Effect of volatile substances fromOriganum majorana andOcimum basilicum on spore respiration and germination of some soil fungi

Oxygen uptake by the spores ofFusarium moniliforme, F. oxysporum, F. semitectum, F. solani, Mucor racemosus andTrichoderma viride was increased in the presence of volatile substances extracted, fromOriganum majorana andOcimum basilicum. This increase was greater in the presence of volatile substances fromO. basilicum thanO. majorana, except in the case ofF. semitectum where the reverse was true. A drop in the RQ of all the germinating spores was observed in the presence of these substances. Volatile substances fromO. majorana reduced the spore germination ofM. racemosus whereas the spores ofT. viride were stimulated to germinate. Volatile substances fromO. basilicum stimulated the spore germination ofM. racemosus whereasT. viride spores were not affected.     

Root specific elicitation and antimicrobial activity of rosmarinic acid in hairy root cultures of Ocimum basilicum

Rosmarinic acid (RA) is a multifunctional caffeic acid ester present in sweet basil (Ocimum basilicum L.). Untransformed normal roots of O. basilicum harbored the maximum titers (0.98% g fresh weight basis) of RA compared to leaves and shoots. Hairy root cultures of O. basilicum transformed with Agrobacterium rhizogenes (ATCC-15834) showed three-fold increases in growth and RA production compared to the untransformed normal roots. Upon elicitation with fungal cell wall elicitors (CWE) from Phytophthora cinnamoni, the production of RA was enhanced 2.67-fold compared with the untreated control. Roots were induced to exude RA by fungal in situ challenge with Pythium ultimum, to our knowledge an undocumented observation. Absence of RA in the root exudates of unchallenged root cultures proves that RA under normal circumstances is not exuded. RA showed antimicrobial activity against a range of soil-borne microorganisms, with its most deleterious effects against Pseudomonas aeruginosa, an opportunistic soil bacterium and human pathogen. Confocal and scanning imaging of Aspergillus niger hyphae treated with RA (250 μM) exhibited damaged cytoskeletons with broken interseptas and convoluted cell surfaces resulting in a multinucleated stage compared to the untreated control. Both strains of P. aeruginosa tested, PAO1 and PA14, showed increased spatial division and condensation of DNA upon RA treatment compared to the untreated control. Our findings suggest that in nature RA is a constitutive antimicrobial compound that may be released into the surrounding rhizosphere upon microbe challenge.     

Effect of seed and root exudates on the growth ofXanthomonas phaseoli var.fuscans

In a medium containing bean, barley and wheat seed exudates,Xanthomonas phaseoli var.fuscans (Burk.) Starr et Burk. grew substantially better than in that containing root exudates of these plants. When the bacteria were cultivated in a medium containing root exudates of bean plants deprived of cotyledons after eleven days of growth, growth was slower than in the presence of root exudates of control plants. On the other hand, the growth was stimulated in a medium containing root exudates of bean plants deprived of leaves. It was found that seed exudates of these plants contained biologically active peptides stimulating the growth of the microorganism. These peptides were not found in root exudates. These findings suggest a relationship between the survival ofXanthomonas phaseoli var.fuscans in the rhizosphere of bean and the exudation of biologically active peptides originating from the stock substances of seeds and cotyledons.     

Positive correlation between VAM-induced changes in root exudation and mycorrhizosphere mycoflora

The influence of vesicular-arbuscular mycorrhizal (VAM) fungi on rhizosphere mycoflora was studied together with the possible mechanism involved in this process. Six combinations of VAM fungi and phosphorus fertilizer treatments were applied to Leucaena leucocephala roots and quantitative and qualitative observations were made periodically of the rhizosphere mycoflora and constituents of root exudates. The results obtained indicate that the presence of specific mycoflora in the rhizosphere of mycrorrhizal roots is mediated through root exudates rather than being an outcome of improved P nutrition.     

Effect of root exudates and extracts on rhizosphere fungi

Summary The effects of root exudates and extracts of Cassia tora L. and Crotalaria medicaginea Lamk., on some dominant rhizosphere fungi isolated from the plants were studied. Root extracts induced a high degree of stimulation in rate of growth of a majority of the fungi tested. Root exudates caused only a marginal effect which was positive on all the fungi except Trichoderma lignorum which was inhibited by the root exudate of C. medicaginea.     

