核桃细菌性疫病菌特性及其与核桃内生菌的竞争作用

为了明确本实验室前期分离得到的一株核桃细菌性疫病菌Xaj DW3F3的相关特性及其与寄主内生菌的竞争能力。对该病原细菌进行了显微观察、生物膜形成测定，不同培养基上多糖和色素含量，以及18项生理生化指标测定。通过毛细管法和滴落法观察其趋化作用，并测试其与寄主内生菌的竞争能力。结果表明，病原菌DW3F3为杆状、极生鞭毛、有运动性并能形成生物膜。其在YPGA、King’B 和NA培养基上生长较好，产生多糖和色素较多。病原菌有较强的耐盐性，致死温度达60℃，对纤维素和淀粉都有较强的分解能力，对NaCl和寄主叶片表现出一定的趋性。在其与核桃叶部内生菌的共培养中表现出一定竞争优势。该病菌菌DW3F3有树生黄单胞菌核桃致病变种（Xanthomonas arboricola pv.juglandis，Xaj）的基本特征及致病相关特性，这些特征可能与其致病性强及病害难以控制有关。     

Root‐associated microbes in sustainable agriculture: models,metabolites and mechanisms

Since the discovery of penicillin in 1928 and throughout the ‘age of antibiotics’ from the 1940s until the 1980s, the detection of novel antibiotics was restricted by lack of knowledge about the distribution and ecology of antibiotic producers in nature. The discovery that a phenazine compound produced by Pseudomonas bacteria could suppress soilborne plant pathogens, and its recovery from rhizosphere soil in 1990, provided the first incontrovertible evidence that natural metabolites could control plant pathogens in the environment and opened a new era in biological control by root‐associated rhizobacteria. More recently, the advent of genomics, the availability of highly sensitive bioanalytical instrumentation, and the discovery of protective endophytes have accelerated progress toward overcoming many of the impediments that until now have limited the exploitation of beneficial plant‐associated microbes to enhance agricultural sustainability. Here, we present key developments that have established the importance of these microbes in the control of pathogens, discuss concepts resulting from the exploration of classical model systems, and highlight advances emerging from ongoing investigations. © 2019 Society of Chemical Industry     

青海高原禾草内生真菌资源的调查和分离

青海高原的高寒特性决定其生境内禾草植物的多样性和特异性,然而有关青海高原地区禾草内生真菌的研究报道较少,本研究对青海高原地区48个样点内禾草植物内生真菌带菌率和内生真菌分离进行了研究.结果表明,14种禾本科植物内生真菌平均感染率低于50％,而12种植物内生真菌平均感染率大于50％.其中,缘毛鹅观草(Roegneria pendulina)、紫花针茅(Stipa purpurea)、醉马草(Achnatherum inebrians)、中华羊茅(Festuca sinensis)、硬质早熟禾(Poa sphondylodes)、微药羊茅(F.nitidula)、毛稃羊茅(F.kiritowii)和紫羊茅(F.rubra) 的平均带菌率均超过80％.同时,同属禾草植物内生真菌带菌率存在差异.禾草内生真菌分离率较低,其中紫花针茅、麦宾草(El ymus tangutorum)、光稃早熟禾(P.psiolepis)和异针茅(S.aliena)内生真菌在菌落和分生孢子形态上存在差异.可见,青海高原禾草内生真菌带菌率较高,同时不同禾草内生真菌可能存在多样性.     

Characterization of Botryosphaeriaceae from plantation‐grown Eucalyptus species in South China

The Botryosphaeriaceae is a species‐rich family that includes pathogens of a wide variety of trees, including Eucalyptus species. Symptoms typical of infection by the Botryosphaeriaceae have recently been observed in Eucalyptus plantations in South China. The aim of this study was to identify the Botryosphaeriaceae associated with these symptoms. Isolates were collected from branch cankers and senescent twigs of different Eucalyptus spp. All isolates resembling Botryosphaeriaceae were separated into groups based on conidial morphology. Initial identifications were made using PCR‐RFLP fingerprinting, by digesting the ITS region of the rDNA operon with the restriction enzymes CfoI and KspI. Furthermore, to distinguish isolates in the Neofusicoccum parvum/N. ribis complex, a locus (BotF15) previously shown to define these species, was amplified and restricted with CfoI. Selected isolates were then identified using comparisons of DNA sequence data for the ITS rDNA and translation elongation factor 1‐alpha (TEF‐1α) gene regions. Based on anamorph morphology and DNA sequence comparisons, five species were identified: Lasiodiplodia pseudotheobromae, L. theobromae, Neofusicoccum parvum, N. ribis sensu lato and one undescribed taxon, for which the name Fusicoccum fabicercianum sp. nov. is provided. Isolates of all species gave rise to lesions on the stems of an E. grandis clone in a glasshouse inoculation trial and on the stems of five Eucalyptus genotypes inoculated in the field, where L. pseudotheobromae and L. theobromae were most pathogenic. The five Eucalyptus genotypes differed in their susceptibility to the Botryosphaeriaceae species suggesting that breeding and selection offers opportunity for disease avoidance in the future.     

