仙草植物内生芽胞杆菌种群多样性研究

为了研究仙草植株内生芽胞杆菌种群的多样性,利用营养琼脂培养基分离仙草根、茎、叶中的内生芽胞杆菌,采用16SrRNA序列鉴定法对分离获得的芽胞杆菌进行鉴定。同一组织内,对16SrRNA序列及形态完全一样的菌株合并冗余,用ClustalX对分离得到的菌株16SrRNA进行排序,构建Neighbor-Joining系统进化树,并计算多样性指数。实验结果显示,共分离得到58株仙草内生芽胞杆菌(根29株、茎28株、叶1株),分属于芽胞杆菌属(Bacillus)、类芽胞杆菌属(Paenibacillus)和赖氨酸芽胞杆菌属(Lysinibacillus),共3个属的11个种,其中与阿氏芽胞杆菌B.aryabhattai、简单芽胞杆菌B.simplex和B.flexus 3个种最接近的内生芽胞杆菌占总分离芽胞杆菌株数的62%;根部内生芽胞杆菌数量及种属均最丰富,其多样性指数高于茎和叶。     

鱼腥草内生微生物的分布特征初探

为了探明鱼腥草中内生菌的分布规律,便于有目的筛选有益菌株,对湖南农业大学附近一农户栽培鱼腥草不同器官中内生菌的数量分布及细菌的一般特性进行了研究.结果 表明,鱼腥草根、茎、叶中都存在大量内生菌,总的数量趋势是根大于茎,茎大于叶.,同时各器官中细菌数大于放线菌数,放线菌数大于真菌数(叶中真菌数大于放线菌数).各器官中的内生细菌以革兰氏阴性芽胞杆菌为主,其中茎表现最为明显.     

香蕉植株内生细菌群落多态性研究

采用平板法对香蕉(Musa nana)植株的内生细菌进行分离纯化,并采用细菌脂肪酸法进行鉴定。结果表明,从香蕉的健康植株和感病植株中共分离得到内生细菌21属24种。从健株分离得到9种内生细菌,其中根、茎和叶分别分离到6种、2种和8种内生细菌。从病株分离得到15属17种内生细菌,其中根、茎和叶分别分离到3种、11种和6种。香蕉健株根部的内生细菌含量最高,达5.195×106cfu g-1,下部叶片内生细菌的含量最低,仅为30 cfu g-1;香蕉病株茎部内生细菌的数量显著高于其他部位,达1.05×107cfu g-1。这说明香蕉在不同生长状态下,其内生细菌的种类和数量存在多样性。     

芽胞杆菌浸种对水稻内生细菌群落结构的影响

水稻内生细菌群落是反映植株内环境是否健康稳定的重要生物学指标,芽胞杆菌是防治水稻病害的重要生防微生物。为揭示芽胞杆菌浸种处理对水稻内生细菌群落结构的影响,采用Illumina MiSeq测序的方法对水稻内生细菌的16S rRNA基因进行测序,剖析了芽胞杆菌浸种处理对不同水稻组织内生细菌的微生态调控作用。结果表明,3种芽胞杆菌浸种处理可以提高水稻根和茎部内生细菌群落的丰富度和均匀度,降低叶部内生细菌群落的丰富度和均匀度,显著增加根部内生细菌群落多样性。变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidetes)是水稻根部和茎部共有优势菌门,厚壁菌门和芽胞杆菌属(Bacillus)是叶部共有优势菌门和属。芽胞杆菌浸种处理显著提高了叶部内生厚壁菌门和芽胞杆菌属的相对丰度,增加了根系和茎部组织内生细菌的分类单元OTU(Operational Taxonomic Units)数量,对叶部组织影响不明显;降低了茎部和叶部中参与各种代谢通路的内生细菌丰度,显著增加了根部参与代谢通路的内生细菌丰度。因此,3种芽胞杆菌浸种处理可以显著改变水稻根部、茎部和叶部内生细菌群落结构,改善水稻生长的微生态环境。     

