Growth of Lucerne (Medicago sativa L.) Exposed to Sulphur Dioxide

The effect of exposure to sulphur dioxide (0 or 96 µgm^–3 SO₂) on the growth and sulphur content of lucerne(Medicago sativa L.) was examined in a period of 135 d duringwhich the plants were harvested four times. The lucerne wasgrown in pots of soil with and without the addition of sulphateand of nitrate. Evidence of sulphur deficiency, including areduction in the weight, the number and the sulphur contentof shoots, was found in plants grown without added sulphate.Deficiency was alleviated through exposure of plants to SO₂.Apart from reducing shoot weight at one harvest and generallyincreasimg the concentration of sulphur in the shoots, exposureto SO₂ had no significant effect on plants grown with addedsulphate. The yield of shoots was greater, and was reduced toa lesser extent with sulphur deficiency, in plants grown withadded nitrate than in those dependent on rhizobia. Whilst thetranspiration coefficient increased greatly in sulphur-deficientplants, it was reduced where otherwise similarly treated plantswere exposed to SO₂; this treatment did not alter the coefficientin plants with an adequate supply of sulphur from the soil.     

Effect of Sodium Chloride Stress and Nitrogen Source on Respiration, Growth and Photosynthesis in Lucerne (Medicago sativa L.)

The effects of salinity at different light intensities on freshweight growth and on carbon dioxide influx and efflux were examinedin young plants of lucerne (Medicago sativa L.) that had beengrown in solution culture with nitrogen supplied either as nitrateor by a symbiotic rhizobium. Although the inoculated plantsgrew more slowly than those supplied with nitrate, NaCl at alevel equivalent to an osmotic stress of –0.3 MPa didnot reduce the growth rate of either type of plant under a 12h day-length in a growth chamber. With a day-length of 5 h saltstress (0.0 to –0.6 MPa) did not greatly affect grossphotosynthesis of plants grown on nitrate but respiration ratereached a maximum at –0.3 MPa and declined at larger saltconcentrations. Salt diminished both gross and net photosynthesisin the inoculated plants at a 5 h day-length without stimulatingrespiration. The relationship between photosynthesis and respiration as thephoton flux density was successively decreased was used to inferthe effect of salt on maintenance respiration of the plantssupplied with nitrate. Growth and maintenance components ofrespiration could not clearly be separated in the inoculatedplants suggesting that these were unable to maintain themselvesunder the combined stresses of salt and low light intensity.This view was supported by chemical analysis of the plant material.We conclude that the failure of the inoculated plants to adaptto these conditions could be attributed to the greater demandfor assimilates by the rhizobium. Key words: Medicago sativa, Nitrogen source, Salt stress     

Microclimate, Photosynthesis and Growth of Lucerne (Medicago sativa L.). II. Canopy Structure and Growth

The growth of lucerne var. Europe was examined in the fieldduring 1976. The annual dry matter production of unirrigatedlucerne during 1976, with no nitrogen fertilizer application,was 82.5 per cent greater than unirrigated S.24 perennial ryegrass,with a nitrogen application of 384 kg ha^–1. The mean aboveground growth rate of lucerne was 7.3 g DM m^–2 day^–1between March and early June, of which stem material contributeda maximum of 76.5 per cent. Significant losses of leaves andstems occurred from the end of April, indicating a loss of potentialforage material. Nitrogen analyses of the above ground crop suggested that in56 days lucerne yielded 10.7 per cent more nitrogen than didS.24 annually with a nitrogen fertilizer addition of 280 kgha^–1. Between 13 and 57 per cent of the daily photosynthate is translocatedbelow ground. Medicago sativaL, lucerne, dry matter production, canopy structure, nitrogen analyses     

The Use of Single Somatic Embryo Culture in Propagating and Regenerating Lucerne (Medicago sativa L.)

