Mycorrhizal inoculation effect on nodulation and N accumulation in Cajanus cajan at soil P concentrations sufficient or inadequate for mycorrhiza-free growth

The interaction of Cajanus cajan with Rhizobium and vesicular-arbuscular mycorrhizal fungi (VAMF) was investigated in a greenhouse experiment. C. cajan was planted in soil that had been inoculated with Glomus aggregatum or treated with benlate to suppress VAMF activity. Initial soil solution P concentrations of 0.06, 0.2, 0.4, and 0.8 mg l^-1 were established to test the interaction at external P levels that ranged from inadequate to nonlimiting for the host plant. At 0.06 and 0.2 mg P l^-1, mycorrhizal inoculation significantly increased plant P concentrations as well as nodule numbers and shoot dry weight. Mycorrhizal inoculation also significantly increased nodule dry weight at a soil P concentration of 0.4 mg l^-1 but did not significantly influence any of the other variables. The mycorrhizal inoculation effect observed at this soil solution P concentration could not be explained by any of the measures of plant P status. At 0.8 mg P l^-1, none of the measured variables were affected significantly by mycorrhizal inoculation. The results indicate that the enhanced nodulation associated with mycorrhizal inoculation at soil P concentrations lower than 0.4 mg l^-1 was explainable by mycorrhizal-mediated P uptake. The small but significant increase in nodule mass due to VAMF inoculation at 0.4 mg P l^-1 suggests that factors not related to plant P nutrition may be involved. On the other hand, the lack of a VAMF inoculation effect at 0.8 mg P l^-1 despite VAMF colonization at a level comparable to that observed at the former P concentration appear to discount this hypothesis. This observation is also supported by the lack of response of plant N status and nodule number to VAMF inoculation at this soil P concentration.Contribution from the Hawaii Institute of Tropical Agriculture and Human Resources Journal Series No.4066     

The unique root-nodule symbiosis between Rhizobium and the aquatic legume,Neptunia natans (L. f.) Druce

We examined the development of the aquatic N₂-fixing symbiosis between Rhizobium sp. (itNeptunia) and roots of Neptunia natans L. f. (Druce) (previously N. oleracea Lour.) under natural and laboratory conditions. When grown in its native marsh habitat, this unusual aquatic legume does not develop root hairs, the primary sites of rhizobial infection for most temperate legumes. Under natural conditions, the aquatic plant floats and develops nitrogen-fixing nodules at emergence of lateral roots on the primary root and on adventitious roots at stem nodes, but not from the stem itself. Cytological studies using various microscopies revealed that the mode of root infection involved an intercellular route of entry followed by an intracellular route of dissemination within nodule cells. After colonizing the root surface, the bacteria entered the primary root cortex through natural wounds caused by splitting of the epidermis and emergence of young lateral roots, and then stimulated early development of nodules at the base of such roots. The bacteria entered the nodule through pockets between separated host cells, then spread deeper in the nodule through a narrower intercellular route, and eventually evoked the formation of infection threads that penetrated host cells and spread throughout the nodule tissue. Bacteria were released from infection droplets at unwalled ends of infection threads, became enveloped by peribacteroid membranes, and transformed into enlarged bacteroids within symbiosomes. In older nodules, the bacteria within symbiosomes were embedded in an unusual, extensive fibrillar matrix. Cross-inoculation tests of 18 isolates of rhizobia from nodules of N. natans revealed a host specificity enabling effective nodulation of this aquatic legume, with lesser affinity for Medicago sativa and Ornithopus sp., and an inability to nodulate several other crop legume species. Acetylene reduction (N₂ fixation) activity was detected in nodules of N. natans growing in aquatic habitats under natural conditions in Southern India. These studies indicate that a specific group of Rhizobium sp. (Neptunia) occupies a unique ecological niche in aquatic environments by entering into a N₂-fixing root-nodule symbiosis with Neptunia natans.We thank J. Whallon for technical assistance, G. Truchet, J. Vasse, S. Wagener, J. Beaman, F. DeBruijn, F. Ewers, and A. Squartini for helpful comments, and N.N. Prasad and G. Birla for assistance in conducting field observations. This work was supported by the Michigan Agricultural Experiment Station and National Science Foundation grants DIR-8809640 and BIR-9120006 awarded to the MSU Center for Microbial Ecology. This study is dedicated to the memory of Dr. Joseph C. Burton, a friend and colleague who made many contributions to the study of the Rhizobiumlegume symbiosis.     

Map order and linkage distances of molecular markers close to the supernodulation ( nts-1 ) locus of soybean

The molecular characteristics of markers in the chromosome region surrounding the supernodulation gene (nts-1) of soybean (Glycine max L. Merr.) were investigated in 187 F₂ plants from a cross of G. max cv. Bragg (nts) and G. soja PI468.397 (wild-type nodulation). RFLP marker pUTG-132a, linked tightly (0.7±0.5 cM) to nts-1, was converted to a PCR marker. The polymorphism resides within a 1.72 kb PstI fragment and consists of an 832 bp insertion in G. max relative to the wild progenitor G. soja. The insertion is flanked by a 35 bp direct duplication that was found only once in G. soja. Data suggest that the pUTG-132a sequence exists only once in the genome, which is compatible with the recessive nature of nts-1. Accordingly, pUTG-132a is a valuable marker for map-based cloning. Another RFLP marker, pA-381, was mapped 4.8 cM distal to nts-1. Marker order, established by Maximum Likelihood Analysis, placed nts-1 between pUTG-132a and pA-381. To generate additional molecular markers, a segregating F₂ population was analysed using bulked segregant analysis (BSA) and single oligonucleotide primer-based PCR (DNA amplification fingerprinting; DAF). PCR marker pcr5-4L was mapped to soybean linkage group H and sequenced. The data revealed (i) recombination events and marker order in the nts-1 region; (ii) the molecular nature and cause of polymorphisms in linked molecular markers; (iii) a low density of polymorphisms around nts-1, and (iv) diploidy of the distal region of linkage group H of soybean. Received: 18 January 1996 / Accepted: 9 October 1996     

