The rhizosphere microflora of wheat grown under controlled conditions II. Influence of the stage of growth of the plant,soil fertility and leaf treatment with urea on the rhizosphere soil microflora

Summary The rhizosphere effect of seminal roots of seedlings and of nodal roots of tillering plants of spring wheat ‘Kaspar’ was investigated under controlled conditions. The total number of micro-organisms recorded in the rhizosphere soil were significantly higher than for the non-rooted soil when investigated with the soil dilution plate method, but lower when fluorescence microscopy was used. Additions of inorganic fertilizer (NPK) decreased their numbers especially in rhizosphere soil of seminal roots and in non-rhizosphere soil, but did not change the ratio between bacteria and actinomycetes (B/A). In the rhizosphere soil the B/A ratio was higher than in the non-rhizosphere soil. An effect of urea leaf treatment was found with the dilution-plate method only in the rhizosphere soil of nodal roots, 3 to 7 days after the first treatment. Increased numbers of actinomycetes were found in this period in NPK fertilized soil, whereas increased numbers of bacteria were found at both fertility levels.     

Changes of microflora of wheat roots after foliar application of urea

Upper parts of wheat grown in containers with brown soil were four-times repeatedly treated with a solution of urea. The foliar application of urea resulted in a considerable increase of the number of bacteria in the roots and a decrease of the number of fungi. Seventeen and 13 genera of fungi could be detected in the rhizosphere of the treated plants, 3 and 5 weeks, respectively, after the application, whereas 23 and 16 genera could be identified in the rhizoplane of the treated plants. Fungi of the generaFusarium andPenicillium predominated in the root region. The foliar application of urea resulted in an increased relative occurrence of the genusFusarium and decreased relative representation of the genusPenicillium. Changes in the relative occurrence of other 18 genera of fungi present in small quantities on wheat roots could also be observed. The foliar application of urea affected not only numbers of fungi determined by the dilution plate method but also the occurrence of active hyphae.     

Impact of azadirachtin, an insecticidal allelochemical from neem on soil microflora, enzyme and respiratory activities

The effect of 10% azadirachtin granules (alcoholic extract of neem seed kernel mixed with China clay) was studied on the population of bacteria, actinomycetes, fungi, Azotobacter and nitrifying bacteria; soil dehydrogenase, phosphatase and respiratory activities on 0, 15th, 30th, 60th and 90th days after application in sandy loam soil collected from the fields. It was observed that baring the Azotobacter sp., azadirachtin at all the doses exerted a suppressive effect on the rest of the microbial communities and enzyme activities in the initial 15 day period. The population of bacteria, actinomycetes besides phosphatase and respiratory activities recovered after 60th day and subsequently increased significantly. The fungi and nitrifiers were most sensitive groups as their numbers were reduced significantly throughout the studies. The two times and five times recommended dose of azadirachtin had very high biocidal effects on the soil microorganisms and its activities. However, analysis of the data by the Shannon Weaver index showed that azadirachtin reduces both the form and functional microbial diversity at all doses.     

Effect of foliar application of urea on the root microflora

The author studied the effect of the foliar application of urea on the microflora of the wheat root surface in relation to time. The effect of this treatment was manifested in the dry weight of the roots and in the parts above the ground seven days after application. The number of bacteria on the plants increased markedly on the first days after application, while the number of fungi showed a pronounced decrease. In time the differences were equalized. Bacteria requiring amino acids and growth factors predominated on the roots of treated plants, while the main fungi were members of the genusFusarium and in the control plants members of the genusPenicillium.     

Changes in the rhizosphere microflora of maize seedlings by pretreatment of roots with hormones and urea

Summary Effect of pretreatment of root of maize seedlings with gibberellic acid, maleic hydrazide and urea was investigated. Gibberellic acid at 1 ppm stimulated the growth of fungi, bacteria, and actinomycetes in the rhizosphere region of maize seedlings while at 5 ppm concentration population of bacteria and actinomycetes were suppressed. Maleic hydrazide treatment stimulated all the three groups at 5 ppm concentration but at 1 ppm there was increase in population of only fungi and actinomycetes. Urea stimulated the growth of fungi, bacteria, and actinomycetes. The order of predominance of different groups also changed with treatments. It was fungi > bacteria > actinomycetes in case of gibberellic acid at 1 ppm and 5 ppm and maleic hydrazide at 1 ppm; and bacteria > fungi > actinomycetes in case of maleic hydrazide at 5 ppm, urea at 0.1M and in control.     

