Symbiotic and saprophytic survival of three unmarked Rhizobium leguminosarum biovar trifolii strains introduced into the field

The symbiotic and saprophytic persistence of three unmarked Rhizobium leguminosarum biovar trifolii (Rlt) strains introduced into a field site in Iceland were followed. This site was free of clover cultivation and initially devoid of clover-nodulating rhizobia as tested by nodulation studies. Nodule occupancy by strains was identified based on their distinct ERIC-polymerase chain reaction (PCR) DNA fingerprint patterns. The survival and persistence of the individual strains in soil were monitored by the quantitative real-time PCR (qRT-PCR) assay, targeting the host-specific nodE gene. The most dominant strain in the nodule population, Rlt 20-15, showed relatively less saprophytic survival ability and maintained high numbers only in the presence of the appropriate host plant. Conversely, the minor nodule occupant, Rlt 32-28, persisted in soil at a relatively higher abundance both in the presence of its host legumes and in the presence of a non-host grass. The qRT-PCR assay was successfully applied to quantify rhizobial strains directly in soil without culturing or nodulation. However, the assay demonstrated less sensitivity compared with the plant infection most-probable-number (MPN) method for estimating the population size of rhizobia in soil. The quantitative detection limit of our qRT-PCR assays was 1 x 10(3) cells per gram of soil, as opposed to the MPN test which has a detection limit of 10 cells per gram of soil.     

Rhizobium trifolii mutants deficient in exopolysaccharide production

Spontaneous mutants of Rhizobium trifolii 24AR5 which did not produce exopoly-saccharide were isolated. The non-mucoid mutants formed small white and ineffective nodules on both red and white clover. These nodules contained infection threads, but only a small number of bacteria were released into nodule cells, and bacteroids were rarely observed. The non-mucoid phenotype was not complemented by the symbiotic plasmid (pJB5JI) of Rhizobium leguminosarum.     

Survival of Rhizobium in monoculture and binary population with Rhizosphere bacteria

The survival of pure cultures of Rhizobium leguminosarum bv. pisum and Rhizobium trifolii and their interaction with associative diazotrophic and phosphate-mobilizing bacteria after inoculation of sterile soil were studied. The viable heterotypical diazotrophic and rhizobial phosphate-mobilizing association was shown to be formed whose efficiency was 14% (clover) and 28% (pea) higher compared to monorhizobial inoculates.     

Identification of a Rhizobium trifolii plasmid coding for nitrogen fixation and nodulation genes and its interaction with pJB5JI, a Rhizobium leguminosarum plasmid

Rhizobium trifolii T37 contains at least three plasmids with sizes of greater than 250 megadaltons. Southern blots of agarose gels of these plasmids probed with Rhizobium meliloti nif DNA indicated that the smallest plasmid, pRtT37a, contains the nif genes. Transfer of the Rhizobium leguminosarum plasmid pJB5JI, which codes for pea nodulation and the nif genes and is genetically marked with Tn5, into R. trifolii T37 generated transconjugants containing a variety of plasmid profiles. The plasmid profiles and symbiotic properties of all of the transconjugants were stably maintained even after reisolation from nodules. The transconjugant strains were placed into three groups based on their plasmid profiles and symbiotic properties. The first group harbored a plasmid similar in size to pJB5JI (130 megadaltons) and lacked a plasmid corresponding to pRtT37a. These strains formed effective nodules on peas but were unable to nodulate clover and lacked the R. trifolii nif genes. This suggests that genes essential for clover nodulation as well as the R. trifolii nif genes are located on pRtT37a and have been deleted. The second group harbored hybrid plasmids formed from pRtT37a and pJB5JI which ranged in size from 140 to ca. 250 megadaltons. These transconjugants had lost the R. leguminosarum nif genes but retained the R. trifolii nif genes. Strains in this group nodulated both peas and clover but formed effective nodules only on clover. The third group of transconjugants contained a hybrid plasmid similar in size to pRtT37b. These strains contained the R. trifolii and R. leguminosarum nif genes and formed N2-fixing nodules on both peas and clover.     