Embryogenic cultures of lucerne (Medicago sativa L.) cv. Robothave been established and propagated on medium containing yeastextract. These cultures consisted of unorganized callus tissuebearing embryogenic centres which increased in size during subculture,yielding new regenerated somatic embryos at the end of each20-d subculture. A development in the propagation of the embryogenic cultureswas the establishment of single embryo culture in hormone-freemedium where, in selected cases, the process of recurrent somaticembryogenesis (RSE) took place on the hypocotyl of explantedembryos. The process was independent of supporting callus tissueand occurred on simple defined medium. Single embryos underwenteither plantlet development or continued RSE on the hypocotyl.One third of the regenerated plantlets showed RSE after thetwo to three trifoliate leaf stage. In these cases shoot developmentstopped and only somatic embryo production took place. In vitrocloning of regenerated plantlets allowed us to reproduce eachparticular genotype before transplantation into soil. Lucerne (alfalfa), Medicago sativa L., somatic embryogenesis, single embryo culture     

The Role of Different Daily Irradiations on Shoot Growth and Root/Shoot Ratio in Lucerne (Medicago sativa L.)

The effect of two levels of daily irradiation on shoot growthand root/shoot partitioning was investigated on two lucernevarieties (Medicago sativa L.). Individual plants were studiedunder constant temperature and optimal water and mineral nutritionconditions. For both lucerne varieties, daily irradiation did not changebiomass partitioning between shoots and roots. It can be shownthat leaf area expansion occurs independently of daily irradiationand that the process of shoot dry matter production is strictlyproportional to daily irradiation, since the ratio leaf area/shootdry matter is inversely proportional to it. From a model of shoot production of the isolated plant, we showthat the relative shoot specific activity is not affected bydaily irradiation. A simple model of partitioning leads us todetermine why partitioning remains the same for the two at thelevels of daily irradiation. Finally, the allocation between shoot and root turns out tobe independent of the main stem extension rate, which is fasterat the higher daily irradiation. Medicago sativa L., lucerne, daily irradiation, shoot production, root/shoot ratio, leaf area expansion, partitioning model     

Microclimate, Photosynthesis and Growth of Lucerne (Medicago sativa L.). I. Microclimate and Photosynthesis

Measurements of microclimate and photosynthesis of lucerne var.Europe were made in the field during the spring of 1976. Themaximum rate of canopy gross photosynthesis (14.3 g CO₂ m^–2h^–1, I = ) was 2.5 times greater than that of S 24 perennialryegrass at the same LAI. This difference was due to differencesin individual leaf photosynthesis. The photosynthetic rate ofthe youngest fully expanded leaf of lucerne remained constantthroughout the experimental period at 3.6 g CO₂ m^–2 h^–1(300 W m^–2). Measurements of soil water potential profiles indicated thatlucerne extracted water from the soil to a depth of at least800 mm, with a region of maximum uptake between 400 and 600mm. This capability, with a moderate mean leaf resistance of460 s m^–1, conferred a high assimilation efficiency onlucerne, with a mean water use efficiency of 34 g H₂O lost pergram of carbohydrate assimilated, compared with 200 g H₂O pergram of carbohydrate for S 24. Medicago sativa L, lucerne, photosynthesis, assimilation efficiency     

Liming of alfalfa (Medicago sativa L.)

Summary A growth-chamber experiment was conducted to study the effect of liming upon growth of alfalfa. The beneficial effects observed were related to changes in soil properties brought about by lime application. Reductions of aluminum and manganese toxicities were the major factors responsible for the increased yields and the decreased growth period required to reach harvest stage. Significant correlations between plant growth parameters and various measures of extractable aluminum were found.     

Photosynthesis, Water Relations, Temperature and Canopy Structure as Factors Influencing the Growth of Sainfoin (Onobrychis viciifolia Scop.) and Lucerne (Medicago sativa L.)

