2株溶磷、解钾与产IAA的内生真菌菌株的筛选、鉴定及促生作用研究

从钾矿区优势蕨类植物-芒萁内生真菌中筛选出具溶磷、解钾、分泌吲哚乙酸(IAA)的功能菌株,并研究其对农作物的促生作用。采用菌株的形态学特性、培养特征及5.8S rDNA、18S rDNA ITS序列分析方法对菌株进行鉴定,并结合液体培养和固体培养方法初步测定菌株的溶磷、解钾能力,采用回接及盆栽试验研究它们对玉米幼苗的促生作用。从42株芒萁内生真菌中筛选得到2株具有高效溶磷、解钾、分泌IAA功能的内生真菌菌株(编号为MQ013和MQ039),经鉴定MQ013菌株为泡盛曲霉(Aspergillus awamori),MQ039菌株为黑曲霉(Aspergillus niger)。促生作用试验结果显示,MQ013、MQ039菌株能有效提高玉米植株体内叶绿素及磷、钾含量。MQ013、MQ039菌株具有一定的溶磷、解钾和分泌IAA活性,并对玉米幼苗的生长有明显的促进作用,该菌株在研制高效生物肥料接种剂方面可能具有较大潜力。     

滇池富磷区土壤中溶磷真菌的筛选及其对油菜的促生作用

从滇池富磷区的89份土样中筛选出48株溶磷真菌,采用钼蓝法测定这些真菌溶解Ca3(PO4)3的能力。结果表明培养液中可溶性磷含量在14.45～64.87?mg/L,其中菌株SPF46、SPF47和Mo-Po的溶磷能力最强,其培养液中可溶性磷含量分别达到55.44、59.78和64.87?mg/L。结合形态特征及ITS rDNA 系统亲缘关系分析,将PSF46鉴定为黄暗青霉（Penicillium citreonigrum）,PSF47菌株鉴定为黑曲霉（Aspergillus niger）,Mo-Po菌株鉴定为草酸青霉（Penicillium oxalicum）。在Ca3(PO4)3、FePO4.4H2O和AlPO4 3种磷源中,这3种溶磷真菌对Ca3(PO4)2的溶磷效果最好。通过盆栽试验表明这3种溶磷真菌对油菜具有较好的促生效果,单一菌株处理,根系增长率在10.16%～294.7%,株高增长率在32.19%～134.5%;叶片直径增长率在35.53%～170.5%;鲜质量和干质量增长率分别在30.71%～189.5%,56%～224%。3种溶磷真菌混合处理,根系增长129.4%,株高增加60.41%,叶片直径增加170.5%;鲜质量和干质量分别增加246.1%、272.2%。     

溶磷菌株组合的溶磷效应及对玉米生长的影响

【目的】 本研究以革兰氏阴性菌Pseudomonas fluorescens CHA0、F113、Phl1c2、PF5与阳性菌Bacillus megaterium X14为研究对象,通过各菌株及菌株组合的溶磷能力评判组合效果,并进一步通过玉米盆栽试验验证其应用效果。【方法】 通过菌株在NBRIP培养基中对难溶磷源Ca₃（PO₄）₂的溶解能力评价各菌株及菌株组合的溶磷能力,测量接菌处理后的玉米干重、株高和全磷含量评价各菌株和菌株组合的盆栽应用效果。【结果】 1）纯培养实验中CHA0、F113、Phl1c2、PF5在NBRIP无机磷液体培养基中可溶磷含量均显著高于X14,且4株阴性菌两两组合后,培养基中可溶磷含量显著高于对应的阴性菌与阳性菌的组合,即阴性菌组合后的溶磷效果高于阴性菌与阳性菌组合后的溶磷效果。2）单菌比较,盆栽试验接种阳性菌X14对玉米生长及全磷吸收累积量的效果与阴性菌的差异不及室内实验显著,说明该阳性菌盆栽应用效果较好。比较菌株组合的处理,发现接种阴性菌与阳性菌组合的处理,其对玉米生长及全磷吸收累积量的效果与接种对应的阴性菌组合的处理效果相近,甚至显著高于一些阴性菌组合。3）综合比较室内实验结果与盆栽试验结果,发现阳性菌X14虽然在培养基中溶磷效果差,但在盆栽试验中,无论是单菌还是与阴性菌组合后的处理均表现出了良好的溶磷效果,且部分阴性菌与该阳性菌配合后,在玉米盆栽应用中促生和溶磷效果最佳。【结论】 与阴性菌组合相比,阳性菌与阴性菌组合室内溶磷效果均较差,但盆栽应用效果良好,甚至优于部分阴性菌组合,表明筛选溶磷菌株组合时,单一的室内纯培养结果不能作为唯一的评价指标,应同时结合盆栽的促生及溶磷效果。     

