Influence of the Size of Indigenous Rhizobial Populations on Establishment and Symbiotic Performance of Introduced Rhizobia on Field-Grown Legumes

Indigenous rhizobia in soil present a competition barrier to the establishment of inoculant strains, possibly leading to inoculation failure. In this study, we used the natural diversity of rhizobial species and numbers in our fields to define, in quantitative terms, the relationship between indigenous rhizobial populations and inoculation response. Eight standardized inoculation trials were conducted at five well-characterized field sites on the island of Maui, Hawaii. Soil rhizobial populations ranged from 0 to over 3.5 × 10⁴ g of soil^-1 for the different legumes used. At each site, no less than four but as many as seven legume species were planted from among the following: soybean (Glycine max), lima bean (Phaseolus lunatus), cowpea (Vigna unguiculata), bush bean (Phaseolus vulgaris), peanut (Arachis hypogaea), Leucaena leucocephala, tinga pea (Lathyrus tingeatus), alfalfa (Medicago sativa), and clover (Trifolium repens). Each legume was (i) inoculated with an equal mixture of three effective strains of homologous rhizobia, (ii) fertilized at high rates with urea, or (iii) left uninoculated. For soybeans, a nonnodulating isoline was used in all trials as the rhizobia-negative control. Inoculation increased economic yield for 22 of the 29 (76%) legume species-site combinations. While the yield increase was greater than 100 kg ha^-1 in all cases, in only 11 (38%) of the species-site combinations was the increase statistically significant (P ≤ 0.05). On average, inoculation increased yield by 62%. Soybean (G. max) responded to inoculation most frequently, while cowpea (V. unguiculata) failed to respond in all trials. Inoculation responses in the other legumes were site dependent. The response to inoculation and the competitive success of inoculant rhizobia were inversely related to numbers of indigenous rhizobia. As few as 50 rhizobia g of soil^-1 eliminated inoculation response. When fewer than 10 indigenous rhizobia g of soil^-1 were present, economic yield was significantly increased 85% of the time. Yield was significantly increased in only 6% of the observations when numbers of indigenous rhizobia were greater than 10 cells g of soil^-1. A significant response to N application, significant increases in nodule parameters, and greater than 50% nodule occupancy by inoculant rhizobia did not necessarily coincide with significant inoculation responses. No less than a doubling of nodule mass and 66% nodule occupancy by inoculant rhizobia were required to significantly increase the yield of inoculated crops over that of uninoculated crops. However, lack of an inoculation response was common even when inoculum strains occupied the majority of nodules. In these trials, the symbiotic yield of crops was, on average, only 88% of the maximum yield potential, as defined by the fertilizer N treatment. The difference between the yield of N-fertilized crops and that of N₂-fixing crops indicates a potential for improving inoculation technology, the N₂ fixation capacity of rhizobial strains, and the efficiency of symbiosis. In this study, we show that the probability of enhancing yield with existing inoculation technology decreases dramatically with increasing numbers of indigenous rhizobia.     

Bradyrhizobium technology: a promising substitute for chemical nitrogen fertilizer in Bangladesh agriculture

Nitrogen is the most limiting element in Bangladesh soils and urea is the fertilizer commonly used for supplying it. Bradyrhizobium/Rhizobium inoculant was tried as a source of N nutrition for grain legumes in a number of field experiments. The inoculants markedly increased nodule number, nodule mass, shoot weight and yield of the crops compared to uninoculated control and urea-N treatments. For soybean (Glycine max), inoculation increased yield 113 percent over the control and 49 percent over the urea treament. For groundnut (Arachis hypogaea), the increases were 36 and 11 percent; for lentil (Lens culinaris), 30 and 13 percent; and for mungbean (Vigna radiata), 47 and 7 percent. The local inoculant strains were suitable for dependable inoculant production. The inoculant technology can be used as a promising and cheap substitute of urea for growing food legume crops in Bangladesh.     