棉花内生细菌数量动态及其对棉花黄、枯萎病菌的拮抗作用

摘要：【目的】了解棉花内生细菌数量动态,从棉花中获得拮抗黄、枯萎病菌的内生细菌资源【方法】对棉花根、茎、叶表面灭菌,采用稀释平板法分离棉花内生细菌;通过对峙培养法体外鉴定分离的棉花内生细菌对棉花黄、枯萎病菌的拮抗作用,并对拮抗棉花黄、枯萎病菌的内生细菌16S rDNA序列进行了分析【结果】棉花根中内生细菌的数量显著高于茎、叶;根的苗期总体内生细菌数量低于开花期、吐絮期,茎、叶中的内生细菌数量在不同生育期呈现一定的波动性,但趋势性不明显;6个棉花品种根中内生细菌平均数量差异并不显著,但茎、叶中内生细菌数量不同品种间呈现一定程度差异。平板对峙鉴定显示:棉花根中具有较高比例的拮抗棉花黄、枯萎病菌的内生细菌,拮抗强致病落叶型黄萎病菌（V107）的内生细菌比例不仅低于枯萎病菌（F108）,而且低于非落叶型黄萎病菌（V396）。同时拮抗棉花黄、枯萎病菌的内生细菌有44株,16S rDNA分子序列分析表明：这些拮抗内生细菌的类群包括了两个门(变形杆菌门、拟杆菌门)8个属,其中10个菌株与已报道菌株相似性<97%,可能是新的种（属）,优势种群为肠杆菌属（18株）、泛菌属（15株）。【结论】棉花的品种、生育期与器官影响棉花内生细菌数量;棉花拮抗黄、枯萎病菌的内生细菌具有优势种群,且具多样性。     

金钗石斛内生细菌的组成及多样性分析

采用9种培养基对贵州省的赤水市旺隆镇(P1)、贵阳市贵州师范大学生命科学学院大棚(P2)和金沙县台金生态农业观光园(P3)内栽培的金钗石斛(Dendrobium nobile Lindl.)根、茎和叶中的内生细菌进行了分离,并采用扩增核糖体DNA限制性分析(ARDRA)法对分离的内生细菌进行了多样性分析。结果表明:从3个样地金钗石斛的根、茎和叶中共分离出1 081株内生细菌,隶属2门5科12属40种1亚种,可分成41个OTUs;其中,芽孢杆菌属(Bacillus)和短芽孢杆菌属(Brevibacillus)为优势属,Bacillus subtilis subsp.subtilis和Brevibacillus invocatus为优势种(亚种)。金钗石斛的内生细菌在不同器官中的分布存在差异,根中内生细菌的菌株数量最多(695),且根中内生细菌的Shannon-Wiener多样性指数(2.20)和Margalef丰富度指数(5.81)均高于茎和叶中内生细菌,而叶中内生细菌的Pielou均匀度指数却最高(0.69);并且,茎和叶间内生细菌的Sorenson相似性系数最高(0.82),显著高于根和茎间以及根和叶间内生细菌的Sorenson相似性系数。3个样地金钗石斛内生细菌的共有OTUs有14个,分离菌株数占金钗石斛内生细菌分离菌株总数的96.5%;并且,3个样地金钗石斛内生细菌的Shannon-Wiener多样性指数和Pielou均匀度指数差异较小,仅P1样地金钗石斛内生细菌的Margalef丰富度指数明显高于P2和P3样地。NA、TSA、LB、KB、R2A和TB培养基从金钗石斛中分离出的内生细菌菌落数和OTU数量均高于YPD、YT和YG培养基。研究结果显示:金钗石斛体内含有丰富的内生细菌资源,并且,其内生细菌的组成和多样性与分离器官、种植地和培养基密切相关。     

番茄附生和内生细菌分离与群落相似性分析

为了溯清番茄生境以及各组织细菌群落间的相关性,以中杂302为试材,采用传统平板培养法结合16S r RNA拷贝数分析、变性梯度凝胶电泳(DGGE)分析,对番茄根区、根际土壤以及根、茎、叶片、果实、种子等组织附生和内生细菌的数量和群落相似性进行分析。结果表明,番茄根际的细菌数量最多,根区土壤次之;番茄组织附生细菌的数量显著低于根区和根际土壤中的细菌数量;组织附生细菌中,以根部最多,茎和叶片次之;组织内生细菌的数量较附生细菌少2-4个数量级,并以根内生细菌数量最多,种子表面胶状物中数量最少。DGGE细菌群落相似度分析表明,根际与组织内生样品间的相似度显著高于根区土壤,各组织内生细菌相互间的相似度最高,尤其是根、茎和叶片,以及种子、胎座、胶状物等相邻组织内生细菌间的相似度达到0.6以上。番茄组织尤其是相邻组织间的内生细菌群落结构相似,并与根际细菌密切相关。     