Effect of foliar application of urea on wheat growth in relation to plant age,soil fertility and light intensity

Summary Treatment of leaves of ‘Kaspar’ wheat seedlings, growing in low inorganic (NPK) fertilized soil, with urea, resulted in reduced growth until 24 days after the first application. Stimulation of growth was observed 31 days after the urea leaf treatment. Addition of NPK to the soil did not obviously change the effect of urea leaf treatment on seedlings, while increase of light intensity from 16.000 lux to 42.000 lux restricted the period of growth reduction. Leaf treatment with urea did not influence growth of one-month-old wheat plants for 14 days after the first application.     

Microbial complexes from apogeotropic roots and from rhizosphere of cycad plants

The microbial complexes of soil, the rhizosphere, and the rhizoplane of the apogeotropic (coralloid) roots of cycad plants were comparatively studied. The aseptically prepared homogenates of the surface-sterilized coralloid roots did not contain bacterial microsymbiont, indicating that it was absent in the root tissues. At the same time, associated bacteria belonging to different taxonomic groups were detected in increasing amounts in the cycad rhizoplane, rhizosphere, and the surrounding soil. The bacterial communities found in the cycad rhizoplane and the surrounding soil were dominated by bacteria from the genus Bacillus. The saprotrophic bacteria and fungi colonizing the cycad rhizosphere and rhizoplane were dominated by microorganisms capable of degrading the plant cell walls. The local degradation of the cell wall was actually observed on the micrographs of the thin sections of cycad roots in the form of channels, through which symbiotic cyanobacterial filaments can penetrate into the cortical parenchyma.     

Microbial Complexes Occurring on the Apogeotropic Roots and in the Rhizosphere of Cycad Plants

The microbial complexes of soil, the rhizosphere, and the rhizoplane of the apogeotropic (coralloid) roots of cycad plants were comparatively studied. The aseptically prepared homogenates of the surface-sterilized coralloid roots did not contain bacterial microsymbiont, indicating that in the root tissues the symbiosis is a two-component one (plant–cyanobacteria). At the same time, associated bacteria belonging to different taxonomic groups were detected in increasing amounts in the cycad rhizoplane, rhizosphere, and the surrounding soil. The bacterial communities found in the cycad rhizoplane and the surrounding soil were dominated by bacteria from the genus Bacillus. The saprotrophic bacteria and fungi colonizing the cycad rhizosphere and rhizoplane were dominated by microorganisms capable of degrading the plant cell walls. The local degradation of the cell wall was actually observed on the micrographs of the thin sections of cycad roots in the form of channels through which symbiotic cyanobacterial filaments can penetrate into the cortical parenchyma.     

Development of rhizosphere and rhizoplane microflora of Aristida coerulescens in the Libyan desert

Summary Microflora of rhizosphere soil, rhizoplane and macerated root-portions of Aristida coerulescens, naturally occurring in the Libyan desert, were different in count and isolates, in the different root zones. A rhizosphere effect characteristic of each zone is shown. The root base contained the lowest numbers of microflora (bacteria and fungi) whilst the root tip included the highest counts. Distribution of most of the individual fungal species in the different root zones and root-surfaces is given in text.     

