Nodulation status of native woody legumes and phenotypic characteristics of associated rhizobia in soils of southern Ethiopia

The nodulation of provenances of Acacia seyal, Acacia tortilis and Faidherbia albida, and other indigenous multipurpose tree species were tested in 14 different soil samples collected from diverse agro-ecological zones in southern Ethiopia. Associated rhizobia were isolated from these and from excavated nodules of field standing mature trees, and phenotypically characterized. Indigenous rhizobia capable of eliciting nodules on at least one or more of the woody legume species tested were present in most of the soils. Tree species were markedly different in nodulation in the different site soils. Sesbania sesban and Acacia abyssinica showed higher nodulation ability across the different sites indicating widespread occurrence of compatible rhizobia in the soils. The nodulation patterns of the different provenances of Acacia spp. suggested the existence of intraspecific provenance variations in rhizobial affinity which can be exploited to improve N fixation through tree selection. Altogether, 241 isolates were recovered from the root nodules of trap host species and from excavated nodules. Isolates were differentiated by growth rate and colony morphology and there were very fast-, fast-, slow-, and very slow-growing rhizobia. The bulk of them (68.5%) were fast-growing acid-producing rhizobia while 25.3% were slow-growing alkali-producing types. Fast-growing alkali-producing (2.9%) and slow-growing acid-producing strains (3.3%) were isolated from trap host species and excavated nodules, respectively. All isolates fell into four colony types: watery translucent, white translucent, dull glistering and milky (curdled) type. The diversity of indigenous rhizobia in growth rate and colony morphology suggested that the collection probably includes several rhizobial genera.     

Nodulation of Medicago truncatula and Medicago polymorpha in two pastures of contrasting soil pH and rhizobial populations

The size of the background rhizobial population can often determine the success of field nodulation and persistence of inoculant rhizobia. Field experiments were conducted to determine the nodulation response of annual medics (Medicago spp.) in a pasture-wheat-pasture rotation when grown in soils of contrasting pH and rhizobial populations. Medicago truncatula Gaertn. and M. polymorpha L. were inoculated with one of three different strains of Sinorhizobium medicae (WSM540, WSM688) or S. meliloti (NA39) or left uninoculated and sown in two fields of pH (CaCl₂) 5.9 and 7.2 of differing soil rhizobial backgrounds (11 and 7.1 × 10⁴ cells/g soil, respectively). Nodulation was assessed in years 1 and 3 of the rotation. At the site with a small rhizobial background, M. polymorpha nodulated poorly when inoculated with the acid-sensitive strain NA39 but nodulated well when inoculated with acid-tolerant strains WSM688 and WSM540. M. truncatula had a similar extent of nodulation with each of the rhizobial inoculants. At the site with a large rhizobial background all treatments had greater than 85% of plants nodulated. Nodule occupancies, assessed by PCR, provided further insight: at the site with a small rhizobial background both medic species successfully nodulated with the acid-tolerant strains WSM540 and WSM688 and these strains persisted to year 3. However, at the site with large rhizobial background, only one strain, WSM688, was identified from M. truncatula nodules in year 3. This study highlights the importance of edaphic constraints and plant–rhizobia interactions to the successful development of nodulation in a field environment.     

Effects of resident rhizobial communities and soil type on the effective nodulation of pulse legumes

Communities of resident rhizobia capable of effective nodulation of pulse crops were found to vary considerably over a range of soil environments. These populations from soils at 50 sites in Southern Australia were evaluated for nitrogen fixing effectiveness in association with Pisum sativum, Vicia faba, Lens culinaris, Vicia sativa, Cicer arietinum and Lupinus angustifolius. The values for nitrogen fixing effectiveness could be related to soil pH as determined by soil type and location. It was found that 33% of paddocks had sufficient resident populations of Rhizobium leguminosarum bv viciae for effective nodulation of faba bean, 54% for lentils, 55% for field pea and 66% for the effective nodulation of the vetch host plant. Mesorhizobium cicer populations were very low with only 7% of paddocks surveyed having sufficient resident populations for effective nodulation. Low resident rhizobial populations (<10 rhizobia g⁻¹ soil) of R. leguminosarum bv viciae and M. cicer were found in acid soil conditions. In contrast, Bradyrhizobium populations increased as soil pH decreased. Inoculation increased faba bean yields from 0.34 to 4.4 t ha⁻¹ and from 0.47 to 2.37 t ha⁻¹ for chickpeas on acid soils. On alkaline soils, where resident populations were large there was no consistent response to inoculation. Observations at experimental field sites confirmed the findings from the survey data, stressing the importance of rhizobial inoculation, especially on the acid soils in south-eastern Australia.     

