Synergistic interactions between the ACC deaminase-producing bacterium Pseudomonas putida UW4 and the AM fungus Gigaspora rosea positively affect cucumber plant growth

Bacteria producing 1-aminocyclopropane-1-carboxylate (ACC) deaminase modulate plant ethylene levels. Decreased ethylene levels increase plant tolerance to environmental stresses and promote legume nodulation. On the contrary, the role of ethylene in mycorrhizal symbiosis establishment is still controversial. In this work, the ACC deaminase-producing strain Pseudomonas putida UW4 AcdS+ and its mutant AcdS(-), impaired in ACC deaminase synthesis, were inoculated alone or in combination with the AM fungus Gigaspora rosea on cucumber. Mycorrhizal and bacterial colonization as well as plant growth and morphometric parameters were measured. The influence of each microorganism on the photosynthetic efficiency was evaluated on the second and fourth leaf. The strain AcdS+, but not the AcdS(-) mutant, increased AM colonization and arbuscule abundance. The mycorrhizal fungus, but not the bacterial strains, promoted plant growth. However, the AcdS+ strain, inoculated with G. rosea, induced synergistic effects on plant biomass, total root length and total leaf projected area. Finally, the photosynthetic performance index was increased by the strain UW4 AcdS+ inoculated in combination with G. rosea BEG9. These results suggest a key role of this enzyme in the establishment and development of AM symbiosis.     

微生物帮助烟草抗旱的机理及其应用

干旱是中国烟草种植业面临的较为严重的非生物胁迫.很多与植物共生或联合的根际微生物能帮助植物避旱和耐旱.微生物能通过菌丝吸水并转运到植物,通过产生植物激素或改变植物内源激素的平衡来促进根发育和伸长,或诱导叶片关闭气孔,促进根吸水和减少叶片散失水分来避旱.微生物能通过调整不同激素介导的信号通路,诱导植物产生系统抗逆性,促进植物细胞产生渗透保护剂、抗氧化物和活性氧清除剂而耐旱.微生物还能帮助植物吸收营养,以支持植物在干旱胁迫下的代谢和生长.本文关注丛枝菌根真菌、模式内生真菌印度梨形孢和根际促植物生长细菌帮助烟草和番茄等植物抗旱的机理,探讨如何在烟草育苗和栽培中应用有益微生物来帮助烟草抗旱.     

Phylogeny of the 1-aminocyclopropane-1-carboxylic acid deaminase-encoding gene acdS in phytobeneficial and pathogenic Proteobacteria and relation with strain biogeography

Deamination of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) is a key plant-beneficial trait found in plant growth-promoting rhizobacteria (PGPR) and phytosymbiotic bacteria, but the diversity of the corresponding gene (acdS) is poorly documented. Here, acdS sequences were obtained by screening putative ACC deaminase sequences listed in databases, based on phylogenetic properties and key residues. In addition, acdS was sought in 71 proteobacterial strains by PCR amplification and/or hybridization using colony dot blots. The presence of acdS was confirmed in established AcdS+ bacteria and evidenced noticeably in Azospirillum (previously reported as AcdS-), in 10 species of Burkholderia and six Burkholderia cepacia genomovars (which included PGPR, phytopathogens and opportunistic human pathogens), and in five Agrobacterium genomovars. The occurrence of acdS in true and opportunistic pathogens raises new questions concerning their ecology in plant-associated habitats. Many (but not all) acdS+ bacteria displayed ACC deaminase activity in vitro, including two Burkholderia clinical isolates. Phylogenetic analysis of partial acdS and deduced AcdS sequences evidenced three main phylogenetic clusters, each gathering pathogens and plant-beneficial strains of contrasting geographic and habitat origins. The acdS phylogenetic tree was only partly congruent with the rrs tree. Two clusters gathered both Betaprotobacteria and Gammaproteobacteria, suggesting extensive horizontal transfers of acdS, noticeably between plant-associated Proteobacteria.     

Effect of plant growth promoting rhizobacteria containing ACC-deaminase on maize (Zea mays L.) growth under axenic conditions and on nodulation in mung bean (Vigna radiata L.)