蚯蚓和秸秆对铜污染土壤微生物类群和活性的影响

试验设置4个Cu浓度水平：0、100、200和400 mg·kg^-1 Cu²⁺,每个Cu浓度水平设置4个处理：对照(CK)、表施秸秆(M)、接种蚯蚓(E)、同时加入蚯蚓和秸秆(ME),研究了在Cu污染土壤中加入蚯蚓和秸秆对土壤微生物数量及活性的影响.结果表明：Cu污染、秸秆和蚯蚓均明显影响土壤微生物类群; Cu污染对细菌、放线菌具有抑制作用,而对真菌没有影响;秸秆显著提高了真菌数量;蚯蚓使土壤细菌、放线菌数量显著增加,而对真菌数量影响不大.Cu污染浓度＞200 mg·kg^-1处理对微生物量碳具有抑制作用;加入秸秆或蚯蚓,可显著提高土壤微生物量碳,而且同时加蚯蚓和秸秆处理土壤微生物量碳增加最显著.加入蚯蚓和秸秆后,土壤呼吸值显著增高.Cu<200 mg·kg^-1时,蚯蚓处理土壤呼吸值最大,平均比对照高3.06～5.58倍;Cu≥200mg·kg^-1时,蚯蚓、秸秆同时加入处理土壤呼吸值最高.4个处理土壤代谢商大小顺序为：ME>E>M>CK.蚯蚓和秸秆处理对土壤NH₄⁺-N没有影响,而对土壤NO₃^--N影响各异.接种蚯蚓,可显著提高土壤NO₃^--N含量;加入秸秆,可显著降低土壤NO₃^--N含量;同时加入蚯蚓和秸秆处理NO₃^--N含量最低.相关分析表明,土壤有效态Cu(DTPA-Cu)与土壤放线菌、细菌呈显著负相关,而与土壤呼吸、土壤NO₃^--N、NH₄⁺-N含量呈显著正相关.引入秸秆和蚯蚓,可在一定程度上减缓Cu污染对微生物数量和活性的影响.     

The effect of foliar application of urea on the root fungi of wheat growing in soil artificially contaminated withFusarium spp.

Repeated treatment of overground parts of wheat with a solution of urea was reflected in an increased count of bacteria and a decreased count of fungi on the roots of plants grown in soil contaminated with the fungusFusarium spp., which is known to be noxious to plants. The occurrence of some fungal genera on the roots of plants growing in contaminated and uncontaminated soil after foliar application of urea was also changed. The roots of treated planted were found to contain more bacteria with a lytic effect and retarding the growth of the fungus used for the contamination. The favourable effect of foliar application of urea on wheat was reflected in an increased dry weight of plants even if growing in a soil contaminated withFusarium.     

3株芽孢杆菌对番茄的促生作用及对番茄根域微生物的影响

通过种子萌发和盆栽促生试验研究3株芽孢杆菌Bs10、Ba12和Bl10对番茄的促生作用及其对番茄根域微生物区系的调节作用.结果表明: 3株芽孢杆菌对番茄种子的胚轴、胚根和番茄植株的生长有明显的促进作用,处理后番茄根系的总长度、总表面积和总体积均显著增加;处理后土壤中细菌数量和比例显著增加,真菌数量和比例明显减少.与对照相比,土壤微生物区系优势菌数量发生改变:优势甲基营养型芽孢杆菌在番茄根区、根表土壤中和根内的数量大幅提高;病原真菌腐皮镰刀菌和尖孢镰刀菌在根区和根表土壤中的数量显著减少.推知芽孢杆菌对根系微生物区系的调节作用是其发挥防病促生作用的重要机制之一.     

Effects of zinc-smelter emissions on forest soil microflora.

Within 2 km of a zinc (Zn) smelter in Palmerton, Pennsylvania, near the Lehigh Water Gap, up to 13.5% Zn by weight has been measured in the O2 horizon of the soil, and up to 8% Zn in the A1 horizon. The total numbers of bacteria, actinomycetes, and fungi (measured by dilution plate counts) were greatly reduced in the most severely Zn-contaminated soils compared with control soils. The reduction of microbial populations may be a partial cause of the decreased rate of litter decomposition at Lehigh Gap. Growth of most bacteria from control sites was reduced by 100 to 200 muM Zn, most actinomycetes by 100 muM Zn, and most fungi by 100 to 1000 muM Zn in thin-Pablum extract agar (TPab). All the tested actinomycetes and non-spore-forming bacteria isolated from Zn-contaminated Lehigh Gap soils were Zn-tolerant, growing normally in media containing 600-2000 muM Zn. Most fungi, regardless of source, were capable of at least 50% of normal growth at 700 muM Zn. Zinc-tolerant bacteria, actinomycetes, and fungi were readily isolated from low-Zn soils, suggesting that selection for Zn tolerance may proceed rapidly. Acidophilic Mortierella species have been selectively eliminated near the smelter, apparently because of elevated soil pH. Peryronellaea glomerata (Corda) Goidanich and Coniothyrium spp. were found only in the high-Zn soils.     

