Effect of calcium concentration in nutrient solution on development of bacterial wilt and population of its pathogen Ralstonia solanacearum in grafted tomato seedlings

We examined the effect of the calcium (Ca) concentration in the nutrient solution on the development of bacterial wilt and the population of its pathogen Ralstonia solanacearum in tomato (Lycopersicon esculentum Mill. cv. Momotarou) seedlings grafted onto the rootstock of a highly resistant cultivar (cv. Hawaii 7998). The grafted seedlings were cultured in a nutrient solution containing Ca at concentrations of 0.4, 4.4, and 20.4 mm, and inoculated with the pathogen by stem puncture at the base of the stem of the rootstock, and the disease incidence was recorded for a period of 21 d. In another experiment, xylem exudates were collected from decapitated scions of the Ca-treated seedlings 5 d after inoculation, and the population of the pathogen in the exudates was counted by plating on a selective medium. The grafted tomato seedlings were highly resistant to bacterial wilt, when cultured in the nutrient solution containing Ca at 20.4 mm. The population of the pathogen in the xylem exudates decreased with increasing concentration of Ca in the solution. However, even in the presence of Ca at a high concentration, infection with the virulent pathogen was observed in the xylem of the scion.     

Effect of calcium concentration in nutrient solution before and after inoculation with Ralstonia solanacearum on resistance of tomato seedlings to bacterial wilt

We previously reported that calcium (Ca) nutrition in tomato (Lycopersicon esculentum Mill.) significantly affected the resistance to bacterial wilt caused by Ralstonia solanacearum Smith. To elucidate the mechanisms underlying the Ca-dependent resistance, the effect of the Ca concentration in the nutrient solution applied before and after inoculation with the pathogen on the resistance of tomato seedlings to bacterial wilt was studied. One week before inoculation, seedlings were transferred to nutrient solutions containing Ca at concentrations of 0.4, 4.4, or 20.4 mM. Soon after inoculation, the seedlings that were treated with each concentration of Ca before inoculation were transferred to solutions containing the same three concentrations of Ca. Although the disease development was not affected by the concentration of Ca in the solution before inoculation, a higher concentration of Ca after inoculation reduced the disease severity. This result suggests that the concentration of Ca in the host, especially in the cell walls, before infection may not be directly involved in the Ca-dependent resistance of tomato seedlings to bacterial wilt.     

Calcium uptake and resistance to bacterial wilt of mutually grafted tomato seedlings

Bacterial wilt of tomato (Lycopersicon esculentum Mill.) caused by Ralstonia solanacearum Smith is a serious disease in Japan. We previously reported that calcium (Ca) nutrition in tomato significantly affected the resistance to the disease, and that highly resistant cultivars were characterized by a high Ca uptake. We examined the relationship between the Ca uptake and resistance using mutually grafted seedlings of tomato cultivars differing in their resistance. A susceptible (‘Ponderosa’) or moderately resistant (‘Zuiei’) cultivar (scion) was grafted on the rootstock of a susceptible, moderately resistant, or highly resistant cultivar (‘Hawaii 7998’). Roots or petioles of the grafted seedlings were inoculated with the pathogen, and the development of bacterial wilt was observed. Although Ca uptake by shoot increased by grafting on the rootstock of a highly resistant cultivar, the development of the disease was not influenced by the difference in Ca uptake, and depended on the resistance of the cultivar to which the inoculated part of the graft belonged. It was concluded that the differences in Ca uptake of the shoot of the grafted tomato seedlings might not be related to the expression of the resistance to bacterial wilt.     

Changes in the nitrogen localized in the lamellae systems and stroma of rice chloroplast accompanying the stages of growth

The change in the nitrogen localized in the lamellae systems and stroma region of the rice chloroplast was pursued throughout the stages of growth and, further, the relationship between the composition of the chloroplast and leaf N content was examined. A common trend accompanying the stages of growth is recognized: that in the early stages of growth the amount of nitrogen existing as a form of stroma N is larger than that of lamellae N. Thereafter, stroma N tends to decrease in relation to lamellae N and finally, lamellae N becomes a dominant nitrogenous fraction in the chloroplast. The change in the composition of the chloroplast seems to be brought about by a change in the nitrogen metabolism of the chloroplast by which the distribution of absorbed nitrogen among the nitrogenous fractions of the chloroplast differs.     