Diagnostic indices of zinc deficiency in tropical legumes

The effect of 0.005 ppm, 0.01 ppm, 0.05 ppm and 0.10 ppm zinc on the growth and chemical composition of Desmodium uncinatum Jacq. cv. Silverleaf; Macroptilium lathyroides L.; Lablab purpureus; and Glycine max. L. cv. Wills were studied in solution culture in a controlled environment room.

Dry matter production of tops of all species was reduced at the lower zinc treatments while that of roots was unaffected by zinc supply.

Zinc concentrations of leaves from three plant positions did not provide a reliable index of zinc nutrition. The lowest zinc treatments often produced greater leaf concentrations of zinc than did higher zinc treatments. The cytoplasmic fractions of the leaf tissue contained the highest zinc concentrations. Cell walls, nuclei, chloroplasts and mitochondria contained extremely low concentrations, and the data were generally variable, reflecting the poor reproducibility of the techniques used.

Phosphorus concentrations in leaves were generally increased at the low zinc treatments, an effect not entirely attributable to the reduction in dry matter production. The proportion of inorganic phosphorus increased and that of organic phosphorus decreased at the lower zinc treatments. Lipoid and residual phosphorus fractions were not affected consistently by zinc treatments.

Leaves of plants receiving the lowest zinc treatment generally had significantly lower nitrogen concentrations than those receiving the higher zinc treatments.

In another experiment, in which Glycine wightii was substituted for G. max., RN‐ase activity increased as the zinc supply was reduced and this was accompanied by a decrease in protein concentration. The data show that RN‐ase activity could be a useful diagnostic index of incipient zinc deficiency.     

几种豆科、禾本科植物对多环芳烃复合污染土壤的修复

通过盆栽试验,研究了几种豆科植物与禾本科植物对多环芳烃(PAHs)复合污染土壤的修复作用。结果显示,90天后8种植物对土壤中PAHs均有不同程度降解效果,其中紫花苜蓿和多年生黑麦草对土壤PAHs的去除率分别达48.4%、46.8%,且对3环PAHs去除较为彻底,对4环及4环以上的PAHs去除效果较差。8种供试植物对PAHs均有一定的吸收、富集与转运的能力,紫花苜蓿和多年生黑麦草对土壤PAHs的生物富集系数分别为0.096、0.085,其提取修复效率为0.017%和0.013%。可见,紫花苜蓿和多年生黑麦草具有较好的根际修复潜力。     

Nodulation and estimation of symbiotic nitrogen fixation by herbaceous and shrub legumes in Guinea savanna in Nigeria