Effect of root exudates on the spore germiantion of rhizosphere and rhizoplane mycoflora of chilli (Capsicum annuum L.) cultivars

Summary From root exudates of three cultivars of chilli (Capsicum annuum L.) 12 amino acids and 7 sugars were detected. Methionine, d-1- phenylalanine, citrulline and d-xylose were detected only from the root exudates of resdistant cultivars. The root exudates of resistant variety inhibited spore germination of the pathogen (Fusarium oxysporum f. sp.capsici), but that of susceptible variety enhanced spore germiantion of the same. Spore germiantion of antagonistic fungi (Trichderma viride andAspergillus sydowi) was also influenced by the root exudates of resistant and susceptible varieties, but the influence was different.Spore germiantion of a number of rhizosphere fungi was studied and in general root exudate of susceptible cultivar enhanced spore germiantion of majority of fungi, but spore germination of antagonistic fungi against the pathogen was inhibited. However, root exudate of resistant cultivar stimulated spore germination of antagonistic fungi.     

Manganese reduction in the rhizosphere of mycorrhizal and nonmycorrhizal maize

The influence of rhizosphere microorganisms and vesicular-arbuscular (VA) mycorrhiza on manganese (Mn) uptake in maize (Zea mays L. cv. Tau) plants was studied in pot experiments under controlled environmental conditions. The plants were grown for 7 weeks in sterilized calcareous soil in pots having separate compartments for growth of roots and of VA mycorrhizal fungal hyphae. The soil was left either uninoculated (control) or prior to planting was inoculated with rhizosphere microorganisms only (MO-VA) or with rhizosphere microorganisms together with a VA mycorrhizal fungus [Glomus mosseae (Nicol and Gerd.) Gerdemann and Trappe] (MO+VA). Mycorrhiza treatment did not affect shoot dry weight, but root dry weight was slightly inhibited in the MO+VA and MO-VA treatments compared with the uninoculated control. Concentrations of Mn in shoots decreased in the order MO-VA > MO+VA > control. In the rhizosphere soil, the total microbial population was higher in mycorrhizal (MO+VA) than nonmycorrhizal (MO-VA) treatments, but the proportion of Mn-reducing microbial populations was fivefold higher in the nonmycorrhizal treatment, suggesting substantial qualitative changes in rhizosphere microbial populations upon root infection with the mycorrhizal fungi. The most important microbial group taking part in the reduction of Mn was fluorescent Pseudomonas. Mycorrhizal treatment decreased not only the number of Mn reducers but also the release of Mn-solubilizing root exudates, which were collected by percolation from maize plants cultivated in plastic tubes filled with gravel quartz sand. Compared with mycorrhizal plants, the root exudates of nonmycorrhizal plants had two fold higher capacity for reduction of Mn. Therefore, changes in both rhizosphere microbial population and root exudation are probably responsible for the lower acquisition of Mn in mycorrhizal plants.     

Root exudates of plants

Summary The release of substances from wheat roots was found to be directly related to the growth of the root system. Plants whose root system did not grow released almost no exudates.When exudate concentration in the vicinity of the roots was lowered by frequent replacements of the nutrient solution or by a simultaneous cultivation of exudate-utilizing bacteria, the release of exudates was enhanced. In axenic wheat cultures, the amount of exudates during a 12-day cultivation with 2- or 4-day intervals between medium replacements represented 50% of root dry weight and 12% of whole plant dry weight.Wheat plants cultivated in the presence of the bacteriumPseudomonas putida released up to double the amount of exudates compared with axenic variants.     

Interactions between<Emphasis Type="Italic"> Trichoderma pseudokoningii</Emphasis> strains and the arbuscular mycorrhizal fungi<Emphasis Type="Italic"> Glomus mosseae</Emphasis> and<Emphasis Type="Italic"> Gigaspora rosea</Emphasis>