Prospects and problems of simple linear models for estimating symbiotic N2 fixation by crop and pasture legumes

Symbiotic dinitrogen (N₂) fixation of crop and pasture legumes is a critical component of agricultural systems, but its measurement is expensive and labour intensive. Simple models which can provide approximations based on crop or pasture dry matter production would be useful for agrononomists and those interested in regional nitrogen (N) cycle fluxes. We investigate meta analysis of published data on legume shoot dry matter production, shoot %N and legume %N fixed (%Ndfa) and look for relationships among these, as a possible way of providing useful approximations of N₂ fixation. We restricted our analysis to Australian studies where we have ready access to the primary data and where cultivars, management and climate are more constrained compared to a universal dataset. Regression analysis between shoot dry matter and amounts of shoot N₂ fixed were strong for all crop and pasture legumes with significant differences in slope and intercept values being obtained between pastures and crops, and between chickpea (Cicer arietinium) and all other crop and pasture legumes. Annual pasture legumes showed the strongest linear relationship between N₂ fixation and shoot dry matter and had the greatest slope (20.2–24.3 kg N₂ fixed/t), compared to 18.7 kg N₂ fixed/t for the perennial pasture legume lucerne (alfalfa, Medicago sativa), and between 10.7 to 23.0 kg N₂/t for crop legumes, depending upon species. It was recognised that the use of such shoot-based relationships would underestimate the total amounts of N₂ fixed since the contributions of fixed N present in, or derived from, roots and nodules are not included. Furthermore there needs to be careful consideration of the validity of an intercept term, which might reflect suppression of N₂ fixation at low dry matter and high soil mineral N availability, or possibly the use of non-linear regression. For chickpea crops grown in north-eastern Australia, multiple regression indicated that N₂ fixation was much more closely correlated with %Ndfa than dry matter production. Evidence presented also indicated that %Ndfa of other crops and lucerne in this region may similarly be influenced by soil mineral N. The regression approach presented provides a statistical basis to approximate N₂ fixation in the first instance. This work highlights some of the dangers of fitting single regressions to aggregated datasets and using these to approximate symbiotic N₂ fixation. The analysis indicates that where pasture legumes are grown in mixtures with non-legumes, and driven to high dependence on N₂ fixation, simple linear regressions may be quite useful, provided that possible differences between species are investigated as the slopes of the regressions between these can be quite different. For crop legumes, where low dependence on N₂ fixation can occur at higher mineral N availability, there is a need to carefully consider the intercept term, obtain estimates of mineral N availability, and/or resort to non-linear models. The gross generalisations presented in scatter plots cannot be reliably applied any more specifically, even within the datasets from which they were generated, and in some cases even within legume species between regions. They cannot substitute for direct measurement where any certainty is required under a particular set of defined conditions.     

Plow down effects of different forage legume species,cultivars, cutting strategies and seeding rates on the yields of subsequent crops

Summary Under some conditions the plow down of forage legumes increases the yield of subsequent crops, which is usually caused by improved soil N. However, better soil structure is also a contributing factor. Three experiments were conducted to measure the effect of legume plow down on the yield of subsequent corn crops grown at the Ottawa Research Station (ORS), Ottawa, Canada. In all experiments, corn yields were not affected by legume species, legume cultivars, and/or planting methods. Corn yields from barley plots receiving 0, 60, or 120 kg N ha⁻¹ did not differ until two years after establishment in one experiment and three years in another. The data from these experiments indicated that soil N was high at the ORS, which may inhibit N₂-fixation by forage legumès in the establishment year. Therefore, legume plow down was not beneficial to subsequent crops under these conditions. Two other experiments were conducted to measure the effect of legume plow down on the yield of subsequent barley crops. In both experiments, barley yields in the field were not affected by legume type or legume seeding density. Greenhouse and field data indicated that the two cuts with removal strategy benefitted the most to succeeding crops. Data from the greenhouse test indicated that soil N levels were not low in the establishment year, and that some cultivars improved soil fertility more than others.     

Indigenous legumes biomass quality and influence on C and N mineralization under indigenous legume fallow systems

