Effects of genetically modified starch metabolism in potato plants on photosynthate fluxes into the rhizosphere and on microbial degraders of root exudates

A high percentage of photosynthetically assimilated carbon is released into soil via root exudates, which are acknowledged as the most important factor for the development of microbial rhizosphere communities. As quality and quantity of root exudates are dependent on plant genotype, the genetic engineering of plants might also influence carbon partitioning within the plant and thus microbial rhizosphere community structure. In this study, the carbon allocation patterns within the plant-rhizosphere system of a genetically modified amylopectin-accumulating potato line (Solanum tuberosum L.) were linked to microbial degraders of root exudates under greenhouse conditions, using (13)C-CO(2) pulse-chase labelling in combination with phospholipid fatty acid (PLFA) analysis. In addition, GM plants were compared with the parental cultivar as well as a second potato cultivar obtained by classical breeding. Rhizosphere samples were obtained during young leaf developmental and flowering stages. (13)C allocation in aboveground plant biomass, water-extractable organic carbon, microbial biomass carbon and PLFA as well as the microbial community structure in the rhizosphere varied significantly between the natural potato cultivars. However, no differences between the GM line and its parental cultivar were observed. Besides the considerable impact of plant cultivar, the plant developmental stage affected carbon partitioning via the plant into the rhizosphere and, subsequently, microbial communities involved in the transformation of root exudates.     

Plants Release Precursors of Histone Deacetylase Inhibitors to Suppress Growth of Competitors

To secure their access to water, light, and nutrients, many plant species have developed allelopathic strategies to suppress competitors. To this end, they release into the rhizosphere phytotoxic substances that inhibit the germination and growth of neighbors. Despite the importance of allelopathy in shaping natural plant communities and for agricultural production, the underlying molecular mechanisms are largely unknown. Here, we report that allelochemicals derived from the common class of cyclic hydroxamic acid root exudates directly affect the chromatin-modifying machinery in Arabidopsis thaliana. These allelochemicals inhibit histone deacetylases both in vitro and in vivo and exert their activity through locus-specific alterations of histone acetylation and associated gene expression. Our multilevel analysis collectively shows how plant-plant interactions interfere with a fundamental cellular process, histone acetylation, by targeting an evolutionarily highly conserved class of enzymes.     

Root Exudation of Phytochemicals in Arabidopsis Follows Specific Patterns That Are Developmentally Programmed and Correlate with Soil Microbial Functions

Plant roots constantly secrete compounds into the soil to interact with neighboring organisms presumably to gain certain functional advantages at different stages of development. Accordingly, it has been hypothesized that the phytochemical composition present in the root exudates changes over the course of the lifespan of a plant. Here, root exudates of in vitro grown Arabidopsis plants were collected at different developmental stages and analyzed using GC-MS. Principle component analysis revealed that the composition of root exudates varied at each developmental stage. Cumulative secretion levels of sugars and sugar alcohols were higher in early time points and decreased through development. In contrast, the cumulative secretion levels of amino acids and phenolics increased over time. The expression in roots of genes involved in biosynthesis and transportation of compounds represented in the root exudates were consistent with patterns of root exudation. Correlation analyses were performed of the in vitro root exudation patterns with the functional capacity of the rhizosphere microbiome to metabolize these compounds at different developmental stages of Arabidopsis grown in natural soils. Pyrosequencing of rhizosphere mRNA revealed strong correlations (p<0.05) between microbial functional genes involved in the metabolism of carbohydrates, amino acids and secondary metabolites with the corresponding compounds released by the roots at particular stages of plant development. In summary, our results suggest that the root exudation process of phytochemicals follows a developmental pattern that is genetically programmed.     