青枯雷尔氏菌Tn5转座子无致病力突变株构建及其生物学特性

青枯雷尔氏菌(Ralstonia solanacearum)是植物细菌性青枯病的病原菌,为研制植物疫苗菌剂提供遗传稳定且突变位点明确的青枯雷尔氏菌无致病力菌株,本研究利用EZ-Tn5转座子随机插入青枯雷尔氏菌(Rs91)构建青枯雷尔氏菌无致病力突变体库,通过电击转化,筛选获得13株具有无致病力菌株形态的突变株。研究了其中5株突变株的插入位点和生物学特性,结果表明,其插入位点分别位于phcA和pchS基因,其生长速率和相对胞外多糖含量显著低于Rs91,而最适pH值和温度未改变。其发酵液经分光光度计扫描,结果显示,突变株在同一波长下的光吸收值均大于Rs91,经聚类分析,显示它们与Rs91可聚为不同的3类,即Rs91为第Ⅰ类,phcA基因突变株为第Ⅱ类,phcS基因突变株为第Ⅲ类。经番茄(Lycopersicon esculentum)盆栽苗致病力检测,15d后均未发病,确定为无致病力青枯雷尔氏菌。本研究为研制防治青枯病的植物疫苗菌剂提供了基础资料。     

插入序列ISRso21在青枯雷尔氏菌中的分布以及插入位点多态性分析

插入序列(insertion sequence,IS)元件可以插入到基因或DNA序列,其可能在生物体进化中起着重要的作用。ISRso21是在青枯雷尔氏菌(Ralstonia solanacearum)FJAT-1458菌株中发现的一个新的插入序列。根据序列分析可知,该插入序列是属于ISL3家族中的一个成员。本研究发现,福建闽北地区菌株ISRso21的阳性率高于其他地区的趋势,不同地区的青枯雷尔氏菌ISRso21的分布有显著性差异,ISRso21的分布可能与菌株分离个体的地理来源有关。不仅如此,由于ISRso21在青枯雷尔氏菌插入位点的不同,从而导致青枯雷尔氏菌致病性的差异。研究表明,由于ISRso21的插入导致表现型转换系统转录调控因子A(phc A)的重排可能在青枯雷尔氏菌的致病性起着极其重要的作用。在所有菌株中,ISRso21在phc A基因上游中插入的检出率达到4.71%,其中无致病力菌株和强致病力菌株中的检出率分别为28.57%和0.00%。而ISRso21在二氨基庚二酸脱羧酶基因中的插入可能是为了更好地适应环境。ISRso21在不同致病性青枯雷尔氏菌中的插入位点的研究,可能为揭示青枯雷尔氏菌致病性机制提供新的研究途径。     

连续施用生物有机肥对烟草青枯病的防治效果

分离获得一株对烟草青枯病病原菌茄科劳尔氏菌(Ralstonia solanacearum,简称RS)具有较强拮抗能力的拮抗菌(SQR11)并制成生物有机肥,研究了连续施用该生物有机肥对烟草青枯病的防治效果。结合生理生化和16SrDNA技术鉴定,菌株SQR11被鉴定为解淀粉芽孢杆菌。施用该生物有机肥后第一季烟草青枯病的生物防治率达到47%以上,第二季为69%以上,第三季达到89%以上。第三批盆栽实验表明,当根际土中病原菌数量达到2×105cfu/g干土时,植株出现发病症状,随着病原菌数量的增加,发病症状加重。当根际土中拮抗菌活菌数量达到2×107cfu/g干土时,病原菌繁殖得到有效抑制,可有效阻止植株染病;若低于107cfu/g干土,则不能有效抑制病原菌增殖,植株表现发病症状。植株各组织内拮抗菌数量检测发现,未发病植株茎部拮抗细菌数量为4×104cfu/g(组织鲜重,下同)左右,而同处理中发病症状的植株茎部拮抗细菌数量仅为6×103cfu/g;相对应的病原菌数量分别为1.5×102cfu/g(健康植株)和3×103cfu/g(发病植株)。SQR11菌株制成的生物有机肥还具有较好的促生作用。总之,利用拮抗菌SQR11菌株制成的生物有机肥对烟草青枯病具有显著的生物防治作用,在根部进行大量定殖后可有效防止病原菌的侵入,能够获得显著的生防效果。     