The interaction between Trichoderma pseudokoningii (Rifai) 511, 2212, 741A, 741B and 453 and the arbuscular mycorrhizal fungi Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe BEG12 and Gigaspora rosea Nicolson & Schenck BEG9 were studied in vitro and in greenhouse experiments. All T. pseudokoningii strains inhibited the germination of G. mosseae and Gi. rosea except the strain 453, which did not affect the germination of Gi. rosea. Soluble exudates and volatile substances produced by all T. pseudokoningii strains inhibited the spore germination of G. mosseae. The germination of Gi. rosea spores was inhibited by the soluble exudates produced by T. pseudokoningii 2212 and 511, whereas T. pseudokoningii 714A and 714B inhibited the germination of Gi. rosea spores by the production of volatile substances. The strains of T. pseudokoningii did not affect dry matter and percentage of root length colonization of soybean inoculated with G. mosseae, except T. pseudokoningii 2212, which inhibited both parameters. However, all T. pseudokoningii strains decreased the shoot dry matter and the percentage of AM root length colonization of soybean inoculated with Gi. rosea. The saprotrophic fungi tested seem to affect AM colonization of root by effects on the presymbiotic phase of the AM fungi. No influence of AM fungi on the number of CFUs of T. pseudokoningii was found. The effect of saprotrophic fungi on AM fungal development and function varied with the strain of the saprotrophic species tested.     

Chemical characterization of root exudates from rice (Oryza sativa) and their effects on the chemotactic response of endophytic bacteria

Root exudates represent an important source of nutrients for microorganisms in the rhizosphere and seem to participate in early colonization inducing chemotactic responses of rhizospheric bacteria. We characterized the root exudates collected from rice plantlets cultured under hydroponic conditions and assessed their effects on the chemotaxis of two strains of endophytic bacteria, Corynebacterium flavescens and Bacillus pumilus, collected from the rice rhizosphere. We compared these chemotactic effects on endophytic bacteria with those on two strains of plant-growth-promoting bacteria, Azospirillum brasilense (isolated from the corn rhizosphere) and Bacillus sp. (from the rice rhizosphere). The root exudates were collected at different time intervals. The highest concentration and diversity of amino acids and carbohydrates were found during the first 2 weeks after seeding. Histidine, proline, valine, alanine, and glycine were the main amino acid residues identified during the 4 weeks of culture. The main carbohydrates identified were glucose, arabinose, mannose, galactose, and glucuronic acid. The chemotactic responses of the analyzed endophytic bacteria to root exudates were 3.9 to 5.1 times higher than those of A. brasilense and 2.2 to 2.8 times higher than Bacillus sp. Our results indicate that rice exudates may induce a higher chemotactic response for endophytic bacteria than for other bacterial strains present in the rice rhizosphere.     

荞麦、高粱根系分泌物对玉米根边缘细胞和根生长的影响

植物的根系分泌物是植物根系与周围环境之间的化学媒介,通过传递特定的信息,调节根际微环境,影响周围植物的生长。玉米(Zea mays L.)和荞麦(Fagopyrum esculentum Moench)是农作物间套作体系中典型的不能搭配的组合,其障碍因素尚不清楚。以玉米为受体植物,采用根悬空培养的方法,研究了荞麦、高粱(Sorghum bicolor(L.) Moench)根系分泌物对玉米根边缘细胞和根生长的影响。结果发现,玉米根边缘细胞离体培养条件下,用荞麦根系分泌物中的小分子物质处理4、8 h显著诱导边缘细胞凋亡、死亡,细胞活率分别比对照降低了71.6%和72.3%;荞麦根系分泌物中的小分子物质对玉米根产生氧化胁迫,诱导根SOD、POD和CAT活性分别比对照高22.6%、33.9%和107.2%,根中超氧阴离子(O₂^-·)和脯氨酸含量分别比对照高33.9%和49.8%;荞麦根系分泌物中小分子物质的胁迫使根细胞膜透性增大,与对照相比升高80.0%,丙二醛(MDA)含量比对照升高31.5%;荞麦根系分泌物中小分子物质诱导根内源激素(IAA)含...     

