樱桃根际促生细菌的筛选与鉴定

【目的】筛选樱桃[Cerasus pseudocerasus(Lindl.)G.Don]生物肥料中应用的优良菌株资源。【方法】通过保绿法和萝卜子叶增重法,结合定量检测细菌分离物产植物激素的能力,从樱桃根际土壤中筛选具有良好应用潜力的细菌分离物。利用盆栽试验,验证其促生效果。对促生效果较好的细菌分离物,进行形态学观察、生理生化测定、16S rRNA基因序列及系统发育树分析以确定其分类地位。【结果】筛选出4株具有良好应用潜力的细菌分离物,盆栽试验结果表明:AI5、AI21、PII17、PI7,尤其是PII17,显著提高樱桃根系及地上部生物量,对樱桃生长有明显的促进作用。分类鉴定结果显示:AI5、AI21为短小芽孢杆菌(Bacillus pumilus),PII17为假单胞菌属(Pseudomonas sp.),PI7为肠杆菌属(Enterobacter sp.)。【结论】AI5、AI21、PII17、PI7,尤其是PII17,对樱桃生长有明显的促进作用,在生物肥料的生产中具有广阔的应用前景。     

Effects of growth-promoting rhizobacteria on maize growth and rhizosphere microbial community under conservation tillage in Northeast China

Conservation tillage in conjunction with straw mulching is a sustainable agricultural approach. However, straw mulching reduces the soil temperature, inhibits early maize growth and reduces grain yield in cold regions. To address this problem, we investigated the effects of inoculation of plant growth-promoting rhizobacteria (PGPR) on maize growth and rhizosphere microbial communities under conservation tillage in Northeast China. The PGPR strains Sinorhizobium sp. A15, Bacillus sp. A28, Sphingomonas sp. A55 and Enterobacter sp. P24 were isolated from the maize rhizosphere in the same area and inoculated separately. Inoculation of these strains significantly enhanced maize growth, and the strains A15, A28 and A55 significantly increased grain yield by as much as 22%–29%. Real-time quantitative PCR and high-throughput sequencing showed that separate inoculation with the four strains increased the abundance and species richness of bacteria in the maize rhizosphere. Notably, the relative abundance of Acidobacteria_Subgroup_6, Chloroflexi_KD4-96, and Verrucomicrobiae at the class level and Mucilaginibacter at the genus level were positively correlated with maize biomass and yield. Inoculation with PGPR shows potential for improvement of maize production under conservation tillage in cold regions by regulating the rhizosphere bacterial community structure and by direct stimulation of plant growth.     

植物根际促生菌的筛选及其对玉米的促生效应

[目的]以不同植物根及根际土壤为研究材料,进行植物根际促生菌(PGPR)的筛选,并探索其植物促生作用机制.[方法]以解磷、固氮、产氨、产IAA和拮抗3种常见病原真菌为筛选标准,测定了初筛菌株的多项促生能力,并通过对这些菌分别单独回接和多菌混接的玉米盆栽试验,测定了其对玉米的促生效应.[结果]从渭南、成阳、安康、商洛和榆林5地分离得到的158株菌中有17株茵具有上述多种植物促生作用的菌株.盆栽试验的测定结果表明:单独接种和多菌混合接种在玉米株高、根长、茎长、茎平均直径和干重方面与对照组相比较都有所增加,尤其是在多个指标上,多菌混合接种所显示出的促生效应均明显优于单菌接种.[结论]所筛选到的具有多种促生能力的菌株,可以为进一步构建植物根际促生菌(PGPR)菌群提供良好的种质资源.     