Measurements of the growth of sainfoin and lucerne were madein the field after cutting on 31 May 1977. Sainfoin reacheda total above-ground dry weight of 408 g m^–2 over thegrowing period of 48 days compared with 598 g m^–2 in lucerne.Final leaf area indices (LAIs) were 2.8 in sainfoin and 6.1in lucerne. The specific leaf areas (SLAs) for sainfoin wereapproximately half those of lucerne throughout the regrowthperiod. The maximum rates of leaf appearance were 0.12 leavesper day in sainfoin and 0.85 leaves per day in lucerne. Themaximum mean rate of plant extension growth for lucerne of 2.12mm h^–1 occurred during the night, whereas, in sainfointhe maximum rate of 1.72 mm h^–1 occurred during the day. Measurements of extinction coefficients for PAR ranged from0.45 to 0.89 in sainfoin and from 0 42 to 0.57 in lucerne. Asthe lucerne crop increased in size leaf water potentials andsolute potentials became more negative. In sainfoin leaf waterpotentials remained remarkably high throughout the growth period,solute potentials decreased and turgor potentials increased.The stomatal conductances of the two species were similar. The photosynthetic capacities and rates of dark respirationper unit leaf area in both species were similar. The rate ofcanopy ‘gross’ photosynthesis at 295 W m^–2was always greater in lucerne than in sainfoin. This was largelya matter of differences between the species in LAI, althoughat higher LAIs the more erect structure of lucerne leads toa better utilization of photosynthetically active radiation. Onobrychis vicifolia Scop, sainfoin, Medicago sativa L., lucerne, photosynthesis, water relations, temperature, canopy structure     

Induced androgenesis in alfalfa (Medicago sativa L.)

Summary Anthers of 10 alfalfa (Medicago sativa L.) lines were used as initial material for the production of androgenic haploids. More than 30 variants of nutrient media were tested. Twenty five different treatments with low temperatures and gamma rays were tried in order to find optimal conditions for callus induction and organogenesis.The genotype, stage of microspore development, phytohormonal composition of the nutrient media and pretreatment with physical agents, alone or in combination, affected the efficiency of organogenesis and regeneration in anther cultures of alfalfa.Plants exhibited a high degree of variability in their chromosome number. Haploids, dihaploids and mixoploids were obtained.Cytological studies of in vitro pollen development revealed the origin of the regenerants from microspores.Abbreviations BAP 6-Benzylaminopurine - 2-ip 6-(,-dimethylallylamino)Purine - IAA Indolylacetic Acid - NAA Naphthaleneacetic Acid - 2,4-D Dichlorophenoxyacetic Acid - CMS Cytoplasmic Male Sterility     

Changes in Carbohydrate Levels in Red Kidney Bean (Phaseolus vulgaris L.) Exposed to Sulphur Dioxide

Red kidney bean (Phaseolus vulgaris L.) was exposed to variousconcentrations of SO² for 24 h (16 h light/8 h dark photoperiod)with continuous monitoring of photosynthetic and respiratoryactivity. Plants were harvested at the end of the dark periodinto samples of mature and immature leaf tissue, stems, androots for determination of sugar and starch levels. In all tissue samples the levels of total sugars were increasedby exposure to the lower concentrations of SO², but decreasedby the higher concentrations. Starch levels in leaves followeda similar trend. Increases in sugar and starch levels precededsymptoms of visible injury. Decreasing rates of photosynthesiswere correlated with increasing rates of respiration, the occurrenceof visible injury, and the depletion of sugar and starch levels.     

Fine root demography in alfalfa (Medicago sativa L.)