缺锌和HCO_3~-处理对诸葛菜和油菜有机酸特征的影响

以诸葛菜和油菜为材料,水培环境下设置4个不同的缺锌和碳酸氢根离子胁迫处理,分别为+Zn0(含Zn且不加HCO3-的处理组),+Zn10(含Zn且加10 mmol·L-1HCO3-的处理组),-Zn0(缺Zn且不加HCO3-的处理组)和-Zn10(缺Zn且加10 mmol·L-1HCO3-的处理组),利用离子色谱法分析了4个处理的两种植物幼苗器官(根、茎、叶)及根系分泌物中的有机酸特征。结果表明:(1)高浓度碳酸氢根离子处理显著增加了两种植物器官及根系分泌的有机酸总量,尤其是在缺锌和高浓度碳酸氢根离子双重胁迫下(-Zn10处理),诸葛菜器官和根系分泌的有机酸比油菜更敏感,草酸、柠檬酸和苹果酸是诸葛菜器官和根系分泌物中的优势酸,这三种有机酸的含量分别占其有机酸总量的75%及以上;(2)叶片是两种植物有机酸产生的主要器官,有机酸的含量和分配比例从地上部分(叶和茎)到地下部分(根)减少;(3)两种植物器官和根系分泌物中的有机酸变化趋势一致,叶片中有机酸主要来源于暗呼吸过程和光呼吸过程,其他器官和根系分泌物中的有机酸主要来源于暗呼吸过程;(4)诸葛菜对缺锌和高浓度碳酸氢根离子的适应能力强于油菜,为诸葛菜的喀斯特适生性和低锌和高浓度碳酸氢根离子环境(如喀斯特环境)的生态修复提供了理论依据。     

Phyllosphere and rhizosphere microflora of pearl millet with reference to downy mildew incited bySclerospora graminicola (Sacc.) Schroet

Summary Analysis of phyllosphere microflora showed that in the resistant cultivar (PHB-14) there was a significantly higher population of fungi, gram positive and gram negative bacteria, compared to susceptible cultivar (NHB-3) under healthy and diseased situations. The cultivars during earhead stage supported maximum phyllosphere fungal and gram negative bacterial populations.The rhizosphere of the susceptible cultivar under downy mildew influence supported maximum fungal and gram negative bacterial populations and the rhizosphere effect was the highest in downy mildew colonized susceptible NHB-3 at the earhead stage (95 days after planting).Among the fungi isolated Cladosporium dominated the phyllosphere of resistant, susceptible and susceptible cultivar under the influence of downy mildew.Among the rhizosphere microflora the population ofAspergillus sp. was found to be greatest in resistant, susceptible and the susceptible cultivar under the influence of downy mildew.     

黑老虎内生真菌及根际土壤真菌的群落结构与生态功能分析

为探讨黑老虎(Kadsura coccinea)根际土壤和组织内生真菌菌群的组成及其生态功能，该研究采用ITS高通量测序技术对成熟黑老虎(根、茎、叶)内生真菌及根际土壤真菌群落结构、多样性和生态功能进行了分析。结果表明：(1)从12个样品中共获得2 241个可操作分类单元(OTU),涉及10门、41纲、95目、212科、367属，内生真菌(根、茎、叶)和根际土壤真菌OTU数分别为386、536、258、1 435个，其中共有的OTU为18个。在门水平上，黑老虎内生真菌及根际土壤真菌优势群落均为子囊菌门和担子菌门，其中子囊菌门在叶和茎中占比分别高达96.99%和95.37%;在属水平上，黑老虎根际土壤真菌中腐生真菌被孢霉属占比较高(为13.5%),叶和茎等生长旺盛的组织中子囊菌门未分类属和痂囊腔菌属占比较高。(2)α多样性分析结果显示，黑老虎根际土壤真菌群落的丰度和多样性明显高于内生真菌，茎中内生真菌丰度显著高于根和叶，而根、茎和叶组织间内生真菌多样性差异不显著；PCoA分析结果显示，叶和茎的真菌群落结构相似性更高。(3)利用FUNGuild数据库进行的功能预测分析结果显示，黑老虎根际土...     

Rhizosphere and phyllosphere fungi of four fern plants growing in Saudi Arabia

Seventy-eight species in addition to 1 variety of A. flavus belonging to 31 genera were collected from the rhizosphere (55 species + 1 variety and 24 genera) and phyllosphere (59 species and 22 genera) of Adiantum capillus-veneris, Ceterach officinarum, Asplenium filare and Cheilanthes catanensis.In the rhizosphere, the most common genera were Aspergillus, Penicillium, Fusarium, Mucor and yeasts. In the phyllosphere, the most prevalent fungi were Cladosporium herbarum, C. sphaerospermum, Alternaria alternata, Drechslera spicifera, Aspergillus spp., Penicillium spp., Cephalosporium roseo-griseum, Myrothecium verrucaria, Mucor racemosus and yeasts.     

