Effect of lucerne saponins and Lotus condensed tannins on survival of grass grub,Costelytra zealandica

Abstract

Crude extracts of lucerne (Medicago sativa) and Lotus pedunculatus root were toxic when administered orally to third-instar Costelytra zealandica larvae. Purified saponins from lucerne root were alone found sufficiently toxic to account fully for the activity of the crude extract. Condensed tannins extracted from L. pedunculatus and L. corniculatus had no effect on larval survival when administered at levels equivalent to those present in crude extracts.     

Olfactory response of Costelytra zealandica (Coleoptera: Melolonthinae) to the roots of several pasture plants

The influence of natural plant odours on the locomotory behaviour of 3rd‐instar larvae of Costelytra zealandica (White) was studied by observing their movement in glass‐sided test chambers. Through an analysis of the paths followed by individual larvae in single‐option choice‐chamber tests it was possible to evaluate the responses of the larvae to different plant materials. The odour of fresh young perennial ryegrass (Loliumperenne) root was more attractive to the insects than was that of older plants. Larvae were also more strongly attracted to the root of lucerne (Medicago sativa), Lotus pedunculatus, red clover (Trifolium pratense), and white clover (T. repens) than to that of perennial ryegrass. The possible role of volatile chemical factors in plant resistance to grass grub attack is discussed.     

Ecology of black beetle Heteronychus arator. Influence of plant species on larval consumption, utilization and growth

The consumotion, utilization and growth of third-instar larvae of black beetle, Heteronychus arator (F.) feeding on roots of ryegrass (Lolium perenne), paspalum (Paspalum dilatatum) and white clover (Trifolium repens) were studied. Consumption was greater on the grasses than on white clover when assessed by gravimetric methods. Larvae had a higher growth rate on the grasses. Utilization of the roots of the three species was similar indicating that poor growth on white clover is the result of low consumption probably induced by the presence of feeding deterrents in the roots.This work is part of a study carried out by the senior author for the degree of D. Phil. at the University of Waikato.     

In vitro propagation ofMedicago andLotus species by node culture

Summary Nodes ofMedicago sativa, Lotus corniculatus, Lotus tenuis, andLotus pedunculatus were cultured on MS basal media with different growth regulators. InM. sativa each node produced one shoot and the apical dominance was unaffected by high levels of cytokinins, and subsequent cycles of culture. Shoot development was stimulated by the presence ofN ⁶-isopentenyl-adenine in the culture medium and was dependent on the genotype of the explant. Shoot development was not affected by the original position of the node on the plant nor by the plant age. Shoots rooted in MS medium gelled with starch and containing 2 mg·liter⁻¹ indol-3-acetic acid. In the threeLotus species, node culture was a more effective technique than inM. sativa. The number of shoots per node increased with the time of culture and with the presence of 0.05 mg·liter⁻¹ of 6-benzylaminopurine. The highest number of shoots derived from one node was achieved inL. pedunculatus and inL. tenuis by culturing single nodes, whereas inL. corniculatus stem segments had to be totally covered by the medium for success. Rooting was easily achieved in MS medium with or without auxins.     

Sitona weevils (Coleoptera: Curculionidae) as agents for rapid transfer of nitrogen from white clover (Trifolium repens L.) to perennial ryegrass (Lolium perenne L.)

The effects of root feeding by larvae of Sitona hispidulus (F.) (a common weevil pest of white clover) on the rate of transfer of nitrogen between plants of white clover (Trifolium repens L.) and perennial ryegrass (Lolium perenne L.) were investigated using a nutrient slant board technique. Clover plants, labelled with ¹⁵N were grown adjacent to ryegrass plants and were either infested with Sitona larvae or not infested. Ryegrass plants associated with the infested clover plants had a significantly higher dry matter yield and nitrogen content (75% and 74% respectively) than the uninvested plants, after 33 days exposure to insect herbivory. It was concluded that root feeding insects could play an important role in the cycling of nitrogen in grass/clover swards.     

