植物内生木霉菌研究进展

内生菌在植物体内广泛存在,内生木霉菌是常见的植物内生真菌,一些内生木霉菌能够通过拮抗、重寄生、产生次生代谢产物、诱导植物系统抗性等方式促进植物生长、提高植物的抗逆性和抗病性,是重要的生防微生物资源,受到广泛关注。本文综述了近年来内生木霉菌的种类、分布、功能、生防作用机理,以及内生木霉菌与宿主植物互作的分子机制等方面的最新研究进展,为促进内生木霉菌作为生防因子的研究和利用提供参考。     

花生内生菌的种群及动态分析

１９９４－１９９７年,先后从山东省主要花生产区监沂、烟台等地采集约１０００株不同类型的花生无病 ,进行了植株内主要微生物类群分析。结果表明,不同品种花生的内生菌都表细菌和真菌,未分离到放线菌和酵母菌,细菌以芽孢杆菌（Ｂａｃｉｌｌｕｓｓｐｐ．）黄单孢菌（Ｘａｎｔｈｏｍｏｎａｓｓｐｐ．）和欧文氏菌（Ｅｒｗｉｎｉａｓｐｐ．）为优势种群。其中芽孢杆菌属以巨大芽孢杆菌（Ｂ．ｍｅｇａｔｅｒｉｕｍ）、蜡样芽孢杆     

Diversity of cultivable endophytic bacteria associated with halophytes in Xinjiang of China and their plant beneficial traits

Endophytic bacteria from halophytes have a wide range of application prospects in various fields, such as plant growth-promoting, biocontrol activity and stress resistance. The current study aimed to identify cultivable endophytic bacteria associated with halophytes grown in the salt-affected soil in Xinjiang Uygur Autonomous Region, China and to evaluate their plant beneficial traits and enzyme-producing activity. Endophytic bacteria were isolated from Reaumuria soongorica (PalL Maxim.), Artemisia carvifolia (Buch.-Ham. ex Roxb. Hort. Beng.), Peganum harmala L. and Suaeda dendroides (C. A. Mey. Moq.) by using the cultural-dependent method. Then we classified these bacteria based on the difference between their sequences of 16S rRNA (16S ribosomal RNA) gene. Results showed that the isolated bacteria from R. soongorica belonged to the genera Brucella, Bacillus and Variovorax. The bacteria from A. carvifolia belonged to the genera Micromonospora and Brucella. The bacteria from P. harmala belonged to the genera Paramesorhizobium, Bacillus and Peribacillus. The bacteria from S. dendroides belonged to the genus Bacillus. Notably, the genus Bacillus was detected in the three above plants, indicating that Bacillus is a common taxon of endophytic bacteria in halophytes. And, our results found that about 37.50% of the tested strains showed strong protease-producing activity, 6.25% of the tested strains showed strong cellulase-producing activity and 12.50% of the tested strains showed moderate lipase-producing activity. Besides, all isolated strains were positive for IAA (3-Indoleacetic acid) production, 31.25% of isolated strains exhibited a moderate phosphate solubilization activity and 50.00% of isolated strains exhibited a weak siderophore production activity. Our findings suggest that halophytes are valuable resources for identifying microbes with the ability to increase host plant growth and health in salt-affected soils.     

Endophytic colonization of white jute (Corchorus capsularis) plants by different Beauveria bassiana strains for managing stem weevil (Apion corchori)

Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales), an entomopathogenic fungus, was introduced through seed inoculation with spore suspension as an endophyte in white jute (Corchorus capsularis L.), a bast fiber crop. Out of nine B. bassiana strains, seven, viz., ITCC 6063, ITCC 4512, ITCC 4563, ITCC 5562, ITCC 4796, ITCC 5408 and ITCC 4705, became established as endophytes. The endophytic colonization was initially detected by cultivation on selective medium. Colonization was confirmed by polymerase chain reaction (PCR) using sequence characterized amplified region (SCAR) marker. Endophytic B. bassiana strains colonized in leaves of all the plants grown from treated seeds. However, the colonization frequency varied among the strains. The highest colonization frequency (70.09%) was recorded in ITCC 6063 followed by ITCC 5562 (67.67%) and ITCC 5408 (64.17%); ITCC 4512 exhibited the lowest (42.54%) colonization. ITCC 4925 and ITCC 4644 did not show any colonization. Endophytic colonization of B. bassiana reduced stem weevil infestation under pot culture. ITCC 5408 and ITCC 6063 were found most efficient, with only 10.44% and 14.06% infested plants, respectively.     