No accession-specific effect of rhizosphere soil communities on the growth and competition of Arabidopsis thaliana accessions

Soil communities associated with specific plant species affect individual plants' growth and competitive ability. Limited evidence suggests that unique soil communities can also differentially influence growth and competition at the ecotype level. Previous work with Arabidopsis thaliana has shown that accessions produce distinct and reproducible rhizosphere bacterial communities, with significant differences in both species composition and relative abundance. We tested the hypothesis that soil communities uniquely affect the growth and reproduction of the plant accessions with which they are associated. Specifically, we examined the growth of four accessions when exposed to their own soil communities and the communities generated by each of the other three accessions. To do this we planted focal accessions inside a ring of six plants that created a "background" soil community. We grew focal plants in this design in three separate soil treatments: non-sterile soil, sterilized soil, and "preconditioned" soil. We preconditioned soil by growing accessions in non-sterile soil for six weeks before the start of the experiment. The main experiment was harvested after seven weeks of growth and we recorded height, silique number, and dry weight of each focal plant. Plants grown in the preconditioned soil treatment showed less growth relative to the non-sterile and sterile soil treatments. In addition, plants in the sterile soil grew larger than those in non-sterile soil. However, we saw no interaction between soil treatment and background accession. We conclude that the soil communities have a negative net impact on Arabidopsis thaliana growth, and that the unique soil communities associated with each accession do not differentially affect growth and competition of study species.     

Achieving similar root microbiota composition in neighbouring plants through airborne signalling

The ability to recognize and respond to environmental signals is essential for plants. In response to environmental changes, the status of a plant is transmitted to other plants in the form of signals such as volatiles. Root-associated bacteria trigger the release of plant volatile organic compounds (VOCs). However, the impact of VOCs on the rhizosphere microbial community of neighbouring plants is not well understood. Here, we investigated the effect of VOCs on the rhizosphere microbial community of tomato plants inoculated with a plant growth-promoting rhizobacterium Bacillus amyloliquefaciens strain GB03 and that of their neighbouring plants. Interestingly, high similarity (up to 69%) was detected in the rhizosphere microbial communities of the inoculated and neighbouring plants. Leaves of the tomato plant treated with strain GB03-released β-caryophyllene as a signature VOC, which elicited the release of a large amount of salicylic acid (SA) in the root exudates of a neighbouring tomato seedling. The exposure of tomato leaves to β-caryophyllene resulted in the secretion of SA from the root. Our results demonstrate for the first time that the composition of the rhizosphere microbiota in surrounding plants is synchronized through aerial signals from plants.Subject terms: Microbial ecology, Soil microbiology     

Stable isotope probing of bacterial community structure and gene expression in the rhizosphere of Arabidopsis thaliana

The rhizosphere is an active compartment where plant and microorganisms establish a molecular dialogue. In this study, we analysed the impact of Arabidopsis thaliana on bacterial community structure and the expression of certain beneficial genes using DNA- and mRNA-SIP in the rhizosphere of plantlets grown under (13)CO(2) for 13, 21 and 27 days. DNA- and rRNA-SIP revealed changes in bacterial communities inhabiting the rhizosphere soil that were probably related to modification of root exudates, while root-colonizing populations were maintained over time suggesting their metabolic versatility and adaptation. The impact of the plant via root exudates on the expression of the noncoding RNAs rsmZ, acdS gene encoding 1-aminocyclopropane-1-carboxylate deaminase and nosZ gene encoding nitrous oxide reductase, in the root-adhering soil and on the roots of A. thaliana was determined using mRNA-SIP. Results showed that these genes were present and expressed by bacteria inhabiting roots and by those that derive nutrients from the breakdown of organic matter in soils or from root exudates. The expression of rsmZ under natural conditions indicates the importance of noncoding RNAs in bacterial adaptation to their ecological niches.     

Bacterial communities associated with the rhizosphere of transgenic Bt 176 maize (Zea mays) and its non transgenic counterpart

The effect of transgenic Bt 176 maize on the rhizosphere bacterial community has been studied with a polyphasic approach by comparing the rhizosphere of Bt maize cultivated in greenhouse with that of its non transgenic counterpart grown in the same conditions. In the two plants the bacterial counts of the copiotrophic, oligotrophic and sporeforming bacteria, and the community level catabolic profiling, showed no significant differences; differences between the rhizosphere and bulk soil bacterial communities were evidenced. Automated ribosomal intergenic spacer analysis (ARISA) showed differences also in the rhizosphere communities at different plant ages, as well as between the two plant types. ARISA fingerprinting patterns of soil bacterial communities exposed to root growth solutions, collected from transgenic and non transgenic plants grown in hydroponic conditions, were grouped separately by principal component analysis suggesting that root exudates could determine the selection of different bacterial communities.     