豆科树种回接根瘤菌的研究

 作者对八种豆科树种的根瘤菌回接进行了研究,不回接根瘤菌的对照植株不结瘤或有少且小的根瘤,而接菌植株的根瘤数量多且个体大;接菌植株的株高、干重及总氮量分别比不接菌的对照植株高出0.7～3.2、1.3～15.8和11.3～14.8倍。根瘤固定的氮量占幼苗生长所需氮的一半以上,固定的氮绝大部分运输到植株其它部位,分配到地上部分的氮素多于根部。固氮量与幼苗生物量显著相关。固氮作用增加了植物对磷、钾元素的吸收和积累。速生树种南洋楹(Albizia falcata)和非速生树种格木(Erythrophloeum fordii)幼苗的结瘤、固氮及生长状况较好,表现出较高的结瘤固氮潜能。     

Identification of Frankia strains in nodules by hybridization of polymerase chain reaction products with strain-specific oligonucleotide probes

A set of oligonucleotides has been developed to study the competitivity of two Frankia strains in the nodulation of the roots of two host plant species: Alnus glutinosa and Alnus incana. Two 20 mer-oligonucleotides, complementary to highly conserved sequences inside the nifH gene, were used as primers for the polymerase chain reaction (PCR) system in order to amplify microsymbiont DNA extracted from actinorhizae. PCR products were analyzed using two strain-specific 15-mer oligonucleotides identified in the amplified region. Hybridization data indicate that strain ACoN24d is more competitive than train ArI3 in the nodulation of both hosts.     

Nodulation,nitrogen fixation and nitrogen uptake in pigeonpea (Cajanus cajan (L.) Millsp) of different maturity groups

Summary The seasonal patterns of nodulation, acetylene reduction, nitrogen uptake and nitrogen fixation were studies for 11 pigeonpea cultivars belonging to different maturity groups grown on an Alfisol at ICRISAT Center, Patancheru, India. In all cultivars the nodule number and mass increased to a maximum around 60–80 days after sowing and then declined. The nodule number and mass of medium- and late-maturing cultivars was greater than that of early-maturing cultivars. The nitrogenase activity per plant increased to 60 days after sowing and declined thereafter, with little activity at 100 days when the crop was flowering. At later stages of plant growth nodules formed down to 90 cm below the soil surface but those at greater depth appeared less active than those near the surface. All the 11 cultivars continued to accumulate dry matter until 140 days, with most biomass production by the late-maturing cultivars (up to 11 t ha⁻¹) and least by the early-maturing determinate cultivars (4 t ha⁻¹). Total nitrogen uptake ranged from 69 to 134 kg ha⁻¹. Nitrogen fixation by pigeonpea was estimated as the difference in total nitrogen uptake between pigeonpea and sorghum and could amount to 69 kg N ha⁻¹ per season, or half the total nitrogen uptake. Fixation by pigeonpea increased with crop duration, but there were differences within each maturity group. The limitations of the methods used for estimating N₂ fixation by pigeonpea are discussed. Submitted as J.A. No. 552 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT).     

An inexpensive cultural system using ash for cultivation of soybean (Glycine max (L.) merrill) on acid clay soils

Several cultural techniques were tested for establishment of nitrogen fixing soybeans on heavy-clay, acid sulfate soils of different fertility at two locations. Holes of 2.5 cm deep were made by inserting a pointed pole into the surface soil. Soybean seeds were dropped into the holes and covered with either (i) Rhizobium inoculant, molybdenum (0.5 kg Mo/ha) and ash from burned rice husks (0.2 t/ha) or (ii) Rhizobium inoculant, molybdenum and local soil instead of the ash. Additional treatments evaluated were the use or absence of tillage and the presence or absence of a rice straw mulch (5 t/ha). Soybean plants treated with ash had the highest number of nodules per plant. The combination of mulching and ash treatments produced the highest soybean grain yields. Without ash all treatments produced low grain yields. Tillage had no effect.     

Acetylene reduction byDaviesia mimosoides under Eucalyptus

Summary Daviesia mimosoides is a common understorey legume in Eucalyptus forests of the Brindabella Range in southeastern Australia, capable of fixing atmospheric nitrogen. Rates of N fixation were measured by the acetylene-reduction technique over a growing season in the field. Pot trials under controlled conditions were also carried out to elucidate effects of soil moisture, temperature, and light. Average rates in the field varied from about 1–5 μ mol C₂H₄/g/h (wet weight of nodule), but rates up to 14 μ mol C₂H₄/g/h were measured in optimum controlled conditions. Annual N-fixation rates approximate 4.5–7.0 kg/ha. In pot trials, rate of acetylene reduction decreased with soil moisture to about−10 MPa tension, with a marked depression at about−6 MPa, but within the normal field range of soil moisture there was little correlation of moisture with average acetylene reduction rate. Rates were similar in the temperature range of 20–30°C, but were depressed by either low or high temperature (<10 or >30°C). Diurnal fluctuations in acetylene reduction rates were not correlated with solar radiation, but rates were limited by high mid-day temperatures.