Endophytic bacteria associated with halophyte Seidlitzia rosmarinus Ehrenb. ex Boiss. from saline soil of Uzbekistan and their plant beneficial traits

Endophytic bacteria of halophytic plants play essential roles in salt stress tolerance. Therefore, an understanding of the true nature of plant-microbe interactions under extreme conditions is essential. The current study aimed to identify cultivable endophytic bacteria associated with the roots and shoots of Seidlitzia rosmarinus Ehrenb. ex Boiss. grown in the salt-affected soil in Uzbekistan and to evaluate their plant beneficial traits related to plant growth stimulation and stress tolerance. Bacteria were isolated from the roots and the shoots of S. rosmarinus using culture-dependent techniques and identified by the 16S rRNA gene. RFLP (Restriction Fragment Length Polymorphism) analysis was conducted to eliminate similar isolates. Results showed that the isolates from the roots of S. rosmarinus belonged to the genera Rothia, Kocuria, Pseudomonas, Staphylococcus, Paenibacillus and Brevibacterium. The bacterial isolates from the shoots of S. rosmarinus belonged to the genera Staphylococcus, Rothia, Stenotrophomonas, Brevibacterium, Halomonas, Planococcus, Planomicrobium and Pseudomonas, which differed from those of the roots. Notably, Staphylococcus, Rothia and Brevibacterium were detected in both roots and shoots, indicating possible migration of some species from roots to shoots. The root-associated bacteria showed higher levels of IAA (indole-3-acetic acid) synthesis compared with those isolated from the shoots, as well as the higher production of ACC (1-aminocyclopropane-1-carboxylate) deaminase. Our findings suggest that halophytic plants are valuable sources for the selection of microbes with a potential to improve plant fitness under saline soils.     

Are roots special? Nematodes have their say

Nematodes are ubiquitous and cosmopolitan parasites of vascular plants, causing substantial crop damage. Although various species exploit all parts of the plant, roots are the major target. Nematodes deploy a broad spectrum of feeding strategies, ranging from simple grazing to the establishment of complex cellular structures (including galls) in host tissues. Various models of feeding site formation have been proposed, and a rôle for phytohormones has long been speculated. Based on recent molecular evidence we present several scenarios involving phytohormones in the induction of giant cells by root-knot nematodes. The origin of parasitism by nematodes, and the rôle of horizontal gene transfer from microbes is discussed. Throughout, parallels with aphid-plant interactions are emphasized.     

植物之间互作效应及其生理机制

在分析不同植物互作效应的基础上,就互作效应在农林复合系统中对植物体内养分、土壤养分有效性及植物生物学产量等的影响进行了概述,(1)互作促进了植物根系生长,使植物根系表现出较强的竞争作用,也有利于根系间的养分竞争;(2)植物之间的他感作用普遍存在于自然生态系统及栽培生态系统中,在植物资源配置时应充分利用这种相生相克作用;(3)互作促使植物体内的养分转移,改善体内的养分状况,达到植物之间养分互补;(4)互作可以有效地提高土壤肥力,改善土壤的养分状况和根际养分吸收环境,维持植物生产力;(5)互作能更有效地利用光、热、水、土和养分等资源,并减轻病虫危害,表现出明显的产量优势。在豆科与非豆科植物互作中,豆科植物的生物固氮作用为其互作植物提供了氮素营养保证,这种形式互作的产量优势通常在缺氮情况下表现更为明显。     

植物内生真菌及其展望

内生真菌在自然界广泛存在,有些内生真菌可以促进植物营养吸收、提高植物抗生物逆境和非生物逆境的能力、刺激植物产生活性物质等。因此,内生真菌是植物病虫害生物防治领域的重要研究内容之一。本文就内生真菌与寄主共生的分子基础、内生真菌对植物影响机制、内生真菌与寄主特异性相互作用、特殊的内生真菌--植物病原真菌、内生真菌形成的分子机制和内生真菌研究展望等6个方面对内生真菌进行了叙述,期望为科学地利用内生真菌资源提供一些参考。     

植物病害生物防治与微生态学

在概述植物病害生物防治生态学意义的基础上,从植物微生态学的角度阐述了植物根围菌及内生菌作为生防菌的依据和意义。特别是内生菌是植物体内的正常菌群,对植物病害生物防治具有重要的生态学实践意义。     