马铃薯内生固氮菌的分离及其促生特性研究

内生固氮菌是一类重要的植物促生菌,占据着植物组织内有利于营养供应和微环境适宜的生态位,能更好的发挥促生功能,促生机制包括固氮、解磷、产植物生长素、产ACC脱氨酶、产铁载体及拮抗病害等。本文从连作3年马铃薯根筛选获得8株内生固氮菌,其中短芽孢杆菌属(Brevibacterium)3株,芽孢杆菌属(Bacillus)3株和泛菌属(Pantoea)2株。促生特性研究发现:菌株Brevibacterium sp.GWR4具有较高固氮酶活性[16.206nmol C2H4/(mg蛋白质·h)],与圆褐固氮菌的固氮酶活性具有极显著性差异(P0.01);菌株Pantoea spp.GWR2、GWR3具有极高的产IAA能力,分别为186.07、162.21μg/m L,GWR2兼具产ACC脱氨酶活性[3.74μmol/(mg蛋白质·h)],GWR3兼具溶解无机磷的能力;菌株Brevibacterium sp.GWR5可以拮抗尖孢镰刀菌MLSOF(Fusarium xysporum)和茄病镰刀菌MLS-QB(Fusarium solani),兼具溶解有机磷和产IAA能力;Bacillus spp.GWR7、GWR8兼具固氮和产IAA能力。盆栽试验显示,分别接种Pantoea sp.GWR3,Brevibacterium spp.GWR4、GWR5和Bacillus spp.GWR7、GWR8后小白菜鲜重显著高于未接菌对照处理。可见,这5株菌兼具多种促生特性且对小白菜具有较好的促生效果,有望进一步研究开发成为微生物肥料生产菌种。     

适应玉米的溶磷细菌筛选及其对玉米生长的影响

从石灰性土壤中分离获得4株高效溶磷细菌X5、X6、Z4和Z8,研究其生物学特征,探索其单独及复合的溶磷促生潜能。研究发现菌株X5、X6、Z4和Z8均可以利用玉米根系分泌物作碳源生长。菌株X6和Z4均能产生吲哚乙酸(IAA)和铁载体,菌株Z8可产生IAA不产生铁载体,菌株X5可产生铁载体不产生IAA。盆栽试验结果表明,接种单一溶磷菌及4株菌复合处理均可促进玉米生长,但复合菌群的溶磷促生效果显著高于单一菌株。通过16S r RNA基因序列分析研究菌株的分类地位,初步鉴定X5、X6、Z4、Z8分别为荧光假单孢菌(Pseudomonas fluorescens)、草假单胞菌(Pseudomonas poae)、巨大芽孢杆菌(Bacillus megaterium)和枯草芽孢杆菌(Bacillus subtilis)。     

pqq基因簇在Escherichia coli DH5α中表达及对其溶磷促生的影响

为丰富植物促生菌的构建模式,本文以Escherichia coli DH5α为供试菌株,通过遗传转化,将水生拉恩氏菌(Rahnella aqautilis)HX2的pqq基因簇导入E.coli DH5α,获得DH5α(pqq)菌株,采用平板溶磷、钼锑抗比色法、HPLC法、温室盆栽等试验分析研究DH5α(pqq)菌株溶磷产酸促生作用机理。DH5α(pqq)菌株与E.coli DH5α相比,溶磷圈直径、有效磷浓度增加,溶解矿质磷的能力显著增强;有机酸代谢显著增加,以产葡萄糖酸为主。DH5α(pqq)处理与对照相比对玉米株高、茎鲜重、植株鲜重、茎干重、植株干重、全磷、土壤有效磷分别提高了5.2%、30.0%、32.5%、18.5%、7.7%、30.8%和24.0%,并呈显著性差异(P0.05)。表明来自HX2菌株的pqq基因簇通过异源表达具有活性,能够实现对溶磷促生的遗传改造。     

