结果表明：5—9月是铁皮石斛茎长的主要生长期,其他月份铁皮石斛的茎长基本处于停止生长的状态,因此,在铁皮石斛大棚栽培过程中,为了提高铁皮石斛的产量和节省种植成本,提高经济效益,可以选择在这段时间进行追施有机肥。研究还表明遮盖1层2针遮阳网对于提高铁皮石斛的茎长效果最好,且追施豆饼肥和花生肥对铁皮石斛的茎长有明显的促进作用,从而可以有效地提高铁皮石斛的产量。     

入侵植物黄顶菊内生细菌对苹果轮纹病菌的拮抗筛选

以衡水湖外来入侵植物黄顶菊Flaveria bidentis为材料,对其不同部位内生细菌分离、纯化,用分离到的内生菌与苹果轮纹病菌Botryosphaeria dothidea进行对峙培养。结果表明:从黄顶菊根、茎、叶、花中分别分离到4,4,4,2株共计14株内生细菌;对峙培养筛选出5株(茎1、茎2、茎4、花1、花2)对苹果轮纹病菌抑制作用明显的细菌,5菌株之间的抑菌差异性不显著;菌株茎4抑制效果最好,真菌菌落直径为(39.96±2.66)mm,抑菌率为33.21%。     

贺兰山地区油松根际固氮菌的多样性研究

对贺兰山北寺自然保护区油松(Pinus tabulaeformis)的根际土壤固氮菌多样性和群落结构进行研究。采用阿须贝培养基对土壤中的固氮菌进行分离和纯化,用通用引物27f/1492r对固氮菌进行16S r DNA扩增并测序,应用DNAMAN 6.0软件进行多序列比对,用MEGA 4.0软件构建聚类树。结果表明:从贺兰山北寺油松根际土壤中分离出的固氮菌属于假单胞菌属(Pseudomonas)、芽孢杆菌属(Bacillus)、叶杆菌属(Phyllobacterium)、节杆菌属(Arthrobacter)、根瘤菌属(Rhizobium)、类芽孢杆菌属(Paenibacillus)、鞘氨醇单胞菌属(Sphingomonas)、柄细菌属(Caulobacter)、土地杆菌属(Pedobacter)等9个不同的类群,其中假单胞菌属(Pseudomonas)、芽孢杆菌属(Bacillus)、叶杆菌属(Phyllobacterium)、节杆菌属(Arthrobacter)是优势类群。这表明贺兰山北寺地区油松根际土壤中固氮菌具有较丰富的多样性。     

狭叶柴胡内生真菌分布特征的研究

从狭叶柴胡(Bupleurum scorzonerifolium Willd.)中共分离得到13株内生真菌,经菌落形态观察及显微观察,鉴定为3目、4科、7属,并未见明显的优势菌群,且没有一个属能够同时从根、茎、叶三种组织中分离得到,表明不同菌属在宿主体内分布存在一定的组织特异性。研究为狭叶柴胡内生真菌的进一步开发奠定了理论与实践基础。     

近野生栽培黔产铁皮石斛生长及品质对不同附生树种的响应

【目的】筛选出适宜贵州兴义地区的铁皮石斛优良附生树种,指导当地铁皮石斛近野生栽培。【方法】在兴义市典型喀斯特次生林内开展了依托9个附生树种实施的铁皮石斛近野生栽培试验,以附生林下石灰岩作为对比,并基于铁皮石斛主要的生长和品质指标对不同附生树种进行了对比评价。【结果】铁皮石斛的新分蘗株（新芽）在5月以后不再增加,茎长、茎粗、叶片数量在大部分附生树种上于5月（盛花期）、7月（生长高峰期）、9月（生长末期）均表现为持续显著增长态势。生长末期,铁皮石斛的新芽数量在不同处理间没有显著差异,茎长在梨、枇杷上表现最好,茎粗在石灰岩、枇杷、李上表现最优,叶片数量在石灰岩、枇杷上相对更多,株丛花朵数量在清香木、天竺桂上相对较少。附生李树的铁皮石斛总黄酮含量显著高于其他处理,附生盐麸木、李、南酸枣以及石灰岩的铁皮石斛总多糖含量显著优于其他处理,总生物碱含量表现较优的是附生李、枇杷、天竺桂处理。根据平均隶属函数值由高到低排序,各附生各树种及石灰岩处理依次为李（0.944）、石灰岩（0.596）、南酸枣（0.558）、枇杷（0.550）、梨（0.498）、盐麸木（0.464）、大叶冬青（0.394）、柏木（0...     

