Quantification of Wautersia [Ralstonia] basilensis in the mycorrhizosphere of Pinus thunbergii Parl. and its effect on mycorrhizal formation

The bacterium Wautersia [Ralstonia] basilensis has been shown to enhance the mycorrhizal symbiosis between Suillus granulatus and Pinus thunbergii (Japanese black pine). However, no information is available about this bacterium under field conditions. The objectives of this study were to detect W. basilensis in bulk and mycorhizosphere soils in a Japanese pine plantation in the Tottori Sand Dunes, determine the density of W. basilensis in soil, and determine the optimal cell density of W. basilensis for mycorrhizal formation in pine seedlings. We designed and validated 16S rRNA gene-targeted specific primers for detection and quantification of W. basilensis. SYBR Green I real-time PCR assay was used. A standard curve relating cultured W. basilensis cell density (10³-10⁸ cells ml⁻¹) to amplification of DNA showed a strong linear relationship (R = 0.9968). The specificity of the reaction was confirmed by analyzing DNA melting curves and sequencing of the amplicon. The average cell density of W. basilensis was >4.8 × 10⁷ cells g⁻¹ of soil in the mycorrhizosphere and 7.0 × 10⁶ cells g⁻¹ in the bulk soil. We evaluated the W. basilensis cell density required for mycorrhizal formation using an in vitro microcosm with various inoculum densities ranging from 10² to 10⁷ cells g⁻¹ soil (10⁴-10⁹ cells ml⁻¹). Cell densities of W. basilensis of >10⁶ cells g⁻¹ of soil were required to stimulate mycorrhizal formation. In vivo and in vitro experiments showed that W. basilensis was sufficiently abundant to enhance mycorrhizal formation in the mycorrhizosphere of Japanese black pine sampled from the Tottori Sand Dunes.     

模拟放牧对高寒草甸丛枝菌根真菌的影响

采用植物样方调查与丛枝菌根（AM）真菌的形态分子鉴定等方法分析了模拟放牧对地上植物及地下AM真菌群落的影响。结果表明：随着放牧程度的增加,地上植物的物种丰富度显著降低,但AM真菌的物种丰富度没有显著变化;AM真菌总侵染率与丛枝侵染率随着放牧强度的增加显著降低,但泡囊侵染与土壤AM真菌的孢子密度均不受放牧强度的显著影响;模拟放牧对植物群落组成具有显著影响,但AM真菌的群落组成不受放牧的影响。以上结果表明较植物群落而言,AM真菌对放牧干扰具有较高的耐受性,凸显了在放牧干扰条件下,AM真菌在维持高寒草甸生物多样性过程中的作用。     

Response of Mycorrhiza,Organic Sources,Secondary and Micro Nutrients on Soil Microbial Activities and Yield Performance of Colocasia (Colocasia esculenta L.) in Alfisols

Response of the integrated use of mycorrhiza, inorganic and organic manures on microbial variables and yield performance of colocasia (Colocasia esculenta L.) was studied in an acid Alfisol. Significantly highest mean cormel yield (14.13 t ha^?1) was recorded due to application of super optimal doses of nitrogen, phosphorus, and potassium (NPK). Integrated application of lime + farmyard manure (FYM) + ½ NPK + zinc sulfate (ZnSO₄) has recorded the highest dehydrogenase activity (2.048 µg TPF hr^?1 g^?1) and Fluorescein Diacetate Hydrolysis assay (1.855 µg g^?1 hr^?1). The highest acid and alkaline phosphatase activities (77.67 and 51.18 µg PNP g^?1 h^?1, respectively) were observed due to Vesicular Arbuscular Mycorrhiza (VAM) inoculation in combination with lime + FYM + ½ NPK. Dehydrogenase and phosphatase activities had a highly significant relationship with cormel yield and the biochemical constituents of colocasia. The study emphasized the conjunctive use of balanced inorganic fertilizers and organic manures to enhance the enzymatic activities and to realize higher crop yields of colocasia in acid Alfisols.     

Amelioration of salinity stress in different basil (Ocimum basilicum L.) varieties by vesicular-arbuscular mycorrhizal fungi

Although the salt resistance mechanisms in plants have received much consideration for many years, varieties’ differences affecting salt resistance are still unsettled. Within the Ocimum genus there occur about 200 species in different varieties and forms. A pot experiment was performed to better understand salt stress responses in crop plants; we compared the impacts of salinity stress on growth and physio-biochemical characteristics in three varieties of basil (Ocimum basilicum) var. odoratus, O. b. var. alba and O. b. var. purpurascens) grown under four levels of salinity stress (0, 50, 100, and 150?mM NaCl) with mycorrhiza (Glomus clarum Nicol. &; Schenck) or without. Results showed significant differences within salinity treatments in all cultivars studied. In this study, the biomass production and physio-biochemical parameters of all cultivars reduced with raised salinity levels except concentration of reducing sugars, sodium, and proline at 150 mM of NaCl, only the variety ‘purpurascens’ didn't show reduction and observed resistant against severe salinity. The colonization of arbuscular mycorrhiza fungi enhanced the biomass production and accumulation of nutrients, reducing sugars, total soluble carbohydrates, photosynthetic pigments, proline, and protein by reducing Na. This study should help understand the function of AMF fungi in basil cultivars’ tolerance to salinity stress.     

