Plant responses to plant growth-promoting rhizobacteria

Non-pathogenic soilborne microorganisms can promote plant growth, as well as suppress diseases. Plant growth promotion is taken to result from improved nutrient acquisition or hormonal stimulation. Disease suppression can occur through microbial antagonism or induction of resistance in the plant. Several rhizobacterial strains have been shown to act as plant growth-promoting bacteria through both stimulation of growth and induced systemic resistance (ISR), but it is not clear in how far both mechanisms are connected. Induced resistance is manifested as a reduction of the number of diseased plants or in disease severity upon subsequent infection by a pathogen. Such reduced disease susceptibility can be local or systemic, result from developmental or environmental factors and depend on multiple mechanisms. The spectrum of diseases to which PGPR-elicited ISR confers enhanced resistance overlaps partly with that of pathogen-induced systemic acquired resistance (SAR). Both ISR and SAR represent a state of enhanced basal resistance of the plant that depends on the signalling compounds jasmonic acid and salicylic acid, respectively, and pathogens are differentially sensitive to the resistances activated by each of these signalling pathways. Root-colonizing Pseudomonas bacteria have been shown to alter plant gene expression in roots and leaves to different extents, indicative of recognition of one or more bacterial determinants by specific plant receptors. Conversely, plants can alter root exudation and secrete compounds that interfere with quorum sensing (QS) regulation in the bacteria. Such two-way signalling resembles the interaction of root-nodulating Rhizobia with legumes and between mycorrhizal fungi and roots of the majority of plant species. Although ISR-eliciting rhizobacteria can induce typical early defence-related responses in cell suspensions, in plants they do not necessarily activate defence-related gene expression. Instead, they appear to act through priming of effective resistance mechanisms, as reflected by earlier and stronger defence reactions once infection occurs.     

Inhibitory effect of ACC deaminase‐producing bacteria on crown gall formation in tomato plants infected by Agrobacterium tumefaciens or A. vitis

This study showed that various rhizosphere bacteria producing the enzyme 1‐aminocyclopropane‐1‐carboxylate (ACC) deaminase (ACCD), which can degrade ACC, the immediate precursor of ethylene in plants, and thereby lower plant ethylene levels, can act as promising biocontrol agents of pathogenic strains of Agrobacterium tumefaciens and A. vitis. Soaking the roots of tomato (Solanum lycopersicum) seedlings in a suspension of the ACCD‐producing Pseudomonas putida UW4, Burkholderia phytofirmans PsJN or Azospirillum brasilense Cd1843 transformed by plasmid pRKTACC carrying the ACCD‐encoding gene acdS from UW4, significantly reduced the development of tumours on tomato plants injected 4–5 days later with pathogenic Agrobacterium strains via wounds on the plant stem. The fresh mass of tumours formed by plants pretreated with ACCD‐producing strains was typically four‐ to fivefold less than that of tumours formed on control plants inoculated only with a pathogenic Agrobacterium strain. Simultaneously, the level of ethylene evolution per amount of tumour mass on plants pretreated with ACCD‐producing bacteria decreased four to eight times compared with that from tumours formed on control plants or plants pretreated with bacteria deficient in ACCD production. Moreover, transgenic tomato plants expressing a bacterial ACCD were found to be highly resistant to crown gall formation relative to the parental, non‐transformed tomato plants. The results support the hypothesis that ethylene is a crucial factor in Agrobacterium tumour formation, and that ACCD‐produced rhizosphere bacteria may protect plants infected by pathogenic Agrobacteria from crown gall disease.     

外源ACC通过激活乙烯合成及信号转导诱导小豆抗锈性

大量研究表明, 乙烯可激发植物对死体营养型真菌的抗性, 但我们前期研究发现, 乙烯合成前体ACC可提高小豆对活体营养型真菌——锈菌的抗性, 为初步明确其机制, 本研究分析了ACC处理对小豆乙烯合成及信号转导的影响, 结果表明, ACC处理显著提高了乙烯合成基因VaACS1及信号通路关键基因VaEIN2?VaEIN3?VaERF5的表达水平?此外, ACC处理后再接种锈菌, 小豆锈病的发病程度显著降低?对接种锈菌后不同时间VaPR2和VaPR4的表达分析表明, 相比ACC处理后不接种对照, ACC处理后再接种锈菌的处理, 接种后1~5 d这两个基因表达量显著升高; 与水处理不接种锈菌相比, 水处理接种锈菌5~8 d后VaPR2和VaPR4的表达量虽显著上调, 但应答时间较ACC处理滞后, 且总体表达水平低, 表明ACC激活乙烯通路进而诱导防卫反应基因上调表达是其诱导小豆抗锈性的关键?     