正交试验法分析环境因子对苦草生长的影响

苦草(Vallisneria natans),是我国长江中下游淡水水体中常见的沉水植物种类。通过室内模拟正交试验的方法,研究不同光照强度、温度和总氮浓度(3光照×3温度×3总氮浓度)对苦草生长的影响。结果表明:(1)苦草在5320lx光照强度、10℃、2—4mg/L水体总氮浓度的条件下生长良好。实验所设100%(12000lx)和50%(5320lx)光照条件下苦草均可正常生长;但对于30℃的高温胁迫耐受性较差;苦草在总氮浓度为4mg/L的水体中各生长指标达到最大值,2mg/L或8mg/L的总氮浓度均会抑制其生长。(2)5320lx的光照强度和10℃的温度对苦草光合色素的合成较为有利;而单纯总氮浓度的变化对苦草光合色素的合成影响不大。(3)苦草的生理活性在高于12000lx或低于1025lx的光强、高于30℃的温度以及8mg/L的总氮浓度下均会受到一定程度的抑制。(4)方差分析结果显示,苦草生长发育的过程中,光照强度和温度是主效环境因子;上述3个环境因子对苦草光合色素的合成均有极显著的影响,并且光强与温度的交互作用对其也有显著影响;光照强度、温度以及这两个环境因子的交互作用为影响苦草生理活性的主效因子。苦草作为不能形成冠层的基生叶莲座型沉水植物,对光强要求不高,对低温的耐受性较好,但较不耐高营养盐浓度,因此,在得到一定修复的富营养化水体中,可以作为秋、冬季水生植物恢复和重建的关键物种。     

Defoliation affects rhizosphere respiration and rhizosphere priming effect on decomposition of soil organic matter under a sunflower species: Helianthus annuus

In the present study, `natural <sup>13</sup>C tracer method' was used to partition the belowground respiration into rhizosphere respiration and soil microbial respiration to test the hypothesis that defoliation affects rhizosphere respiration and rhizosphere priming effect on decomposition of soil organic matter (SOM). A C<sub>3</sub> plant species, Helianthus annuus (sunflower), was grown in `C₄' soil in microcosms so that the CO₂ evolved from plant-soil system can be partitioned. Four levels of defoliation intensities were established by manual clipping. CO₂ evolved from plant-soil system was trapped during 0–4 h after defoliation (HAD), 5–22 HAD and 23–46 HAD using a closed circulating system, respectively. We found that both rhizosphere respiration and soil microbial respiration of the clipped plants were either unchanged or significantly enhanced compared to unclipped plants at 45% defoliation level during all sampling intervals. Soil microbial respiration increased significantly at all defoliation levels during 0–4 HAD, however, both rhizosphere respiration and soil microbial respiration decreased significantly during 5–22 HAD or 23–46 HAD when 20% or 74 clearly demonstrated that the defoliation treatments modified the rhizosphere priming effect on SOM decomposition. The total cumulative rhizosphere priming effects on SOM decomposition during 0–46 HAD were 146%, 241%, 204% and 205% when 0%, 20%, 45% and <74%.     

Plant diversity shapes microbe‐rhizosphere effects on P mobilisation from organic matter in soil

Plant species richness (PSR) increases nutrient uptake which depletes bioavailable nutrient pools in soil. No such relationship between plant uptake and availability in soil was found for phosphorus (P). We explored PSR effects on P mobilisation [phosphatase activity (PA)] in soil. PA increased with PSR. The positive PSR effect was not solely due to an increase in C_org concentrations because PSR remained significant if related to PA:C_org. An increase in PA per unit C_org increases the probability of the temporal and spatial match between substrate, enzyme and microorganism potentially serving as an adaption to competition. Carbon use efficiency of microorganisms (C_mic:C_org) increased with increasing PSR while enzyme exudation efficiency (PA:C_mic) remained constant. These findings suggest the need for efficient C rather than P cycling underlying the relationship between PSR and PA. Our results indicate that the coupling between C and P cycling in soil becomes tighter with increasing PSR.     