Isolation and identification of endophytic bacteria with plant growth promoting and biocontrol potential from oak trees

Samples of roots, leaves and stems of healthy oak trees were collected from various locations in the Baneh and Marivan regions, Iran. In total, 63 endophytic bacteria were isolated and grouped according to phenotypic properties. Seven selected isolates were further identified by partial sequencing of the 16S rRNA gene. Isolates Pp54, Pp88, Pp95 and Pp177 were Pseudomonas spp., isolates Sm59 and Sm79 were Stenotrophomonas spp., and isolate Bf 172 was a Bacillus sp. The ability of these isolates for plant hormone production such as auxin and gibberellin was evaluated, along with siderophore production, phosphate solubilization, atmospheric nitrogen fixation, protease and hydrogen cyanide production. All strains produced auxin and gibberellin in different amounts. Atmospheric nitrogen fixation ability was positive for strains Sm79, Pp54, Pp88 and Pp95. All strains except Sm79 solubilized phosphate. Strains Sm59, SM79, Pp88, Pp177 and Bf172 produced protease. Pp88, Pp95, Bf172 and Pp177 were able to produce siderophore. Strains Sm79 and Pp95 released low concentrations of hydrogen cyanide. Amongst the strains tested, Pp95, Pp88, Pp177 and Sm79 had different inhibitory effects on the bacterial plant pathogen, Pseudomonas syringae pv. syringae under in vitro conditions. This is the first reported case of endophytic Bacillus, Pseudomonas and Stenotrophomonas strains from oak trees in Iran.     

Non-mycorrhizal fungal endophytes in two orchids of Kaiga forest (Western Ghats), India

We used standard isolation protocols to explore the endophytic fungal communities in three tissue types of two dominant orchids (Bulbophyllum neilgherrense and Vanda testacea) of the Kaiga forest ofthe Western Ghats. We surface sterilized and assessed 90 segments ofeach orchid for the occurrence and diversity of endophytic fungal taxa.The 118 fungal isolates were obtained from root, bulb and leaves of B.neilgherrense, consisting of 17 anamorphic taxa (range, 10 15 taxa) with 1.3 fungal taxa per segment (range, 1.2 1.4 taxa). Four taxa (Aspergillus flavus, A. niger, Penicillium sp. and morpho sp. 1) belonged to the coregroup (11.1% 32.2%). The relative abundance of A. flavus and morpho sp.1 was more than 10%. A total of 130 fungal isolates from roots, stems and leaves of V. testacea yielded 20 anamorphic taxa (range, 11 15 taxa)with 1.4 fungal taxa per segment (range, 1.4 1.5 taxa). Aspergillus flavus,A. niger, A. ochraceus, Gliocladium viride, Penicillium sp. and morphosp. 1 belonged to the core group. Relative abundance exceeded 10% for A. flavus, A. niger, and morpho sp. 1. The Simpson and Shannon diversity indices were higher in leaf than root or bulb/stem of both orchids.Jaccard’s similarity coefficient was higher between root and leaf in both orchids (56.3% 60%) than between other pairs. Our study revealed thatthe endophytic fungal assemblage and diversity of B. neilgherrense and V.testacea of Kaiga forest of the Western Ghats were relatively similar between orchids and their tissues.     

Characterization of endophytic bacteria with plant growth promotion and biological control potential isolated from walnut trees

In this study, a total of 68 endophytic bacteria were isolated from different tissues of walnut trees. About 55% and 22% of the strains had the ability to produce indole acetic acid and gibberellic acid, respectively. The capability of isolates to solubilize phosphate, growth on N‐free medium, siderophore, protease and lipase production was varied. Based on phenotypic grouping and plant growth promotion properties, twelve isolates were selected and 16S rRNA gene‐based phylogenetic analysis revealed that strains showed 99%–100% similarity to Pseudomonas, Bacillus, Arthrobacter, Roseomonas and Streptomyces genera. Amongst the selected strains, PS12, KS54, JS66 and KS74 showed root and shoot growth enhancement of poplar cutting. NS70, KS54 and PL36 strains showed antagonistic activity against Pseudomonas syringae pv. syringae;RR47, KS74 and NR69 strains had inhibition effects against Brenneria nigrifluens; and JS66 and RR26 strains had antagonistic activity against both phytopathogens under in vitro conditions. This is the first reported study to elucidate the endophytic bacterial diversity associated with walnut trees with beneficial attributes.     