Development of a real-time PCR assay for detection and quantification of Rhizobium leguminosarum bacteria and discrimination between different biovars in zinc-contaminated soil

Primers were designed to target 16S rRNA and nodD genes of Rhizobium leguminosarum from DNA extracted from two different soil types contaminated with Zn applied in sewage sludge. Numbers of rhizobia estimated using 16S rRNA gene copy number showed higher abundance than those estimated by both nodD and the most-probable-number (MPN) enumeration method using a plant trap host. Both 16S rRNA gene copies and the MPN rhizobia declined with increased levels of Zn contamination, as did the abundance of the functional gene nodD, providing compelling evidence of a toxic effect of Zn on R. leguminosarum populations in the soil. Regression analysis suggested the total Zn concentration in soil as a better predictor of rhizobial numbers than both NH4NO3-extractable and soil solution Zn. R. leguminosarum bv. viciae nodD gene copies were generally less sensitive to Zn than R. leguminosarum bv. trifolii nodD. The latter were generally below detection limits at Zn levels of >250 mg kg(-1). Although there were differences in the actual numbers estimated by each approach, the response to Zn was broadly similar across all methods. These differences were likely to result from the fact that the molecular approaches assess the potential for nodulation while the MPN approach assesses actual nodulation. The results demonstrate that the use of targeted gene probes for assessing environmental perturbations of indigenous soil rhizobial populations may be more sensitive than the conventional plant bioassay and MPN methods.     

华南热带人工林土壤有机碳含量及其稳定性特征

研究华南热带多雨地区小良人工森林生态系统定位站不同人工林的碳固持特征,测定了林地表土有机碳含量、有机碳随土层深度的变化以及有机碳在不同粒级团聚体上的分布。结果表明：无植被覆盖的光裸地储存了极低水平的有机碳,在光裸地上种植桉树林后,如果地表枯落物长期人为取走,则土壤有机碳积累大大低于未受干扰桉林。当桉林进一步改造为阔叶混交林后,30-40a林龄混交林土壤有机碳含量均高于16.5gkg^-1,但仍只有邻近天然林土壤有机碳水平的76%,因而,这些人工混交林土壤具有进一步积累碳的潜力。光裸地在1m剖面上的含碳量均极低,并无显著的深度变化,约2-3gkg^-1,未保护的桉林含量水平在4-5gkg^-1,同样表土与心土层没有明显差别,通过地表输入的凋落物对土壤有机碳的影响主要集中在表层0.5m范围内。以豹皮樟Litsea rotundifolia,阴香Cinnamomum burmanni,潺槁木姜子Litsea glutinosa,陈氏钓樟Lindera chuni等为优势种的混交林,深层土壤含碳量显著高于其它混交林,并与村边林相似。土壤有机碳储量与土壤团聚体发育有一定的相关性,非水稳性团聚体以最小粒径（〈0.25mm）的含碳浓度最高,其它粒级的有机碳浓度相似,未成熟人工林,小团聚体的碳浓度较高,成熟林则在不同粒级中较均匀地分配有机碳,但实际碳储量于最大粒级的非水稳性团聚体中最多。成熟森林土壤水稳性团聚体含碳浓度大大高于非水稳性团聚体,水稳性团聚体在0.25-0.5mm及0.5-1.0mm两个粒级上有最大的碳积累量。结果表明,建立特定类型的混交林是增加林地碳汇功能的一个有效途径,非水稳性的大团聚体与水稳性的小团聚体的有效发育可能是成熟森林高固碳量的重要机理。     

Effects of inoculation with Rhizobium leguminosarum biovar trifolii on wheat cultivated in clover crop rotation agricultural soil in Morocco

One hundred strains of Rhizobium leguminosarum bv. trifolii were isolated from roots of wheat cultivated in rotation with clover in two different regions of Morocco. The isolates were first screened for their effect on the growth of the cultivar Rihane of wheat cultivated in an agricultural soil under greenhouse conditions. After 5 weeks of growth, 14 strains stimulating the fresh or dry matter yield of shoots were selected and used in a second pot inoculation trial performed with two different agricultural soils. The results show that the strains behaved differently according to the soil used. In the loamy sand Rabat, strain IAT 168 behaved potentially like a plant growth promoting rhizobacteria (PGPR), as indicated by the 24% increases (P < 0.1) observed in wheat shoot dry matter and grain yields. In the silty clay Merchouch, no PGPR activity was observed, and 6 strains showed a significant deleterious effect on yields. These observations suggest that it is very important in a crop rotation system to choose a R. leguminosarum bv. trifolii strain that is effective with clover and shows PGPR activity with wheat to avoid deleterious effects on wheat yields.     