高抗虫玉米内生真菌对玉米蚜生长、繁殖及取食选择的影响

植物内生菌普遍存在于健康植物组织或器官内,可通过自身的代谢产物或信号传导作用促进植物生长、增强宿主植物抗病虫性、抗胁迫性等。为明确前期筛选出的高抗虫玉米内生真菌YC1、GX5、YB8、YB15对玉米蚜生长繁殖的影响,室内测定了玉米蚜取食不同菌株种子处理的玉米后存活、繁殖以及对接种不同菌株玉米的取食选择。结果显示,取食菌株处理玉米后,玉米蚜产雌历期显著低于对照;产雌量、净增殖率、世代平均寿命、内禀增长力、周限增长率均低于对照,但差异不显著。取食选择试验结果表明,选择YC1、GX5、YB8、YB15处理组玉米的玉米蚜比例分别为17.52%、19.02%、19.34%和18.84%,均显著低于对照组25.28%。说明这4株玉米内生真菌对玉米蚜的生长繁殖有不利影响,且对玉米蚜有趋避作用。     

Ecological weed management by cover cropping: effects on weed growth in autumn and weed establishment in spring

Cover crops grown in the period between two main crops have potential as an important component of a system‐oriented ecological weed management strategy. In late summer and autumn, the cover crop can suppress growth and seed production of weeds, whereas the incorporation of cover crop residues in spring may reduce or retard weed emergence. Based on these two criteria, six cover crop species were evaluated for their weed suppressive potential in 2 years of experimentation in the Netherlands. Fodder radish, winter oilseed rape and winter rye had the strongest competitive ability in autumn; the competitive strength of Italian ryegrass was intermediate and white lupin and lucerne were poor competitors. Competitiveness was strongly correlated to early light interception. Surprisingly, doubling the recommended sowing density did not increase weed suppressive ability. Although a poor competitor in the fall, after incorporation in spring, lucerne had the strongest inhibitory effect on seedling establishment, followed by winter oilseed rape and white lupin. Winter rye and fodder radish did not affect seedling establishment, whereas Italian ryegrass was not evaluated because of re‐growth after incorporation. Competition in autumn and subsequent residue‐mediated suppression of weed establishment in spring varied among the cover crop species, with winter oilseed rape offering relatively strong effects during both periods.     

堆肥茶生防作用影响因素及应用前景探讨

农业有机废弃物堆肥化处理是目前常用的资源化再循环技术，所得的堆肥产品兼具营养、促生和抗病作用，是促进“减肥减药”工作的重要产品。将腐熟的堆肥加水进行二次好氧或厌氧发酵制备的堆肥茶，可最大化地提取堆肥中功能性物质和养分，并促进有益微生物菌群的扩增繁殖，表现出更好的抗病、促生和营养功能。同时，由于堆肥茶是液态生防制品可以通过喷灌、滴灌等技术进行根层滴灌或叶面喷施，实现药肥一体的作用。本文旨在通过总结堆肥茶抗病效果、作用机理及影响其抗病效果的制备工艺、施用方法，分析其在生态农业中的应用价值，以期为后续开展应用技术研究提供理论依据。     

柠檬桉内生真菌的分离及其提取物的抗细菌活性

采用组织块分离法分离柠檬桉中的内生真菌,通过菌落观察和分子生物学相结合的方法对分离到的内生真菌进行鉴定;进一步采用薄层层析-生物自显影法测定了柠檬桉内生真菌次生代谢产物的抗细菌活性。从柠檬桉中共分离鉴定得到10株内生真菌,主要分布于附球菌属Epicoccum、镰刀菌属Fusarium、炭疽菌属Colletotrichum、Rhytidhysteron、半壳霉属Leptostroma、Campylocarpon、Neofusicoccum、白腐菌属Phlebia和Dinemasporium等9个属中。抗细菌活性的测定结果表明,内生真菌Ecf-4(Rhytidhysteron sp.,MK211261)和Ecf-1(Epicoccum sp.,MK211258)表现出最强的抑菌活性,两菌对桉树青枯病菌的抑菌斑最大直径均大于10 mm;依据其R_f值的范围和抑菌斑的最大直径,内生真菌Ecf-4的抗菌活性高于Ecf-1,表现出更强的抗细菌活性,可作为候选菌株,进一步分离和鉴定其中的活性成分。     

Endophytic Beauveria bassiana activates expression of defence genes in grapevine and prevents infections by grapevine downy mildew Plasmopara viticola