Plant growth-promoting rhizobacteria do not pose any deleterious effect on cowpea and detectable amounts of ethylene are produced

Summary The use of trap crops such as cowpea could reduce the effects of the root parasitic weed, Striga hermonthica and its subsequent constraints on the growth of cereals. Certain bacteria could augment the trap crop stimulatory effect. We studied the effect of three bacteria introduced to the rhizosphere of three cowpea varieties at planting. Number of days to cowpea flowering was noted and at harvest, data were collected on pod characteristics and biomass. Means of data subjected to ANOVA were compared using Tukey’s Studentized Range Test. We analysed bacterial headspace volatiles for ethylene by gas chromatography and gas chromatography–mass spectrometry. Bacterial type significantly influenced the cowpea varieties with better performance over the non-inoculated control. Average pod weight (g) with bacterial treatment was 37.97 for Enterobacter sakazakii 8MR5, 34.38 for Pseudomonas 44MS8 and 27.46 for Pseudomonas 10M3. Non-inoculated control had an average weight of 20.98 g. Bacteria promoted a significant increase in pod weight (≥30.89%), fresh biomass (≥24.22%), and improved pod number (≥20.54%) and pod wall thickness (≥7.33%) with no deleterious effect on plant health. Ethylene released by the bacteria ranged from trace concentrations in Pseudomonas sp. to 210 nmoles/10⁸ c. f. u./ml in Ent. sakazakii 8MR5.     

Effects of plant architecture and water velocity on sediment retention by submerged macrophytes

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Influence of submerged macrophytes on sediment composition and near-bed flow in lowland streams

1. Submerged macrophytes have important physical and structural effects on lowland streams. This study investigated the ability of submerged macrophytes to modify the near-bed flow and to retain mineral and organic particles in patches of four common macrophytes in shallow Danish streams during mid-summer. 2. In dense patches of Callitriche cophocarpa and Elodea canadensis, where near-bed velocity was reduced, the sediment surface was markedly raised and enriched with fine particles. In dense patches of Ranunculus peltatus, fine sediments were deposited among rooted shoots in the upstream part of the patches, while erosion and coarse sediments prevailed in the downstream part of the patches because of the strong vortices that formed at the rear and moved up under the trailing canopy. The open canopy of Sparganium emersum, with its streamlined leaves, had little effect on flow and sediment. 3. Patterns of sediment deposition and composition were closely related to the morphology and canopy structure of plant species and the presence of low velocity above the sediment among the rooted shoots. The mineral particles retained probably originate from bed-load, and the enrichment with finer particles within the patches probably results mainly from size-selective processes during erosion and transport of particles rather than during deposition. The mixed sediment composition within patches suggests that the flow-resistant shoots generate an environment conducive to deposition of all transported particles. 4. Fine sediments within macrophyte beds contained high concentrations of organic matter, carbon, nitrogen and phosphorus. The wide scatter in the relationships between mineral grain size and the content of organic matter and nutrients reflects the spatial and temporal complexity of erosion, transport and sedimentation of mineral and organic particles. 5. Enrichment of sediment within macrophyte beds relative to the surrounding substratum ranged from 780 g organic matter m^–2, 30 g N m^–2 and 25 g P m^–2 for the flow-resistant dense canopies af Callitriche cophocarpa to 150 g organic matter m^–2, 6.6 g N m^–2 and 3.4 g P m^–2 for the open canopies of Sparganium emersum. Retention of nutrient-rich particles within the macrophyte beds is probably of limited importance for plant growth in most lowland European streams, because macrophyte growth is rarely nutrient limited.     

叶片损害强度与基质营养水平对苦草补偿性生长的影响

通过室外双因子控制试验,研究了叶片损害强度与基质营养水平对沉水植物苦草补偿性生长的影响.结果表明：叶片损害与基质类型对苦草的累积生物量、生物量的分配（根叶比）及分株数均有显著影响.与未受损植物相比,强受损使苦草的累积生物量及无性分株数显著降低,根叶比明显升高,而弱受损与未受损处理之间没有明显差别.与营养贫瘠的岸泥相比,生长在肥力较高湖泥中的苦草的累积生物量较高,而根叶比与分株数较低;叶片损害与基质类型对苦草的相对生长率也有显著影响,受损使苦草的生长率明显提高,而两受损处理之间无显著差别.未受损与强受损时,湖泥中苦草的生长速率明显高于岸泥,弱受损时则相近.并初步探讨了水生植物受损后的补偿生长机理.     