Ecology of black beetle,Heteronychus arator (Coleoptera: Scarabaeidae) — relative consumption of pasture plant roots by larvae

Abstract

The relative consumption by third-instar black beetle larvae of roots of paspalurn, ryegrass, and white clover was determined from the rate of passage of ingested glass beads through the gut Consumption of grasses exceeded that of white clover, indicating their greater acceptance as a food source.     

Effect of oxygen availability on nitrogen fixation by two Lotus species under flooded conditions

The pasture legumes Lotus uliginosus (Schk.) and Lotus corniculatus (L.), known to differ in their tolerance to flooding, were inoculated with Rhizobium loti and flooded for 60 d while subjected to two levels of dissolved pO2: 0.241 and 0.094 mol ml^-1. L. uliginosus showed significantly greater growth (shoot and root) and N2 fixation under both pO2s, compared to L. corniculatus, although growth and N2 fixation by L. corniculatus was not affected by the low pO2. Surprisingly, in L. uliginosus., growth, nodulation and N2 fixation were all increased by low pO2 while nodulation of L. corniculatus where low pO2 plants showed a significant increase over that of the higher pO2 plants while L. uliginosus plants showed a decline. Root porosity of L. uliginosus doubled in the low pO2-treatment from a mean of 14.5% in high pO2 roots to 28.5%, whereas that of L. corniculatus was relatively unaffected by pO2, being 7% and 9% for high and low pO2 plants, respectively. The structure of nodules differed little between species and treatments, although nodules/nodulated roots from the L. uliginosus plants had particularly profuse lenticels and aerenchyma. However, L. corniculatus nodules, especially those grown in the lower pO2 showed signs of early senescence with vacuolation of infected cells and green coloration when cut open. Leghaemoglobin (Lb) concentrations in nodules from both species were unaffected by low pO2, although that of L. corniculatus nodules, regardless of pO2, was significantly greater than L. uliginosus. Concentrations of the intercellular glycoprotein recognized by the monoclonal antibody MAC265 were significantly reduced in nodules from the low pO2 treatment in both species. Immunogold labelling showed that the MAC265 antigen was localized primarily within intercellular spaces within nodule cortices from both Lotus species. A marked decrease in deposition of the MAC265 antigen within the cortices of L. uliginosus nodules grown in the lower pO2, is discussed in terms of the relative abilities of the two Lotus spp. to maintain an O2 supply to the N2-fixing bacteroids within submerged nodules.Keywords: Lotus uliginosus, Lotus corniculatus, N2 fixation, flooding, oxygen.      

The comparative phosphorus requirements of some temperate perennial legumes

The phosphorus (P) responses of seven temperate perennial pasture legumes and two species of lupins were compared in a field trial over a range of nine P rates, from 0 to 800 kg ha^-1. The two lupins produced more than 5 t ha^-1 of dry matter in the absence of added P and showed no response to the fertiliser. In contrast, the pasture legumes initially failed to grow without added P and responded to applications of between 200 and 800 kg ha^-1. At the higher rates of P, dry matter production of the pasture legumes was equivalent to that of the lupins.In the first 2 years of the trial; the most productive pasture legume species at the higher rates of added P were also the most productive at the lower rates. Phosphorus requirements for 90% of maximum yield varied greatly between species, but were closely related to maximum yield. Thus species with low P requirements for maximum yield were not necessarily P-efficient species. In the third and subsequent years of the trial Lotus corniculatus performed better than the other pasture legumes at the lower rates of added P. In contrast to other studies Lotus pedunculatus showed no ability to outyield Trifolium repens at low rates of P. Critical P concentrations of the pasture species for the late spring-early summer period declined in the order Trifolium repens (0.34%) > Lotus pedunculatus (0.30%) > Triofolium pratense (0.28%) > Trifolium hybridum (0.27%) > Trifolium ambiguum (0.26%) > Lotus corniculatus (0.23%).Mineralisable nitrogen (N) levels were determined in soils under three species in the 7th year of the trial. At the lowest rates of added P, mineralisable N levels were much higher under Lupinus polyphyllus than under Trifolium repens or Lotus corniculatus. With increasing P rate, levels under the latter species increased, and at 100 kg P ha^-1 were equivalent to those under the lupin with no added P.     