宁南山区植被恢复对土壤团聚体水稳定及有机碳粒径分布的影响

土壤团聚作用和土壤有机碳固定之间密切相关.对宁南山区不同植被恢复措施和年限下土壤团聚体粒径分布及稳定性、土壤团聚体中有机碳及其组分分布进行了研究,探讨了有机碳及其组分对植被恢复的响应.结果表明,不同植被恢复措施下,土壤团聚体粒径分布表现为“V”字分布:＞5 mm和＜0.25 mm这两个粒径的团聚体含量最多,5-2 mm、1-0.25 mm团聚体的含量次之,2-1 mm粒径的团聚体含量最少.坡耕地的平均重量直径(MWD)最低,为1.4,其他植被恢复措施下土壤的平均重量直径MWD在1.9-3.1之间.不同的植被恢复措施下,0-20 cm土层和20-40 cm土层全土有机碳含量在7.4-17.7 g/kg之间、微生物碳含量分布在50.3-664.7 mg/kg之间、腐殖质碳含量在0.9-2.5g/kg之间.胡敏酸碳含量分布在0.2-0.6 g/kg,富里酸碳含量在0.6-1.9 g/kg之间.全土有机碳、微生物碳、腐殖质碳、富里酸碳均为坡耕地最低,其他植被恢复措施的有机碳、微生物碳、腐殖质碳、富里酸碳含量分别是坡耕地的1.1-2.3倍、2.0-8.4倍、1.0-2.0倍、1.2-2.4倍.不同粒径团聚体有机碳相比较,大多呈现中间高两边低的变化趋势,最大值出现在中间粒径,即5-2 mm、2-1 mm、1-0.25 mm这3个粒径.逐步回归表明,5-2 mm团聚体和1-0.25 mm团聚体有机碳含量的提高有助于土壤水稳性团聚体的形成.研究结果表明,植被恢复提高了土壤团聚体有机碳含量,在碳形态上,富里酸碳和微生物生物量碳对不同植被恢复措施的敏感度较高,胡敏酸碳含量则相对稳定.     

Characterization of the anomalous infection and nodulation of subterranean clover roots by Rhizobium leguminosarum 1020

Anomalous nodulation of Trifolium subterraneum (subterranean clover) roots by Rhizobium leguminosarum 1020 was examined as a model of modified host-specificity in a Rhizobium-legume symbiosis. Consistent with previous reports, these nodules (i) appeared most often at sites of secondary root emergence, (ii) were ineffective in nitrogen fixation and (iii) were as numerous as nodules formed by an effective Rhizobium trifolii strain. R. leguminosarum 1020, grown on agar plates or in the clover root environment, did not bind the white clover lectin, trifoliin A. This strain did not attach in high numbers, and did not induce shepherd's crooks or infection threads, in subterranean clover root hairs. However, R. leguminosarum 1020 did cause branching, moderate curling and other deformations of root hairs. The bacteria probably entered the clover root through breaks in the epidermis at sites of lateral root emergence. The anomalous nodulation was inhibited by nitrate. Only trace amounts of leghaemoglobin were detected in the nodules by Western blot analysis. The nodules were of the meristematic type and initially contained well-developed infection, bacteroid and senescent zones. Infection threads were readily found in the infection zone of the nodule. However, the bacteroid-containing tissue senesced more rapidly than in the effective symbiosis between subterranean clover and R. trifolii 0403. This anomalous nodulation of subterranean clover by R. leguminosarum 1020 suggests a naturally-occurring alternative route of infection that allows Rhizobium to enlarge its host range.     

Natural endophytic association between Rhizobium leguminosarum bv. trifolii and rice roots and assessment of its potential to promote rice growth