Stimulated saprotrophic fungi in arable soil extend their activity to the rhizosphere and root microbiomes of crop seedlings

Saprotrophic fungi play an important role in ecosystem functioning and plant performance, but their abundance in intensively managed arable soils is low. Saprotrophic fungal biomass in arable soils can be enhanced with amendments of cellulose-rich materials. Here, we examined if sawdust-stimulated saprotrophic fungi extend their activity to the rhizosphere of crop seedlings and influence the composition and activity of other rhizosphere and root inhabitants. After growing carrot seedlings in sawdust-amended arable soil, we determined fungal and bacterial biomass and community structure in roots, rhizosphere and soil. Utilization of root exudates was assessed by stable isotope probing (SIP) following ¹³CO₂-pulse-labelling of seedlings. This was combined with analysis of lipid fatty acids (PLFA/NLFA-SIP) and nucleic acids (DNA-SIP). Sawdust-stimulated Sordariomycetes colonized the seedling's rhizosphere and roots and actively consumed root exudates. This did not reduce the abundance and activity of bacteria, yet higher proportions of α-Proteobacteria and Bacteroidia were seen. Biomass and activity of mycorrhizal fungi increased with sawdust amendments, whereas exudate consumption and root colonization by functional groups containing plant pathogens did not change. Sawdust amendment of arable soil enhanced abundance and exudate-consuming activity of saprotrophic fungi in the rhizosphere of crop seedlings and promoted potential beneficial microbial groups in root-associated microbiomes.     

健康与患病刺梨植株可培养叶际真菌菌群差异比较

本研究比较了健康与患叶斑病刺梨植株叶际真菌群落特征差异,以期探索病原菌的潜在来源,为人工构建具拮抗功能群落和刺梨叶斑病的生物防控提供参考。通过可培养方法对不同健康状况的刺梨叶际真菌进行分离培养,结合形态学和分子系统学对菌株进行综合鉴定;利用FUNGuild平台对真菌进行功能注释;结合根际、根部真菌作拆分网络分析探索病原菌的潜在来源。本研究结果表明：1）刺梨叶际真菌具有丰富的多样性。从刺梨叶际8个样品中共分离到真菌266株,其隶属于3门、6纲、13目、30科、46属中的61个种。其中,健康植株叶际内生真菌（LHE）包括8属10种27株,附生真菌（LHS）包括33属37种77株。患病叶际内生真菌（LDE）分离到7属10种38株;附生真菌（LDS）分离到31属35种124株。2）不同样品的真菌优势属和特有类群有差异。不同健康状况下叶际附生真菌的优势属均为拟盘多毛孢属Pestalotiopsis,但二者的相对多度存在差异,LHS为11.49%,LDS为32.26%;内生真菌优势属二者均为链格孢属Alternaria,但相对多度各异,LHE为33.33%,LDE为63.16%。其中,LHE特有类群为盘长孢状刺盘孢 Colletotrichum gloeosporioides和果生刺盘孢Colletotrichum fructicola等8种;LDE特有类群为茄链格孢 Alternaria solani和Didymella sinensis 等8种;LHS特有类群是草酸青霉Penicillium oxalicum和Peniophora crassitunicata等21种;LDS特有类群是尖孢镰刀菌Fusarium oxysporum、赭绿青霉Penicillium ochrochloron和易脆毛霉Mucor fragilis等19种。3）不同样品叶际真菌功能不同。经FUNGuild解析表明,LHS、LHE和LDE的叶际真菌功能群主要以腐生型为主,LDS则主要以植物病原菌群为主。本研究结果初步揭示刺梨植株健康与患叶斑病叶际间真菌多样性、群落组成及营养功能群存在差异,植株健康状况与其真菌群落特征密切相关;叶斑病病原菌主要源于刺梨叶际的附生微生物群。     

Comparison of Survival of Campylobacter jejuni in the Phyllosphere with That in the Rhizosphere of Spinach and Radish Plants