一株分离自葛藤根际高效溶磷细菌特性研究及菌株鉴定

对分离自葛藤(Pueraria lobata L.)根际的一株高效溶磷细菌GTR15进行促生特性、主要生理生化指标测定和16S rDNA序列分析。结果表明,菌株GTR15的HD/CD值(溶磷圈直径HD,菌落直径CD)为2.22,28℃液体振荡培养7 d后对磷酸钙的溶解量为138.72 mg.L-1,分泌IAA(3-吲哚乙酸)及有机酸量分别为14.44 mg.L-1、46.00 mmol.L-1。菌株革兰氏染色为阴性,细胞短杆状;淀粉水解、吲哚、V-P(二乙酰试验)、苯丙氨酸脱氨酶、明胶液化及M-R(甲基红)试验呈阴性;柠檬酸盐、过氧化氢酶、硫化氢及硝酸盐还原等试验呈阳性,结合菌株16S rDNA序列分析结果,初步鉴定为肠杆菌(Enterobacter sp.)。该菌株在研制高效微生物磷肥接种剂方面可能具有较大潜力。     

溶磷真菌的筛选及配施难溶态磷对土壤磷素有效性的影响

为研究溶磷真菌菌群对土壤磷素有效性的影响,首先在室内对3株不同种类溶磷真菌(1株属于被孢霉属Z1,1株为青霉属Z2,1株为黑曲霉Z3)的组合效应进行了研究,确定了最佳的菌株组合Z1+Z2+Z3,试验选用Z1、Z2、Z3组成菌群作为试验菌株;然后通过盆栽油菜试验研究溶磷真菌配施难溶态磷(磷酸三钙和磷矿粉)对土壤磷素有效性的影响。结果表明:溶磷真菌处理土壤有效磷、有机质、碱性磷酸酶、蔗糖酶含量和油菜产量分别比基质处理显著增加了60.00%,20.21%,56.45%,53.81%,14.38%,溶磷真菌配施难溶态磷上述各指标都高于单施溶磷真菌处理;单施溶磷真菌对土壤最大吸磷量的影响与基质无差异,溶磷真菌配施难溶态磷可以显著降低土壤最大吸磷量,溶磷真菌+磷酸三钙和溶磷真菌+磷矿粉处理土壤最大吸磷量比溶磷真菌处理显著减少158.7,47.6 mg/kg,溶磷真菌各处理土壤吸附常数都低于对应的基质处理,溶磷真菌可以降低土壤对磷的吸附。在土壤上溶磷真菌应与难溶态磷配合施用,对提高土壤磷素有效性有积极的作用。     

一株红壤花生根际溶磷真菌的分离、鉴定及溶磷能力的研究

为提高磷素利用率,从花生根际土壤样品中筛选到一株溶磷能力强的菌株B1-A,结合菌落形态特征和ITS rDNA序列分析将其鉴定为黑曲霉(Aspergillus niger)。菌株B1-A的不同接种量(1%,3%,5%)在两种难溶性磷酸盐(FePO4,AlPO4)培养液的溶磷量和pH随时间动态变化表明溶磷量均与pH呈显著负相关,B1-A的溶磷量与接种量、培养时间、磷矿粉浓度有关,实验结果表明菌株B1-A在接种量为3%～5%、培养时间168 h、云南磷矿粉浓度为3 g L-1、江西磷矿粉浓度为5 g L-1时溶磷效果较好。菌株B1-A对磷酸三钙、磷酸铝、磷酸铁、磷矿粉有较强的溶解能力,最高溶磷量分别为418.7、942.3、242.2、177.4 mg L-1,有望为开发高效红壤微生物磷肥提供种质资源。     