海南粗榧内生真菌F127发酵液的次级代谢产物研究

对海南粗榧(Cephalotaxus hainanensis Li.)内生真菌F127发酵液化学组分进行研究。用多种色谱技术对F127代谢产物进行分离纯化,以LC-MS、ESI-MS和超导核磁共振分析鉴定化合物结构,最后采用MTT法测定化合物的抗肿瘤活性。从海南粗榧内生真菌F127的发酵液中分离得到5个单体化合物,分别鉴定为oblongolide T(1)、sorbicillin(2)、邻苯二甲酸二丁酯(3)、phomopsolide B(4)、6,8-二羟基-3-甲基-3,4-二氢异香豆素(5),且化合物1、4、5对肿瘤细胞K562、NB4、HL-60、Hep G-2和Lovo表现出不同抑制活性。化合物4对5株肿瘤细胞均表现出抑制活性,对K562、NB4、HL-60抑制效果显著,IC50值分别为3.35、0.014和0.16μg/m L,对Hep G-2和Lo Vo只有温和抑制作用;化合物1对K562、NB4、HL-60抑制作用良好,IC50值分别为51.82、54.25和29.31μg/m L;化合物5对NB4和Hep G-2则具有一定抑制活性;化合物2和3对测试细胞株未表现出抑制活性。5个化合物均是首次从海南粗榧内生真菌中分离得到,并首次报道了化合物1、4对K562、NB4、HL-60的优良细胞毒活性,为进一步研究海南粗榧内生真菌中的活性天然产物奠定了基础。     

Interaction of Cd-hyperaccumulator Solanum nigrum L. and functional endophyte Pseudomonas sp. Lk9 on soil heavy metals uptake

Bioaugmentation is a promising method for assisting phytoextraction of heavy metals from contaminated soil, and the development of bioaugmentation-assisted phytoextraction requires the understanding of the mechanism involved in the interaction between plants and inocula. In this study, a pot study was conducted to evaluate the effect of bacterial endophyte Pseudomonas sp. Lk9 which can produce biosurfactants, siderophores and organic acids on the growth and metal uptake of Cd-hyperaccumulator Solanum nigrum L. growing in multi-metal-contaminated soil. The results revealed that Lk9 inoculation could improve soil Fe and P mineral nutrition supplies, enhance soil heavy metal availability, and affect host-mediated low-molecular-weight organic acids secretion, thereby significantly increasing S. nigrum shoot dry biomass by 14% and the total of Cd by 46.6%, Zn by 16.4% and Cu by 16.0% accumulated in aerial parts, compared to those of non-inoculated control. The assessment of phytoextraction showed that Lk9 inoculation elevated the bioaccumulation factor of Cd (28.9%) and phytoextraction rates of all metals (17.4%, 48.6% and 104.6% for Cd, Zn and Cu, respectively), while the translocation factors had negligible difference between Lk9 inoculation (3.30, 0.50 and 0.40 for Cd, Zn and Cu, respectively) and non-inoculated control (2.95, 0.53 and 0.42 for Cd, Zn and Cu, respectively). It was also found that the symbiotic association between S. nigrum and Lk9 significantly increased the soil microbial biomass C by 39.2% and acid phosphatase activity by 28.6% compared to those in S. nigrum without Lk9. This study would provide a new insight into the bioaugmentation-assisted phytoextraction of heavy metal-contaminated soils.     