Effects of plant growth-promoting rhizobacteria on nodulation of <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L. are dependent on plant P nutrition

Several plant growth-promoting rhizobacteria (PGPR) have shown potential to enhance nodulation of legumes when coinoculated with Rhizobium. To optimize the efficiency of these Rhizobium-PGPR-host plant interactions, unravelling the underlying mechanisms and analyzing the influence of specific environmental conditions is crucial. In this work the effect of four PGPR strains on the symbiotic interaction between Rhizobium and common bean (Phaseolus vulgaris) was studied under deficient versus sufficient phosphorus supply. It was observed that the effect on nodulation of three out of four PGPR tested was strongly dependent on P nutrition. Further, the use of specific PGPR mutant strains indicated that bacterial indole-3-acetic-acid production (IAA) and 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity play an important role in the host nodulation response, particularly under low P conditions. Moreover, it was shown that the differential response to PGPR under low versus high P conditions was associated with changes in the host hormone sensitivity for nodulation induced under P deficiency. These findings contribute to the understanding of the interplay between Rhizobium, PGPR and the plant host under different environmental settings.     

Molecular and genetic characterization of ethylene involvement in mycotoxin-induced plant cell death

AAL-toxin and the structurally related mycotoxin, fumonisin B₁(FB₁), cause interveinal cell death in susceptible lines of tomato with morphological characteristics of apoptosis. We observed previously that concomitant with cell death, an increase in 1-aminocyclopropane-1-carboxylic acid (ACC) and ethylene occurs, while the addition of inhibitors of ethylene biosynthesis or action significantly reduce the amount of dead tissue. Using a molecular approach, we have identified the primary ACC synthase gene family member involved, LE ACS2, and have observed an interesting aspect of its regulation. Fumonisin B₁caused the accumulation of LE ACS2 mRNA with a similar time course but to a lower level than did AAL-toxin. Nevertheless, ACC levels were similar for AAL- and FB₁-treated tissue. ACC oxidase mRNA was also induced by both mycotoxins, and again AAL-toxin induction was greater than that with FB₁. The induction of ACC synthase and ACC oxidase mRNA observed here represents the earliest changes in gene expression noted in this cell death system to date. The effects of theNever ripemutation of tomato, which affects ethylene perception, on toxin-induced cell death also were determined. TheNever ripemutant showed significantly less necrosis or chlorosis in response to the mycotoxins than did its wild type counterpart. These results indicate that alteration in ethylene perception can markedly reduce the amount of tissue damage during this susceptible response. These findings extend our understanding of ethylene-associated signal transduction during plant cell death and suggest strategies for manipulation of cell death to enhance plant disease resistance.     

Striga asiatica seed conditioning and 1-aminocyclopropane-1-carboxylate oxidase activity

Conditioned seeds of Striga asiatica (L.) Kuntze release ethylene, which elicits germination. We investigated the activity of 1-aminocyclopropane-1-carboxylate (ACC) oxidase and respiration during conditioning. Seeds incubated in vivo with ACC, the substrate for ACC oxidase, produced negligible ethylene at the beginning of conditioning or if they were dormant (i.e. would not germinate after conditioning and treatment with stimulant). Non-dormant seeds produced 3000 ηL of ethylene/600 seeds/24 h after 12 days of conditioning. In vitro ACC oxidase activity at day 0 of conditioning produced 500 ηL of ethylene/μg protein/h and 8000 ηL of ethylene/μg protein/h after 12 days of conditioning. Incubation of seeds in strigol before protein extraction did not enhance enzyme activity. Seeds released 4000 μL/L CO₂ in the first 24 h of conditioning, with the rate increasing to 15 000 μL/L/24 h on day 4 and then remaining roughly unchanged. Maximum in vitro activity of ACC oxidase required ACC, catalase, O₂, Fe²⁺, ascorbate and CO₂. In vivo activity of ACC oxidase required ACC and/or germination stimulant(s), suggesting that stimulants may be involved in providing substrates for the ACC oxidase. No difference was observed in the separation of extracted proteins, which suggests that ACC oxidase is activated during conditioning, perhaps as a result of changes in co-factor concentration. Application of these findings to Striga control is discussed.     