Non-cultivated N₂-fixing indigenous legumes can be harnessed to enhance soil fertility replenishment of smallholder farms. Understanding N release patterns of biomass generated by such legumes is key in managing N availability to crops. Nitrogen and C mineralization patterns of indigenous legume species, mainly ofTephrosia andCrotalaria genera, and of soils sampled at termination of 1- and 2-year indigenous legume fallows (indifallows)were investigated in leaching tube incubations under laboratory conditions. With the exception ofTephrosia longipes Meisn (2.4%) andCrotalaria cylindrostachys Welw.ex Baker (1.8%), all indigenous legumes had >2.5% N. Total polyphenols and lignin were <4% and 15%, respectively, for all species.Crotalaria pallida (L.) andEriosema ellipticum Welw.ex Baker mineralized >50% of the added N in the first 30 days of incubation. Similar to mixed plant biomass from natural weed fallow,C. Cylindrostachys immobilized N during the 155-day incubation period. Indifallow fallow biomass reached peak N mineralization 55 days after most legumes had leveled off. Carbon release by legume species closely followedN release patterns,with mostCrotalaria species releasing >500 mg CO₂-C kg^?1 soil. Soils sampled at termination of fallows reached peak N mineralization in the first 21 days of incubation, with indifallows mineralizing significantly (P<0.05) more N than natural fallows. Application of mineral P fertilizer to indifallows and natural fallows increased C and N mineralization relative to control treatments. It was concluded that (i) indigenous legumes generate biomass of high quality within a single growing season, (ii) the slow N release of biomass generated under indifallow systems suggests that such fallows can potentially be manipulated to enhance N availability to crops, and (iii) N and C mineralization of organic materials in sandy soils is likely controlled by availability of P to the soil microbial pool.     

Pod shattering in grain legumes: emerging genetic and environment-related patterns

A reduction in pod shattering is one of the main components of grain legume domestication. Despite this, many domesticated legumes suffer serious yield losses due to shattering, particularly under arid conditions. Mutations related to pod shattering modify the twisting force of pod walls or the structural strength of the dehiscence zone in pod sutures. At a molecular level, a growing body of evidence indicates that these changes are controlled by a relatively small number of key genes that have been selected in parallel across grain legume species, supporting partial molecular convergence. Legume homologs of Arabidopsis thaliana silique shattering genes play only minor roles in legume pod shattering. Most domesticated grain legume species contain multiple shattering-resistance genes, with mutants of each gene typically showing only partial shattering resistance. Hence, crosses between varieties with different genes lead to transgressive segregation of shattering alleles, producing plants with either enhanced shattering resistance or atavistic susceptibility to the trait. The frequency of these resistance pod-shattering alleles is often positively correlated with environmental aridity. The continued development of pod-shattering-related functional information will be vital for breeding crops that are suited to the increasingly arid conditions expected in the coming decades.

Recent genetic, genomic, and phenotypic studies of pod shattering in grain legumes lay the foundation for breeding crops suited for increasingly arid conditions.     

Nitrogen Relationships of Vigna mungo in Pure Stands and in Celosia argentea Mixtures

Dynamics of the nitrogen relationships of Vigna mungo in pureand in mixed stands with Celosia argentea, a common weed ofleguminous crops in certain regions of India, were investigated.The weed significantly depressed nodulation and reduced thedry matter and nitrogen yield of the legume which was lowestat the highest density of the weed. A significant amount ofnitrogen transfer occurred from the legume in the weed mixtureswhich appeared to be taken up by the weed plants. Therefore,weed plants benefited from their association with the legumeby accumulating more dry matter and nitrogen in the legume mixtures.The possible competitive or biochemical interference of theweed with nitrogen relationships of the legume in mixtures isdiscussed. Vigna mungo, Celosia argentea, pure stands, mixed stands, total nitrogen, interference     

Lablab purpureus—A Crop Lost for Africa?

In recent years, so-called ‘lost crops’ have been appraised in a number of reviews, among them Lablab purpureus in the context of African vegetable species. This crop cannot truly be considered ‘lost’ because worldwide more than 150 common names are applied to it. Based on a comprehensive literature review, this paper aims to put forward four theses, (i) Lablab is one of the most diverse domesticated legume species and has multiple uses. Although its largest agro-morphological diversity occurs in South Asia, its origin appears to be Africa. (ii) Crop improvement in South Asia is based on limited genetic diversity. (iii) The restricted research and development performed in Africa focuses either on improving forage or soil properties mostly through one popular cultivar, Rongai, while the available diversity of lablab in Africa might be under threat of genetic erosion. (iv) Lablab is better adapted to drought than common beans (Phaseolus vulgaris) or cowpea (Vigna unguiculata), both of which have been preferred to lablab in African agricultural production systems. Lablab might offer comparable opportunities for African agriculture in the view of global change. Its wide potential for adaptation throughout eastern and southern Africa is shown with a GIS (geographic information systems) approach.     