Stability and succession of the rhizosphere microbiota depends upon plant type and soil composition

We examined succession of the rhizosphere microbiota of three model plants (Arabidopsis, Medicago and Brachypodium) in compost and sand and three crops (Brassica, Pisum and Triticum) in compost alone. We used serial inoculation of 24 independent replicate microcosms over three plant generations for each plant/soil combination. Stochastic variation between replicates was surprisingly weak and by the third generation, replicate microcosms for each plant had communities that were very similar to each other but different to those of other plants or unplanted soil. Microbiota diversity remained high in compost, but declined drastically in sand, with bacterial opportunists and putative autotrophs becoming dominant. These dramatic differences indicate that many microbes cannot thrive on plant exudates alone and presumably also require carbon sources and/or nutrients from soil. Arabidopsis had the weakest influence on its microbiota and in compost replicate microcosms converged on three alternative community compositions rather than a single distinctive community. Organisms selected in rhizospheres can have positive or negative effects. Two abundant bacteria are shown to promote plant growth, but in Brassica the pathogen Olpidium brassicae came to dominate the fungal community. So plants exert strong selection on the rhizosphere microbiota but soil composition is critical to its stability. microbial succession/ plant–microbe interactions/rhizosphere microbiota/selection.     

Lorenz Hiltner, a pioneer in rhizosphere microbial ecology and soil bacteriology research

Lorenz Hiltner is recognized as the first scientist to coin the term “rhizosphere” in 1904. His scientific career and achievements are summarized in this essay. Most of his research he performed in the Bavarian Agriculture–Botanical Institute (later named the “Bavarian Institute of Plant Growth and Plant Protection”) in Munich, where he was the director from 1902 to 1923. Beginning with intensive and thorough investigations on the germination and growth of different crop plants (legumes and non-legumes) Hiltner became convinced, that root exudates of different plants support the development of different bacterial communities. His definition of the “rhizosphere” in the year 1904 centered on the idea, that plant nutrition is considerably influenced by the microbial composition of the rhizosphere. Hiltner observed bacterial cells even inside the rhizodermis of healthy roots. In analogy with fungal root symbionts, Hiltner named the bacterial community that is closely associated with roots “bacteriorhiza.” In his rhizosphere concept, Hiltner also envisioned, that beneficial bacteria are not only attracted by the root exudates but that there are also “uninvited guests,” that adjust to the specific root exudates. Based on his observations he hypothesized that “the resistance of plants towards pathogenesis is dependent on the composition of the rhizosphere microflora.” He even had the idea, that the quality of plant products may be dependent on the composition of the root microflora. In addition to his scientific achievements, Hiltner was very dedicated to applied work. Together with F. Nobbe he had the first patent on Rhizobium inoculants (Nitragin). He continuously improved formulations and the effectivity of the Rhizobium preparations and he also initiated seed dressing with sublimate for plant protection of seedlings. Thus, Hiltner tightly linked breakthroughs in basic research to improved rhizosphere management practices. In addition, he wrote a pioneering monograph on plant protection for everybody’s practical use. His emphasis on understanding microbes in the context of their micro-habitat, the rhizosphere, made him a pioneer in microbial ecology. Even now, in the era of genome and postgenome analysis with our better understanding of plant nutrition and soil bacteriology, his ideas and contributions are as fresh as they were more than 100 years ago.     