Improvement of early growth of two tropical peat-swamp forest tree species Ploiarium alternifolium and Calophyllum hosei by two arbuscular mycorrhizal fungi under greenhouse conditions

Tropical peat-swamp forests are one of the largest near-surface reserves of terrestrial carbon. However, many peat-swamp forest tree species have resulted in the reduction due to over-exploitation, forest fires and conversion into agricultural land in Indonesia. The objective of this study was to determine the effects of two arbuscular mycorrhizal (AM) fungi, Glomus clarum and G. aggregatum, on the early growth of two slow-growing peat-swamp forest tree species, Ploiarium alternifolium and Calophyllum hosei, under greenhouse conditions. Cuttings of P. alternifolium and C. hosei were uninoculated or inoculated with G. clarum and G. aggregatum and grown under greenhouse conditions for 6 months. Percentage AM colonization, plant growth, phosphorus (P) concentration and survival rate were measured. The AM colonization of P. alternifolium and C. hosei ranged from 27 to 32% and 18 to 19%, respectively. Colonization by G. clarum and G. aggregatum increased shoot height, stem diameter, leaf number, and shoot and root dry weights. Cutting shoot P content were increased by AM fungal colonization. The survival rates of inoculated plants were higher (100%) than those of control plants (67%). The results suggest that inoculation with AM fungi improves early growth of P. alternifolium and C. hosei in a tropical peat-swamp forest and can therefore contribute to rehabilitation of peat-swamps.     

孟加拉国Dinajpur农林复合区内不同树种和作物种中丛枝菌根分布和孢子群落多样性的研究

从孟加拉国Dinajpur地区采集了不同农林复合区内树种和作物种,对采集植物的丛枝菌根多样性进行了研究.在3个取样地采集了18种植物的根和根际土,取样地和被采取植物分别为：取样地Dashmail为黄豆树 （Albizia procera Benth.）、小指椒 （Capsicum frutescens L）、姜黄（Curcuma domestica Vahl）、印度黄檀 （Dalbergia sissoo Roxb.） 和大叶桃花心木（Swietenia macrophylla）5种植物; 取样地Kantaji为姜黄（C.Domestica）、印度檀 （D.Sissoo）、赤桉 （Eucalyptus camaldulensis Dehnn.）、石梓（Gmelina arborea （Roxb） DC） 和水稻 （Oryza sativa L.） 5种植物; 取样地Ramsagar为姜黄（C.Domestica）, 印度檀 （D.Sissoo）, Litchi chinensis Sonn.和水稻.在取样地Dashmail,植物丛枝的分布范围在36%-79%,其中分布最高的是小指椒,达到79%,而C.Domestica仅占36%.取样地 Kantaji丛枝的分布范围在33%-70%之间,其中分布最高的是石梓,达到79%,而O.Sativa分布最低,仅36%.取样地 Ramsagar丛枝的分布范围在35%-70%之间,印度檀丛枝分布在该地最高, 达70%,而水稻丛枝在该地的分布最小,仅35%.在采样地Dashmail,每100克干土中所含采样植物丛枝菌根孢子群落54-140个;孢子群落最多的是印度檀（140/100g dry soil）,最低的是姜黄（C.Domestica） （40/100g dry soil）.在采样地Kantaji,孢子群落在63-221/100g dry soil,石梓具有的群落最高,为221/100g dry soil;印度檀为63/100g dry soil.在采样地Ramsagar, 所具有的最高和最低群落分别是 160/100g dry soil （D.Sissoo）和69/100g dry soil（L.Chinensis）.土壤Ph值与土壤中丛枝分布和菌根孢子群落无显著相关性.Simpson多样性指数和Shannon多样性指数在取样地Kantaji 的印度檀土壤中最高;而在取样地Ramsagar的水稻土壤中最低.研究表明孟加拉国Dinajpur地区丛枝菌根的存在、树种和作物种中的营养性质、丛枝菌根的必要性和贡献以及复合农林植物生长与丛枝?     