Twelve herbaceous and shrub legume species were grown in pot and field experiments in five sites representing three agroecological zones in moist savanna in Nigeria. The objectives were to: (1) assess natural nodulation of the legumes and characterize their indigenous rhizobia, (2) determine their need for rhizobia inoculation and (3) estimate the amount of N₂ fixed by each of these legumes. At 4 weeks after planting (WAP), Crotolaria verrucosa was not nodulated at any of the sites while Centrosema pascuorum had the highest number of nodules in all sites. At 8 WAP, all legumes were nodulated, with Mucuna pruriens having the least number of nodules and Stylosanthes hamata the highest. The number of nodules, however, was inversely correlated to the mass of nodules. Significant differences in nodulation of the legume species grown in the field also occurred between and within sites. Mucuna pruriens and Lablab purpureus produced more shoot and nodule biomass than the other legumes in all sites. Growth of most of these legumes responded to fertilizer application, except for C. verrucosa and Aeschynomene histrix. Except for C. verrucosa, average proportion of N₂ fixed was about 80% and this was reduced by about 20% with N fertilizer application. The majority of rhizobia isolates (60%) were slow growing, belonging to the Bradyrhizobia spp. group. Selected rhizobia isolates evaluated on Cajanus cajan, C. pascuorum, M. pruriens and Psophocarpus palustris varied from ineffective to highly effective in Leonard jar conditions. However, only growth of M. pruriens responded to inoculation in potted soils, whereas it was lower than that obtained with N fertilizer application. This indicated the need to screen more rhizobia in order to improve N₂ fixation and growth of legume species such as M. pruriens when it is introduced in soils deficient in N.     

Biodegradation kinetics of monosaccharides and their contribution to basal respiration in tropical forest soils

Low molecular weight (LMW) organic compounds in soil solution are easily biodegradable and could fuel respiration by soil microorganisms. Our main aim was to study the mineralization kinetics of monosaccharides using ¹⁴C-radiolabelled glucose. Based on these data and the soil solution concentrations of monosaccharides, we evaluated the contribution of monosaccharides to basal respiration for a variety of tropical forest soils. Further, the factors controlling the mineralization kinetics of monosaccharides were examined by comparing tropical and temperate forest soils. Monosaccharides comprised on average 5.2 to 47.7% of dissolved organic carbon in soil solution. Their kinetic parameters (V _max and K_M ), which were described by a single Michaelis-Menten equation, varied widely from 11 to 152?nmol?g^?1?h^?1 and 198 to 1294?µmol?L^?1 for tropical soils, and from 182 to 400?nmol?g^?1?h^?1 and 1277 to 3150?µmol?L^?1 for temperate soils, respectively. The values of V _max increased with increasing microbial biomass-C in tropical and temperate soils, while the K_M values had no correlations with soil biological or physicochemical properties. The positive correlation between V _max values and microbial biomass-C indicates that microbial biomass-C is an essential factor to regulate the V _max values in tropical and temperate forest soils. The biodegradation kinetics of monosaccharides indicate that the microbial capacity of monosaccharide mineralization far exceeds its rate at soil solution concentration. Monosaccharides in soil solution are rapidly mineralized, and their mean residence times in this study were very short (0.4–1.9?h) in tropical forests. The rates of monosaccharide mineralization at actual soil solution concentrations made up 22–118% of basal respiration. Probably because of the rapid and continuous production and consumption of monosaccharides, monosaccharide mineralization is shown to be a dominant fraction of basal respiration in tropical forest soils, as well as in temperate and boreal forest soils.     

Evaluation of different agroforestry tree species for their suitability in the phytoremediation of seleniferous soils

Seven agroforestry tree species were grown in a clay loam soil treated with different levels of selenate‐Se, viz. 0, 1.25, 2.5 and 5.0 mg/kg supplied through sodium selenate. After 1 year of growth, a progressive decrease in dry matter of leaves, stem and roots was observed with increasing levels of applied Se. However, a significant decrease in dry matter yield was observed only at or above 2.5 mg Se per kg soil and shisham (Dalbergia sissoo) proved to be highly sensitive to the presence of selenate‐Se in the soil. On average, the largest above‐ground and below‐ground biomass was accumulated by arjun (Terminalia arjuna) and the lowest by the acacia tree (Acacia tortillas). The selenium content of leaves, stem and roots of all the tree species increased significantly with increasing levels of applied Se, although a large variation within species was observed. In the stem portion of different trees, the highest concentration of Se was found in dek (Melia azedarach) (5.1 mg/kg) and the lowest in mulberry (Morus alba) (2.6 mg/kg). The efficiency of selenium removal (including leaves, stem and roots) was the highest in arjun followed by eucalyptus (Eucalyptus hybrid) – Clone 10, mulberry, jambolin (Syzygium cumini), dek, shisham and acacia. Effective removal of Se takes place through the stem portion of different trees where it constitutes 30–50% of total Se. Large variation in Se uptake by different tree species suggests that trees vary in their potential for phytoremediation of seleniferous soils. In one growing season, shisham aged 24 years, poplar (Populus deltoides)– Clone G 48 (10 years old) and eucalyptus – Clone 10 (10 years old) could remove 2385, 1845 and 1407 g Se per hectare respectively. Corresponding reductions in Se capital of the soil varied between 24 and 37, 19 and 29 and 14 and 32%, respectively, in the surface layer (0–15 cm) alone or 7–11, 6–9 and 4–7% for the whole soil profile (0–120 cm). Removal further increased to 4207 g Se per hectare under an agroforestry farming system of poplar–mentha/wheat with Se being reduced from 43 to 65% for the surface layer and from 13 to 20% for the whole profile.     