Quantitative Variation in Responses to Root Spatial Constraint within Arabidopsis thaliana

Among the myriad of environmental stimuli that plants utilize to regulate growth and development to optimize fitness are signals obtained from various sources in the rhizosphere that give an indication of the nutrient status and volume of media available. These signals include chemical signals from other plants, nutrient signals, and thigmotropic interactions that reveal the presence of obstacles to growth. Little is known about the genetics underlying the response of plants to physical constraints present within the rhizosphere. In this study, we show that there is natural variation among Arabidopsis thaliana accessions in their growth response to physical rhizosphere constraints and competition. We mapped growth quantitative trait loci that regulate a positive response of foliar growth to short physical constraints surrounding the root. This is a highly polygenic trait and, using quantitative validation studies, we showed that natural variation in EARLY FLOWERING3 (ELF3) controls the link between root constraint and altered shoot growth. This provides an entry point to study how root and shoot growth are integrated to respond to environmental stimuli.     

Aromatic compounds degradation plays a role in colonization of Arabidopsis thaliana and Acacia caven by Cupriavidus pinatubonensis JMP134

Plant rhizosphere and internal tissues may constitute a relevant habitat for soil bacteria displaying high catabolic versatility towards xenobiotic aromatic compounds. Root exudates contain various molecules that are structurally related to aromatic xenobiotics and have been shown to stimulate bacterial degradation of aromatic pollutants in the rhizosphere. The ability to degrade specific aromatic components of root exudates could thus provide versatile catabolic bacteria with an advantage for rhizosphere colonization and growth. In this work, Cupriavidus pinatubonensis JMP134, a well-known aromatic compound degrader (including the herbicide 2,4-dichlorophenoxyacetate, 2,4-D), was shown to stably colonize Arabidopsis thaliana and Acacia caven plants both at the rhizoplane and endorhizosphere levels and to use root exudates as a sole carbon and energy source. No deleterious effects were detected on these colonized plants. When a toxic concentration of 2,4-D was applied to colonized A. caven, a marked resistance was induced in the plant, showing that strain JMP134 was both metabolically active and potentially beneficial to its host. The role for the β-ketoadipate aromatic degradation pathway during plant root colonization by C. pinatubonensis JMP134 was investigated by gene inactivation. A C. pinatubonensis mutant derivative strain displayed a reduced ability to catabolise root exudates isolated from either plant host. In this mutant strain, a lower competence in the rhizosphere of A. caven was also shown, both in gnotobiotic in vitro cultures and in plant/soil microcosms.     

植物根系分泌物对土壤污染修复的作用及影响机理

生物修复是一种经济环保的土壤修复技术。根系分泌物是利用生物修复污染土壤过程中的关键物质,也是植物与土壤微生物进行物质交换和信息传递的重要载体,在植物响应污染物胁迫中扮演重要角色。研究植物根系分泌物对土壤污染修复的作用和影响机理,是深入理解植物和微生物环境适应机制的重要途径,对促进生物修复污染土壤有重要指导意义。从污染物胁迫对根系分泌物的影响、根系分泌物对土壤污染物环境行为的影响、根系分泌物在调控污染土壤中根际微生物群落结构和多样性中发挥的作用等几个方面综述了根系分泌物对土壤污染修复的影响及内在机制。研究结果表明,根系分泌物在降低重金属对植物的毒性、加速有机污染物降解等方面有非常重要的作用。根系分泌物对土壤微生物的丰度和多样性均有显著影响,其与根际微生物互作在土壤污染物的消减中发挥了重要的调控作用。在此基础上,提出了以往研究中的不足,并对污染物胁迫下根系分泌物未来研究的方向和趋势进行了展望。     

根系分泌物介导的根际效应及在水体生态修复中的应用潜力

文章主要以根系分泌物为核心, 综述了根系分泌物的分类、发生机理及影响因素; 围绕着植物-土壤-微生物三者的关系, 阐述了根系分泌物介导的植物与植物之间的化感作用、植物与根际微生物之间的协同作用以及植物微生物相互作用对土壤物质循环的影响。水生植物以其生境的特殊性和功能的不可替代性, 对沉积物污染物去除和水体生态修复产生显著的影响。在水体生态问题较为严峻的今天, 充分认识水生植物根系分泌物介导的根际过程将为水生植物生态学和水体生态修复领域的发展提供基础, 并为学科应用潜力的开发提供依据。     