Fungal entomopathogens like Beauveria bassiana (Ascomycota: Hypocreales) are known as antagonists of insects with multiple functional and ecological roles, and have attracted increased attention as biocontrol agents in integrated pest management programmes. For some crop plants, it has been proven that endophytic B. bassiana, besides its entomopathogenic habit, can provide protection against plant pathogens or limit their damaging effects. However, for grapevine, limited knowledge is available on the influence of endophytic B. bassiana on fungal pathogens and about the mechanisms underlying putative protection effects. This study assessed the protective potential of endophytic B. bassiana against grapevine downy mildew Plasmopara viticola in greenhouse experiments. Three and seven days after a B. bassiana treatment, potted grapevine plants were inoculated with P. viticola and the evolving disease severity was assessed. Disease severity was significantly reduced in B. bassiana-treated plants compared to control plants, depending on the age of leaves. Furthermore, a microarray and an RT-qPCR analysis were performed to work out fundamental aspects of genes involved in the interaction between grapevine and B. bassiana. The results indicate an up-regulation of diverse defence-related genes in grapevine as a response to endophytic establishment of B. bassiana. Thus, endophytic establishment of an entomopathogenic fungus such as B. bassiana in grapevine plants would represent an alternative and sustainable plant protection strategy, with the potential for reducing pesticide applications in viticulture.     

Development of herbicide resistance in weeds in a crop rotation with acetolactate synthase‐tolerant sugar beets under varying selection pressure

The development of acetolactate synthase (ALS) tolerant sugar beet provides new opportunities for weed control in sugar beet cultivation. The system consists of an ALS?inhibiting herbicide (foramsulfuron + thiencarbazone‐methyl) and a herbicide‐tolerant sugar beet variety. Previously, the use of ALS‐inhibitors in sugar beet was limited due to the susceptibility of the crop to active ingredients from this mode of action. The postulated benefits of cultivation of the ALS‐tolerant sugar beet are associated with potential risks. Up to now, with no relevant proportion of herbicide‐tolerant crops in Germany, ALS‐inhibitors are used in many different crops. An additional use in sugar beet cultivation could increase the selection pressure for ALS‐resistant weeds. To evaluate the impact of varying intensity of ALS‐inhibitor use on two weed species (Alopecurus myosuroides and Tripleurospermum perforatum) in a crop rotation, field trials were conducted in Germany in two locations from 2014 to 2017. Weed densities, genetic resistance background and crop yields were annually assessed. The results indicate that it is possible to control ALS‐resistant weeds with an adapted herbicide strategy in a crop rotation including herbicide‐tolerant sugar beet. According to the weed density and species, the herbicide strategy must be extended to graminicide treatment in sugar beet, and a residual herbicide must be used in winter wheat. The spread of resistant biotypes in our experiments could not be attributed to the integration of herbicide‐tolerant cultivars, although the application of ALS‐inhibitors promoted the development of resistant weed populations. Annual use of ALS‐inhibitors resulted in significant high weed densities and caused seriously yield losses. Genetic analysis of surviving weed plants confirmed the selection of ALS‐resistant biotypes.     

Influence of canopy structure on sheath blight epidemics in rice

Sheath blight (caused by Rhizoctonia solani) is one of the most important constraints in achieving high grain yield in intensive rice production systems. Canopy structure can influence the development of sheath blight epidemics. The objective of this study was to determine the effect of canopy structure parameters such as shoot number, leaf area index, biomass production, contact frequency, light transmittance and plant height on the development of sheath blight epidemics in commercial fields. Field experiments were conducted in both early and late seasons of 2009 and 2010 in Wuxue, Hubei province, China. The effects of nitrogen (N) rate and hill density on structure and production parameters and sheath blight severity were investigated. Sheath blight severity was recorded as a sheath blight index or relative lesion height on inoculated and uninoculated plants in each crop. Lesion length was measured on inoculated plants in 2010. The results showed that the sheath blight index increased with an increase of N rate and hill density in uninoculated plots in each trial. Stepwise multiple regression analysis demonstrated that contact frequency was consistently related to sheath blight. Lesion length on inoculated plants was not correlated with canopy structure. These results indicate that canopy structure influences sheath blight epidemics. A ‘healthy’ canopy resulting from appropriate crop management practices can be used to suppress sheath blight epidemics in rice.     

银杏内生真菌No.1028代谢产物的抗真菌活性及其培养生物学特性

采用无菌技术从银杏茎中分离到一株内生真菌(No.1028),经形态分类学和rDNAITS序列分析研究鉴定为黑孢属Nigrosporasp.,菌丝生长速率法测定结果表明,其代谢产物对重要作物真菌病害病原菌具有明显的抑制作用。离体条件下内生真菌发酵液对番茄早疫病菌、黄瓜枯萎病菌、菜豆炭疽病菌、葡萄炭疽病菌、水稻纹枯病菌、黄瓜立枯病菌均有较强的抑制作用,对上述不同病原菌的抑制率分别为66.7%、48.3%、64.6%、36.5%、57.1%和23.0%。经提取分离得到纯的白色晶体状活性物质。同时也研究了不同碳源、氮源、无机离子对内生真菌No.1028生物学特性和生长的影响,发现玉米粉、黄豆饼粉比较适合其生长和抑菌代谢产物的合成,Na 对抑菌代谢产物的合成有较明显的促进作用。     