赤峰地区杨树人工林碳储量研究

以赤峰市杨树人工林为研究对象,对不同立地、不同林龄、不同林种杨树生物量、碳储量进行研究,结果表明:杨树人工林生物量随着树龄、胸径、树高的增长而增加,各个器官的生物量也不相同,干的生物量最大,叶的生物量最小,生物量依次顺序为:干根枝叶。树干的平均碳含量最大,为471.00 g/kg,根为461.14 g/kg,枝为468.00 g/kg,叶最小,为446.20 g/kg,不同器官的含碳率大小排序为树干树根树枝树叶。杨树人工林胸径与单株碳储量,树高、胸径与生物量、单株碳储量模型均符合乘幂模型,模型拟合率均大于80%。杨树人工林乔木层平均碳储量为37.783 t/hm2,赤峰地区杨树人工林乔木层碳储总量为26 539 627.38 t。     

Isolation of endophytic streptomycetes from above- and belowground organs of Quercus serrata

The occurrence and localization of endophytic actinomycetes within diverse organs of host plants provide ecological information that can be used to evaluate the significance of their spatial habitats. The aim of this study was to isolate and characterize endophytic actinomycetes in different organs of Quercus serrata. For this purpose, actinomycete isolates were obtained from surface-sterilized tissues of both symptomless seedlings and aged trees of Q. serrata and rhizosphere soil of the sampled seedlings. Thirty-five isolates with the ability to sporulate, including 4 from leaves of the aged trees, 10, 6, and 15 from leaves, stems, and roots of the seedlings, respectively, and 8 soil-derived isolates, were selected and characterized. The 16S rDNA nucleotide sequence analyses revealed that all of them belonged to the genus Streptomyces. According to a neighbor-joining phylogenetic tree constructed based on the results, the isolates of plant origin were divided into three major clades with high bootstrap values of 98 or 99 %, whereas eight soil-derived isolates were located at different positions from those of the endophyte isolates. Moreover, two larger clades were formed, one of which contained isolates derived only from aboveground parts, while the other contained isolates from all of the organs. These results suggest that the endophytic streptomycetes in Q. serrata may differ in their habitat positions (i.e., either above- or belowground parts).     

Phillyrin produced by Colletotrichum gloeosporioides, an endophytic fungus isolated from Forsythia suspensa

To investigate the phillyrin-producing endophytic fungi from the medicinal plant Forsythia suspensa, a total of 24 strains of endophytic fungi were isolated from the healthy stems, leaves and fruits of the plant, 9 from stems, 5 from leaves, and 10 from fruits respectively. All fungal isolates were fermented in liquid PDA medium and their extracts were preliminary analyzed by TLC. One isolated strain G10, which was from the fruit of F. suspensa, had the same R_f value as authentic phillyrin. The potential phillyrin-producing fungus G10 was further analyzed by HPLC and HPLC-MS, and the results showed that the isolate G10 possessed of a retention time and ion peaks identical with the authentic compound phillyrin. The isolate G10 was identified as Colletotrichum gloeosporioides based on the morphological characteristics and ITS sequence analysis. The current research indicates that the numerous endophytic fungi inside the medicinal plants are precious resource for the pharmaceutical natural products that are originally from the plants.     

杨树冰核细菌溃疡病寄主树栖细菌种群相互拮抗关系的研究

“杨树树栖细菌种群”的构成成员之中有引起树木病害的病原细菌。而病原细菌在引起杨树病害时，并非单独起作用，而是有与其他伴生微生物发生着种群演替、协同或排斥等过程，也可能长期潜伏而不致病。     

Fungal endophytes associated with Sonneratia (Sonneratiaceae) mangrove plants on the south coast of China

The aim of this study was to describe endophytic fungal communities residing in five mangrove species (Sonneratia caseolaris, Sonneratia hainanensis, Sonneratia ovata, Sonneratia Paracaseolaris, Sonneratia apetala) collected at the south coast of China. The colonization frequency (CF) of endophytes in roots, stems and leaves was measured and the structures of endophytic communities were investigated, using endophytic fungal diversity indices and similarity coefficients. We found high biodiversity and tissue specificity of endophytic fungi. A total of 391 fungal isolates were identified using morphological and molecular methods. We recovered a total of 39 distinct endophytic species, of which Cytospora, Diaporthe, Fusarium, Glomerella, Mycosphaerella, Phoma, Phomopsis and Stemphylium were the dominant fungal taxa. Most recovered species were found on more than one host species but the dominant species differed according to host and tissue type.     

杉木人工林各植物组分含碳率研究

分别采集会同县广坪林区第2代杉木人工林的杉木树干(去皮)、树皮、枝、叶、根系样品和灌木的干、枝(叶)和根样品与草本植物的地上、地下部分样品以及林内的枝、叶、果和碎屑凋落物和枯死根,用干烧法测定其含碳率。结果表明:杉木各器官含碳率大小的顺序是:树皮>树叶>树根>树干>树枝。树皮、树叶、树干、树枝含碳率随着树木年龄的增大而增加,树根含碳率随着树木年龄增大出现波动。林冠下草本植物的平均碳含量比木本植物低,且草本植物间碳含量差异要比木本植物间大。林龄对凋落物同种组分碳含量影响不显著。不同凋落物的碳含量即使在同一龄级也存在较大差异。枯死根系的碳含量要低于地上凋落物各组分的碳含量,林木各器官活有机体内的碳含量均大于相应死有机体(凋落物)内的碳含量。