Host-plant resistance to Pseudomonas solanacearum in tomato germplasm

Seedlings of two hundred and thirty-three accessions of the tomato collection maintained at the Centro Agronómico Tropical de Investigación y Enseñanza, Turrialba, Costa Rica (CATIE) and 7 cultivars used as controls were evaluated for host-plant resistance to 4 virulent strains of Pseudomonas solanacearum representing race 1 biovars 1 and 3. In general, biovar 3 strains wilted seedlings faster than biovar 1 strains but, after 20 days post-inoculation, no significant differences were noted in susceptible control ratings. Significant differences for disease index were noted, but no line with complete resistance was found. For the USA biovar 1 strain UW-25, only 5 accessions, CATIE 17331, 17334, 17349, 17739, 17740, and 2 of the control cultivars, Hawaii 7998 and UC-82B showed some degree of resistance. Conversely, both the frequency and the degree of resistance were high for Costa Rican biovar 1 strain UW-256. For biovar 3, the Costa Rican strain UW-255 was more virulent than the Peruvian strain UW-130. Eight CATIE accessions, 5539, 17331, 17333, 17334, 17345, 17349, 17742, and MIP-CH1, were as resistant as the resistant control Hawaii 7998 to 3 strains and accession 17740 was as resistant as Hawaii 7998 to all 4 strains.Journal Paper No. J-16011 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Project 3123.     

Survival of,and root colonization by,alginate-encapsulated Pseudomonas fluorescens cells following introduction into soil

Summary The survival of Pseudomonas fluorescens cells encapsulated in alginate beads and colonization of wheat roots was studied in soil microcosms inoculated with the cells in alginate beads of varying composition. Cells encapsulated in beads and introduced into a non-sterile loamy sand survived better than cells added directly to the same soil. A recovery/growth step for the bead-encapsulated cells was added before they were introduced into the soil, in an attempt to obtain optimal population levels in the soil. Further, bacterial populations that grew to the highest density in the beads subsequently showed the highest survival levels in soil. The addition of 3% skim milk, or 3% skim milk and 3% bentonite clay to all bead types consistently resulted in the highest survival of the encapsulated cells in soil. Root colonization by P. fluorescens was generally not impaired by the encapsulation in alginate. One week after inoculation into the soil, encapsulated cells in the various bead types were able to colonize the wheat rhizoplane at high population levels, similar to or exceeding those found when free cells were inoculated. In a second root colonization experiment the wheat rhizoplane was also efficiently colonized 7 weeks after the inoculant cells had been introduced into the soil in different bead types. In both assays, the cells encapsulated in beads amended with skim milk plus bentonite clay showed the highest root colonization rates. It is clear, therefore, that alginate-mediated establishment of inoculants can improve inoculant effectiveness.     

Suppression of other soil microorganisms by mycelium of arbuscular mycorrhizal fungi in root-free soil

The influence of mycelium of two arbuscular mycorrhizal (AM) fungi, Glomus intraradices and Glomus mosseae, on other soil microorganisms, was examined in root-free soil with and without organic substrate amendment in terms of cellulose. The AM fungi were grown in symbiosis with cucumber in a compartmented growth system, which allowed AM fungal external mycelium to grow into root-free compartments. The fungicide Benomyl was applied to the root-free compartments to create an alternative non-mycorrhizal control treatment. Whole cell biomarker fatty acids were employed to quantify different groups of soil microorganisms including the two AM fungi. Abundance of most microbial groups were reduced by external mycelium of both AM fungi, though differential effects on the microbial community composition were observed between the two AM fungi as revealed from principal component analysis. Inhibition of other soil microorganisms was more pronounced in root-free soil with mycelium of G. mosseae than with mycelium of G. intraradices. In general, cellulose increased the amount of biomarker fatty acids of most groups of soil microorganisms, but cellulose did not affect the influence of AM fungi on other soil microorganisms. Benomyl suppressed growth of the external mycelium of the two AM fungi and had limited non-target effects on other microbial groups. In conclusion, our results show differential effects of external mycelium of AM fungi on other soil microbial communities, though both AM fungi included in the study overall inhibited most microbial groups as examined using whole cell biomarker fatty acids.     