AIMS: This study was conducted to test the hypothesis that the bacterial strains possessing 1-aminocyclopropane-1-carboxylic acid (ACC)-deaminase activity may also promote growth of inoculated plants and could increase nodulation in legumes upon co-inoculation with rhizobia. METHODS AND RESULTS: Several rhizobacteria were isolated from maize rhizosphere through enrichment on ACC as a sole N source. Purified isolates were screened for growth promotion in maize under axenic conditions and for in vitro ACC-deaminase activity. A significant positive correlation was observed between in vitro ACC-deaminase activity of bacterial cells and root elongation. None of the isolates produced auxins. Bradyrhizobium japonicum produced less amount of auxins but did not carry ACC-deaminase activity. Results of pot experiment revealed that co-inoculation with Bradyrhizobium and plant growth promoting rhizobacteria (PGPR) isolates enhanced the nodulation in mung bean compared with inoculation with Bradyrhizobium alone. CONCLUSIONS: It is highly expected that inoculation with rhizobacteria containing ACC-deaminase hydrolysed endogenous ACC into ammonia and alpha-ketobutyrate instead of ethylene. Consequently, root and shoot growth as well as nodulation were promoted. SIGNIFICANCE AND IMPACT OF THE STUDY: The ACC-deaminase trait could be employed as an efficient tool to screen effective PGPR, which could be successfully used as biofertilizers to increase the growth of inoculated plants as well as nodulation in legumes.     

The impact of Pseudomonas putida UW3 and UW4 strains on photosynthetic activities of selected field crops under saline conditions

Abstract

This research was aimed to assess the photosynthetic activities of barley (Hordeum valgare L.), clover (Trifolium repens L.), and pearl millet (Pennisetum glaucum (L.) R. Br.) under different saline conditions with two strains of Pseudomonas putida (UW3 and UW4) treatments. An exceptional observation was revealed on barley biomass ratio (288.8%) that irrigated with brackish saline water (10,000?mg/L) with the presence of P. putida UW4 strain. In general, P. putida UW3 strain was significantly increased crops biomass ratio (249.4%, 202.1%, and 212.5%) for barley, pearl millet, and clover, respectively, which were irrigated with 10,000?mg/L brackish saline water. Plant root and shoot systems were significantly increased in their length and weight reflecting the improvement of plants’ photosynthetic activities under salt stress conditions with the presence of P. putida strains. The results from pulse amplitude modulation fluorometry showed that the plants were recovered from the saline stress effect once P. putida strains were applied. The outcome of this research was highly recommended to apply P. putida strains (UW3 and UW4) with field crops for phytoremediation, in particular, where salinity (soil and/or brackish water) was environmentally challenging.     

Effects of arbuscular mycorrhizal fungi on tree growth, leaf water potential, and levels of 1-aminocyclopropane-1-carboxylic acid and ethylene in the roots of papaya under water-stress conditions

 Seedlings of papaya (Carica papaya L. var. Solo) were transplanted to pots with or without an arbuscular mycorrhizal (AM) fungus (Gigaspora margarita Becker and Hall). After 3 months, half the plants were subjected to water stress by withdrawing irrigation. The leaf water potential (LWP) was measured during 20 days of water-stress treatment and then the plants were harvested. Root ethylene and 1-aminocyclopropane-1-carboxylic acid (ACC) concentrations were measured and plant fresh weight determined. The LWP decreased during the water-stress treatment and this decrease was more severe in the non-AM plants. Plant fresh weight was higher for AM than non-AM plants under both conditions. Under well-irrigated conditions, the ethylene concentration in the roots was increased by the presence of AM, although there was no significant difference between AM and non-AM roots in ACC levels. ACC increased in both AM and non-AM roots under water-stress conditions. The water-stress treatment resulted in a marked increase in ethylene concentration in non-AM roots but the concentration in AM roots was slightly lower than under normal conditions. Accepted: 7 July 2000     

Application of bio-fertilizers for enhancing growth and yield of common bean plants grown under water stress conditions