Promotion of Peanut Growth by Co-inoculation with Selected Strains of <Emphasis Type="Italic">Bradyrhizobium</Emphasis> and <Emphasis Type="Italic">Azospirillum</Emphasis>

The ability of inoculated rhizobial strains to increase root nodulation of host legumes often depends on their competitiveness with existing native soil strains. Results of studies to date on rhizobial inoculation for improvement of peanut (Arachis hypogaea L.) production in Argentina have been inconsistent and controversial. In many cases, nodulation and yield of peanut crops have been increased by inoculation of specific rhizobial strains. Native peanut-nodulating strains are generally present in soils of agricultural areas, but their growth-promoting effect is often lower than that of inoculated strains. Many species of the genus Bradyrhizobium interact in a host-specific manner with legume species and form nitrogen-fixing root nodules. Other free-living rhizobacteria such as species of the genus Azospirillum are facultatively capable of interacting with legume roots and promoting plant growth. We evaluated and compared the effects of various single inoculation and co-inoculation treatments on peanut growth parameters in greenhouse and field experiments. In the greenhouse studies, co-inoculation with various Bradyrhizobium strains (native 15A and PC34, and recommended peanut inoculant C145), and Azospirillum brasilense strain Az39 generally resulted in increases in the measured parameters. The growth-promoting effect of 15A was similar to or higher than that of C145. In the field studies, 15A-Az39 co-inoculation had a greater promoting effect on measured growth parameters than did C145-Az39 co-inoculation. Our findings indicate that careful selection of native rhizobacterial strains adapted to peanut soils is useful in strategies for growth promotion, and that 15A in particular is a promising candidate for future inoculant formulation.     

Estimation of residual N effect of faba bean and pea on two succeeding cereals using15N methodology

A field experiment was conducted using¹⁵N methodology to study the effect of cultivation of faba bean (Vicia faba L.), pea (Pisum sativum L.) and barley (Hordeum vulgare L.) on the N status of soil and their residual N effect on two succeeding cereals (sorghum (Sorghum vulgare) followed by barley). Faba bean, pea and barley took up 29.6, 34.5 and 53.0 kg N ha^–1 from the soil, but returned to soil through roots only 11.3, 10.8 and 5.7 kg N ha^–1, respectively. Hence, removal of faba bean, pea and barley straw resulted in a N-balance of about –18, –24, and –47 kg ha^–1 respectively. A soil nitrogen conserving effect was observed following the cultivation of faba bean and pea compared to barley which was of the order of 23 and 18 kg N ha^–1, respectively. Cultivation of legumes resulted in a significantly higher A_N value of the soil compared to barley. However, the A_N of the soil following fallow was significantly higher than following legumes, implying that the cultivation of the legumes had depleted the soil less than barley but had not added to the soil N compared to the fallow. The beneficial effect of legume cropping also was reflected in the N yield and dry matter production of the succeeding crops. Cultivation of legumes led to a greater exploitation of soil N by the succeeding crops. Hence, appreciable yield increases observed in the succeeding crops following legumes compared to cereal were due to a N-conserving effect, carry-over of N from the legume residue and to greater uptake of soil N by the succeeding crops when previously cropped to legumes.     

Soil profile gravel concentration and its effect on rainfed crop yields

Summary Soil characteristics in the crop root zone are critical to soil water and nutrient availability to rainfed crops and determine crop production in coarse textured soils. A four-year field study was conducted in the foot-hills of North Himalayas near Chandigarh (India) on a coarse textured soil (Gravelly udic ustocrepts) to evaluate the effect of varying soil profile gravel concentration on the yield of rainfed crops of Taramira (Eruca sativa Mill.) in winter followed by maize (Zea mays L.), sorghum (Sorghum vulgare Pers.), cowpea (Vigna unguiculata L.) and sesamum (Sesamum indicum L.) in summer. Taramira gave a mean grain yield of 683, 410 and 275 kg ha^–1 at gravel concentration (GC) of 18, 28 and 40 percent by volume in the surface one metre soil depth. The grain and forage yield of summer crops decreased with the increasing GC. The gross monetary returns decreased in the order: Sorghum fodder, cowpea, sesamum and maize. The dilution of soil mass with increasing GC and corresponding decrease in nutrient and water holding capacity of the soil appears to have depressed the crop yields. The results indicated that the legume which can also conserve rainwater with dense canopy like cowpea or crops having vigorous fibrous root system and are relatively drought tolerant like sorghum may provide better economic returns in light textured soil containing gravel upto 40 percent.     