Effects of nitrogen source and aluminum on growth of tropical tree seedlings adapted to low pH soils

The effects of N-source and Al on the growth of seedlings of Melastoma malabathricum, Acacia mangium, and Melaleuca cajuputi, which are tropical woody plants and are very tolerant to Al, and barley (Hordeum vulgare), which is a typical Al-sensitive plant, were investigated. The Al and N treatments consisted of the application of either 0 or 0.5 mM Al, and 2 mM NH₄ ⁺ or N0₃ ^-, respectively. Growth of the tropical plants was enhanced by Al and NH₄ application. In all the plant species, the pH of the culture solution decreased and the concentrations of soluble Al and P increased with the + NH₄ treatment, which positively affected the growth of the tropical plant species. Excised roots of M. malabathricum dissolved insoluble Al with NH₄ application and absorbed Al mainly from root tips. Al did not affect the leaf N concentration except in the case of barley. Roots of M. cajuputi exuded a large amount of citrate, which slightly increased by the + Al treatment. In A. mangium, the reactivity of soluble Al to PCV (pyrocatecholviolet) decreased in the culture solution of the + Al + NH₄. treatment and Al concentration of roots in this treatment was very low. Roots of M. malabathricum released H+ along with Al uptake as well as NH₄ ⁺ uptake. It is concluded that Al and NH₄ ⁺ exert beneficial effects on the growth of tropical tree seedlings.     

Root-nodule bacteria from indigenous legumes in the north-west of Western Australia and their interaction with exotic legumes

Bacteria were isolated from root-nodules collected from indigenous legumes at 38 separate locations in the Gascoyne and Pilbara regions of Western Australia. Authentication of cultures resulted in 31 being ascribed status as root-nodule bacteria based upon their nodulation of at least one of eight indigenous legume species. The authenticated isolates originated from eight legume genera from 19 sites. Isolates were characterised on the basis of their growth and physiology; 20 isolates were fast-growing and 11 were slow-growing (visible growth within 3 and 7 d, respectively). Fast-growers were isolated from Acacia, Isotropis, Lotus and Swainsona, whilst slow-growers were from Muelleranthus, Rhynchosia and Tephrosia. Indigofera produced one fast-growing isolate and seven slow-growing isolates. Three indigenous legumes (Swainsona formosa, Swainsona maccullochiana and Swainsona pterostylis) nodulated with fast-growing isolates and four species (Acacia saligna, Indigofera brevidens, Kennedia coccinea and Kennedia prorepens) nodulated with both fast- and slow-growing isolates. Swainsona kingii did not form nodules with any isolates. Fast-growing isolates were predominantly acid-sensitive, alkaline- and salt-tolerant. All slow-growing isolates grew well at pH 9.0 whilst more than half grew at pH 5.0, but all were salt-sensitive. All isolates were able to grow at 37 °C. The fast-growing isolates utilised disaccharides, whereas the slow-growing isolates did not. Symbiotic interactions of the isolates were assessed on three annual, one biennial and nine perennial exotic legume species that have agricultural use, or potential use, in southern Australia. Argyrolobium uniflorum, Chamaecytisus proliferus, Macroptilium atropurpureum, Ononis natrix, Phaseolus vulgaris and Sutherlandia microphylla nodulated with one or more of the authenticated isolates. Hedysarum coronarium, Medicago sativa, Ornithopus sativus, Ornithopus compressus, Trifolium burchellianum, Trifolium polymorphum and Trifolium uniflorum did not form nodules. Investigation of the 31 authenticated isolates by polymerase chain reaction with three primers resulted in the RPO1 primer distinguishing 20 separate banding patterns, while ERIC and PucFor primers distinguished 26 separate banding patterns. Sequencing the 16S rRNA gene for four fast- and two slow-growing isolates produced the following phylogenetic associations; WSM1701 and WSM1715 (isolated from Lotus cruentus and S. pterostylis, respectively) displayed 99% homology with Sinorhizobium meliloti, WSM1707 and WSM1721 (isolated from Sinorhizobium leeana and Indigofera sp., respectively) displayed 99% homology with Sinorhizobium terangae, WSM1704 (isolated from Tephrosia gardneri) shared 99% sequence homology with Bradyrhizobium elkanii, and WSM1743 (isolated from Indigofera sp.) displayed 99% homology with Bradyrhizobium japonicum.     