For over 7 centuries, production of rice (Oryza sativa L.) in Egypt has benefited from rotation with Egyptian berseem clover (Trifolium alexandrinum). The nitrogen supplied by this rotation replaces 25- 33% of the recommended rate of fertilizer-N application for rice production. This benefit to the rice cannot be explained solely by an increased availability of fixed N through mineralization of N- rich clover crop residues. Since rice normally supports a diverse microbial community of internal root colonists, we have examined the possibility that the clover symbiont, Rhizobium leguminosarum bv. trifolii colonizes rice roots endophytically in fields where these crops are rotated, and if so, whether this novel plant-microbe association benefits rice growth. MPN plant infection studies were performed on macerates of surface-sterilized rice roots inoculated on T. alexandrinum as the legume trap host. The results indicated that the root interior of rice grown in fields rotated with clover in the Nile Delta contained 10⁶ clover-nodulating rhizobial endophytes g fresh weight of root. Plant tests plus microscopical, cultural, biochemical, and molecular structure studies indicated that the numerically dominant isolates of clover-nodulating rice endophytes represent 3 – 4 authentic strains of R. leguminosarum bv. trifolii that were Nod Fix on berseem clover. Pure cultures of selected strains were able to colonize the interior of rice roots grown under gnotobiotic conditions. These rice endophytes were reisolated from surface-sterilized roots and shown by molecular methods to be the same as the original inoculant strains, thus verifying Koch's postulates. Two endophytic strains of R. leguminosarum bv. trifolii significantly increased shoot and root growth of rice in growth chamber experiments, and grain yield plus agronomic fertilizer N-use efficiency of Giza-175 hybrid rice in a field inoculation experiment conducted in the Nile Delta. Thus, fields where rice has been grown in rotation with clover since antiquity contain Fix strains of R. leguminosarum bv. trifolii that naturally colonize the rice root interior, and these true rhizobial endophytes have the potential to promote rice growth and productivity under laboratory and field conditions.     

Direct amplification of nodD from community DNA reveals the genetic diversity of Rhizobium leguminosarum in soil

Sequences of nodD , a gene found only in rhizobia, were amplified from total community DNA isolated from a pasture soil. The polymerase chain reaction (PCR) primers used, Y5 and Y6, match nodD from Rhizobium leguminosarum biovar trifolii , R. leguminosarum biovar viciae and Sinorhizobium meliloti . The PCR product was cloned and yielded 68 clones that were identified by restriction pattern as derived from biovar trifolii [11 restriction fragment length polymorphism (RFLP) types] and 15 clones identified as viciae (seven RFLP types). These identifications were confirmed by sequencing. There were no clones related to S. meliloti nodD . For comparison, 122 strains were isolated from nodules of white clover ( Trifolium repens ) growing at the field site, and 134 from nodules on trap plants of T. repens inoculated with the soil. The nodule isolates were of four nodD RFLP types, with 77% being of a single type. All four of these patterns were also found among the clones from soil DNA, and the same type was the most abundant, although it made up only 34% of the trifolii -like clones. We conclude that clover selects specific genotypes from the available soil population, and that R. leguminosarum biovar trifolii was approximately five times more abundant than biovar viciae in this pasture soil, whereas S. meliloti was rare.     

云雾山草原区不同植被恢复阶段土壤团聚体活性有机碳分布特征

以宁夏云雾山草原自然保护区不同植被群落为研究对象,对0～20 cm土层土壤团聚体活性有机碳分布特征进行分析.结果表明:(1)不同植被群落土壤团聚体活性有机碳含量顺序为退耕草地<百里香群落<铁杆蒿群落<大针茅群落<长芒草群落,与退耕草地相比,封育草地各粒径团聚体活性有机碳含量均显著提高 (P<0.05),表明随着植被的恢复土壤团聚体活性有机碳含量提高并趋于稳定,土壤碳汇效应有可能增强.(2)植被恢复主要影响大团聚体(>0.25 mm)中活性有机碳含量,其中0.5～0.25 mm粒径团聚体中活性有机碳含量最高,微团聚体(<0.25 mm)中活性有机碳含量最低.(3)植被恢复前期(退耕草地-铁杆蒿群落)0～10 cm土层>0.5 mm粒径团聚体中活性有机碳含量较10～20 cm土层有所增加,<0.5 mm粒径团聚体活性有机碳含量变化不大,恢复至后期到长芒草阶段时,0～10 cm土层<0.5 mm粒径团聚体中活性有机碳含量也开始提高, 各粒径0～10 cm土层团聚体活性有机碳含量均比10～20 cm土层有所提高.(4)相关性分析表明,土壤团聚体活性有机碳含量与土壤团聚体总有机碳含量呈极显著线性正相关关系(r=0.9394),团聚体活性有机碳含量可以作为衡量土壤团聚体有机碳动态的一个敏感性指标.     