溶磷复合菌剂筛选及其对山核桃幼苗的生物学效应

为探究不同溶磷复合菌剂对山核桃幼苗生物学效应的影响,本研究从山核桃根际土壤中分离获得3株溶磷菌:果胶杆菌Pectobacterium cypripedii(R7)、伯克霍尔德氏菌Burkholderia sp.(R8)、肠杆菌Enterobacter sp.(R9)和4株自生固氮菌株:解聚糖类芽孢杆菌Paenibacillus glycanilyticus(G2)、葡萄球菌Staphylococcus sp.(G3)、假单胞菌Pseudomonas sp.(G4)、芽孢杆菌Bacillus sp.(G5),分别组合筛选出4种复合菌剂(R7G5、R8G4、R8G5、R9G4),并以蒸馏水为对照(CK),将其接种于山核桃幼苗的土壤中,通过测定接种处理后幼苗各项生长和生理指标,评价复合菌株对山核桃幼苗的作用效果。结果显示,R9G4和R8G5复合菌剂在蒙金娜无磷培养基(PVK)连续培养5 d后溶磷量显著高于其他组合。所有复合菌剂对山核桃幼苗均有促生作用;其中R8G5复合菌剂比其他复合菌的总体促生效果更为明显,施用后山核桃幼苗的根长、根面积、总生物量、全磷含量和可溶性糖含量较CK分别增加了105.53%、89.07%、52.94%、55.23%和91.99%;而施用R9G4复合菌剂后山核桃幼苗净光合速率(Pn)和全磷含量的提升效果最佳,较CK分别增加了79.47%和58.16%。本研究结果为进一步研制山核桃苗期促生菌剂提供了理论依据。     

Agricultural potential of Mediterranean grain and forage legumes: Key differences between and within Vicia species in terms of phenology, yield, and agronomy give insight into plant adaptation to semi-arid environments

In order to identify key adaptive traits which affect productivity in Mediterranean grain and forage legumes and simultaneously determine the agricultural potential of a wide range of Vicia species, germplasm collected from the wild throughout the eastern Mediterranean was grown under semi-arid conditions in Tel Hadya, Syria (313 mm growing season rainfall). These included species currently in use in Mediterranean agriculture, such as V. sativa L., as well as those more widely used in the past-such as V. ervilia L., but also a broad selection from Section Narbonensis (B. Fedtsch. ex Radzhi) Maxted, including V. narbonensis L, V. johannis Tamamsch., V. hyaeniscyamus Mout., V. serratifolia Jacq., and V. kalakhensis Khattab et al. V. faba, a near relative of the taxa in Section Narbonensis, was included as a domesticated control. Where applicable, a representative range of subspecies was used. Accessions were chosen from a wide range of habitats in terms of latitude (31.02–40.72 decimal^o), longitude (27.1–43.17 decimal^o), altitude (20–1510 m), rainfall (180–1700 mm/yr) and soil depth (5–50 cm) in order to maximise diversity within species. Agricultural potential was determined by measuring seed, hay and biological yield, as well as agronomic traits such as harvest index, standing crop height, and seed size. The comparative influence of phenology and key agronomic traits such as plant habit and seed size on productivity varied tremendously between species, depending on their reproductive strategies. In V. sativa and V. ervilia, the smaller seed species which rely on long vegetative phases and growing seasons to accumulate sufficient biomass to set seed, and in which there was comparatively little agronomic variation, phenology had a large impact on yield. In early emerging taxa such as V. ervilia and V. s. subsp. sativa, with built-in long vegetative phases and growing seasons, seed yield was negatively correlated with flower ing (r = –0.86 to –0.88), whereas the opposite was the case for later emerging taxa such as V. s. subsp. nigra (L.) Ehrh. (r = 0.95). Within V. narbonensis and relatives, the larger seeded Vicia species which rely on more conservative reproductive strategies where high seedling vigour associated with large seeds enables the species to enter reproductive phases relatively early, phenology had a much smaller impact on yield than did variation of key traits such as seed weight, plant habit and pod shattering. Among the undomesticated germplasm harvest indices ranged from 0.09–0.31, hay yields from 0.1–3.4 t/ha, seed yield from 0–2.0 t/ha, and dry matter at maturity from 1.6–6.5 t/ha. Sub-specific taxonomy was crucial in determining agronomic potential. V. narbonensis var. aegyptiaca Kornhuber ex Asch. et Schweinf. showed the most potential, combining an upright habit, large seeds (212 mg) and tendency to retain intact pods after maturity, with the highest yield, harvest index and crop height of all the wild Vicia species. V. sativa subsp. sativa, V. ervilia and V. narbonensis var. narbonensis were less productive, but still showed agricultural potential. The smaller seeded V. narbonensis, var. affinis, var. jordanica H. Schäf. and var. salmonea (Mout.) H. Schäf., and their close relatives V. johannis, V. hyaeniscyamus, V. serratifolia and V. kalakhensis have little to offer Mediterranean agriculture on the basis of poor agronomy.     