Plant growth-promoting and non-promoting rhizobacteria from avocado trees differentially emit volatiles that influence growth of Arabidopsis thaliana

Microbial volatile organic compounds (mVOCs) play important roles in inter- and intra-kingdom interactions, and they are also important as signal molecules in physiological processes acting either as plant growth-promoting or negatively modulating plant development. We investigated the effects of mVOCs emitted by PGPR vs non-PGPR from avocado trees (Persea americana) on growth of Arabidopsis thaliana seedlings. Chemical diversity of mVOCs was determined by SPME–GC–MS; selected compounds were screened in dose–response experiments in A. thaliana transgenic lines. We found that plant growth parameters were affected depending on inoculum concentration. Twenty-six compounds were identified in PGPR and non-PGPR with eight of them not previously reported. The VOCs signatures were differential between those groups. 4-methyl-2-pentanone, 1-nonanol, 2-phenyl-2-propanol and ethyl isovalerate modified primary root architecture influencing the expression of auxin- and JA-responsive genes, and cell division. Lateral root formation was regulated by 4-methyl-2-pentanone, 3-methyl-1-butanol, 1-nonanol and ethyl isovalerate suggesting a participation via JA signalling. Our study revealed the differential emission of volatiles by PGPR vs non-PGPR from avocado trees and provides a general view about the mechanisms by which those volatiles influence plant growth and development. Rhizobacteria strains and mVOCs here reported are promising for improvement the growth and productivity of avocado crop.

     

衰亡期沉水植物对水和沉积物磷迁移的影响

通过室内模拟的方式,分别研究了秋季黑藻、苦草和春季菹草的衰亡过程,分析了沉水植物在衰亡期间水、沉积物中磷与环境因子[pH、氧化还原电位(Eh)和溶解氧(DO)]的相互作用.结果表明:沉水植物黑藻和菹草在衰亡期间能显著提高水中总磷(TP)、溶解性总磷(DTP)、颗粒磷(PP)、溶解性活性磷(SRP)和溶解性有机磷(DOP)的含量;苦草在衰亡期对水体各形态磷含量影响不显著,且各形态磷含量的变化相对较小(TP,0.04-0.06 mg/L);环境因子的变化对水中磷含量影响显著,黑藻和菹草水体中TP的含量和环境因子pH、DO和Eh均呈负相关,而苦草组水中各形态磷的含量受环境因子影响不显著.实验期间不同植物组沉积物中总磷(TP)、NaOH提取磷(NaOH-P)、HCl提取磷(HCl-P)、无机磷(IP)和有机磷(OP)的含量均呈上升趋势.沉积物IP的含量主要受NaOH-P的影响,OP对TP的影响要大于IP,沉积物OP与OM(有机质)的含量存在显著正相关.     

Effect of inoculation with putative plant growth-promoting rhizobacteria isolated from Pinus spp. on Pinus pinea growth, mycorrhization and rhizosphere microbial communities