Relation between plant nutrition, hormones, insecticide applications, bacterial endophytes, and Candidatus Liberibacter Ct values in citrus trees infected with Huanglongbing

Intensive insecticide and nutrient management have been attempted worldwide to reduce citrus huanglongbing (HLB) symptom development and yield loss. However, effects of insecticide and nutrient applications on HLB have been poorly understood. Leaf nutrients, jasmonic and salicylic acid contents, cycle threshold (Ct) values of Ca. Liberibacter asiaticus (Las), and community structure of endophytic α-proteobacteria were evaluated after insecticide treatment, ‘nutrition’ treatment (including systemic resistance inducing agents), or both in comparison with a control in a two-factor field experiment in 2008–2012. Leaf N, Mn, Zn and B significantly increased whilst Cu decreased after nutrient applications. Salicylic acid significantly increased in old leaves treated with insecticides, nutrients or both, and in young leaves treated with nutrients only. The jasmonic acid concentration was highest after the nutrition treatment in both old and young leaves. Ct values of Las and leaf area and weight significantly increased after long-term nutrient applications in 2011 and/or 2012. Redundancy analysis of the endophytic α-proteobacteria community structure indicated that the communities were mainly separated according to nutrient applications, which were positively associated with Ct values of Las and Ca, Mn, Zn, B, Mg, and Fe contents in leaf samples collected in 2012. Thus, effects of insecticides on HLB were significant in the early 2-year period whilst nutrients had significant effects on Las content and leaf size and weight after at least 3 years of application.     

Plant-virus interactions and the agro-ecological interface

As a result of human activities, an ever-increasing portion of Earth’s natural landscapes now lie adjacent to agricultural lands. This border between wild and agricultural communities represents an agro-ecological interface, which may be populated with crop plants, weeds of crop systems, and non-crop plants that vary from exotic to native in origin. Plant viruses are important components of the agro-ecological interface because of their ubiquity, dispersal by arthropod vectors, and ability to colonize both crop and wild species. Here we provide an overview of research on plant-virus dynamics across this interface and suggest three research priorities: (1) an increased effort to identify and describe plant virus diversity and distribution in its entirety across agricultural and ecological boundaries; (2) multi-scale studies of virus transmission to develop predictive power in estimating virus propagation across landscapes; and (3) quantitative evaluation of the influence of viruses on plant fitness and populations in environmental contexts beyond crop fields. We close by emphasizing that agro-ecological interfaces are dynamic, influenced by the human-mediated redistribution of plants, vectors, and viruses around the world, climate change, and the development of new crops. Consideration of virus interactions within these environmentally complex systems promises new insight into virus, plant, and vector dynamics from molecular mechanisms to ecological consequences.     

Endophytic bacterial communities in the periderm of potato tubers and their potential to improve resistance to soil-borne plant pathogens

To evaluate whether the location of bacterial endophyte communities contributes to disease resistance in potato tubers ( Solanum tuberosum ), the population density, biodiversity and antibiotic activity of endophytic bacteria was examined from the tuber peel (periderm plus top 3 mm of tissue) of four cultivars (Russet Burbank, Kennebec, Butte and Shepody). There were no significant differences for population density of bacteria among the layers of peel examined and no cultivar × peel layer interaction. Endophytic bacteria from several layers of peel were challenged in in vitro bioassays to the soil-borne plant pathogens Fusarium sambucinum , Fusarium avenaceum , Fusarium oxysporum and Phytophthora infestans (mating types A1 and A2). In general, antibiosis of bacterial endophytes against these pathogens was significantly higher ( P  = 0.01) in isolates recovered from the outermost layer of tuber peel and decreased progressively toward the centre of the tuber. Antibiosis against P. infestans was variable, with a progressive decrease in antibiotic activity from outer to inner layers of peel occurring in cvs Russet Burbank and Kennebec only. For antibiosis there were significant cultivar × peel, and cultivar × pathogen interactions ( P  = 0.01). In all cases the inhibitory activity of endophytic bacteria was significantly greater ( P  = 0.01) against the A1 than the A2 mating type of P. infestans . In four of seven cases, where the same species of bacteria were recovered from all three peel layers, antibiosis to pathogens decreased significantly ( P  = 0.01) with depth of recovery (from the periderm to inside the tuber), indicating that in certain communities of endophytic bacteria, defence against pathogens may be related to bacterial adaptation to location within a host and may be tissue-type and tissue-site specific.