产气克雷伯氏杆菌 Klebsiella aerogenes S2对植物病原真菌的抑菌活性及番茄促生效果

本研究以广西红树林的根际土壤为材料, 利用选择性培养基分离筛选多功能菌株, 并利用生理生化和分子生物学相结合的方法进行鉴定?采用平板对峙法测定筛选获得的菌株对几种植物病原真菌的拮抗效果, 同时通过盆栽试验和发芽试验研究其对番茄幼苗和种子的促生效果, 从而挖掘兼具促生防病功能的植物根际促生菌(plant growth-promoting rhizobacteria, PGPR)?经筛选获得一株具有溶磷及分泌生长素(indole-3-acetic acid, IAA)能力的细菌菌株S2, 经鉴定该菌株属于克雷伯氏菌属Klebsiella, 其对禾谷镰孢Fusarium graminearum, 玉蜀黍平脐蠕孢Bipolaris maydis等3种植物病原真菌具有一定的拮抗效果, 菌株发酵液浓度为1×104?2×104 cfu/mL时对番茄种子和幼苗的促生效果最好?综上, 菌株S2 是一株新型且兼具促生防病效果的多功能PGPR菌株, 该结果为绿色现代农业开发土壤微生物菌肥和植物生长调节剂提供优质菌源参考?     

小豆抗锈病诱导剂的筛选

<正>由豇豆单胞锈(Uromyces vignae)引起的小豆锈病是我国小豆生产中的重要病害之一[1,2],在中国华北及东北地区发生严重,可造成严重的产量损失[3]。当前小豆锈病的防治主要依赖化学药剂,但长期使用化学药剂存在残留、破坏生态等弊端。因此寻找绿色可持续的防控措施越来越受到人们的关注。利用外源诱导剂诱导植物抗性是一种潜在的病害防控措施[4]。目前,已有超过30种的植物被证实可被诱导产生抗病性[5]。但在诱导小豆抗锈病方面的研究报道尚     

小豆抗锈病诱导剂的筛选

Four common inducers, salicylic acid (SA), 1-aminocyclopropane-1-carboxylic acid (ACC), benzothiadiazole (BTH) and methyl jasmonate (MeJA), were selected to evaluate the effects of inducers on the resistance of adzuki bean to rust by investigating urediospores germination and the adzuki bean resistance to Uromyces vignae. The results showed that four inducers had no effects on the urediospores germination. However, they can significantly induce adzuki bean resistance to U. vignae. Among these four inducers, 0.25 mg·mL^-1 ACC can significantly reduce the disease index, which was 45.06% lower than the control. Aditionally, seedlings treated with ACC exhibited a typical "triple reaction", which indicated that this induced resistance may relate with the endogenous ethylene biosynthesis in adzuki bean. Then the adzuki bean seedlings were treated with an ethylene receptor inhibitor, 1-methylcyclopropene (1-MCP) alone or combined with ACC. The results showed that 1-MCP alone had no significant effect on the growth and rust resistance of adzuki bean. However, 1-MCP could eliminate the "triple reaction" and did not affect the induced resistance caused by ACC when it used combined with ACC. Results obtained in current study suggested that ACC can significantly induce the resistance of adzuki bean to the rust. Moreover, we speculate that ACC-induced rust resistance of adzuki bean may not depend on ethylene signaling pathway.     