Investigations of the host feeding preferences ofSitona weevils found commonly on white clover (Trifolium repens) in the UK

A series of laboratory and field studies were done to evaluate a range of leguminous plant species for their feeding potential by adult weevils of the genusSitona Germar. (Coleoptera: Curculionidae). Three species ofSitona, S. lineatus L.,S. flavescens Marsh. andS. hispidulus F. all of which are found commonly on white clover (Trifolium repens L.) in the UK were offered a range of 11 legume species,T. repens (white clover, cv. Olwen),T. pratense L. (red clover, cv. Marcom),T. fragiferum L. (strawberry clover, cv. Palestine),T. hybridum L. (hybrid clover, cv. Tetra),T. incarnatum L (crimson clover),T. dubium Sibth. (lesser yellow trefoil),Lotus corniculatus L. (birdsfoot-trefoil, cv. Leo),L. uliginosus Schkuhr. (large birdsfoot-trefoil),Melilotus alba Desr. (white melilot),Medicago sativa L. (lucerne, cv. Europe) andM. lupulina L. (black medick) in two laboratory experiments. The weevils were offered a choice of these legumes in one experiment whilst in the other they did not have a choice of food material. These legumes were also sown in the field and a number of measurements of damage, together with counts ofSitona spp., were made. In the laboratoryS. lineatus andS. hispidulus favoured some of the legumes to a greater or lesser extent than white clover.S. flavescens was more restricted in its feeding than the other two weevil species. In the field studyS. lineatus invaded the experimental area quickly and tended to favourMedicago spp. andMelilotus spp. Later in the yearS. flavescens dominated the sitona fauna on the experiment, with the exception of aggregations ofS. lineatus onM. sativa andM. alba. In a separate screen of 5 varieties of white clover (cvs Donna, Menna, Kersey, Olwen and Grasslands Huia), cv. Olwen appeared to be the most susceptible to sitona attack.     

Stability of CaMV 35S-gus gene expression in (bird's foot trefoil) hairy root cultures under different growth conditions

Lotus corniculatus is an agronomically important forage legume. Genetic engineering offers opportunities both to improveL. corniculatus as a crop and to increase basic understanding of plant biochemistry and metabolism. Biosynthesis of secondary products and nitrogen fixation are two areas in which gene expression has been studied using hairy root cultures ofL. corniculatus. The stability of foreign gene expression in these cultures is critically important. TwoL. corniculatus root culture lines containing a reporter gene (CaMV 35S-gus) were used to investigate the stability of expression of a foreign gene under a range of conditions likely to be encountered in experiments. The hairy root culture lines were grown under varying conditions of light, temperature, nutrient supply, and in the presence of the auxin 2,4-D, or the elicitor glutathione. Expression of thegus gene, detected by measuring GUS activity, was found to be relatively stable under all of the conditions investigated.     

Physiological and Anatomical Basis of Differential Tolerance to Soil Flooding of Lotus corniculatus L. and Lotus glaber Mill

