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。     

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。     

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.     

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     

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.     

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.     

内生真菌与高羊茅之间的共生关系

从内生真菌的涵义及其基本生物学特征出发,着重对国内外近10 年来有关内生真菌与高羊茅关系的研究情况作了综述,主要包括内生真菌对高羊茅生长发育及其抗逆性的影响、内生真菌对高羊茅植物群落及生态系统的影响等,并对内生真菌研究的实际意义及其应用前景作了总结。     

叶圈微生物的群落结构与生态功能研究进展

植物叶圈和根际是两个独特、复杂的生态系统,含有丰富的微生物种类,在促进植物生长,增强植物抗病性、抗逆性等方面有着非常重要的作用。在近几十年内的研究中,根际一直是微生物学家和植物学家研究和关注的焦点,特别是在根际微生物促进植物生长和增强植物抗病性的效果和机理方面取得了重要的研究进展。相对而言,对于叶圈微生物的研究远远滞后于对根际微生物的研究。叶圈微生物最初主要是对植物致病微生物进行研究,但是由于技术条件(主要是采用传统可培养方法进行研究)的限制,叶圈微生物研究进展缓慢。2000年以后,随分子生物学技术的发展,特别是高通量测序技术的发展,叶圈微生物研究进入了快速发展段。研究发现,叶圈微生物中只有少数致病,而大多数能够为植物提供养分、促进植物生长、提高植物抗病性和抗逆性,在植物群落组成和生态功能方面发挥了重要作用。通过介绍叶圈定义的提出及发展过程,阐述了叶圈微生物在研究方法、生物学功能及作用机理方面的研究进展,并对叶圈微生物未来发展方向进行了展望。     

金龟子绿僵菌在草原羊草和克氏针茅根际的种群动态和根内宿存鉴定

绿僵菌防治草原蝗虫效果显著,其在草原田间的存活能力影响其持续控害效果。除了侵染昆虫,绿僵菌还具有在植物根际宿存和根内生的生活方式,但相关宿存规律和内生性的研究报道很少。本文研究了绿僵菌在内蒙古草原两种优势种植物羊草和克氏针茅根际的种群数量变化并对其在这两种草的根内宿存进行了鉴定。结果表明,在干旱的内蒙古草原,绿僵菌施用后种群数量在30 d内快速下降,但能够以低密度在羊草、克氏针茅根际土壤中至少延续宿存75 d,羊草根际环境比较利于绿僵菌生存。对菌株egfp基因标记的特异PCR检测证明了绿僵菌在羊草和克氏针茅根内宿存。试验数据为指导植保生物防治中充分利用绿僵菌的昆虫病原性和植物内生特性提供理论基础,也将成为绿僵菌物种生活方式多样性、与植物互作及共进化研究的重要基础。     

中国西北地区披碱草属植物所带内生真菌的培养特征

内生真菌与植物所形成的共生体普遍存在于冷季禾草中,共生体有利于提高宿主植物的生物和非生物抗性。虽然目前研究者们已对禾草内生真菌做了大量的研究,但是尚未发现对分布于中国西北地区披碱草属植物无性世代Neotyphodium内生真菌培养特征的研究。本研究从中国西北地区分离出15株来自不同地理种群披碱草属植物无性世代Neotyphodium内生真菌。通过分离培养,测定了这15株无性世代Neotyphodium内生真菌的菌落生长速度,结果显示,这15株无性世代Neotyphodium内生真菌菌落的平均生长速度为(0.41~0.85) mm/d,且生长速度与海拔具有相关性。分布在海拔低于3000 m的无性世代Neotyphodium内生真菌的菌落生长速度比生长于海拔高于3000 m菌落的生长速度快。观察菌落发现,分布在海拔高于3000 m的菌落正面特征较为一致,而分布在海拔低于3000 m的菌落正面特征出现不同。另外,通过多基因联合建树发现,分布在海拔高于3000 m的无性世代Neotyphodium内生真菌聚在一个分支,而分布在海拔低于3000 m的无性世代Neotyphodium内生真菌呈现星状分布。由于目前尚未对中国西北地区披碱草属植物无性世代Neotyphodium内生真菌命名。因此,希望本研究结果能为该菌的命名提供理论依据。