玉米丛枝菌根真菌根外菌丝表面定殖细菌解磷功能鉴定

【目的】在田间原位条件下研究丛枝菌根(Arbuscular mycorrhizal, AM)真菌根外菌丝表面有无解磷细菌定殖,并对存在的解磷细菌的种类进行鉴定,对其活化有机磷的能力进行检测,从而为更好地认识菌丝际土壤有机磷的周转和磷的生物地球化学循环过程提供依据。【方法】利用河北省曲周县中国农业大学实验站的玉米长期定位试验,采用田间埋膜方式从玉米根系周围收集AM真菌的根外菌丝,用蒙金娜有机磷固体培养基筛选菌丝表面具有矿化植酸钙能力的细菌,对筛选出的细菌进行分离、 培养,然后提取细菌DNA,通过16S rDNA测序分析来确定解磷细菌的种类。分离鉴定的菌株先用蒙金娜有机磷固体培养基通过测定菌落直径(d)及溶磷圈直径(D)初步鉴定其活化植酸钙的能力,再用无菌的蒙金娜有机磷液体培养基确定每株解磷细菌矿化植酸磷的能力,并对溶液的pH进行测定,每个菌株重复3次。最后采用两室隔网根盒将分离纯化的解磷细菌回接至AM真菌根外菌丝,鉴定回接成功率,确定分离出的解磷细菌能否成功定殖于菌丝表面。【结果】从AM真菌根外菌丝表面分离得到了29株具有活化有机磷能力的细菌,分属于芽胞杆菌、 假单胞菌、 沙雷氏菌、 葡萄球菌和肠杆菌5个不同的属。通过有机磷液体培养进一步检测这些菌株活化植酸磷的能力,发现它们对植酸磷的矿化率为1.9%~21.9%。其中假单胞菌属细菌的解磷能力相对较强,对植酸磷的矿化率达14%以上,液体培养基的pH值下降2~4个单位。将分离纯化的细菌回接至两室隔网根盒的菌丝室,培养30 d后,从菌丝表面再次检测到除假单胞菌属外的芽胞杆菌属(Bacillus)、 沙雷氏菌属(Serratia)、 葡萄球菌属(Staphylococcus)和肠杆菌属(Enterobacter)细菌,另外还检测到贪铜菌属(Cupriavidus)细菌。【结论】在田间原位条件下,与玉米共生的AM真菌的根外菌丝表面有多种解磷细菌定殖,它们活化有机磷能力存在差异,其中以假单胞菌属细菌的解磷能力相对较强。     

类芦的生长与VA菌根的关系分析

类芦是我国南方快速绿化水蚀荒漠化地区的优良草种。对类芦根蔸土壤中VA菌根真菌的调查表明，栽植类芦后，土壤中的VA菌根真菌数量大幅度增加，并与类芦根系形成一种协调的共生体系，极显著地促进了类芦的生长。新栽植类芦时，可在基肥中增施少量VA菌根土与磷肥，以促进土壤中VA菌根菌的快速繁衍。     

Biological nitrogen fixation by common beans (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.) increases with bio-char additions

This study examines the potential, magnitude, and causes of enhanced biological N₂ fixation (BNF) by common beans (Phaseolus vulgaris L.) through bio-char additions (charcoal, biomass-derived black carbon). Bio-char was added at 0, 30, 60, and 90 g kg⁻¹ soil, and BNF was determined using the isotope dilution method after adding ¹⁵N-enriched ammonium sulfate to a Typic Haplustox cropped to a potentially nodulating bean variety (CIAT BAT 477) in comparison to its non-nodulating isoline (BAT 477NN), both inoculated with effective Rhizobium strains. The proportion of fixed N increased from 50% without bio-char additions to 72% with 90 g kg⁻¹ bio-char added. While total N derived from the atmosphere (NdfA) significantly increased by 49 and 78% with 30 and 60 g kg⁻¹ bio-char added to soil, respectively, NdfA decreased to 30% above the control with 90 g kg⁻¹ due to low total biomass production and N uptake. The primary reason for the higher BNF with bio-char additions was the greater B and Mo availability, whereas greater K, Ca, and P availability, as well as higher pH and lower N availability and Al saturation, may have contributed to a lesser extent. Enhanced mycorrhizal infections of roots were not found to contribute to better nutrient uptake and BNF. Bean yield increased by 46% and biomass production by 39% over the control at 90 and 60 g kg⁻¹ bio-char, respectively. However, biomass production and total N uptake decreased when bio-char applications were increased to 90 g kg⁻¹. Soil N uptake by N-fixing beans decreased by 14, 17, and 50% when 30, 60, and 90 g kg⁻¹ bio-char were added to soil, whereas the C/N ratios increased from 16 to 23.7, 28, and 35, respectively. Results demonstrate the potential of bio-char applications to improve N input into agroecosystems while pointing out the needs for long-term field studies to better understand the effects of bio-char on BNF.     

Litter decomposition in beech forests — effects of excluding tree roots

Summary The hypothesis proposed by Gadgil and Gadgil (1971) that mycorrhizal tree roots suppress the decomposition of forest litter was examined in trenching experiments. The tests were conducted in two beech forest sites in southern Sweden, one with mor and one with mull. The exclusion of tree roots had no apparent influence on litter mass loss at any of the sites, which differed markedly in soil fertility and root distribution. Nor did trenching have any significant effect on total organic matter over the following 6 years, although trenched plots tended to be wetter than undisturbed soil. The present results as well as results from other similar studies suggest that the suppression of litter turnover by mycorrhizal tree roots is not an important mechanism in northern temperate and boreal forests.