Lotus corniculatus L. and Lotus glaber Mill. are warm-season legume species adapted to many kinds of environmental stress, including flooding conditions, whereas other popular forage legumes, like alfalfa or white clover, cannot thrive. This study evaluates the relationship between root aerenchyma, water relations and leaf gas exchange and the differential tolerance to soil flooding of L. corniculatus and L. glaber. Adult plants of these species, established independently in grasslands mesocosms, were subjected to 40 days of early spring flooding at a water depth of 6 cm. Both species presented constitutive aerenchyma tissue in the roots. Under flooding conditions, this parameter was 26.2% in L. glaber and 15.3% in L. corniculatus. In addition, flooded plants of L. glaber presented a leaf biomass 47.5% higher above water while L. corniculatus showed a leaf biomass 59.6% lower in the same layer, in comparison to control plants. Flooded plants of L. glaber maintained the stomatal conductance (g _s) and transpiration rate (E) for 25 days, although these parameters reduce slightly to 40–60% in comparison to controls after 40 days of flooding. In this species, a reduction in photosynthesis (A) in flooding conditions was detected only on the last day of measurement. In L. corniculatus, the same parameters (g _s, E and A) were affected by flooding since day 18 of treatment, and values reached 25–40% in comparison to control plants after 40 days of flooding. Flooding did not affect above-ground biomass in L. glaber; while in L. corniculatus, above-ground biomass was 35% lower than in control plants. Our results confirmed that L. glaber is more able to cope with flooding stress than L. corniculatus, even in the presence of natural competitors. On the whole, this experiment provides information that can aid in the identification of anatomical and physiological parameters associated with flood-tolerance in this forage legume species, with economic potential for the agricultural areas subject to periodic flooding.     

Performance of the legume-feeding herbivore, Colias philodice (Lepidoptera: Pieridae) is not Affected by Elevated CO2

Presumably due to their association with nitrogen-fixing bacteria, the nutritional quality of legumes decreases less than that of non-legume C₃ plants when grown under elevated atmospheric CO₂. Therefore, it seems likely that legume-feeding herbivores will be less adversely affected than herbivores of non-legume C₃ plants by anthropogenic increases in atmospheric CO₂. When the legumes Medicago sativa (alfalfa), Trifolium repens (white clover), and Lotus corniculatus (birdsfoot trefoil) were grown under elevated (756 ppm) CO₂, leaf nitrogen remained the same or increased, and C:N ratio did not change. Unlike most insects fed non-legume C₃ plants, Colias philodice (sulfur butterfly) larvae fed elevated-grown M. sativa and T. repens did not exhibit reduced relative growth rate (RGR), and larvae fed elevated-grown L. corniculatus exhibited a nearly significant 37% increase in RGR. Pupal weight was unaffected by growth of host plants under elevated CO₂. Relative nitrogen growth rate (RGRN) did not change for larvae fed elevated-grown M. sativa or T. repens, but increased by 34% for larvae fed elevated-grown L. corniculatus. These results suggest that legume-feeding herbivores will be relatively buffered against the adverse effects of elevated CO₂ typically experienced by herbivores of non-legume C₃ plants.     

Germination and Initial Root Growth of Four Legumes as Affected by Landfill Biogas Atmosphere

The most important problem in the restoration of closed landfills is the production of toxic gases by decomposition of refuse. Such gases affect the root system of plants growing on these sites. The aim of the present study was to assess the effects induced by landfill biogas contamination on germination and initial root growth of Vicia villosa (hairy vetch), Lotus corniculatus (birdsfoot trefoil), Trifolium pratense (red clover), and Trifolium repens (white clover). In laboratory conditions, simulated landfill and control gas were supplied to the seedlings. The composition of the simulated landfill gas used was: 16% O₂, 8% CO₂, 3% CH₄, and 73% N₂; a control gas was also tested (21% O₂, 0. 035% CO₂, and 78% N₂). Percentage of germinated seeds was determined after 6 and 12 days from the starting date; at the same time qualitative assays of metabolic root functionality were also performed by using an agar technique in order to visualize changes in rhizosphere pH. At the end of the experiment, the length of the primary and secondary root was measured. Germination after 6 days was affected by the gas treatment; the landfill biogas caused a delay in germination with respect to the control in seeds of V. villosa and L. corniculatus. Root fresh weight and dry weight were significantly decreased by biogas treatment in V. villosa and T. repens. In contrast, root dry weight was higher in gas treated L. corniculatus and T. pratense compared to control seedlings. Total root system was significantly higher in treated T. pratense. The qualitative assay suggests, with the exception of T. pratense, a metabolic adjustment of the treated seedlings. Key words: restoration, landfill biogas, legumes.     