This study was planned to enhance the growth and productivity of common bean plants (Phaseolus vulgaris L.) grown under different water stress level by using different microorganisms as bio-fertilizer agents. Water stress is a international problem that effects on morphological, functional and chemical processes of plants occasioning in altering growth, yield and water relations of economic plants like common bean plants. The interaction effect between water stress (WW as recommended irrigation after 6 days, WS1 after 12 days and WS2 after 18 days) and inoculation with different microorganisms [AMF (Glomus mosseae) and endophytic bacteria, (Bacillus amyloliquefaciens)] used alone or in mixed was examined on the development and productivity of common bean plants. Mutual application of AMF and endophytic bacteria significantly increased the average values of most of growth, water relations (photosynthetic rate, transpiration rate and stomatal conductance) and yield parameters of common bean plants grown at WS1 and WS2 comparing with non-colonized plants. In this connection, colonization with AMF and endophytic bacteria with WS1 are the greater pods number, pod length, pods weight, 100 seeds weight, Yield by ton /Fed and water-use efficiency (WUE) by ton/ m³ than other treatments. Common bean yielded seeds had significantly increased nutrients content (nitrogen, potassium, phosphorus, magnesium and calcium), vitamin B₁, Folic acid, crude protein and crude fibers at AMF + endophytic bacteria under second water stress (WS1) when compared to other treatments.     

The plant growth‐promoting fungus Fusarium equiseti and the arbuscular mycorrhizal fungus Glomus mosseae stimulate plant growth and reduce severity of anthracnose and damping‐off diseases in cucumber (Cucumis sativus) seedlings

To alleviate the environmental contamination due to persistent chemical usage, approaches to integrated pest management were conceived. In this perspective, microbe–microbe interactions such as mycorrhizal relationships with other soil microbiota in the rhizosphere like the plant growth‐promoting fungi (PGPF) are particularly important. Better understanding of the interactions between beneficial microbial groups is imperative in the identification of possible synergistic or antagonistic effects to improve their practical usage as biocontrol agents or biofertilizers. In this study, the consequence of co‐inoculation of the arbuscular mycorrhizal fungus (AMF) Glomus mosseae (Gm) and the PGPF Fusarium equiseti (isolates GF18‐3 and GF19‐1) in terms of plant growth enhancement, root and rhizosphere colonisation, and development of anthracnose (Colletotrichum orbiculare) and damping‐off (Rhizoctonia solani AG‐4) diseases in cucumber plants was investigated under controlled conditions. The amendment of either GF18‐3 or GF19‐1 singly or in combination with Gm indicated a general tendency to significantly enhance the shoot dry weight (SDW) of cucumber plants at 4 weeks after planting (WAP). Similarly, Gm alone significantly enhanced SDW at 4 WAP. Gm showed a tendency to depress root colonisation by F. equiseti but such antagonistic effect was not observed in the rhizosphere soil. Both GF18‐3 and GF19‐1 significantly reduced percent root colonisation of Gm. However, these general tendencies may vary with the inoculum densities of AMF and PGPF. Both F. equiseti and Gm inoculated singly significantly increased percent of protection against anthracnose, but the combined inoculation was more effective in controlling the disease compared to single inoculation. The inoculation of the cucumber seedlings with GF18‐3, GF19‐1 or Gm, 6 or 12 days prior to damping‐off pathogen inoculation, increased percent of protection against damping‐off disease. This study shows that the co‐inoculation of F. equiseti and Gm resulted in additive effect on the suppression of anthracnose disease in cucumber.     

丛枝菌根真菌与根围促生细菌相互作用的效应与机制

丛枝菌根(arbuscular mycorrhiza,AM)真菌是植物活体营养专性共生菌,广泛存在于陆地各生态系统中.研究表明,AM真菌与根围促生细菌(plant growth promoting rhizobacteria,PGPR)之间的相互作用,尤其是它们之间的协同作用不仅影响植物养分吸收利用、病原物发生发展、土壤理化特性与生物修复等,而且对于可持续农、林、牧业生产、稳定生态系统都具有十分重要的意义.因此,近年来给予众多关注和研究.综述了AM真菌与PGPR之间的相互影响及其可能的作用机制,以及AM真菌与PGPR协同改善植物营养和生长、协同抑制病原菌、协同修复土壤方面的作用,旨在总结AM真菌与PGPR相互作用的效应与机制方面的最新研究进展,为今后研究发展提供依据.     