In perennial forages like alfalfa (Medicago sativa L.), repeated herbage removal may alter root production and mortality which, in turn, could affect deposition of fixed N in soil. Our objective was to determine the extent and patterns of fine-diameter root production and loss during the year of alfalfa stand establishment. The experiment was conducted on a loamy sand soil (Udorthentic Haploboroll) in Minnesota, USA, using horizontally installed minirhizotrons placed directly under the seeded rows at 10, 20, and 40 cm depths in four replicate blocks. We seeded four alfalfa germplasms that differed in N₂ fixation capacity and root system architecture: Agate alfalfa, a winter hardy commercially-available cultivar; Ineffective Agate, which is a non-N₂-fixing near isoline of Agate; a new germplasm that has few fibrous roots and strong tap-rooted traits; and a new germplasm that has many fibrous roots and a strongly branched root system architecture. Video images collected biweekly throughout the initial growing season were processed using C-MAP-ROOTS software.More than one-half of all fine roots in the upper 20 cm were produced during the first 7 weeks of growth. Root production was similar among germplasms, except that the highly fibrous, branch-rooted germplasm produced 29% more fine roots at 20 cm than other germplasms. In all germplasms, about 7% of the fine roots at each depth developed into secondarily thickened roots. By the end of the first growing season, greatest fine root mortality had occurred in the uppermost depth (48%), and least occurred at 40 cm (36%). Survival of contemporaneous root cohorts was not related to soil depth in a simple fashion, although all survivorship curves could be described using only five rates of exponential decline. There was a significant reduction in fine root mortality before the first herbage harvest, followed by a pronounced loss (average 22%) of fine roots at the 10- and 20-cm depths in the 2-week period following herbage removal. Median life spans of these early-season cohorts ranged from 58 to 131 days, based on fitted exponential equations. At all depths, fine roots produced in the 4 weeks before harvest (early- to mid-August) tended to have shorter median life spans than early-season cohorts. Similar patterns of fine root mortality did not occur at the second harvest. Germplasms differed in the pattern, but not the ultimate extent, of fine root mortality. Fine root turnover during the first year of alfalfa establishment in this experiment released an estimated 830 kg C ha^–1 and 60 kg N ha^–1, with no differences due to N₂ fixation capacity or root system architecture.     

Growth of Perennial Ryegrass (Lolium perenne L.) Exposed to a Low Concentration of Sulphur Dioxide

The influence of a low concentration of SO₂ on the growth andsulphur status of perennial ryegrass (Lolium perenne L.) cv.S23 was examined in a system of specially designed growth chambers.For plants grown without additions of sulphate to the soil,erposure to ifitered air containing 50 µg m^–3 SO₂alleviated sulphur deficiency that developed during a sequenceof successive harvests, and gave increased yield. The plantshad higher contents of organic and inorganic sulphur than thosegrown with neither sulphate nor SO₂. Plants grown with addedsulphate showed no signs of sulphur deficiency and althoughexposure to SO₂ increased their sulphur content it had no effecton yield. The coefficient of transpiration (g water transpiredg^–1 dry wt. of shoots) was reduced when plants which otherwisehad an inadequate supply of sulphur were exposed to SO₂; forplants with adequate supplies of sulphate, it was similar whetherthey were grown with or without the addition of SO₂ to the air.The results are discussed in relation to other known reactionsof plants to SO₂.     

Hairy root transformation in alfalfa (Medicago sativa L.)

Summary The widely cultivated forage legume alfalfa (Medicago sativa L.) was transformed with the agropine type Agrobacterium rhizogenes NCPPB 1855. Sterile root and callus cultures were derived from tumorous hairy roots which were easily obtained independent of the plant variety or genotype. Plant regeneration, via somatic embryogenesis, was achieved only when a selected alfalfa line, characterized by high regenerative capability, was utilized. Genetic transformation was confirmed by the presence of agropine and T-DNA. Phenotypic alterations, mainly affecting the root system, were observed in transformed plants. The possibility that T-DNA-induced variations could be useful in the improvement of M. sativa is discussed.Research work was partially supported by Progetto Strategico Agrobiotecnologia C.N.R., Italy     

Plantlet Regeneration via Multiple Shoot Induction in Indian Cultivars of Lucerne (Medicago sativa L)

An efficient protocol has been developed for in vitro plant regeneration via multiple shoot induction in lucerne (Medicago sativa L). Shoot tips from in vitro grown 5–6 days old seedlings of 3 cultivars, LLC-3, Chetak and RL-88 were used as explants for multiple shoot induction on MS medium supplemented with cytokinins. Maximum of 14 shoots per apical meristem were observed in case of cv Chetak on MS medium supplemented with BAP (12.6 μM) and KN (9.3 μM). Shoot elongation on MS medium supplemented with GA (5.8 μM), while root induction was achieved on MS medium supplemented with IAA (11.4 μM) and activated charcoal (2.0 g l^?1). Tissue raised plants showed 75% survival after transfer to soil under field conditions.     