Interaction between Sitona lepidus and red clover lines selected for formononetin content

Adult clover root weevil Sitona lepidus show a feeding preference for white clover Trifolium repens over red clover Trifolium pratense. The effects on S. lepidus of three red clover T. pratense lines, selected for high, medium, or low levels of the isoflavone formononetin in foliage, were compared in three experiments using white clover as a control. In a no‐choice slant board experiment, weevil larval weights were greater for larvae feeding on white clover roots than those feeding on roots of the red clovers. The effect of larval root herbivory on plant growth was similar for all four clovers. Following root herbivory, a large increase in root and shoot formononetin levels was observed in the high‐formononetin selection of red clover but little change in the low‐formononetin red clover. In a no‐choice experiment with sexually mature female adult weevils feeding on foliage of the four clovers, all the red clovers had increased weevil mortality. Female weevils eating the high‐formononetin red clover laid fewer eggs than weevils eating white clover. The red clover diet caused a large accumulation of abdominal fat and/or oil in the weevils, whereas weevils feeding on white clover did not accumulate fat/oil. When sexually immature adult weevils were given a choice of foliage from all four clovers, white clover was eaten preferentially, and the low‐formononetin red clover was preferred to the high‐formononetin red clover. The results suggest that formononetin and associated metabolites in red clover may act as chemical defences against adult S. lepidus and that distribution in forage legumes can be manipulated by plant breeding to improve root health.     

Growth and ion relations in response to combined salinity and waterlogging in the perennial forage legumes Lotus corniculatus and Lotus tenuis

Lotus tenuis (Wadst. & Kit.) is a perennial legume widely grown for pasture in the flood-prone and salt affected Pampa region of Argentina. The physiology of salt and waterlogging tolerance in L. tenuis (four cultivars) was evaluated, and compared with Lotus corniculatus (three cultivars); the most widely cultivated Lotus species. Overall, L. tenuis cultivars accumulated less Na⁺ and Cl⁻, and more K⁺ in shoots than L. corniculatus cultivars, when exposed to 200 mM NaCl for 28 days in aerated or in stagnant solutions. Root porosity was higher in L. tenuis cultivars due to greater aerenchyma formation. In a NaCl dose–response experiment (0–400 mM NaCl in aerated solution), L. tenuis (cv. Chaja) accumulated half as much Cl⁻ in its shoots than L. corniculatus (cv. San Gabriel) at all external NaCl concentrations, and about 30% less shoot Na⁺ in treatments above 250 mM NaCl. Ion distributions in shoots were determined for plants at 200 mM NaCl. L. tenuis (cv. Chaja) again accumulated about half as much Cl⁻ in old leaves, young leaves and stems, compared with concentrations in L. corniculatus (cv. San Gabriel). There were not, however, significant differences between the two species for Na⁺ concentrations in the various shoot tissues. The higher root porosity, and maintenance of lower shoot Cl⁻ and Na⁺ concentrations in L. tenuis, compared with L. corniculatus, contributes to the greater tolerance to combined salt and waterlogging stress in L. tenuis. Moreover, significant variation for tolerance to combined salinity and waterlogging stress was identified within both L. tenuis and L. corniculatus.     

The effect of source,concentration and time of application of nitrogen on the growth,nodulation and nitrogen fixation ofLotus pedunculatus andTrifolium repens

Summary Studies under growth cabinet conditions investigated the effect of source and concentration of nitrogen and timing of nitrogen application on the growth and nitrogen fixation byLotus pedunculatus cv. Maku andTrifolium repens cv. S184. KNO₃, NaNO₃ and NH₄NO₃ were added at transplanting at the following rates: 3.33, 7.78 and 13.33 mg N/plant. KNO₃ was added at 3.33 and 7.78 mg N/plant at 0, 6, 12, 18, 24 or 30 days after transplanting.Lotus shoot weight increased with all increasing nitrogen sources but clover only responded to KNO₃ and NaNO₃. The root weight of both species increased with increasing KNO₃ and NH₄NO₃. The percentage increase in lotus and clover shoot growth was greater than that of root growth when KNO₃ was added within a week of transplanting. Increases in growth by both species resulted from added nitrogen except with lotus when NaNO₃ was applied where increased nitrogen fixation also contributed to increased growth.Weight and number of effective nodules on both species were increased with 3.33 mg N per plant as KNO₃ but nitrogen fixation was not affected. Addition of 13.33 mg N as NaNO₃ reduced weight and number of effective nodules in both species and also nitrogen fixation by lotus.KNO₃ increased growth and nodulation of both species when applied within one week after transplanting. Nodulated lotus plants responded to KNO₃ by increasing growth but not nodulation.KNO₃ appeared to affect infection and development of nodules on lotus and may affect the growth of existing nodules on clover.     