Beyond landraces: developing improved germplasm resources for underutilized species – a case for Bambara groundnut

The potential for underutilized crops (also known as minor, neglected or orphan crops) to improve food and nutrition security has been gaining prominence within the research community in recent years. This is due to their significance for diversified agricultural systems which is a necessary component of future agriculture to address food and nutritional security concerns posed by changing climate and a growing world population. Developing workable value chain systems for underutilized crop species, coupled with comparative trait studies with major crops, potentially allows us to identify suitable agricultural modalities for such species. Bambara groundnut (Vigna subterranea L. Verdc.), an underutilized leguminous species, is of interest for its reported high levels of drought tolerance in particular, which contributes to environmental resilience in semi-arid environments. Here, we present a synopsis of suitable strategies for the genetic improvement of Bambara groundnut as a guide to other underutilized crop species. Underutilized crops have often been adapted over thousands of years in particular regions by farmers and largely still exist as landraces with little or no genetic knowledge of key phenotypic traits. Breeding in these species is fundamentally different to breeding in major crops, where significant pedigree structures and history allow highly directed improvement. In this regard, deploying new integrated germplasm development approaches for variety development and genetic analysis, such as multi-parent advance generation inter-crosses (MAGIC), within breeding programmes of underutilized species will be important to be able to fully utilize such crops.     

Green manure technology: Potential,usage, and limitations. A case study for lowland rice

The growing concern about the sustainability of tropical agricultural systems stands in striking contrast to a world-wide decline in the use of soil-improving legumes. It is timely to assess the future role that soil-improving legumes may play in agricultural systems. This paper reviews recent progress, potential, and limitations of green manure technology, using lowland rice cropping systems as the example.Only a few legume species are currently used as green manures in lowland rice. Sesbania cannabina is the most widely used pre-rice green manure for rice in the humid tropics of Africa and Asia. Astragalus sinicus is the prototype post-rice green manure species for the cool tropics. Stem-nodulating S. rostrata has been most prominent in recent research. Many green manure legumes show a high N accumulation (80–100 kg N ha^-1 in 45–60 days of growth) of which the major portion (about 80%) is derived from biological N₂ fixation. The average amounts of N accumulated by green manures can entirely substitute for mineral fertilizer N at current average application rates. With similar N use efficiencies, green manure N is less prone to loss mechanisms than mineral N fertilizers and may therefore contribute to long-term residual effects on soil productivity.Despite a high N₂-fixing potential and positive effects on soil physical and chemical parameters, the use of green manure legumes for lowland rice production has declined dramatically world-wide over the last 30 years. Land scarcity due to increasing demographic pressure and a relatively low price of urea N are probably the main determining factors for the long-term reduction in pre-rice green manure use. Post-rice green manures were largely substituted for by high-yielding early-maturing grain legumes. Unreliability of green manure performance, non-availability of seeds, and labor intensive operations are the major agronomic constraints. The recognition and extrapolation of niches where green manures have a comparative advantage may improve an often unfavorable economic comparison of green manure with cash crop or fertilizer N. Socio-economic factors like the cost of land, labor, and mineral N fertilizer are seen to determine the cost-effectiveness and thereby farmers' adoption of sustainable pre-rice green manure technology. Hydrology and soil texture determine the agronomic competitiveness of a green manure with N fertilizers and with alternative cash crops. In general, the niches for pre-rice green manure are characterized by a relatively short time span available for green manure growth and a soil moisture regime that is unfavorable for cash crops (flood-prone rainfed lowlands with coarse-textured soils).Given the numerous agronomic and socio-economic constraints, green manure use is not seen to become a relevant feature of favourable rice-growing environments in the foreseeable future. However, in environments where soil properties and hydrology are marginal for food crop production, but which farmers may be compelled to cultivate in order to meet their subsistence food requirements, green manures may have a realistic and applicable potential.     

The effectiveness and competitiveness of some Indonesian Rhizobium strains on tropical legumes grown in four soil types of Java

Several Indonesian and some imported Rhizobium strains were assessed for their effectiveness in nodulating four legume species in four soil types of Java. Naturally occurring Rhizobia formed effective symbioses onVigna unguiculata, Macroptilium atropurpureum andDesmodium heterocarpon in all four soils and the applied strains, with some exceptions, did not infect a majority of nodules of these legumes.Centrosema pubescens was more specific in its Rhizobia requirements and applied strains formed effective symbioses in two clay soils, but not in two sandy loam soils.     