Ferric Phosphate-dissolving Compound,Alfafuran, from Alfalfa (Medicago sativa L.) in Response to Iron-deficiency Stress

A ferric phosphate-dissolving compound was isolated from alfalfa (Medicago sativa L.) and characterized as 2-(3,5-dihydroxyphenyl)-5,6-dihydroxybenzofuran, which was named alfafuran.     

Microgametophytic selection in two alfalfa (Medicago sativa L.) clones

Summary Microgametophytic selection was investigated using two ecologically diverse autotetraploid clones of alfalfa. Several selection pressures (drying, aging, freezing, and high and low temperatures) were applied to microgametophytes at three stages of the life cycle, 1) during microsporogenesis, 2) post-anthesis, and 3) pollen tube growth. Pollen aging produced a progeny population with a greater mean plant size and a lower coefficient of variation than the control progeny. High temperature (29.5 °C) applied both during microsporogenesis and pollen tube growth resulted in progeny populations which were significantly taller and, in one case, had a larger leaf number than the control populations. In contrast, air dried pollen resulted in a progeny population which had significantly smaller character means and larger coefficients of variation than the control population. Also, low temperature (15 °C) during pollen tube growth yielded progeny with reduced branch number and a larger coefficient of variation than the control progeny. In cases where progeny derived from selected microgametophytes were found to differ from the control offspring, corresponding shifts in the reciprocal cross were not observed. For the temperature stress treatments, the lack of reciprocal differences may be related to the different temperature adaptations of the two ecotypes. These results suggest that microgametophytic selection can be effective in shifting the mean of the progeny generation; however, the results obtained will vary depending upon the selection pressure, stage of selection, and the parents used.     

Tolerance to manganese toxicity among cultivars of lucerne (Medicago sativa L.)

Two sand culture experiments were carried out to identify commercial cultivars of lucerne or alfalfa (Medicago sativa L.) which contain elite, Mn-tolerant plants for use in a selection programme to increase the acid-soil tolerance of this perennial legume. Differences in Mn tolerance, both within and between cultivars, were observed when a range of cultivars were exposed to regular waterings with dilute nutrient solution containing 20 or 25 mg Mn L^–1. Under these moderately toxic regimes, the winter dormant cultivars Cimmaron and WL 318 were found to contain elite plants that had greater dry matter yields than their mean cultivar yield under non-toxic Mn conditions.Cultivars which contained elite, Mn-tolerant plants could not be identified by phenotypic characteristics such as their height or their toxicity symptom score, nor by their winter dormancy class. Possible reasons for the occurrence of elite plants in these cultivars are discussed. The elite, high yielding Mn-tolerant plants could not be identified from the other plants within their cultivar population by their Mn toxicity symptoms nor by their height.     

Role of Potassium in Carbon Dioxide Assimilation in Medicago sativa L

Alfalfa was grown hydroponically in 0, 0.6, and 4.8 millimolar K in order to determine the influence of tissue level of K on photosynthesis, dark respiration, photorespiration, stomatal and mesophyll resistance to CO₂, photosystem I and II activity, and synthesis and activity of ribulose 1,5-bisphosphate carboxylase (RuBPc).     

Purification of beta-amylase from alfalfa (Medicago sativa L.) seeds

An amylase from alfalfa (Medicago sativa L. c.v. Moapa) seeds was purified by column chromatography and gel filtration, followed by chromatofocusing on Mono P HR 5/20. The last step was effective for separation of the alfalfa amylase to a homogeneous state. The purified amylase was identified as beta-amylase from the fact that only beta-maltose was formed by the enzymatic degradation of soluble starch. The molecular weight and specific activity of the beta-amylase (E1%(280 nm) = 18.3) were determined to be 61,000 and 1,077 A.U./mg, respectively. The beta-amylase activity was inhibited by the modification of sulfhydryl groups with p-chloromercuribenzoic acid. The optimum pH and isoelectric point of alfalfa beta-amylase were 7.0 and 4.8, respectively, which were different from other plant beta-amylases.