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调查了中国西北和东部地区不同生境的拂子茅属Calamagrostis植物中Neotyphodium属内生真菌的分布。采样范围包括新疆、青海、甘肃、江苏、安徽、浙江、上海7省（市、区）,共采集拂子茅属植物样品426株。调查发现西北3省（区）所有276个拂子茅属植物样品中都不含有内生真菌;而采集自东部的样品中,南京的无性系拂子茅属植物和黄山的拂子茅C. epigeios含菌率为100%。但是,采自南京抽穗的拂子茅却完全不含内生菌。通过研究分离菌株的形态学特征,发现南京与黄山的菌株都具有Neotyphodium属内生真菌的典型特征,而它们之间在菌落形态、生长速度、孢子大小等方面又存在较明显差异。以上结果说明我国拂子茅属植物及其中的禾本科植物内生真菌具有一定的生物多样性,拂子茅属植物中的内生真菌含量受地理环境以及宿主植物种类的影响十分明显。     

内生真菌对高寒草地紫花针茅凋落物分解的影响

禾草内生真菌对于提高宿主植物对生物和非生物胁迫的耐性发挥着积极作用,然而,禾草内生真菌共生体凋落物分解对于草地生态系统养分循环的影响仍不清楚。本研究以紫花针茅为研究对象,比较带内生真菌（E+）与不带内生真菌（E-）植株凋落物分解过程中重量及全氮、木质素和纤维素含量,以及木质素∶N、纤维素∶N与全氮、木质素和纤维素残留率的变化,以期揭示内生真菌在紫花针茅凋落物分解过程中发挥的作用。主要结果如下：E+紫花针茅凋落物分解速率高于E-,具有更短的分解周期;随着时间的延长,紫花针茅凋落物木质素含量和木质素∶N由E+显著高于E-逐渐变为二者之间差异不显著,纤维素含量、纤维素∶N和纤维素残留率则逐渐变为E+显著低于E-;另外,随着分解时间延长,E+和E-凋落物全氮含量呈上升趋势,而全氮残留率呈现出下降-上升-下降的趋势,且后期E+显著低于E-。因此,内生真菌不同程度地促进了紫花针茅凋落物全氮、木质素和纤维素的降解。     

红树林内生真菌1403的形态与分子鉴定(英文)

红树林内生真菌1403是从中国南海分离的一株镰刀菌,在液体发酵条件下能够产生灰黄霉素和葸醌类的抗生素。基于1403在形态学方面具有以下特征:产小型分生孢子的分生孢子梗较长,小型分生孢子呈椭圆形,大型分生孢子线形或略微弯曲,本文将它与有类似形态特征的镰刀菌进行了比较,发现1403与植物病原菌Fusarium verticillioides(有性态为Gibberella moniliforme)表面上很相似,气生菌丝均可产生大量假头状或链状的小型分生孢子,但深入的研究却发现1403与F.verticillioides具有不同的性状,如产生的多生分生孢子梗有时与单生分生孢子梗长度相当,产生环状菌丝,以及菌丝发生融合等现象,进一步以最大简约法和贝叶斯分析法将1403与其它相似镰刀菌进行系统发育比较,综合18S、28S、ITS三水平上的分析结果可以将1403归于镰刀菌亚组Gibberella fujikuroi,并最终在EF—1α水平上将其定为Fusarium proliferatum.     

First report of an endophyte (Diaporthe phaseolorum var. sojae) from Kandelia candel

Mangrove endophytic fungus 1893 was isolated from Kandelia candel from an estuarine mangrove on the South China Sea Coast. Two new lactones 1893A and 1893B, together with other known compounds, have been isolated from its fermentation broth. To classify the endophyte correctly for further industrial application, a combination of morphological and molecular techniques was used to approach its identity. The endophyte was compared with similar species having trichogynes or trichogyne-like hyphae which apparently fused with antheridium-like hyphae, and perithecia initials developing from an ascogonial coil surrounded by enveloping hyphae in early developmental stages on pure culture. Further morphological characteristics on host and non-host were used for comparison with similar species when the endophyte was cultivated on leaves of Kandelia candel and Mangifera indica, respectively, which resulted in classifying the endophyte as a Phomopsis species. The ITS sequence of rDNA was used to infer its phylogenetic relationships with Phomopsis species that resembled the strain in morphology or ecology. Finally, the endophyte was identified as Diaporthe phaseolorum var. sojae based on morphological and molecular evidence. Our study is a first report of Diaporthe phaseolorum var. sojae isolated from mangrove Kandelia candel. Foundation item: The search was supported partly by the Guangzhou Natural Science Foundation (Grant No. 2007Z3-EO581), the Guangdong Provincial Natural Science Foundation (Grant No. 2007A0200300001-7), the Chinese High-Tech 863 Project (Grant No. 2006AA09Z422), and the National Natural Science Foundation of China (Grant No. 20572136). The first two authors have the same contribution.