The invasion and development of the cereal cyst-nematode, Heterodera avenae in the roots of autumn-and spring-sown cereals

Observations were made at 2 or 4 wk intervals from December to harvest on all stages of Heterodera avenae in winter oats growing on infested land. Second-stage larvae were present in all soil samples except on 5 and 20 July. Invasion and development of larvae was slow during winter. The nodal and seminal roots of winter oats were both heavily invaded by the nematode; larvae which invaded seminal roots tended to become male whereas those in nodal roots tended to become female. There was a small second invasion in August. Females were first observed on the roots of winter oats on 17 May, 214 days after the crop was sown and 62 days after the first fourth-stage larva was observed. The nodal roots of spring barley contained few H. avenae larvae whereas these roots were heavily invaded in winter wheat and oats. In spring barley the nodal roots were developing in June and July when few second-stage larvae were in the soil whereas in winter oats and wheat the nodal roots were growing rapidly in April when larvae were most numerous, and so were heavily invaded.     

小蓬竹根际土壤微生物及内生真菌多样性分析

为了解喀斯特典型物种-小蓬竹根际土壤微生物及不同部位内生真菌多样性,采用沿等高线等距离取样法采集小蓬竹根际土壤及健康植株,通过可培养对根际土微生物及内生菌进行分离,利用分子技术对其进行鉴定,根据鉴定结果构建系统发育树,并计算小蓬竹根际土壤微生物和根茎叶内生真菌多样性。结果如下：（1）共从根际土壤、根、茎、叶分离得到139个真菌菌株,隶属于27属,其中根际土壤分离得到34个真菌菌株隶属于12属,根部分离得到的63个内生真菌菌株隶属于17个属,茎部分离得到的14个内生真菌菌株隶属于8个属,叶部分离得到28个内生真菌菌株隶属于9个属;（2）根际土壤共分离得到41株细菌菌株,隶属于7个属26个种,20株放线菌菌株,隶属于1属15种;从Shannon-Wiener多样性指数、均匀度指数、Simpson指数排序来看,真菌主要表现为根 > 根际土壤 > 茎 > 叶,细菌和放线菌多样性均较低。（3）按层次聚类分析可分别将真菌、细菌、放线菌聚为3支。小蓬竹根际土壤、根、茎和叶具有丰富的微生物多样性,不同部位菌群组成存在差异性（P<0.05）,且存在以假单胞菌属、芽孢杆菌属等为优势属的抗盐耐旱菌群,这有助于揭示小蓬竹对喀斯特生境的适应性,以及为微生物-植物群落之间相互关系提供一定基础数据,为后期寻找小蓬竹相关耐性功能菌奠定基础。     

放线菌制剂对人参生长及根域土壤微生物区系的影响

以小兴安岭地区人参为研究对象,探索放线菌制剂对人参的促生效应及对人参根区、根表土壤微生物区系的影响.结果表明： 经放线菌制剂Streptomyces pactum（Act12）处理后,人参药用部分产量增加,品质改善;叶片诱导酶活性提高,根系活力增强;土壤中细菌、放线菌的数量和比例显著增加,真菌的数量和比例减少.与对照相比,土壤微生物区系结构改变:优势菌荧光假单胞菌、韩国假单胞菌和氧化微杆菌在根区、根表土壤中的数量大幅提高;病原真菌烟色织孢霉在根区土壤中减少,在根表土壤中消失.表明施用放线菌制剂Act12能够改善土壤微生物区系,提高人参植株的抗性和根系活力,增加产量并改善品质.     