Aims: In this study, 10 putative plant growth-promoting rhizobacteria (PGPR) were assayed for their ability to improve Pinus pinea growth and mycorrhization. Methods and Results: After an inoculation assay, except for two, all strains stimulated plant growth. All bacteria altered rhizosphere microbial communities as revealed by phospholipid fatty acid analysis; associating plant growth promotion with a decrease in biological diversity. Three strains were tested for their ability to enhance pine mycorrhization with wild fungi species. Only strain BB1 increased the total number of mycorrhizal root tips. Mycorrhizas present in the roots of each treatment were identified by ribosomal RNA sequencing and denaturing gradient gel electrophoresis analysis, detecting specificity between mycorrhizal species colonizing the roots and the inoculated PGPR. Conclusions: In conclusion, BB1 appears to be a good candidate to be developed into a biofertilizer directed to enhance pine growth and mycorrhization, which should result in a better establishment rate for plants used in reforestation. Significance and Impact of the Study: This study shows the potential of PGPR to improve fitness of forest tree specie. Moreover, the specificity between the bacteria inoculated and the mycorrhiza that the plant selects involve a potential biotechnological use in production of value-added fungi.     

Effects of retrogressive permafrost thaw slumping on sediment chemistry and submerged macrophytes in Arctic tundra lakes

1. Global warming is predicted to cause changes in permafrost cover and stability in the Arctic. Zones of high ion concentration in regions of ice‐rich permafrost are a reservoir of chemicals that can potentially be transferred to fresh waters during thawing. Consequently, input of enriched runoff from the thaw and sediment and vegetation from the landscape could alter lakes by affecting their geochemistry and biological production. 2. Three undisturbed lakes and five lakes disturbed by retrogressive permafrost thaw slumps were sampled during late summer of 2006 to assess the potential effects of thermokarst shoreline slumping on water and sediment chemistry, the underwater light regime, and benthic macrophyte biomass and community structure. 3. Undisturbed lakes had sediments rich in organic material and selected micronutrients, while disturbed lakes had sediments richer in calcium, magnesium and strontium, greater transparency of the water column, and a well‐developed submerged macrophyte community. 4. It is postulated that enriched runoff chemistry may alter nutrient availability at the sediment–water interface and also the degradation of organic material, thus affecting lake transparency and submerged macrophytes. The results suggest that retrogressive permafrost slumping can significantly affect food webs in arctic tundra lakes through an increase in macrophyte biomass and development of a more complex benthic habitat.     

Low-molecular-weight organic acids in rhizosphere soils of durum wheat and their effect on cadmium bioaccumulation

Cadmium (Cd) accumulation has been found to vary between cultivars of durum wheat (Triticum turgidum var. durum), and it is hypothesized that low-molecular-weight organic acids (LMWOAs) produced at the soil-root interface (rhizosphere) may play an important role in the availability and uptake of Cd by these plants. The objective of this study, therefore, was to (1) investigate the nature and quantity of LMWOAs present in the rhizosphere of durum wheat cultivars Arcola (low Cd accumulator) and Kyle (high Cd accumulator) grown in three different soils: Yorkton, Sutherland and Waitville, and (2) determine the relationship between Cd accumulation in these plants and LMWOAs present in the rhizosphere. Plants were grown for two weeks in pot-cultures under growth chamber conditions. Oxalic, fumaric, succinic, L-malic, tartaric, citric, acetic, propionic and butyric acids were found and quantified in the water extracts of rhizosphere soil, with acetic and succinic acids being predominant. No water extractable LMWOAs were identified in the bulk soil. Total amount of LMWOAs in the rhizosphere soil of the high Cd accumulator (Kyle) was significantly higher than that for the low Cd accumulator (Arcola) in all three soils. Furthermore, large differences in amounts of LMWOAs were found in the rhizosphere soil for the same cultivars grown in different soils and followed the pattern: Sutherland > Waitville > Yorkton. Extractable soil Cd (M NH4Cl) and Cd accumulation in the plants also followed the same soil sequence as LMWOA production. Cadmium accumulation by the high and low Cd accumulating cultivars was proportional to the levels of LMWOAs found in the rhizosphere soil of each cultivar. These results suggest that the differing levels of LMWOAs present in the rhizosphere soil played an important role in the solubilization of particulate-bound Cd into soil solution and its subsequent phytoaccumulation by the high and low Cd accumulating cultivars.     