Studies on the Characterization of Root Morphology of White Clover Infected by Transconjugant of Rhizobium leguminosarum

White clover plants were inoculated with transconjugant strain' 290 which was obtained from introduction of host specific nodulation genes of wild-type Rhizobium trifolii strain ANU 843 to Rhizobium leguminosarum strain 300. The characterization of root morphology of white clover induced by the transconjugant was observed and compared to the plants induced by the parent strains. White clover started tO form a typical root hair curling inoculated with transconjugant strain 290 24h after inoculation, at 48h a part of cell wall of root hair was degradated, infection thread was observed in the infected root hair cell, cortical cell divisions occurred extensively. All these characterizations were similar to that infected by strain ANU 843. Plant inoculation test indicated that no nodule was formed when inoculated by R. leguminosarum strain 300, while plants nodulated when inoculated with transconjugant strain 290 as well as R. trifolii ANU 843. This suggests that introduction of host specific nodulation genes of R. trifolii results in conferring the nodulation ability of R. leguminosarum on white clover.     

Root colonization of different plants by plant-growth-promoting Rhizobium leguminosarum bv. trifolii R39 studied with monospecific polyclonal antisera.

Monospecific polyclonal antisera raised against Rhizobium leguminosarum bv. trifolii R39, a bacterium which was isolated originally from red clover nodules, were used to study the colonization of roots of leguminous and nonleguminous plants (Pisum sativum, Lupinus albus, Triticúm aestivum, and Zea mays) after inoculation. Eight weeks after inoculation of soil-grown plants, between 0.1 and 1% of the total bacterial population in the rhizospheres of all inoculated plants were identified as R. leguminosarum bv. trifolii R39. To characterize the associative colonization of the nonleguminous plants by R.leguminosarum bv. trifolii R39 in more detail, a time course study was performed with inoculated roots of Z. mays. R. leguminosarum bv. trifolii R39 was found almost exclusively in the rhizosphere soil and on the rhizoplane 4 weeks after inoculation. Colonization of inner root tissues was detected only occasionally at this time. During the process of attachment of R. leguminosarum bv. trifolii R39 to the rhizoplane, bacterial lipopolysaccharides were overexpressed, and this may be important for plant-microbe interaction. Fourteen weeks after inoculation, microcolonies of R. leguminosarum bv. trifolii R39 were detected in lysed cells of the root cortex as well as in intracellular space of central root cylinder cells. At the beginning of flowering (18 weeks after inoculation), the number of R. leguminosarum bv. trifolii R39 organisms decreased in the rhizosphere soil, rhizoplane, and inner root tissue.     

植物篱-农作坡地土壤团聚体组成和稳定性特征

基于植物篱控制水土流失的长期定位试验,研究植物篱-农作坡地土壤团聚体组成和稳定性特征.结果表明： 与常规等高农作模式相比,植物篱-农作复合农业模式下土壤>0.25 mm机械稳定性和水稳性团聚体含量分别显著增加13.3%~16.1%和37.8%~55.6%,明显提高了各坡位粒级>0.25 mm土壤水稳性团聚体含量,改善了粒级>0.25 mm土壤水稳性团聚体在下坡位的相对富集和上坡位相对贫乏的状况.植物篱显著提高了土壤团聚体平均质量直径和几何平均直径,降低了土壤团聚体分形维数和>0.25 mm土壤团聚体破坏率,进而增强了土壤团聚体的稳定性和抗蚀性.坡度与植物篱类型对土壤团聚体组成、稳定性和坡面变化无显著影响.     

植物篱-农作坡地土壤团聚体组成和稳定性特征

基于植物篱控制水土流失的长期定位试验,研究植物篱-农作坡地土壤团聚体组成和稳定性特征.结果表明： 与常规等高农作模式相比,植物篱-农作复合农业模式下土壤>0.25 mm机械稳定性和水稳性团聚体含量分别显著增加13.3%~16.1%和37.8%~55.6%,明显提高了各坡位粒级>0.25 mm土壤水稳性团聚体含量,改善了粒级>0.25 mm土壤水稳性团聚体在下坡位的相对富集和上坡位相对贫乏的状况.植物篱显著提高了土壤团聚体平均质量直径和几何平均直径,降低了土壤团聚体分形维数和>0.25 mm土壤团聚体破坏率,进而增强了土壤团聚体的稳定性和抗蚀性.坡度与植物篱类型对土壤团聚体组成、稳定性和坡面变化无显著影响.     