Acacia,climate, and geochemistry in Australia

Background

Nitrogen-fixing legumes are key species in grassland ecosystems, as their ability to fix atmospheric nitrogen can facilitate neighboring plants. However, little is known about the fate of this legume effect in the face of extreme weather events, which are increasingly expected to occur.

Methods

Here, we examined experimentally how the presence of a legume modifies above-ground net primary production (ANPP) and nitrogen supply of neighboring non-legumes under annually recurrent pulsed drought and heavy rainfall events by comparing responses of three key species in European grassland versus without legume presence over 4 years.

Results

Legume presence facilitated community productivity of neighboring non-legumes under ambient weather conditions and also under experimental heavy rainfall. However, no facilitation of community productivity by the legume was found under experimental drought. Productivity of the three target species responded species-specifically to legume presence under different weather conditions: Holcus lanatus was facilitated only under control conditions, Plantago lanceolata was facilitated only under heavy rainfall, and Arrhenatherum elatius was facilitated irrespective of climate manipulations. The legume effects on δ ¹⁵N, leaf N concentration, and N uptake were also species-specific, yet irrespective of the climate manipulations. The data suggest that the missing legume effect on community productivity under the pulsed drought was rather caused by reduced N-uptake of the target species than by reduced N-fixation by the legume.

Conclusions

In contrast to heavy rain, the presence of a legume could not effectively buffer community ANPP against the negative effects of extreme drought events in an experimental temperate grassland. Facilitation also depends on the key species that are dominating a grassland community.     

Bruchid pest management in pulses: past practices,present status and use of modern breeding tools for development of resistant varieties

Bruchids (Callosobruchus spp.) are recognised as the most detrimental storage pest of pulses, especially in the tropics and subtropics. They invade matured pods as well as seeds during storage and, to some extent, farming fields, in turn reducing the net yield of the crops. Several approaches including cultural, biological, physical and chemical control measures have been implemented with the aim of managing these pests, but none of these have been successful across time and space. Recently, transgenic‐ and marker‐assisted breeding approaches have appeared as promising tools for the successful management of these pests. Although some efforts have been made on the development of bruchid‐resistant transgenic crops, the cultivars developed are yet to be commercialised worldwide because of various limitations. In contrast, marker‐assisted breeding involving the identification of DNA‐based markers linked to host resistance against bruchids, have shown some success in the quest for the development of bruchid‐resistant cultivar(s). DNA markers linked to bruchid resistance have been identified in various grain legumes, particularly in the genus Vigna, and include mung bean (Vigna radiata), azuki bean (Vigna angularis), rice bean (Vigna umbellata), cowpea (Vigna unguiculata) and black gram (Vigna mungo). After their validation in different genetic backgrounds, these markers could be utilised for marker‐assisted selection and breeding ventures to protect pulse crops. The present study discusses the pros and cons of different approaches for the successful management of the bruchid pests in pulses. The review also highlights about the integrative approach aided with molecular interventions to improve productivity by avoiding losses incurred due to bruchids, and to attain sustainable yields for major pulse crops.     

Monovalent cation transporters; establishing a link between bioinformatics and physiology

Toxic aluminum (Al) ion is a major constraint to plant growth in acid soils. Aluminum tolerance in wheat (Triticum aestivum L.) is strongly related to the Al-triggered efflux of malate from root apices. A role of the secreted malate has been postulated to be in chelating Al and thus excluding it from root apices (malate hypothesis), but the actual process has yet to be fully elucidated. We measured Al content and root growth during and after Al exposure using seedlings of near-isogenic lines [ET8 (Al tolerant) and ES8 (Al sensitive)] differing in the capacity to induce Al-triggered malate efflux. Aluminum doses that caused 50% root growth inhibition during 24-h exposure to Al in calcium (Ca) solution (0.5 mM CaCl₂, pH 4.5) were 50 μM in ET8 and 5 μM in ES8. Under such conditions, the amount of Al accumulated in root apices was approximately 2-fold higher in ET8 than ES8. Al-treated seedlings were then transferred to the Al-free Ca solution for 24 h. Compared to control roots (no Al pretreatment), root regrowth of Al-treated roots was about 100% in ET8 and about 25% in ES8. The impaired regrowth in ES8 was observed even after 24-h exposure to 2.5 μM Al which had caused only 20% root growth inhibition. The addition of malate (100 μM) during exposure to 50 μM Al in ES8 enhanced root growth 1.6 times and regrowth in Al-free solution 7 times, resulting in similar root growth and regrowth as in ET8. Short-term Al treatments of ES8 for up to 5 h indicated that the Al-caused inhibition of root regrowth started after 1-h exposure to Al. The stimulating effect of malate on root regrowth was observed when malate was present during Al exposure, but not when roots previously exposed to Al were rinsed with malate, although Al accumulation in root apices was similar under these malate treatments. We conclude that the malate secreted from root apices under Al exposure is essential for the apices to commence regrowth in Al-free medium, the trait that is not related to the exclusion of Al from the apices.     