Changes in trace element speciation in kola north surface waters during snow melt

The powerful mass-flow of melt water during the brief period of spring thaw provides a surge of pollutants and causes extremely toxic short term conditions. Together with the increase in total metal concentrations during this period, a redistribution in metal speciation occurs that increases the proportion of more toxic forms. Concentrations of ion-exchangeable fractions of nickel (Ni), copper (Cu), aluminium (Al), and manganese (Mn) show strong negative correlations with pH. The sum of complex metal species shows a strong positive correlation with total organic carbon (TOC). A data analysis model based on relationships observed between key parameters and metal behaviour together with gidrological data allowed an assessmrnt of the metal impact on waterbodies from accumulated airborne pollution (Ni, Cu) as well as from accelerated leaching from watersheds (Al, Mn).     

On the Taxonomy of the Members of ‘Saccharum Complex’

The difficulty and problems encountered in the study of cultivated plants, in general and sugarcane, in particular has been indicated. In order to understand these problems, a brief review on the taxonomy of Saccharum and closely related taxa, namely, Erianthus, Sclerostachya, Narenga and Miscanthus (generally known as ‘Saccharum complex’) has been given. A short account on the important morphological features that are specific to sugarcane has also been stated as classification is commonly based on such morphological characters. A note has been added on the chromosome number, origin, and distribution of the species of ‘Saccharum complex’ members. Taxonomic keys have been devised for identification of the genera of Saccharinae and for the species of Saccharum and Erianthus occurring in India. A new combination, Sclerostachya fallax (Balansa) Amalraj et Balasundaram, has also been proposed.     

Polymorphism of high molecular weight glutenin subunits in three species of Aegilops

A collection of 136 accessions of Aegilops umbellulata (39), Ae. comosa (75) and Ae. markgrafii (22) was analysed for high-molecular-weight (HMW) glutenin subunits composition. The homogeneity of the accessions was studied and 55.1% of the collection was homogeneous for HMW glutenin subunits (29 Ae. umbellulata, 33 Ae. comosa and 14 Ae. markgrafii). The HMW glutenin subunits of Ae. umbellulata are encoded by the Glu-U1 locus; in Ae. comosa results showed that this proteins are encoded at the 1M chromosome, and the locus was named Glu-M1. In Ae. markgrafii it was assumed that HMW glutenin subunits were encoded by an homoeologous locus and it was named Glu-C1. All the accessions of Ae. umbellulata and Ae. markgrafii expressed both, x-type and y-type subunits. Among the Ae. comosa accessions, only one expressed an x-type subunit alone. All the accessions of Ae. umbellulata and some of Ae. comosa had x-type glutenins of higher molecular weights than those commonly present in bread wheat. A total of 8 alleles were detected at the Glu-U1 locus, 11 at the Glu-M1 and 4 at the Glu-C1. The new HMW glutenin variation found in this work suggests their possible utilisation in breeding for wheat quality.     

The major features of an infestation by the invasive weed legume gorse (Ulex europaeus) on volcanic soils in Hawaii

Gorse (Ulex europaeus) infestation occupies over 4,000 ha of agriculture and conservation lands on the southeastern slope of Mauna Kea on the Island of Hawaii. The aim of this investigation is to identify ecological features associated with this weed invasion by comparing the gorse-infested areas to the surrounding uninfested areas of this landscape. The soils within the gorse infestation are more acidic, resulting in higher levels of KCl-extractable Al and lower levels of Mehlich III-extractable Ca, Mg, Mn, and Zn. Yet, gorse accumulates higher concentrations of Ca, Zn and, Cu than the kikuyu grass (Pennesitum clandestinum), which is ubiquitous throughout the site. The Ca:Al and Mg:Al molar charge ratios of the soils are lowest within the epicenter of the gorse infestation, while the molar ratios are highest in the gorse apical stem tissues. All gorse plants are nodulated and have higher nitrogen contents than the surrounding kikuyu grass. Furthermore, the δ¹⁵N of the gorse stem tissues approaches 0‰, suggesting that nitrogen is being symbiotically fixed from the atmosphere. Characterization of the Bradyrhizobium isolated from gorse nodules shows similarities and distinctions to Bradyrhizobium isolated from the endemic legume koa (Acacia koa) within the same location. Population densities of the indigenous Bradyrhizobium are higher within the gorse rhizosphere than the kikuyu grass. Soil acidification, nutrient depletion, and symbiotic nitrogen fixation distinguish gorse-infested areas from the surrounding uninfested areas. These observations suggest that gorse has a competitive advantage over kikuyu grass under conditions of soil nutrient deficiency.     