Fluazifop-butyl causes membrane peroxidation in the herbicide-susceptible broad leaf weed bristly starbur (Acanthospermum hispidum)

In order to clarify the action mechanism of fluazifop-butyl, an aryloxyphenoxypropionate (AOPP) herbicide in bristly starbur (Acanthospermum hispidum DC.), a unique fluazifop-butyl-susceptible broad-leaved weed, ethylene evolution and membrane lipid peroxidation in the plant seedlings were investigated. Foliar application of fluazifop-P-butyl induced ethylene evolution only from bristly starbur, but not from oat (Avena sativa L.), another fluazifop-butyl-susceptible species, and two tolerant species, pea (Pisum sativum L.) and hairy beggarticks (Bidens pilosa L.). The other AOPP herbicides, quizalofop-ethyl and fenoxaprop-ethyl, and a cyclohexanedione (CHD) herbicide, sethoxydim, did not enhance ethylene production from bristly starbur. This fluazifop-butyl-induced ethylene production in bristly starbur was completely suppressed by aminoethoxyvinylglycine (AVG), a 1-aminocyclopropane-1-carboxylic acid (ACC) synthase inhibitor, but not by p-chlorophenoxyisobutyric acid (PCIB), an anti-auxin compound, suggesting this evolved ethylene was not auxin-induced. Phytotoxic action by fluazifop-P-butyl (5 μM) in bristly starbur was reduced markedly by two lipid-soluble antioxidants, vitamin E, and ethoxyquin. The ethylene production from the plant was also inhibited by these two antioxidants. Content of malondialdehyde, an indicator of lipid peroxidation, increased only by fluazifop-P-butyl in bristly starbur seedlings but not in oat, and this increase was inhibited by ethoxyquin. These results strongly suggest that the primary site of action for fluazifop-butyl in bristly starbur is on the membranes and active oxygen species and/or free radicals are involved in peroxidation. Ethylene evolution is probably induced by these reactive oxygen species.     

Effects of fosthiazate and aldicarb on populations of plant-growth-promoting bacteria, root-lesion nematodes and bacteria-feeding nematodes in the root zone of potatoes

A 2-year study investigated the relationship between the nematicides aldicarb and fosthiazate and populations of plant growth-promoting (PGP) and plant-growth-inhibiting (PGI) bacteria, and root-lesion nematodes and bacteria-feeding nematodes in the root-zone soils of potatoes. Fewer ( P  < 0.05) bacterial genera and species were recovered from aldicarb-treated soils than from the fosthiazate and untreated soils but bacterial population densities were greatest in the aldicarb-treated soils. In potato plantlet bioassays using root-zone bacteria from the three soil sources, bacteria from the aldicarb-treated soils reduced ( P  < 0.01) the mean wet and dry weight of shoots and roots compared with those from the fosthiazate and untreated soils. In the field, fosthiazate (but not aldicarb) significantly increased tuber yields and reduced nematode population densities of Pratylenchus penetrans in roots and soil. Population densities of bacteria-feeding nematodes (primarily Diplogaster lheritieri ) were not affected by either nematicide. Aldicarb appeared to suppress the populations of plant-growth-promoting bacteria that contribute to enhanced growth in potatoes. This is attributed to the long-term use of aldicarb at the trial site, which may have encouraged the selection, or adaptation, of soil microorganisms that degrade carbamate pesticides but do not enhance potato growth.     

Beta-cyanoalanine synthase as a molecular marker for induced resistance by fungal glycoprotein elicitor and commercial plant activators

ABSTRACT The biocontrol agent Pythium oligandrum produces glycoprotein elicitor in the cell wall fraction, designated CWP, and induces resistance to a broad range of pathogens. To understand the mechanism of CWP-induced resistance to pathogens, gene expression at the early stage of CWP treatment in tomato roots was analyzed using a cDNA array. At 4 h after CWP treatment, 144 genes were up-regulated and 99 genes were down-regulated. In the 144 up-regulated genes, nine genes exhibited about eightfold increased expression. Analysis of the response of these nine genes to three commercial plant activators indicated that a high level of one gene, beta-cyanoalanine synthase gene (LeCAS) encoding hydrogen cyanide (HCN) detoxification enzyme, was stably induced in tomato roots by such treatment. However, expression of LeCAS was not significantly induced in tomato roots at 4 h by abiotic stresses, whereas only a very low level of induction of such expression by cold stress was observed. This LeCAS expression was also induced after exogenous treatment with a low level of 1-amino-cyclopropane-1-carboxylate as the precursor of ethylene, but not with either salicylic acid or methyl jas-monate. The induction of LeCAS expression in CWP-treated and plant activator-treated roots is likely to be caused by the detoxification of HCN during ethylene production. Transient activation of LeCAS expression caused by ethylene production in tomato roots may be a general phenomenon in fungal elicitor-induced and synthetic plant activator-induced resistance. LeCAS seems to be useful for screening possible novel plant activators for plant protection against pathogens.     