Nutrient composition and in vitro ruminal fermentation of tropical legume mixtures with contrasting tannin contents

Various combinations of a low-tannin herbaceous legume (Vigna unguiculata) and foliage of tanniniferous shrub legumes (Calliandra calothyrsus, Flemingia macrophylla and Leucaena leucocephala) or a low-tannin shrub legume (Cratylia argentea), all mixed together with a low-quality tropical grass (Brachiaria humidicola), were tested in vitro for differences in the effects on ruminal fermentation. Two experiments with the gas transducer technique were carried out, where each forage mixture was tested either with or without polyethylene glycol in order to be able to identify tannin-related effects (n = 3). In Experiment 1, a stepwise replacement of V. unguiculata by C. calothyrsus (5:0, 4:1, 3:2, 2:3, 1:4, 0:5) at a legume proportion of 1/3 or 2/3 in the mixture was evaluated. Together with two grass-alone and four pure legume treatments this added up to 30 treatments. In Experiment 2, V. unguiculata was gradually replaced by each of the four shrub legumes (3:0, 2:1, 1:2, 0:3) in grass–legume ratios of 2:1, adding up, together with two grass-alone treatments, to 28 treatments. When added alone, V. unguiculata resulted in high fermentative activity as measured by gas production and kinetics as well as low proportion of undegraded crude protein. When V. unguiculata was replaced by the low-tannin C. argentea in Experiment 2, there was no noticeable difference (P>0.05) in fermentative activity. In both experiments, the effect of the substitution of V. unguiculata by tanniniferous shrub legumes resulted in a declining gas production and an increasing proportion of undegraded crude protein (P<0.001). However, the extent of these changes depended on the level of replacement and the shrub legume species (P<0.001). The results of Experiment 2 illustrate that this was the consequence not only of different tannin contents (less adverse effects with L. leucocephala than with C. calothyrsus) but also differences in the chemical properties of the tannins present in these shrub legume species (much less adverse effects with L. leucocephala than with F. macrophylla despite similar tannin contents). Furthermore these results indicate that, once the extent of the effects of a tanniniferous legume is known, one may calculate the maximal level of replacement of a low-tannin legume in a grass diet possible without negative effects on ruminal fermentation. This allows to improve dry season grass-based diets with as few as possible of the expensive and less well growing low-tannin legume.     

Genome re-sequencing of two accessions and fine mapping the locus of <Emphasis Type="Italic">lobed leaflet margins</Emphasis> in mungbean

Mungbean (Vigna radiata L.) is a fast-growing warm-season legume crop widely distributed in Asia. A large amount of self-shading of mungbean plants could reduce its seed yield. Thus, modification of the leaflet type could affect the leaf canopy and improve yield. In this study, a classical locus, lobed leaflet margins (lma) in mungbean, was investigated, which was controlled by a semi-dominant allele. In order to map the lma, the whole genomes of AL127 carrying lma and another accession Sulu with oval leaflets were re-sequenced; 236,998 single nucleotide polymorphisms and 8896 insertion/deletions (InDels) were identified between the two accessions. Using the validated InDels and SNP markers, lma was mapped to a syntenic region about 376 kb on chromosome 3 in mungbean and on chromosome 1 in common bean, respectively. Our results provided a framework for map-based gene cloning in mungbean. Cloning the lma should shed light on the underlying molecular mechanism controlling leaf shape in legumes and further provide the molecular basis for genetic improvement on legume crops.