番茄对酸性黄壤中铅的吸收特性与富集效果

通过盆栽试验,研究番茄对酸性黄壤中铅(Pb)的吸收和积累效应。试验结果表明,当土壤中加入不同比例的铅时(0～200mg/kg),基本没有观察到铅对番茄的毒害病症,铅对作物的生长发育没有明显的影响;但番茄具有强烈吸收土壤中铅的特性,并在番茄体内积累。随土壤中铅添加量的提高,番茄铅吸收量增加,其中根系的吸收最强,明显高于叶、茎和果实,特别是当土壤中铅添加量较高时,差异更为显著。在土壤铅浓度较低时,番茄对土壤铅的富集率较高(4.38%),当酸性黄壤中铅的浓度大于50mg/kg时,可引起番茄中铅含量超标。     

Bacterial 16S rDNA sequences in immature volcanic ash soil on volcanoes Mt. Sakurajima and Mt. Fugen in Japan determined by PCR amplification

Volcanogenous soils are widely distributed in Japan. Andosols, a group of volcanogenous soil, are known to show several physicochemical characteristics such as high porosity, presence of allophane, and high content of organic carbon (FitzPatrick 1980). The formation of Andosols is a very rapid process resulting from the large surface area of the volcanic ash-derived parent materials.     

Use of the fluorescent antibody technique to characterize equilibrium survival concentrations of Bradyrhizobium japonicum strains in soil

Summary The ability of Bradyrhizobium strains to survive saprophytically in soil was studied by means of fluorescent antibodies (FA). It was found that the recovery rate may be considered a constant value although the limit of detection by the FA technique is approximately 10³ bacteria g^–1 soil. By studying the survival kinetics of B. japonicum strains introduced into soils, we observed that whichever soil-strain combination was tested in a given soil during the incubation all the different populations of a strain reached the same survival balance level, generally about 10³–10⁴ Bradyrhizobium g^–1 soil. When we reintroduced strains into a soil containing rhizobia of the same specificity, the new inhabitants reached the same equilibrium level as that of the initial population. The balance threshold level does not appear to be a very sensitive way to classify, strains on their saprophytic ability. We suggest that survival kinetics should be characterized by the rate at which the population reaches equilibrium.     

Influence of the earthworm Aporrectodea trapezoides (Lumbricidae) on the colonization of alfalfa (Medicago sativa L.) roots by Rhizobium meliloti L5-30R and the survival of R. meliloti L5-30R in soil

A greenhouse study was performed to examine the ability of the earthworms Microscolex dubius and Aporrectodea trapezoides to transport Rhizobium meliloti L5-30R through soil. When R. meliloti L5-30R was inoculated into either ezi-mulch (a pelleted formulation of cereal-pea straw), oat hay, pea hay, or sheep dung and placed on the soil surface together with either A. trapezoides or M. dubius, >10⁴ colony-forming units (CFU) of R. meliloti L5-30R g^-1 soil were detected at 90 mm soil depth after 18 days. In the absence of earthworms, R. meliloti L5-30R was not detected at 90 mm soil depth after this time. In a second experiment using ezi-mulch as the inoculant material and in the presence of A. trapezoides (in a number equivalent to 471 or 785 m^-2), ca. 10³ CFU of R. meliloti L5-30R per 10 mm of alfalfa root were detected at 0–30, 30–60, and 60–90 mm soil depth after 18 days, while <3 CFU were detected per 10 mm of root in the absence of A. trapezoides. In a third experiment in which R. meliloti L5-30R was distributed evenly through soil at the start of the experiment, A. trapezoides (in a number equivalent to 157, 471, or 785 m^-2) significantly decreased the survival of L5-30R in soil after 40 days of incubation at 15°C, but not after 20 days. The decrease in survival of R. meliloti L5-30R was correlated with the density of A. trapezoides. These results demonstrate that A. trapezoides can increase root colonization of alfalfa by R. meliloti L5-30R, but may also reduce the ability of R. meliloti L5-30R to survive in soil.     

Suppression of chlorate degradation in longan plantation soil after multiple applications of chlorate and accelerated degradation by sugar amendment

Potassium chlorate is widely used as an active substance for flower induction in longan plantations for off-season production of fruits in northern Thailand. Contamination of groundwater with residual chlorate in soil is a cause for concern because of its toxicity to human health. Based on the assumption that potassium chlorate might adversely affect chlorate-reducing soil microorganisms due to its disinfecting nature, the effect of repeated applications of chlorate on chlorate decomposition in soil was examined in a laboratory study. In the longan plantation field soil which had received an annual application of potassium chlorate for three years, chlorate decomposition was slower than that in the soil without application. The suppression of chlorate decomposition was confirmed in the soil to which potassium chlorate was applied repeatedly in the laboratory. However, even in such soils that had received multiple applications of potassium chlorate, chlorate decomposition was remarkably enhanced by soil amendment with glucose or sucrose. Sugar amendment seemed to be an effective way of cleaning-up the residual chlorate in longan plantation field soils.     