南方红豆杉产IAA内生芽胞杆菌的分离、鉴定及产脂肽类化合物研究

通过组织研磨法对南方红豆杉(Taxus wallichiana var. mairei)新鲜组织进行内生细菌的分离，利用16S rRNA测序技术对分离的内生细菌进行鉴定分析，使用高效液相色谱-高分辨飞行质谱测定菌液提取物IAA含量，利用HPLC-TOF-MS对提取物脂肽类化合物进行分离、鉴定，结合数据库筛选菌株产脂肽类化合物种类及含量差异。结果表明，从南方红豆杉根茎叶中共分离鉴定出内生细菌31株，筛选出产IAA菌株20株，其中编号为KLBMPTC10的芽胞杆菌属菌株产IAA含量高达210.3955 μg·L^-1。在对产IAA菌株所产脂肽类化合物的研究中发现，菌株KLBMPTC10是所有产IAA内生细菌中脂肽类化合物含量最丰富的，其产Iturin A2相对含量为26.06%，明显优于其他产IAA菌株。芽胞杆菌属菌株KLBMPTC10同时具备高产IAA和脂肽类化合物的能力，可作为研究菌株进一步开发。     

烟草内生真菌多样性和种群结构

从云南“世界烟草品种园”10种烟草的根、茎、叶中分离得到199株内生真菌,根据rDNA-ITS系统发育分析鉴定为17属25种,其中格孢腔菌目Pleosporales内生真菌的种类和数量最多;茎点霉属Phoma、链格孢属Alternaria和镰孢菌属Fusarium为主要优势属,相对频率分别为25.1%、24.6%和11.6%,优势度Y值分别为0.251、0.172和0.104。烟草不同组织内生真菌的种群结构存在显著差异,分离自根的内生真菌的主要优势属为Fusarium和Phoma,Y值分别为0.235和0.123;分离自茎的内生真菌的主要优势属为Phoma和Alternaria,Y值分别为0.186和0.155;分离自叶片的内生真菌的主要优势属为Alternaria,Y值为0.286。Phoma从烟草根茎叶中均可分离得到,而Alternaria只分布在地上部茎叶中,Fusarium只分布在根茎中,表明这3个优势属真菌对根茎叶组织的专化性不同。     

基于可培养法分析亚热带植物内生与根际芽胞杆菌种类分布异质性

【背景】芽胞杆菌是农业上重要的微生物菌剂,对植物具有促生、防病、防虫等作用。【目的】了解亚热带植物内生和根际芽胞杆菌的种群分布,为其功能挖掘提供科学依据。【方法】采用可培养手段对甘蔗(Saccharum officinarum)、红麻(Hibiscus cannabinus L.)、黄麻(Corchorus capsularis L.)、黄秋葵(Abelmoschus esculentus)和玫瑰茄(Hibiscus sabdariffa)根际土壤和根内生芽胞杆菌进行分离,利用16S rRNA基因测序对分离菌株进行分类鉴定,并分析系统发育地位。【结果】共获得了可培养芽胞杆菌菌株144株,其中根内生芽胞杆菌82株,根际土壤62株。经16S rRNA基因测序鉴定为芽胞杆菌4个属37个种,分别为芽胞杆菌属、短芽胞杆菌属、类芽胞杆菌属和赖氨酸芽胞杆菌属。芽胞杆菌菌落数量和种类在根部及其根际土壤中差异较大,根际土壤中芽胞杆菌菌落数量远远大于根部,根际土壤中芽胞杆菌菌落含量范围为(0.2-370.0)×10~5CFU/g,而根部为(0.1-81.0)×10~3 CFU/g。根部分离获得的芽胞杆菌种类远大于根际土壤中,从作物根际土壤中共获得了芽胞杆菌19个种,从根部获得内生芽胞杆菌共32个种。阿氏芽胞杆菌(Bacillus aryabhattai)、仙草芽胞杆菌(Bacillus mesonae)和假蕈状芽胞杆菌(Bacillus pseudomycoides)同时存在于4种作物的根际土壤和根部,其他芽胞杆菌种类仅存在于1种作物的根际或者根部。【结论】亚热带作物根际土壤和根部芽胞杆菌种类和数量极为丰富,而且还存在可分离培养的芽胞杆菌潜在新物种,这为了解植物与根际微生物相互作用、根际环境生态平衡提供了理论基础和科学依据。     