Application of plant growth-promoting rhizobacteria to soybean (Glycine max [L.] Merr.) increases protein and dry matter yield under short-season conditions

We previously reported that application of plant growth-promoting rhizobacteria (PGPR) increased soybean growth and development and, specifically, increased nodulation and nitrogen fixation over a range of root zone temperatures (RZTs) in controlled environment studies. In order to expand on the previous studies, field experiments were conducted on two adjacent sites, one fumigated with methyl bromide and one nonfumigated, in 1994. Two experiments were conducted at each site, one involving combinations of two soybean cultivars and two PGPR strains, the other involving the same factors, but also in combination with two strains Bradyrhizobium japonicum. Soybean grain yield and protein yield were measured. The results of these experiments indicated that co-inoculation of soybean with B. japonicum and Serratia liquefaciens 2-68 or Serratia proteamaculans 1-102 increased soybean grain yield, protein yield, and total plant protein production, compared to the nontreated controls, in an area with low spring soil temperatures. Interactions existed between PGPR application and soybean cultivar, suggesting that PGPRs applied to cultivars with higher yield potentials were more effective. PGPRs applied to the rhizosphere without addition of B. japonicum also increased only leaf area and seed number at the fumigated site. Overall, inoculation of soybean plants with PGPRs in the presence of B. japonicum increased soybean grain yield, grain protein yield, and total plant protein production under short season conditions.     

The effect of deposition of organic matter on phosphorus dynamics in experimental marine sediment systems

The effect of deposition of organic matter on phosphorus dynamics in sandy marine sediments was evaluated using an experimental system (boxcosms) and three different strategies: (1) no supply (2) one single addition (3) weekly additions of a suspension of algal cells (Phaeocystis spec.). Macrofauna (3 species, 6 individuals of each) were added to half of the boxes. Both in the case of the single and weekly additions a clear effect of increased organic matter loading on phosphorus dynamics was found. Following the organic matter addition, porewater phosphate concentrations in the upper sediment layer increased, phosphate release rates from the sediment increased by a factor 3–5 and in the boxes to which a single addition was applied NaOH-extractable phosphorus increased substantially. The increase in phosphate release rates from the sediment was attributed to mineralization of the added material and to direct release from the algal cells. No clear effect of the presence of macrofauna on sediment-water exchange of phosphate could be discovered. The macrofauna were very effective at reworking the sediment, however, as illustrated by the organic carbon profiles. It is hypothesized that the sediment-water exchange rates of phosphate were regulated by the layer of algal material which was present on the sediment surface in the fed boxes. In the boxes to which the single addition was applied porewater phosphate concentrations were lower and NaOH-extractable phosphorus was higher in the presence of macrofauna, suggesting that macrofauna can stimulate phosphate binding in the sediment.Publication no. 40 of the project Applied Scientific Research Netherlands Institute for Sea Research (BEWON)     

Effect of sediment depth and sediment type on the survival of Vallisneria americana Michx grown from tubers

Sedimentation resulting from storms may have been one of the reasons for the elimination of submersed aquatic vegetation from the tidal Potomac River in the late 1930's. Laboratory studies were conducted to investigate the effects of different depths of overlying sediment and composition of sediment on the survival of Vallisneria americana Michx (wildcelery) grown from tubers. Survival of plants grown from tubers decreased significantly with increasing sediment depth. Survival of tubers declined from 90% or more when buried in 10 cm to no survival in greater than 25 cm of sediment. Survival with depth in sand was significantly lower than in silty clay.Field investigation determined that the majority of tubers in Vallisneria beds are distributed between 10 and 20 cm in depth in silty clay and between 5 and 15 cm in depth in sand. Based on the field distribution of tubers and on the percent survival of plants growing from tubers at each depth in the laboratory experiment, we suggest that the deposition of 10 cm or more of sediment by severe storms such as occurred in the 1930s could contribute to the loss of vegetation in the tidal Potomac River.