Genotype and rhizobium inoculation modulate the assembly of soybean rhizobacterial communities

Rhizosphere bacterial communities are vital for plants, yet the composition of rhizobacterial communities and the complex interactions between roots and microbiota, or between microbiota, are largely unknown. In this study, we investigated the structure and composition of rhizobacterial communities in two soybean cultivars and their recombinant inbred lines contrasting in nodulation through 16S rRNA amplicon sequencing in two years of field trials. Our results demonstrate that soybean plants are able to select microbes from bulk soils at the taxonomic and functional level. Soybean genotype significantly influenced the structure of rhizobacterial communities and resulted in dramatically different co‐occurrence networks of rhizobacterial communities between different genotypes of soybean plants. Furthermore, the introduction of exogenous rhizobia through inoculation altered soybean rhizobacterial communities in genotype‐dependent manner. Rhizobium inoculation not only stimulated the proliferation of potential beneficial microbes but also increased connections in rhizobacterial networks and changed the hub microbes, all of which led to the association of distinctive bacterial communities. Taken together, we demonstrated that the assembly of soybean rhizobacterial communities was determined by both genotype and the introduction of exogenous rhizobia. These findings bolster the feasibility of root microbiome engineering through inoculation of specific microbial constituents.     

Phytotoxic Allelochemicals From Roots and Root Exudates of Leafy Spurge (Euphorbia esula L.)

Invasive plants are a widespread problem but the mechanisms used by these plants to become invasive are often unknown. The production of phytotoxic natural products by invasive weeds is one mechanism by which these species may become successful competitors. Here we conducted a bioactivity-driven fractionation of root extracts and exudates from the invasive plant leafy spurge (Euphorbia esula L.), and structurally characterized jatrophane diterpenes and ellagic acid derivatives. Ellagic acid derivatives and one of the jatrophane diterpenes, esulone A, have been previously reported from leafy spurge, but another of the jatrophane diterpenes, kasuinine B, has not. We show that these compounds are phytotoxic but affect plants in different ways, either inducing overall plant necrosis or reducing root branching and elongation.Key Words: phytotoxicity, allelochemicals, roots, root exudates, jatrophane diterpenes, kansuinine B, ellagic acid derivatives, leafy spurge, Euphorbia esula, Arabidopsis thaliana     

Intraspecific interactions in the annual legume Medicago minima are shaped by both genetic variation for competitive ability and reduced competition among kin

Documenting if plants exhibit kin competition avoidance in intraspecific plant interactions is relevant both to improve crop growth, and to understand diversity and composition in natural plant communities. However, a number of confounding mechanisms complicates detecting kin competition avoidance from experiments comparing plants growing with kin and non-kin neighbors. We conducted complementary greenhouse experiments using genotypes from four populations of the annual Medicago minima, which in a previous study showed higher survival when interacting with kin relative to non-kin. We show that genotypes vary in kin competition avoidance, and in competitive ability, but find no indication of complementary resource use. Importantly, from our first experiment of root growth behavior, we know that some genotypes exhibit kin competition avoidance. Yet, the variation in competitive ability we find in our second experiment, where plants grow in mini communities together with either kin or unrelated genotypes, can alone explain the variation we observe in growth and biomass among communities. In our case, the genotypes with highest competitive ability were also those that showed kin competition avoidance. This confounding effect obscured the disentangling of mechanisms underlying difference in growth between kin and non-kin interactions. When silencing root exudates by adding activated carbon to a subset of our genotype combinations, we found increased size asymmetry of plants grown together, and mostly in kin communities. This suggests that plants recognize the identity of neighbors via root exudates, and compete less with neighbors recognized as kin. To detect kin competition avoidance we suggest designing experiments that pair unrelated genotypes with similar competitive abilities. Such design, combined with silencing root exudates would be powerful to detect whether plants show kin competition avoidance or not.