丛枝菌根真菌对盐胁迫下黄瓜植株生长、果实产量和品质的影响

采用有机基质栽培,选用盐敏感黄瓜品种‘津春2号’为试验材料,研究了丛枝菌根真菌(AMF)对盐胁迫下黄瓜植株生长、矿质营养吸收、果实品质和产量的影响.结果表明：接种AMF可以有效促进黄瓜植株生长和对矿质营养的吸收,提高果实产量和改善蔬菜营养品质;盐胁迫下,黄瓜生长受到抑制,植株体内N、P、K、Cu、Zn含量减少和K⁺/Na⁺降低,果实产量和可溶性蛋白、总糖、Vc、硝酸盐含量下降;接种AMF可缓解盐胁迫对黄瓜生长的抑制作用,使植株体内N、P、K、Cu和Zn含量分别比对照提高7.3%、11.7%、28.2%、13.5%和9.9%,K⁺/Na⁺、果实产量、可溶性蛋白、总糖、Vc含量明显提高,果实硝酸盐含量显著降低.表明AMF可通过促进盐胁迫下黄瓜植株对矿质营养的吸收,促进植株生长,增强植株对盐胁迫的耐性,进而提高其产量和改善营养品质.     

模拟岩溶旱钙土壤基质中AM真菌对玉米幼苗光合生长的影响

利用盆栽试验,探讨了AM真菌在模拟岩溶区干旱、高钙及其双重胁迫的土壤基质中对玉米幼苗光合生长的影响。结果表明:玉米幼苗的菌根侵染率在不同处理下的大小顺序为对照干旱双重胁迫高钙。无论接种与否,干旱、高钙及其双重胁迫均导致玉米幼苗生物量、净光合速率下降。未接种AM真菌条件下,玉米幼苗生物量在干旱、高钙及其双重胁迫下较对照分别低3.2%、63.7%、76.0%,净光合速率较对照分别低33.4%、86.9%、98.8%;接种AM真菌条件下,玉米幼苗生物量在干旱、高钙及其双重胁迫下较对照分别低16.3%、78.4%、80.2%,净光合速率较对照分别低9.7%、92.8%、91.7%。与同种条件下的非菌根植株相比,干旱及双重胁迫下的菌根植株生物量、叶绿素含量、光合蒸腾速率、最大光化学效率,以及P吸收均呈上升趋势;高钙胁迫下的菌根植株叶绿素含量、最大光化学效率有所增加,但生物量、光合蒸腾速率以及N、P的吸收未体现菌根促进效应。AM真菌与干旱及双重胁迫的交互作用对玉米幼苗的净光合速率影响显著,与高钙交互作用对玉米幼苗净光合速率无显著影响。AM真菌能够通过促进玉米幼苗N、P吸收及叶绿素含量增加,光化学效率、气孔导度增大,从而提高玉米幼苗光合作用能力促进生长。实验结果对岩溶生态系统中合理利用菌根技术及制定合理的农业生产措施具有重要的理论和实践意义。     

水分胁迫下AM真菌对柠条锦鸡儿(Caragana korshinskii)生长和抗旱性的影响

利用盆栽实验研究了水分胁迫条件下AM真菌对柠条锦鸡儿(Caragana korshinskii)生长和抗旱性的影响.在土壤相对含水量为80%、60%和40%条件下,分别接种摩西球囊霉(Glomus mosseae)和柠条锦鸡儿根际土著菌,结果表明,水分胁迫对AM真菌的接种效果有显著影响.不同水分条件下,接种AM真菌显著提高了宿主植物根系菌根侵染率.土壤相对含水量为40%～60%时,接种株的株高、茎粗、生物干重和叶片保水力明显高于不接种株;接种AM真菌提高了植株对土壤有效N和有效P的利用率,增加了植株全P、叶片叶绿素和可溶性糖含量以及SOD、POD、CAT等保护酶活性.土壤相对含水量为40%时,叶片MDA含量明显下降.水分胁迫条件下,以接种柠条锦鸡儿根际土著菌的效果最佳.AM真菌增强宿主植物的抗旱性可能源于促进宿主植物根系对土壤水分和矿质元素吸收的直接作用和改善植物体内生理代谢活动、提高保护酶活性的间接作用.     