不同开垦年限黑土溅蚀与团聚体分选特征

分别以开垦年限为8 a、30 a、50 a的典型黑土耕地和未开垦的天然次生林林地表层土壤为对象,采用人工模拟降雨的方法,对比分析了不同开垦年限土壤的溅蚀规律及溅蚀过程中团聚体分选、分布特征,并通过析因分析研究了开垦对黑土溅蚀的影响机制.结果表明：不同开垦年限黑土耕地土壤的溅蚀量明显高于未开垦次生林林地土壤,且随开垦年限的延长而增加,变化范围为0.95～7.06 g·cm^-1;土壤溅蚀量与溅蚀距离表现出紧密的指数函数关系,而且随着水平空间距离的增加,小粒级水稳性团聚体比例逐渐增加;溅蚀对黑土团聚体产生富集作用与损耗作用的临界粒级为1.0 mm;粒级＞2 mm、＜0.25 mm水稳性团聚体及有机质含量是典型黑土土壤溅蚀量及其变化特征的主要影响因子.     

Distribution of carbon, nitrogen and phosphorus in water-stable aggregates of an organic waste amended Ultisol in southern Nigeria

This study was undertaken to evaluate the effects of different organic and inorganic amendments on nutrient (C, N and P) distribution in water-stable aggregates (WSA) of a degraded tropical Ultisol at Nsukka in southern Nigeria. The treatments were control (C), inorganic fertilizer (F) (N-P-K, 12-12-17), rice-mill wastes (RW), RW + F, poultry manure (PM) and RW + PM. These were arranged in a randomized complete block design (RCBD) with four replications. At 0, 3, 6 and 12 months after application of the amendments, topsoil (0-20 cm) samples were collected from all the treatments, dried and separated into 4.75-2.0, 2.0-1.0, 1.0-0.5, 0.50-0.25 and <0.25 mm aggregates classes by wet sieving. The mesoaggregates fraction (1.0-0.50 mm) made up the highest percentage of the whole soil irrespective of the treatments. From the sixth month, the treatments generally reduced the proportion of these mesoaggregates as well as those of the microaggregates (<0.25 mm diameter) while increasing those of the other sizes. The largest (>2.0 mm) and the smallest (<0.25 mm) aggregates were preferentially enriched in C and N relative to the 2.0-0.25 mm aggregates in all the treatments, whereas P distribution within the aggregates was random. There was a low, but significant correlation between organic carbon and mean-weight diameter (MWD) of the aggregates. The correlation between C and N within the aggregates was also positive (r = 0.917) and highly significant (P < 0.01). The implication of these results for management of these soils is discussed.     

不同轮作制度对淮北白浆土团聚体及其有机碳的积累与分布的影响

淮北白浆土是苏鲁交界地区主要低产土壤 ,同时也是我国黄淮地区商品粮生产基地的重要土壤资源。该土壤除了剖面发生分异强烈、土壤理化性质不良外 ,有机碳匮乏是其主要肥力限制因子 [1]。土壤有机碳不但是维持和培育土壤质量的关键组成成分 ,而全球土壤有机碳每年分解释放大气 CO2 而且达到 0 .1～ 5.4C Pg·年 -1,土壤碳 0 .1 %的变化将导致大气圈 CO2 浓度 1 mg· L - 1的变化。因而其存储和释放的变化与大气 CO2 动态有密切的关系 [9,10 ] 。农业土壤对大气 CO2 的截存贡献是研究陆地系统对大气 CO2 的汇效应 (sink effect)的焦点 …     

The construction, detection and use of bioluminescent Rhizobium leguminosarum biovar trifolii strains

A. CRESSWELL, L. SKØT AND A.R. COOKSON. 1994. The gene encoding the firefly luciferase enzyme ( luc ) was introduced to Rhizobium leguminosarum biovar trifolii strains with a view to using the resulting bioluminescent strains to study the survival of genetically-engineered rhizobia in soil microcosms. The genetically-engineered micro-organisms (GEMs) behaved similarly to their parent strains with respect to growth rate in laboratory media and in their symbiotic performance with their host plants. No gene transfer could be detected in laboratory mating experiments. When inoculated onto a non-sterile soil the population of the GEM declined sharply from an initial cell density of 2 times 107⁷ g^-1 soil to approach a stable cell density of approximately 3 times 10² g^-1 after 150 d. Direct photography of bioluminescent rhizobia enabled the detection of colonies as small as 0.1 mm in diameter without the need for transferring colonies onto filter paper. When a Rhizobium strain carrying the luc marker on a plasmid was used as inoculant it was possible to visualize differences in colonization of the rhizosphere of white clover and ryegrass by contact print and colour transparency films. The photographic detection methods described here demonstrate the possibilities of using bioluminescent rhizobia for assessing their survival in soil, and for looking at rhizosphere populations which may be an important site for potential gene transfer.