增强的UV—B辐射对春小麦植株化学成分、真菌定殖和分解的影响

在UV－B辐射下,叶可溶性糖含量显著降低,叶可溶性蛋白含量和粗纤维含量以及茎粗纤维含量显著增加,而根粗纤维含量没有显著变化,在生长期接受UV－B辐射的叶和茎上,赭绿青霉和黑曲霉的定殖率显著增加,康宁木霉和出芽短梗霉的定殖率明显降低,而土曲霉的定殖率未受明显影响,这些叶和茎经过60d和100d的分解,分解率均显著增加,叶分解率与粗纤维含量和可溶性蛋白含量呈显著正相关,而与可溶性糖含量呈显著负相关,茎分解率与粗纤维含量呈显著正相关,在增强的UV－B辐射下,春小麦植株化学成分的变化,真菌定殖率的改变,分解率的增加,可能会导致麦田生态系统营养周转加快,土壤库中营养贮量增加。     

Differences in growth and physiological traits of Populus cathayana populations as affected by enhanced UV-B radiation and exogenous ABA

During one growing season, the effects of enhanced ultraviolet-B (UV-B) radiation, exogenous abscisic acid (ABA) and their combination on biomass accumulation, gas exchange, endogenous ABA, the concentration of UV-absorbing compounds, antioxidant system and on the carbon (C) and nitrogen (N) content and C/N ratio were investigated in two contrasting Populus cathayana Rehd. populations, originating from high and low altitudes in south-west China. Exogenous ABA was sprayed to the leaves, and enhanced UV-B treatments were applied using a square-wave system to expose the seedlings to ambient (1×) or twice ambient (2×) doses of biologically effective UV-B radiation (UV-B_BE). Enhanced UV-B radiation significantly decreased height, basal diameter, total leaf area, total biomass, net CO₂ assimilation rate (A), stomatal conductance (g_s), transpiration rate (E) and carbon (C) content in leaves, and significantly increased the activities of superoxide dismutase (SOD) and guaiacol peroxidase (GPx), and the contents of hydrogen peroxide (H₂O₂) and malonaldehyde (MDA), as well as the accumulation of UV-absorbing compounds and endogenous ABA concentrations among different organs in both populations. In contrast, exogenous ABA induced a significant decrease in A and significant increases in the activities of SOD and GPx, in the content of H₂O₂ and MDA, and in the endogenous ABA concentrations. Compared with the low altitude population, the high altitude population was more tolerant to enhanced UV-B and exogenous ABA. Significant interactions between UV-B and ABA were observed in A, E, and in the activities of SOD and GPx, as well as in endogenous ABA in the leaves and roots of both populations. Across all treatments, the C and N contents of leaves were strongly correlated with their contents in stems and roots. Additionally, the N content of leaves and stems were significantly correlated with the C content of stems.     

增强的UV-B辐射对春小麦植株化学成分、真菌定殖和分解的影响

在UVB辐射下,叶可溶性糖含量显著降低,叶可溶性蛋白含量和粗纤维含量以及茎粗纤维含量显著增加,而根粗纤维含量没有显著变化.在生长期接受UVB辐射的叶和茎上,赭绿青霉和黑曲霉的定殖率显著增加,康宁木霉和出芽短梗霉的定殖率明显降低,而土曲霉的定殖率未受明显影响.这些叶和茎经过60d和100d的分解,分解率均显著增加.叶分解率与粗纤维含量和可溶性蛋白含量呈显著正相关,而与可溶性糖含量呈显著负相关.茎分解率与粗纤维含量呈显著正相关.在增强的UVB辐射下,春小麦植株化学成分的变化,真菌定殖率的改变,分解率的增加,可能会导致麦田生态系统营养周转加快,土壤库中营养贮量增加.     

Reduction of ambient UV-B radiation does not affect growth but may change the flowering pattern of Rosmarinus officinalis L.