Nematofauna associated with exotic and native leguminous plant species in West Africa: effect of<Emphasis Type="Italic"> Glomus intraradices</Emphasis> arbuscular mycorrhizal symbiosis

Ten leguminous trees, four exotic species (Australian Acacia) and six indigenous species (three Sahelian Acacia spp. and three Sesbania spp.), were grown for 4 months in a natural Sahelian soil inoculated with or without the endomycorrhizal fungus, Glomus intraradices. In control trials, the determinant factor structuring the soil nematode fauna was the plant species, related plants having a similar influence on the nematode community in the soil. Soil nematode abundance increased from exotic acacias (3.3 g^-1 dry soil) to native acacias (11.5 g^-1 dry soil) and Sesbania species (17.6 g^-1 dry soil). Plant feeding nematodes (mainly Scutellonema and Tylenchorhynchus) were significantly less abundant under exotic acacias (1.4 g^-1 dry soil) than under native acacias (7.2 g^-1 dry soil) or Sesbania species (7.3 g^-1 dry soil). Bacterial feeding nematode density increased from exotic acacias (1.2 g^-1 dry soil) to native acacias (3.0 g^-1 dry soil) and Sesbania species (7.7 g^-1 dry soil) as total densities. However, the differences in the structure of the nematode communities between plant groups were suppressed in the presence of the mycorrhizal fungus. In fact, no difference in nematode densities remained between plant groups when G. intraradices developed in several dominant taxa belonging to different trophic groups, particularly: Tylenchorhynchus, Apelenchina, Cephalobus and Dorylaimoidea. This study clearly indicated that inoculation with the arbuscular mycorrhizal fungus G. intraradices diminished the plant-specific effect on the structure of the soil nematode community.     

The effects of feeding by Oniscus asellus on leaf litter sulfur constituents

Summary Feeding by the isopod, Oniscus asellus, produced changes in the sulfur constituents of leaf litter substrates (Acer negundo, A. saccharum, Fagus grandifolia, Picea rubens, and Tsuga canadensis). Isopod consumption of leaf litter generally accelerated the mineralization of carbon-bonded S and increased the formation of ester sulfate in all substrates. After the isopod egestion of A. negundo leaves, fecal decomposition over 6 weeks increased total S concentration from 68 to 120 mol S/g due to the catabolism of organic carbon. During the same period sulfate decreased from 34 to 20 mol S/g and carbon-bonded S increased from 34 to 100 mol S/g. Thus the total S pool in aged feces became enriched with organic S (83% of total S). Macroinvertebrate consumption accelerated the transformation of S constituents and the carbon-bonded S concentration approached that of the Oa organic horizon of a northern hardwood forest.Dedicated to the late Prof. Dr. M.S. Ghilarov     

Genetic diversity of rhizobia associated with indigenous legumes in different regions of Flanders (Belgium)

We investigated the diversity of rhizobia isolated from different indigenous legumes in Flanders (Belgium). A total of 3810 bacterial strains were analysed originating from 43 plant species. Based on rep-PCR clustering, 16S rRNA gene and recA gene sequence analysis, these isolates belonged to Bradyrhizobium, Ensifer (Sinorhizobium), Mesorhizobium and Rhizobium. Of the genera encountered, Rhizobium was the most abundant (62%) and especially the species Rhizobiumleguminosarum, followed by Ensifer (19%), Bradyrhizobium (14%) and finally Mesorhizobium (5%). For two rep-clusters only low similarity values with other genera were found for both the 16S rRNA and recA genes, suggesting that these may represent a new genus with close relationship to Rhodopseudomonas and Bradyrhizobium. Primers for the symbiotic genes nodC and nifH were optimized and a phylogenetic sequence analysis revealed the presence of different symbiovars including genistearum, glycinearum, loti, meliloti, officinalis, trifolii and viciae. Moreover, three new nodC types were assigned to strains originating from Ononis, Robinia and Wisteria, respectively. Discriminant and MANOVA analysis confirmed the correlation of symbiosis genes with certain bacterial genera and less with the host plant. Multiple symbiovars can be present within the same host plant, suggesting the promiscuity of these plants. Moreover, the ecoregion did not contribute to the separation of the bacterial endosymbionts. Our results reveal a large diversity of rhizobia associated with indigenous legumes in Flanders. Most of the legumes harboured more than one rhizobial endosymbiont in their root nodules indicating the importance of including sufficient isolates per plant in diversity studies.