Yield,root morphology and chemical composition of two pasture legumes as affected by lime and phosphorus applications to an acid soil

Summary Effects of increasing rates of lime (0, 900, 1725, and 3000 kg Ca(OH)₂/ha producing soil pH of 4.0, 4.7, 5.1 and 5.6) and P (50, 150, 250 and 350 kg P/ha) on top and root yield, root morphology and chemical composition of lotus (Lotus pedunculatus Cav.) and white clover (Trifolium repens L.), were studied, using an acid soil in a greenhouse experiment. Increasing rates of applied lime and phosphate resulted in substantial increases in top yields of both species but concomitant increases in root yield were small. In the unlimed soil, lotus out-yielded (tops and roots) white clover at all P levels. However, in the three limed treatments, white clover clearly out-yielded lotus. Yield response curves to applied P levelled off at the two highest lime rates for lotus but not for white clover. Nodulation and N content of white clover increased significantly with increasing lime applications, but for lotus there was a significant decrease in nodulation at the highest lime rate. Increased P rates had a small stimulatory effect on nodulation in both species. Of the total root weight, the percentage contribution of the tap and primary lateral root fractions was smaller and that of the secondary plus tertiary lateral roots was greater for lotus than for white clover although root length per unit weight tended to be larger for white clover at the two highest lime rates. Furthermore, lotus possessed longer and more numerous root hairs than white clover. Lime applications significantly decreased the percentage contribution of the tap and primary lateral roots to the total root weight and increased the percentage contribution of the secondary plus tertiary lateral roots. Al and Mn contents of tops and roots of both species decreased with increasing lime rates. There was a highly significant negative correlation between relative yield and Al content of lotus and white clover tops. In comparison with the limed treatments, in the unlimed treatments a greater percentage of total P, Al, Mn and N content accumulated in the roots of both species. In addition, lotus accumulated a much greater percentage Al in its roots than white clover.     

Nitrate reductase and nitrite reductase activity in free-living cells and bacteroids of Rhizobium loti

Cells of Rhizobium loti strains T1 and U226 cultured in defined medium with glutamate as the only nitrogen source and bacteroids isolated from root nodules of Lotus corniculatus, L. pedunculatus and L. tenuis did not show constitutive (non-nitrate induced) nitrate reductase activity (NRA). In contrast, nitrite reductase activity (NiRA) was present in both free-living cells and bacteroids of either strain T1 or U226. Constitutive NRA and NiRA were detected in the cytosol fraction from nodules of all three symbioses examined. An induced NRA was expressed in bacteroids after a 10 h incubation in the presence of nitrate.     

The origin of Lotus corniculatus

Summary Earlier students of the origin of Lotus corniculatus suggested that this tetraploid species arose as an autotetraploid of the closely related diploid species L. tenuis or L. alpinus. More recent studies suggested that L. alpinus and L. japonicus could be ancestral forms. The present study of tannin content, phenolic content, cyanide production, morphology, cytogenetics, Rhizobium specificity and self-incompatibility in the corniculatus group virtually excludes the possibility that L. corniculatus could have arisen through autopolyploidy of L. tenuis or L. alpinus, and suggests that L. corniculatus arose through hybridization of L. alpinus and/or L. tenuis (probably as female parent) with L. uliginosus (probably as male parent), followed by chromosome doubling in the hybrid.