The effects of sub-ambient levels of UV-B radiation on the shrub Rosmarinus officinalis L. were investigated in a field filtration experiment in which the ambient UV-B was manipulated by a combination of UV-B transmitting and UV-B absorbing filters. As a result, the plants were receiving near-ambient or drastically reduced UV-B radiation doses. Drastic reduction of UV-B radiation had no effect on mean, total and maximum stem length, number of stems per plant, dry mass of leaves, stems and roots and leaf nitrogen and phenolic contents. However, flowering was more pronounced under reduced UV-B radiation during the winter period which coincides with ascending ambient UV-B radiation. In contrast, during autumn and early winter, a period which coincides with descending ambient UV-B radiation, flowering was unaffected by reduced UV-B radiation. We can conclude that natural UV-B radiation does not affect growth of Rosmarinus officinalis, but its reduction could influence the flowering pattern of the species.     

Inhibitory effects of ambient levels of solar UV-A and UV-B radiation on growth of cucumber

The influence of solar UV-A and UV-B radiation at Beltsville, Maryland, on growth and flavonoid content in four cultivars of Cucumis sativus L. (Ashley, Poinsett, Marketmore, and Salad Bush cucumber) was examined during the summers of 1994 and 1995. Plants were grown from seed in UV exclusion chambers consisting of UV-transmitting Plexiglas, lined with Llumar to exclude UV-A and UV-B, polyester to exclude UV-B, or cellulose acetate to transmit UV-A and UV-B. Despite previously determined differences in sensitivity to supplemental UV-B radiation, all four cultivars responded similarly to UV-B exclusion treatment. After 19–21 days, the four cultivars grown in the absence of solar UV-B (polyester) had an average of 34, 55, and 40% greater biomass of leaves, stems, and roots, respectively, 27% greater stem height, and 35% greater leaf area than those grown under ambient UV-B (cellulose acetate). Plants protected from UV-A radiation as well (Llumar) showed an additional 14 and 22% average increase, respectively, in biomass of leaves and stems, and a 22 and 19% average increase, respectively, in stem elongation and leaf area over those grown under polyester. These findings demonstrate the extreme sensitivity of cucumber not only to present levels of UV-B but also to UV-A and suggest that even small changes in ozone depletion may have important biological consequences for certain plant species.     

Response of 19 cultivars of soybeans to ultraviolet-B irradiance

Nineteen soybean cultivars were grown for four weeks in controlled environmental chambers with artificial daylight supplemented by five UV-B irradiance regimes to determine the range of growth and development responses of seedlings. Data from nine plant characteristics were assessed: leaf area, dry weight of leaves, stems and roots, total plant dry weight, height, ratio of roots to shoots and leaf area to weight and rating of leaves for damage. Significant differences were observed in the responses noted. Stunting, leaf chlorosis and loss of apical dominance were three general symptoms apparent on all cultivars which received UV-B irradiance. Varying degrees of reduced leaf area and dry weight of the plants and altered ratios of weights of leaves per unit area and weight of roots to shoots were also found. It was concluded that different soybean cultivars demonstrate a marked difference in sensitivity to UV-B radiation under the artificial conditions of controlled environmental growth chambers and this may indicate a genetic basis for variability in sensitivity of soybean cultivars to this waveband. However, the sensitivity to UV-B radiation was increased by the lower than normal photon fluence of photosynthetically active radiation (225 μE m⁻² s⁻¹).     

Effects of enhanced ultraviolet-B radiation on plant nutrients and decomposition of spring wheat under field conditions

Spring wheat (Triticum aestivum) was grown in the field under ambient and supplemental levels of ultraviolet-B (UV-B, 280–315 nm) radiation to determine the potential for alteration in plant nutrients, decomposition, leaf quality and dry matter yield. Supplemental UV-B radiation simulating a 12, 20 and 25% stratospheric ozone depletion significantly decreased dry matter yield, but had no significant impact on harvest index. UV-B radiation resulted in an increase of the concentrations of N and K in all plant parts; changes of the concentrations of P, Mg, Fe and Zn varied in a tissue-dependent manner, as the decrease of P in leaves and stems, and its increase in spikes and grains. The mass of N, P, K, Mg, Fe and Zn in various plant parts and whole plant was generally decreased except leaf N mass was increased by enhanced UV-B radiation. Enhanced UV-B radiation decreased the concentrations of soluble carbohydrates in leaves and increased that of holocellulose and soluble proteins. After 60 and 100 days of decomposition of leaves and stems in the field, enhanced UV-B radiation stimulated the loss of organic C. As a consequence, the nutrient content of soils might be less diminished under enhanced UV-B radiation.     