Selective effects of five pesticides on soil and cotton-rhizosphere and-rhizoplane fungus flora

In two field sowings, the effect of five pesticides on soil, rhizosphere and rhizoplane fungi was tested. Ceresan and Orthocid (fungicides used as seed dressing) after 3 days were very toxic to the total count of soil fungi and to almost all fungal genera and species. After 40 days their toxicity was almost alleviated in the soil, but persisted in the rhizosphere. The herbicide VCS 438 did not significantly affect the total count of soil fungi but was initiative to the total rhizosphere fungi. Some fungal species were significantly promoted in the soil and rhizosphere and others were depressed. Dipterex (Insecticide) was promotive to the total count of soil fungi after 3 days and to some fungal species but this effect was almost alleviated after 40 days. In the rhizosphere, it exerted a promotive effect on the total count of fungi and on some fungal genera and species. Dursban (Insecticide) was of no significant effect on the total count of soil and rhizosphere fungi, but few species were significantly promoted.The rhizoplane fungi were the least sensitive, and none of the five pesticides induced a significant effect on the total count, but some species responded significantly, being either promoted or inhibited.     

Relationships between plant roots,proteolytic organisms and activity of protease

Proteolytic bacteria represented 18–58% of the bacterial population isolated from the rhizoplane of different crops. The activity of protease was considerably higher on roots of wheat growing in the soil than in the rhizosphere or free soil. However, only a slightly positive rhizosphere effect in the relative occurrence of casein-hydrolyzing bacteria could be observed. An indirect relationship between numbers of bacteria hydrolyzing casein and the activity of the enzyme could be found. The activity of protease related to a unit of culturable proteolytic bacteria was considerably higher on the root than in the rhizosphere and in the soil. A relationship between characteristics of the production of the enzyme by proteolytic bacteria and the protease activity on the surface of roots was demonstrated. The resulting enzyme activity on the surface of roots depended apparently on growth conditions of the plant and nature of root exudates and was influenced both by inactivation and protection due to adsorption of the enzyme by roots.     

Changes in soil microflora associated with control of Sclerotium Rolfsii by Furfuraldehyde

The efficacy of 2‐furfuraldehyde for control of Sclerotium rolfsii was studied in laboratory and greenhouse experiments. Mycelial growth of the fungus was reduced proportionally with concentrations of 0.1–0.5 ml furfuraldehyde l^‐1 agar medium, and viability of sclerotia diminished on exposure to 2‐furfuraldehyde vapours. Detectable populations of bacteria and fungi, including Trichoderma spp., were reduced significantly (9=0.05) when furfuraldehyde was added to the agar used for soil dilution plates of untreated soil. Repeated treatments of natural soil with the fumigant significantly increased populations of Trichoderma spp. and bacteria, but diminished numbers of actinomycetes. Increasing dosages applied to soil artificially infested with S. rolfsii caused a reduction of disease on lentil, Lens culinaris. Results indicate that the compound, when applied to field soil, changes the composition of soil microflora and has potential for integrated control of S. rolfsii.     

Micro-organisms in the rhizosphere of plants inoculated withAzotobacter chroococcum

Summary The hypothesis that inoculation withAzotobacter chroococcum affects the growth of plants indirectly through changing the rhizosphere microflora was investigated. Inoculated and uninoculated wheat and tomato plants were grown in the glasshouse in two different soils, and total bacteria, chitinolytic bacteria, actinomycetes, glucosefermenting bacteria, aerobic cellulose-decomposing bacteria, and anaerobes were determined in intervals in the rhizosphere and in the soil. Root-surface fungi were studied using the Harley and Waid's root-washing technique¹⁰. Azotobacter became established in the rhizosphere of wheat and tomato plants and stimulated their growth. All the bacterial groups examined were more abundant in the rhizosphere than in the soil. Inoculation with Azotobacter delayed the colonization of roots by bacteria, actinomycetes, and fungi in the rhizosphere, but had no effect on other organisms. Inoculation did not affect the dominant root-surface fungi, and minor changes were not consistent.Part of a thesis accepted by the University of London for the degree of Ph.D. in Microbiology.