Bioactive compounds in legumes and their germinated products

Nineteen domestic legume varieties, including 6 soybeans, 7 black soybeans, 4 azuki beans, and 2 mung beans, were evaluated for contents of dietary fiber, total phenolics, and flavonoids. Nine varieties of legumes (black soybean TN6, TN3, BM, and WY; soybean KS1, KS2, and KS8; azuki bean AKS5 and AKS6) were good sources of bioactive compounds and were selected for germination tests. After short- and long-term germinations, the bioactive compounds were determined and compared with compositions of isoflavones in soybeans. The reducing power of legumes correlated well with their total flavonoid contents (r (2) = 0.9414), whereas less correlation was found between reducing power and total phenolics contents (r (2) = 0.6885). The dark-coat seeds, such as azuki beans and black soybeans, contained high amounts of phenolic compounds and contributed to high antioxidative ability, whereas their phenolics content and antioxidative abilities significantly decreased after short-term germination due to losses of pigments in the seed coats. After long-term germination, the contents of bioactive compounds (total phenolics and flavonoids) increased again and the ratio of aglycones to total isoflavones significantly increased in black soybeans. TN3 and TN6 seeds and their long-term germinated seeds and AKS5 seeds were identified as the legume samples that might have the highest antioxidant ability according to the results of chemometric analysis. Selection of the right legume varieties combined with a suitable germination process could provide good sources of bioactive compounds from legumes and their germinated products for neutraceutical applications.     

Diversity and phylogeny of rhizobial strains isolated from Lotus uliginosus grown in Uruguayan soils

Lotus uliginosus is generally nodulated by rhizobia of the genus Bradyrhizobium when used for improvement of Uruguayan pastures. The genetic diversity and phylogenetic relationships of 111 isolates from nodules of L. uliginosus collected from four fields with or without prior inoculation history were analyzed in this study. Genetic diversity estimated by ERIC-PCR revealed 75 different genomic fingerprints, and showed a relatively greater value compared with other methods and varied by soil type. 16S ribosomal RNA gene RFLP analysis revealed three different ribogroups, A, B and C, with 71 isolates in ribogroup A, three isolates in ribogroup B and only one in ribogroup C. Phylogenetic analyses based on 16S RNA gene sequences, ITS, as well as atpD, recA and glnII gene sequences indicated that ribogroup A strains were affiliated with B. japonicum bv. genistearum strains. The three isolates in ribogroup B did not clearly associate with any Bradyrhizobium species described previously and could represent a novel species within this genus. Unlike B. japonicum strains these isolates were able to nodulate and fix nitrogen with other Lotus species as well as with Spartium, a leguminous shrub. The unique isolate in ribogroup C clustered with Mesorhizobium and appeared genetically and phenotypically related to broad host-range Mesorhizobium sp. NZP2037. Our data suggest that Uruguayan soils contain native or naturalized bradyrhizobia that are able to nodulate L. uliginosus as efficiently as the commercial strain NZP2309 but could have adaptive advantages making them more suitable for inoculant purposes.     