Effects of UV-B radiation on seed yield of Glycine max and an assessment of F1 generation progeny for carryover effects

Glycine max (L.) Merr plants were grown outdoors in potted sand exposed to elevated ultraviolet-B (UV-B) radiation provided by filtered fluorescent lamps to determine the effects of UV-B on seed yield and UV-B-induced carryover effects in the F1 generation. Increased UV-B radiation had no detectable effects on reproductive parameters except for a reduction on seed number per plant and an increase in the number of unseeded pods per plant and dry weight of unseeded pods per plant in the field supplemental UV-B experiment. Studies on carryover effects in the greenhouse progeny growth trial also showed no effect of parental treatment with UV-B on biomass production, and most symbiotic-N traits and plant metabolite measured. However, the concentrations of N in nodules and starch in roots were significantly increased in the F1 generation progeny from elevated UV-B radiation relative to their F1 counterparts from ambient radiation. Assessing the effects of seed size on plant growth and symbiotic function in the F1 progeny showed that total biomass, dry matter yield of individual organs (leaves, stems, roots and nodules), total plant N and fixed-N rose with increasing seed size. Seed concentration of flavonoids was also enhanced with increasing seed size. These findings suggest that subtle changes did occur in the F1 generation progeny of parental plants exposed to elevated UV-B with potential to accumulate with further exposure to elevated UV-B radiation.     

增强的UV-B辐射对麦田生态系统中种群数量动态的影响

研究了大田栽培和自然光条件下,模拟UV-B辐射(UV-B,280～315nm)增强对麦田生态系统杂草、大型土壤动物和麦蚜种群数量动态的影响。在UV-B辐射下,杂草和大型土壤动物的种类和数量降低,物种多样性改变,杂草总生物量也降低。UV-B辐射降低麦蚜复合种群数量,并与麦叶粗纤维、可溶性蛋白、可溶性糖、Mg和Zn含量有显著的相关性。UV-B辐射还导致麦蚜与麦叶Mg、Fe和Zn含量均显著增加。     

Quantitative changes in secondary metabolites of dark-leaved willow (Salix myrsinifolia) exposed to enhanced ultraviolet-B radiation

This is a study of the impact of increased ultraviolet-B (UV-B) radiation on the secondary chemistry of Salix myrsinifolia (dark-leaved willow). For nearly two decades, the loss of stratospheric ozone above the high latitudes of the Northern Hemisphere has increased UV-B radiation (280–320 nm) over the long-term mean. Willows (Salicaceae) are widely distributed in these northern regions. To determine the effects of increased UV-B radiation on willows, the plantlets of three clones of S. myrsinifolia were grown under ambient (3.6 kJ m⁻² day⁻¹) or enhanced (7.18 kJ m⁻² day⁻¹) UV-B irradiance. After the 2-week indoor experiment, the concentrations of UV-B-screening phenolics (flavonoids and phenolic acids) and low-UV-B-screening phenolics (salicylates and condensed tannins) in fresh leaves were investigated and the biomass of leaves, stems and roots was determined. As expected, the total amount of flavonoids in willow leaves clearly increased when plantlets were exposed to higher UV-B irradiation. However, the degree of increase of individual compounds varied: luteolin-7-glucoside, monomethyl-monocoumaryl-luteolin-7-glucoside and one myricetin derivative increased significantly, while the apigenin-7-glucuronide increased only slightly. The enhanced UV-B also increased the amount of p -hydroxycinnamic acid derivative. The UV-B effects on other phenolic acids and tannins were minor. In contrast to the other phenolics, the amounts of two salicylates, salicin and saligenin, decreased under enhanced UV-B irradiation. Our results indicate that the concentrations of both UV-B-screening and low-UV-B-screening phenolic compounds in leaves of S. myrsinifolia may vary in response to elevated UV-B radiation. However, while the UV-B protective flavonoids and phenolic acids accumulate during UV-B exposure, the concentrations of certain salicylates decrease.