Arbuscular mycorrhizal fungi improve the growth and drought tolerance of Cinnamomum migao by enhancing physio‐biochemical responses

Drought is the main limiting factor for plant growth in karst areas with a fragile ecological environment. Cinnamomum migao H.W. Li is an endemic medicinal woody plant present in the karst areas of southwestern China, and it is endangered due to poor drought tolerance. Arbuscular mycorrhizal fungi (AMF) are known to enhance the drought tolerance of plants. However, few studies have examined the contribution of AMF in improving the drought tolerance of C. migao seedlings. Therefore, we conducted a series of experiments to determine whether a single inoculation and coinoculation of AMF (Claroideoglomus lamellosum and Claroideoglomus etunicatum) enhanced the drought tolerance of C. migao. Furthermore, we compared the effects of single inoculation and coinoculation with different inoculum sizes (20, 40, 60, and 100 g; four replicates per treatment) on mycorrhizal colonization rate, plant growth, photosynthetic parameters, antioxidant enzyme activity, and malondialdehyde (MDA) and osmoregulatory substance contents. The results showed that compared with nonmycorrhizal plants, AMF colonization significantly improved plant growing status; net photosynthetic rate; superoxide dismutase, catalase, and peroxidase activities; and soluble sugar, soluble protein, and proline contents. Furthermore, AMF colonization increased relative water content and reduced MDA content in cells. These combined cumulative effects of AMF symbiosis ultimately enhanced the drought tolerance of seedlings and were closely related to the inoculum size. With an increase in inoculum size, the growth rate and drought tolerance of plants first increased and then decreased. The damage caused by drought stress could be reduced by inoculating 40–60 g of AMF, and the effect of coinoculation was significantly better than that of single inoculation at 60 g of AMF, while the effect was opposite at 40 g of AMF. Additionally, the interaction between AMF and inoculum sizes had a significant effect on drought tolerance. In conclusion, the inoculation of the AMF (Cl. lamellosum and Cl. etunicatum) improved photosynthesis, activated antioxidant enzymes, regulated cell osmotic state, and enhanced the drought tolerance of C. migao, enabling its growth in fragile ecological environments.     

Effects of simultaneous infections of endophytic fungi and arbuscular mycorrhizal fungi on the growth of their shared host grass Achnatherum sibiricum under varying N and P supply

Grasses can be infected by endophytic fungi and arbuscular mycorrhizal (AM) fungi simultaneously. Here, we investigated the interactions of a native grass, Achnatherum sibiricum, with both endophytic and AM fungi (Glomus mosseae, GM and Glomus etunicatum, GE) at different nitrogen (N) and phosphorus (P) levels. The results showed that endophyte infection significantly suppressed the colonization rates and spore density of GE, but had no effect on those of GM. Endophyte infection increased shoot biomass regardless of the nutrient conditions. The effects of AM fungi on host growth were dependent on mycorrhizal species. There was no significant interaction between endophytic fungi and GE on host growth; however, a significant interaction between endophytic fungi and GE occurred in total phenolic concentrations and P uptake. As for GM, a significant interaction among endophytic fungi, AM fungi and nutrient availability occurred in shoot growth. Under sufficient N and P conditions, endophyte infection alleviated the detrimental effects of GM colonization on host growth.     