Variations in infectivity of indigenous rhizobial isolates of some soils in the rainforest zone of Nigeria

Nodule formation in legumes is a process that starts with root infection by rhizobia. The present study assessed the population and infectivity of the indigenous rhizobial strains in rainforest soils of Nigeria. Soils were collected from three sites – Idi-Ayunre, Orile-Ilugun (OI) and the University of Ibadan Teaching and Research Farm (UITRF) – and analysed for physico-chemical properties and rhizobial population. Soybean varieties TGx1448-2E and TGx1456-2E and a cowpea variety IT89KD-288 were planted as trap crops on each of the soils, and rhizobia were isolated from their nodules. Infectivity assay was conducted using eight varieties of soybean and a cowpea variety. Most probable number estimate of the rhizobial population showed that the UITRF had significantly higher rhizobial population than the other two locations. OI and the UITRF soils planted with TGx1448-2E had significantly higher nodules and number of strains than other treatments. Among the 70 slow-grower strains isolated, only nine were infective. Three of the nine strains – IDC8, TRC2 and OISa-6e – nodulated at least seven of the eight soybean varieties used for infectivity test. Indigenous rhizobial infectivity of the studied locations was low, and cultivation of grain legume may require rhizobial inoculation for high productivity.     

Decomposition of rice residue in tropical soils

Nitrogen mineralization and immobilization of rice residue in Maahas clay soil under lowland and upland conditions were investigated by using ¹⁵N-labelled rice straw. The mineralization of residue-nitrogen was taking place even though the net mineralization was depressed by incorporation of rice residue.

There were some significant differences in the pattern of nitrogen transformation between lowland and upland soil conditions. The nitrogen transformation measured by mineralization of soil nitrogen and rice-residue nitrogen and the nitrogen immobilization into rice residue were more active under lowland conditions than under upland conditions, during the earlier period of residue decomposition.     

Rhizobia in tropical legumes—XIII. Biochemical basis of acid and alkali reactions

Three fast-growing rhizobia (Rhizobium meliloti isolated from Medicago saliva, R. trifolii from Trifolium subterraneum, and Rhizohium sp. from Leucaena leucocephala) and three slow-growing rhizobia R. japonicum from Glycine max, Rhizobium spp from Centrosema pubescens and Crotolaria anagyroides) were grown in defined media. The mean generation times of the fast-growing and slowgrowing strains were 3.8 h and 8.6 h respectively. Slow-growing organisms raised the initial pH of the defined medium while the fast-growing organisms lowered it. Rates of oxygen consumption tended to be higher in the slow-growing organisms.UMKL 19 (isolated from L. leucocephala) possessed all the normal reactions of fast-growing rhizobia but had a single sub-polar flagellum, similar to the three slow-growing strains studied.Certain combinations of amino acids and sugars (e.g. glutamine and galactose) induced an acidic reaction in the fast-growing organisms while the slow-growing ones changed the media to alkaline. Fast-growing organisms utilized more galactose for growth compared to slow-growing ones. Both types of organisms synthesized and released a wide range of amino acids into the medium.We suggest that pH changes produced by rhizobia growing on yeast-extract mannitol media are caused by the preferential utilization of sugars by fast-growing organisms and nitrogenous compounds by slow-growing ones.     

濒危植物生殖生态学研究进展

濒危植物生殖生态学是濒危植物保护生物学研究的热点。本文综述了国内外濒危植物生殖生态学研究进展,包括濒危植物植物学及物候学特性、生殖系统、种子及幼苗生态学、生殖值及生殖分配、种群统计、生命表以及无性繁殖等方面,并指出当前研究中存在的问题及今后发展方向。