Campylobacter jejuni has been isolated previously from market produce and has caused gastroenteritis outbreaks linked to produce. We have tested the ability of this human pathogen to utilize organic compounds that are present in leaf and root exudates and to survive in the plant environment under various conditions. Carbon utilization profiles revealed that C. jejuni can utilize many organic acids and amino acids available on leaves and roots. Despite the presence of suitable substrates in the phyllosphere and the rhizosphere, C. jejuni was unable to grow on lettuce and spinach leaves and on spinach and radish roots of plants incubated at 33°C, a temperature that is conducive to its growth in vitro. However, C. jejuni was cultured from radish roots and from the spinach rhizosphere for at least 23 and 28 days, respectively, at 10°C. This enteric pathogen also persisted in the rhizosphere of spinach for prolonged periods of time at 16°C, a temperature at which many cool-season crops are grown. The decline rate constants of C. jejuni populations in the spinach and radish rhizosphere were 10- and 6-fold lower, respectively, than on healthy spinach leaves at 10°C. The enhanced survival of C. jejuni in soil and in the rhizosphere may be a significant factor in its contamination cycle in the environment and may be associated with the sporadic C. jejuni incidence and campylobacteriosis outbreaks linked to produce.     

Effects of Ozone and Sulfur Dioxide on Phyllosphere Fungi from Three Tree Species

Short-term effects of ozone (O₃) on phyllosphere fungi were studied by examining fungal populations from leaves of giant sequoia (Sequoiadendron giganteum (Lindl.) Buchholz) and California black oak (Quercus kelloggii Newb.). Chronic effects of both O₃ and sulfur dioxide (SO₂) were studied by isolating fungi from leaves of mature Valencia orange (Citrus sinensis L.) trees. In this chronic-exposure experiment, mature orange trees were fumigated in open-top chambers at the University of California, Riverside, for 4 years with filtered air, ambient air plus filtered air (1:1), ambient air, or filtered air plus SO₂ at 9.3 parts per hundred million. Populations of Alternaria alternata (Fr.) Keissler and Cladosporium cladosporioides (Fres.) de Vries, two of the four most common fungi isolated from orange leaves, were significantly reduced by chronic exposure to ambient air. In the short-term experiments, seedlings of giant sequoia or California black oak were fumigated in open-top chambers in Sequoia National Park for 9 to 11 weeks with filtered air, ambient air, or ambient air plus O₃. These short-term fumigations did not significantly affect the numbers of phyllosphere fungi. Exposure of Valencia orange trees to SO₂ at 9.3 parts per hundred million for 4 years reduced the number of phyllosphere fungi isolated by 75% compared with the number from the filtered-air treatment and reduced the Simpson diversity index value from 3.3 to 2.5. A significant chamber effect was evident since leaves of giant sequoia and California black oak located outside of chambers had more phyllosphere fungi than did seedlings within chambers. Results suggest that chronic exposure to ambient ozone or SO₂ in polluted areas can affect phyllosphere fungal communities, while short-term exposures may not significantly disturb phyllosphere fungi.     

Rhizospheric exudation of Eriophorum vaginatum L. — Potential link to methanogenesis

Root exudates are a direct link between primary production in higher plants and methanogenesis. The relationship has been widely studied on rice paddies, but less is known about its role in wetlands populated by naturally occurring species. This study provides information about the amount and composition of root exudates produced by a widespread mire plant, Eriophorum vaginatum L. For this purpose, E. vaginatum plants were grown in quartz sand in pots from April to October, and root exudates were collected once a month by percolation of the cultivation substrate. In June and October, a set of plants was labelled with ¹⁴CO₂ for two days and subsequently harvested for the determination of dry weight and for root exudates collected by the dipping method. The study supports earlier findings that natural wetland plants can enhance methanogenesis in their rhizosphere via active and seasonally varying exudation, but that the amount of exuded carbon (C) is many times lower than that delivered via litter formation. At both harvests in June and October, the proportion of incorporated radioactivity in shoots, roots and exudates was 92–96%, 4–8%, and 0.2%, respectively. New C was primarily fixed in the metabolically important carbohydrates, as well as acid anions that composed the main compounds of the new exudates. However, microbes seemed to rapidly metabolise the exudates into other substances like acetate. This was the dominant compound in the rhizoplane and rhizosphere, and it was the only detected substance that occurred in higher amounts outside the roots than inside them. Further studies in the field, including the quantification of gaseous end products, are necessary to complete our understanding of the carbon cycling in E. vaginatum-soil-microbe-system.