Lysobacter gummosus OH17 induces systemic resistance in Oryza sativa ‘Nipponbare’

Although rice is one of the most cultivated, consumed, and essential crops worldwide, it is highly susceptible to a wide range of bacterial and fungal pathogens that significantly reduce the production and quality of rice. Recently, our research group reported that the plant growth-promoting rhizobacterium Lysobacter gummosus OH17 was able to enhance the ethylene levels in Oryza sativa ‘Nipponbare’ plants at the late interaction stages. In this work, L. gummosus OH17 was found to be capable of inducing the overexpression of relevant genes of the jasmonic acid and ethylene transduction pathways in Nipponbare plants, such as OsACC, OsACO, OsERF3, and OsLOX, which resulted in the up-regulation of a number of pathogenesis-related proteins. The observed metabolic effects enhanced the disease resistance of rice against the three most devastating rice pathogens: Magnaporthe oryzae causing rice blast, Rhizoctonia solani causing rice sheath blight, and Xanthomonas oryzae pv. oryzae causing bacterial leaf blight. Furthermore, it was shown that L. gummosus OH17 also enhanced ethylene production levels in other O. sativa varieties from both the japonica and indica subspecies. Here, we report for the first time the metabolic alterations produced by plant growth-promoting rhizobacterium L. gummosus OH17 at the late interaction stages and how these alterations induce systemic resistance.     

Studies on the mechanism of selectivity of the auxin herbicide quinmerac

Investigations were conducted to elucidate the mechanism of selectivity of the auxin herbicide, quinmerac, in cleavers (Galium aparine) and the tolerant crops sugarbeet (Beta vulgaris), oilseed rape (Brassica napus) and wheat (Triticum aestivum). After root treatment with the herbicide, the selectivity has been quantified as approximately 400-fold between oilseed rape and Galium and 1000-fold between sugarbeet or wheat and the weed species. When 1 and 10 μM [¹⁴C]quinmerac were applied for 4 h, no significant differences between root absorption and translocation of ¹⁴C by Galium and the crop species were found. After 16 h, metabolism of [¹⁴C]quinmerac to the biologically inactive hydroxymethyl and dicarboxylic acid derivatives was more rapid in wheat and sugarbeet than in Galium. In oilseed rape, a lower rate of herbicide metabolism was observed. In Galium, accumulations of abscisic acid (ABA), triggered by quinmerac-stimulated ethylene biosynthesis, were found to cause the herbicidal growth inhibition which develops during 24 h of application. Within 1 h of treatment, quinmerac stimulated 1-aminocyclopropane-1-carboxylic acid (ACC) synthase activity and ACC concentration specifically in Galium shoot tissue. During the next 4 h, ACC synthase activity was increased up to 50-fold, relative to the control. Within 3 h of exposure to quinmerac, increased ethylene formation followed by higher ABA levels was detected. In sugarbeet, oilseed rape and wheat, quinmerac did not stimulate ACC synthase activity and ACC and ABA levels. It is suggested that (i) the selectivity of quinmerac is primarily based upon the lower sensitivity to the herbicide of the tissue/target in the crop species, (ii) the induction process of the ACC synthase activity in the shoot tissue is the primary target of herbicidal interference. In wheat and sugarbeet, tolerance to quinmerac is additionally increased by a more rapid metabolism. © 1998 SCI.     