Use of soil nonexchangeable potassium by paddy rice with clay structural changes under long-term fertilizer management

To prove the hypothesis that paddy rice utilizes soil nonexchangeable potassium (neK) and causes associated structural changes in clay minerals, K status and clay mineralogy of 22 surface soils from three paddy fields under long-term fertilizer management for 51–93 years were investigated. Soil neK content was determined as the difference between 1 mol L⁻¹ hot HNO₃ extractable K and 1 mol L⁻¹ ammonium acetate exchangeable K. Clay mineralogy was identified by X-ray diffraction (XRD). The radiocesium interception potential (RIP), an index of frayed edge sites in the interlayer sites of 2:1 type clay minerals, was also determined. The neK contents under the -K and NPK treatments were considerably lower than those under the unfertilized treatment in all the fields, indicating the exploitation of soil neK by rice. XRD analysis of the clay samples revealed 7% shift from the 1.0 peak to 1.4 nm one under the -K treatment compared with the unfertilized one, and the amounts of neK were negatively correlated with those of RIP (p < .01), suggesting the expansion of interlayer spaces of the 2:1 type phyllosilicates such as mica due to the release of neK. In addition, the neK content positively correlated with K balance of the long-term experiments (p < .05). The differences of neK between unfertilized K and -K treatments corresponded to 22–157 kg K ha⁻¹, or 0.42–1.68 kg K ha⁻¹ year⁻¹. In conclusion, utilization of considerable amount of soil neK under K depleted conditions should be considered to establish sustainable K management for paddy rice.     

Evaluation of the different soil organic matter (SOM) pools stability in long-term field experiments of germany by physical fractionation

Samples of Ap horizons of different soil types with different clay content selected from nil plot and different fertilized plots of the long-term field experiments Thyrow, Groß Kreutz, Seehausen, Methau, Bad Lauchstädt, Straußfurt located in closely related climatic conditions had been analyzed by method of granulo-densimetric fractionation developed in Dokuchaev Soil Science Institute (Moscow). For comparison mountainous soil from long-term experiment Lauterbach located in more moist and cold conditions had been studied. Three groups of functional significant SOM fractions had been isolated: (1) light fraction (d < 2g·cm^-3), clay fraction (< 1 μm) and fraction rest. The partition of C and N accumulated in these fractions related to the level of soil fertilization and clay content had been estimated. Light fraction is the least stable: it is sensitive to the level and forms of fertilizers as well as to the changing ecological conditions. SOM interacting with clay minerals is a main factor of soil carbon level stabilization. Light fraction is responsible for decomposable SOM pool dynamics and may be appropriate criteria of its estimation.     

Speciation of aluminum in soil environments

Although aluminum (Al) is abundant in soil environments, it is not an essential element and it is toxic to most organisms. Since the toxicity of Al depends on their chemical forms, the importance of Al speciation has been recognized worldwide. Difficulties in Al speciation are caused by the complex coordination chemistry of Al for the hydrolysis and formation of polynuclear species with a variable degree of solubility in aqueous solution. Nondestructive analyses, such as nuclear magnetic resonance (NMR) spectroscopy, could supply primarily im-portant information on the chemical forms of Al and may enable to evaluate the results ob-tained by other methodologies. In the present report, NMR spectral characteristics of environmentally important Al-containing components, such as hydoxyaluminum ions, Alinorganic complexes, Al-organic complexes, and primary and secondary minerals, are summarized for the nuclei of ²⁷Al and ²⁹Si determined by solution NMR and solid-state magic angle spinning (MAS) NMR. Applications of NMR techniques to soil science, including speciation of phytotoxic Al in soil environments and whole soil NMR studies, are described.     

土壤中微量钒的分光光度测定方法研究 ——1 （2 吡啶偶氮） 2 萘酚-过氧化氢-十二烷基苯磺酸钠法

对钒(V)和1-(2-吡啶偶氮)-2-萘酚(PAN)-过氧化氢-十二烷基苯磺酸钠(SDBS)显色反应进行了试验,结果表明:在1 mol.L-1盐酸介质中,在SDBS存在下,钒与1-(2-吡啶偶氮)-萘酚和过氧化氢生成红色的多元络合物,络合物最大吸收波长为575 nm,表观摩尔吸收率为3.64×104L.mol-1.cm-1,钒的浓度在0.2～1.2μg.mL-1内符合比耳定律。络合物的组成比为n钒∶nPAN∶n过氧化氢=1∶1∶1。该方法用于土壤中钒的测定,测定值的相对偏差(n=20)均小于6%,回收率为100.4%～105.6%。