超积累植物与丛枝菌根真菌共生及其联合吸收积累重金属的效应

利用超积累植物进行重金属污染土壤修复是应对全球大面积分布无机污染问题的重要解决方法之一。超积累植物虽然具有超量积累重金属的能力,但其定植、生长和提取功能的发挥都受到重金属胁迫的显著影响。利用丛枝菌根真菌 (Arbuscular mycorrhizal fungi,AMF) 强化超积累植物功能可联合发挥二者的功能优势,提升修复效率、缩短修复周期、保持修复效果的稳定性和长期性,在日益复杂、严峻的重金属污染治理领域具有重要的研究价值和广阔的应用前景。文中首先给出了超积累植物的概念、中国本土首次报道的典型重金属元素超积累植物和能与AMF形成共生体系的超积累植物名录,系统深入地探讨了AMF对超积累植物生长和吸收累积重金属的影响,以及超积累植物+AMF联合吸收积累重金属的效应与作用机制,认为AMF可通过调节根围理化与生物条件、元素平衡状况、生理代谢和基因表达等途径,增强超积累植物吸收积累重金属的效应,超积累植物+AMF构建的共生体系具备联合修复重金属污染生境的潜力。最后指出了超积累植物+AMF共生联合修复技术当前面临的关键问题、发展方向和应用前景。     

Arbuscular mycorrhizal inoculation improves growth and antioxidative response of Jatropha curcas (L.) under Na2SO4 salt stress

This study investigated the effect of arbuscular mycorrhizal (AM) fungal consortia on growth, photosynthetic pigments, solutes concentration (e.g., sugars and proline), and antioxidant responses at different levels of Na₂SO₄ stress (0–0.5%, w:w) in potted culture of Jatropha. Results showed that increasing salt levels caused a significant reduction in survival (%), growth parameters, leaf relative water content (LRWC) (%), and chlorophyll content with an increase in electrolyte leakage (%) and lipid peroxidation of membranes of Jatropha. AM inoculation improved biomass yields as well as other physiological parameters (LRWC (%), chlorophyll, proline, and soluble sugar) of salt-stressed Jatropha over noninoculated plants. Tolerance index of Jatropha was higher with AM fungi than without at all salt levels; however, a decline in its value was recorded with increased salinity levels. AM inoculation also enhanced the activities of antioxidant enzymes (e.g., superoxide dismutase, peroxidase, ascorbate peroxidase, and glutathione reductase) and decreased oxidative damage to lipids. In conclusion, results indicate that AM inoculation was capable of alleviating the damage caused by salinity stress on Jatropha plants by reducing lipid peroxidation of membrane and membrane permeability and increasing the accumulation of solutes and antioxidant enzyme activity.     

Effect of plant growth promoting bacterium; Pseudomonas putida UW4 inoculation on phytoremediation efficacy of monoculture and mixed culture of selected plant species for PAH and lead spiked soils

Plant growth promoting bacteria (PGPB) enhanced phytoremediation (PEP) is an attractive remedial strategy for the remediation of polycyclic aromatic hydrocarbon (PAH) and heavy metal (HM) contaminated sites. The effect of PGPB; Pseudomonas putida UW4 inoculation on the phytoremediation efficiency of Medicago sativa, Festuca arundinacea, Lolium perenne, and mixed plants (L. perenne and F. arundinacea) was assessed. This involved two contaminant treatments; “PAH” (phenanthrene; 300?mg·kg^?1, fluoranthene; 200?mg·kg^?1, and benzo[a]pyrene; 5?mg·kg^?1) and “PAH?+?HM” (‘PAH’ treatments +100?mg of Pb/kg). PGPB inoculation significantly enhanced root biomass yield of F. arundinacea in PAH treatment, and the mixed plant shoot biomass and L. perenne root biomass yields of the PAH?+?HM treatment. PGPB significantly enhanced dissipation of phenanthrene and fluoranthene for M. sativa-PAH?+?PGPB treatment and fluoranthene for F. arundinacea-PAH?+?HM?+?PGPB treatment. In others, PGPB inoculation either had no impact or inhibited PAH dissipation. PAH dissipation for the single and mixed plant treatments with PGPB inoculation were not different. The efficiency of PEP is dependent on different factors such as PGPB inoculum biomass, plant species, plant–microbe specificity and type of contaminants. Exploiting PEP technology would require proper understanding of plant tolerance and growth promoting mechanisms, and rhizosphere activities.