O-酰基化丝氨酸的合成及其对樱桃番茄乙烯合成量和果实硬度的影响

通过L-丝氨酸与不同的酰氯在室温下反应,合成了8个未见文献报道的乙烯合成抑制剂O-酰基化丝氨酸（2a~2h）,其结构通过核磁共振氢谱、碳谱及高分辨质谱确证。以氨乙氧乙烯基甘氨酸（aminoethoxyvinylglycine,AVG）为阳性对照,测定了目标化合物对樱桃番茄Lycopersivon esculentum Mill.乙烯合成量和果实硬度的影响,同时运用分子对接的方法分析了目标化合物与番茄1-氨基环丙烷羧酸（1-aminocyclopropane-1-carboxylic acid, ACC）合成酶（ACS, 1IAY）可能的结合模式。结果显示:大部分目标化合物具有延缓樱桃番茄果实软化和抑制乙烯合成的作用,其中化合物2c和2h效果较为突出。用2c和2h处理番茄后第5天,与空白对照相比,果实硬度分别提高27.62%和40.04%,均与AVG处理无显著性差异;2c和2h对乙烯合成量的抑制率分别达71.89%和53.28%,其中2c处理的抑制效果显著优于AVG处理。分子对接方法分析结果表明:化合物2c与番茄ACS酶活性空腔的氨基酸残基有较好的相互作用,2c的羧基可与Ala54和Arg412的氨基形成氢键,从而占据ACS酶的活性空腔。O-酰基化丝氨酸类化合物结构简单,容易获取,对乙烯合成抑制剂的开发具有推动作用。     

Rhizobacteria-mediated Induced Systemic Resistance: Triggering, Signalling and Expression

Selected strains of rhizosphere bacteria reduce disease by activating a resistance mechanism in the plant named rhizobacteria-mediated induced systemic resistance (ISR). Rhizobacteria-mediated ISR resembles pathogen-induced systemic acquired resistance (SAR) in that both types of induced resistance render uninfected plant parts more resistant towards a broad spectrum of plant pathogens. Some rhizobacteria trigger the salicylic acid (SA)-dependent SAR pathway by producing SA at the root surface. In other cases, rhizobacteria trigger a different signalling pathway that does not require SA. The existence of a SA-independent ISR pathway has been demonstrated in Arabidopsis thaliana. In contrast to pathogen-induced SAR, ISR induced by Pseudomonas fluorescens WCS417r is independent of SA accumulation and pathogenesis-related (PR) gene activation but, instead, requires responsiveness to the plant hormones jasmonic acid (JA) and ethylene. Mutant analyses showed that ISR follows a novel signalling pathway in which components from the JA and ethylene response are successively engaged to trigger a defensive state that, like SAR, is controlled by the regulatory factor NPR1. Interestingly, simultaneous activation of both the JA/ethylene-dependent ISR pathway and the SA-dependent SAR pathway results in an enhanced level of protection. Thus combining both types of induced resistance provides an attractive tool for the improvement of disease control. This review focuses on the current status of our research on triggering, signalling, and expression of rhizobacteria-mediated ISR in Arabidopsis.     

病菌侵染对番茄果实乙烯生成,ACC合成酶和脂氧合酶活性的影响

 本文以红熟期的番茄(Lycopersicon esculentum Mill.)果实为材料,主要研究病菌侵染与果实中乙烯生成,ACC合成酶和脂氧合酶(LOX)的关系,以及不同乙烯处理对感病果实ACC合成酶和脂氧合酶(LOX)活性的影响。结果表明:用葡萄孢菌(Botrytis cinerea)接种采后番茄果实在25℃下存放7d后,感病果实的乙烯释放量显著增加,ACC合成酶和脂氧合酶(LOX)活性也明显提高。在贮藏环境中增加乙烯吸收剂可明显地延缓葡萄孢菌的发病,而且ACC合成酶和LOX酶的活性也相对较低。由此证明病菌侵染与果实乙烯释放量,以及与ACC合成酶和LOX酶之间存在明显的正相关     

Induced Systemic Resistance by Fluorescent Pseudomonas spp

ABSTRACT Fluorescent Pseudomonas spp. have been studied for decades for their plant growth-promoting effects through effective suppression of soilborne plant diseases. The modes of action that play a role in disease suppression by these bacteria include siderophore-mediated competition for iron, antibiosis, production of lytic enzymes, and induced systemic resistance (ISR). The involvement of ISR is typically studied in systems in which the Pseudomonas bacteria and the pathogen are inoculated and remain spatially separated on the plant, e.g., the bacteria on the root and the pathogen on the leaf, or by use of split root systems. Since no direct interactions are possible between the two populations, suppression of disease development has to be plant-mediated. In this review, bacterial traits involved in Pseudomonas-mediated ISR will be discussed.