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Biological Nitrogen Fixation in Pachyrhizus Ahipa (Wedd.) Parodi

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Biological Nitrogen Fixation in Pachyrhizus Ahipa (Wedd.) Parodi Annals of Botany 70 11-17, 1992 Biological Nitrogen Fixation in Pachyrhizus ahipa (Wedd.) Parodi S0REN KJ/ER Botanical Section, Department of Botany, Dendrology and Forest Genetics, Royal Veterinary and Agricultural University, Rolighedsvej 23, DK-1958 Frederiksberg C (Copenhagen), Denmark Accepted 15 January 1992 Downloaded from https://academic.oup.com/aob/article/70/1/11/129022 by guest on 29 September 2021 The patterns of plant growth and N, fixation capability in Pachyrhizus ahipa (Wedd) Parodi inoculated with Bradyrhuobium ' Pachyrhizus Spec 1' strains (LiphaTech) were investigated in a zero-N culture system under greenhouse conditions The P ahipa plant is day-neutral with respect to reproductive development Competition occurred between the two storage organs (legume and tuber) and prevented high tuber yield in P ahipa The symbiotic effectiveness of the association was high, as the profuse nodulation provided the inoculated plants with adequate amounts of N Nodules were present throughout the cycle of P ahipa The change in rate of N, fixation (RNF) and relative growth rate (RGR) was almost paraJlel dunng ontogenesis The developmental pattern of N, fixation activity revealed that 65 % of total N, fixation occurred after N began to accumulate in the reproductive (pod wall plus seed) tissue Dunng pod filling allocation of N compounds to the seeds exceeded N, fixation, the pod walls being the pnmary source of redistnbuted N, followed by the leaves Key words: Pachyrhizus ahipa (Wedd ) Parodi, ahipa, tuber crop, dinitrogen fixation, dry matter and N partitioning, reproductive growth (1989) observed that in P erosus nodules were present INTRODUCTION through flowering and pod maturation Furthermore, they Leguminous root crops in Third World countries have observed that remobihzation of vegetative tissue N to the received renewed attention as potentially high-yielding root tuber occurred some time after pod maturation The capacity crops of good nutritional value (National Research Council, to sustain a high N, fixation capability dunng pod filling 1979) The tuber-bearing genus Pachyrhizus Rich ex DC may be the cause for these observations (Fabaceae, tnbus Phaseoleae) includes five species native to The objectives of the present study were to investigate the Central America and northern to central South America ontogenetic patterns of growth, N2 fixing capability, and (Serensen, 1988) Unlike the other members of the genus, P mineral N uptake by P ahipa Such information would ahipa is bushy, non-climbing, fast maturing and insensitive provide a basis for investigations on the interspecific hybrid to photopenod (National Research Council, 1989) P ahipa characters of Pachyrhizus spp regarding patterns of growth was known and cultivated by the Incas in pre-Columbian and increased reliance on biologically fixed N in agricultural time according to Yacovleff(1933) and Ugent, Pozorski and plant production Pozorski (1986) The cultivation of P ahipa is very limited today, restricted to Andean valleys in Bolivia and Peru MATERIALS AND METHODS between 1500 and 2500 m above sea level (Sorensen, 1990) A closely related species, Pachyrhizus erosus (L ) Urban is P ahipa plants were cultured in greenhouse from 7 Apr to native to Mexico and Central America and has many 21 Oct 1990 at the Royal Veterinary and Agricultural morphological characteristics in common with P ahipa University in Copenhagen Daily maximum and minimum Interspecific breeding between cultivars of P ahipa and P greenhouse air temperature were on average 28 °C and erosus was studied by Serensen (1990) for the stability of 18 °C, respectively The climate computer system was the various genetic characters and valuable agricultural hybrids DGT/Volmatic system LCC1220 The symbiotic association between the legume P erosus Seeds of a uniform size (mean d wt 370 ±20 mg), cultivar and indigenous strains of Bradyrhizobium was investigated AC 102 from Bolivia, were immersed in 70% ethanol with by Vansuyt and Zinsou (1989) The Bradyrhizobium strains 0-01% Tween 20, then surface sterilized by using 3% were infective in vertisols (pH 8 4) and neutral ferralhtic Korsohn (Ferrosan A/S) for 30 min, while the stenlant soils (pH 6 9), but not in acid ferralhtic soils (pH 4 6) The solution was agitated The seeds were nnsed three times in influence of favourable soil fertility on the nodule func- sterilized water and pregerminated in Petn dishes 3—4 d tioning with P erosus as host plant was studied by Lynd and at 25 °C in the dark and transferred to 145 mm diameter Purcino (1987) They found that adequate soil levels of pots Seedlings were thinned to one plantlet per pot 2 weeks phosphorus, calcium and potassium increased nodule mass after sowing and nitrogenase activity considerably Vansuyt and Zinsou Nutrient solutions were prepared with deminerahzed 0305-7364/92/070011 +07 S03 00/0 © 1992 Annals of Botany Company 12 Kjar—Nitrogen Fixation in Pachyrhizus ahipa Downloaded from https://academic.oup.com/aob/article/70/1/11/129022 by guest on 29 September 2021 FIG 1 Development of underground parts in Pachvrhizus ahipa (Wedd ) Parodi plants growing in N-free culture solution with Bradyrhizobium 'Pachyrhizus Spec 1' (LiphaTech) ( —N + I) A, Plantlet growing in N-free culture solution without Bradyrhizobium (treatment — N —I) 20 d after emergence (DAE) Bar = 11-0 mm B, Plantlet showing 'root curling' originating from the infection of roothairs (20 DAE) Bar = 11-Omm C, Apical part of the root showing the profuse nodulation (45 DAE) Bar = 5 5 mm D, Necrosed nodule together with young spherical nodules (85 DAE) Bar = 55 mm E, Indeterminate nodules with terete branches (148 DAE) Bar = 55 mm F, Representative tubers in plants (185 DAE), that may develop to 0-5 kg fresh weight Bar = 27 4 mm water A basal nutnent solution was used containing 1 0 mM solution with mineral N Control plants (designated -N-I) KHjPO4,3 8mMCaSO4 2H2O,0 9 mMMgSO4 7H,O, 1/tM without Bradyrhizobium and exogenous N were cultivated KC1, 54 fiM Fe-III-Na-EDTA, 46 ;tM H,BO3, 12 fm MnSO4, as a reference The pH was adjusted to 6 5 with 01 M K.OH 0 8 fiM ZnSO4, 0 3 /tM CuSO4 and 0 5 /tM Na2MoO4 2H,O On 7 Apr 1990, pregerminated seeds of the —N + I The test plants were grown in three different N regimes N- treatment were inoculated with a peat-based culture control plants received the basal nutnent solution adding containing two Bradyrhizobium strains (120B2 and 120B6) 2 5 mM Ca(NO3)2 and no inoculum (designated +N —I) isolated from P erosus in Thailand (Dr R Stewart Smith, Inoculated plants with Bradyrhizobium 'Pachyrhizus Spec LiphaTech, pers comm ) Each pot received approx I x I08 1' (LiphaTech) were fed with the N-free basal nutnent viable Bradyrhizobia solution (designated -N + I), 2 5 mM CaSO4 2H;O was The +N —I plants were cultured without mineral N till +I added to equalize the Ca concentration of the nutnent 19 d following seedling emergence On 6 May 1990, 19 d Kjcer—Nitrogen Fixation in Pachyrhizus ahipa 13 after seedling emergence (DAE) nodules were observed on to the root was narrow and non-girdling, occasionally the — N + I plants, and the nutrient solution containing clusters of nodules developed Nodules were present and 2 5 CaCNOj);, was added to the +N-I plants active (red colouring by leghaemoglobin) through flowering The artificial growth medium vermiculite (grade 4 and pod setting Senescence of the first developed nodules 3-8 mm) was immersed in nutrient solutions for 48 h The was observed during pod development 85 DAE (Fig 1 D) pots were placed on 05 1 nutrient solution reservoirs, and a The root system of the control plants ( —N —I) developed 7-mm thick and 120-mm long wick made of synthetic extended primary roots without nodules (Fig 1 A) ceramic fibre (Fred Johansen A/S) was provided to obtain Patterns of development The initiation of fertile shoots a capillary nse of the nutrient solution from the reservoir occurred during early vegetative growth (fourth trifoliate The reservoirs were covered with non-transparent white leaf) After floral initiation, a rapid formation of leaves polyethylene to exclude light Autoclaved coarse granite occurred during the development of inflorescences, which gravel serving as a biological barrier was used to cover the led to the accumulation of leaf and stem matter until the growth medium to a depth of 20 mm The nutrient solution beginning of pod setting (Fig 2 A, B) Anthesis was lengthy reservoirs were adjusted daily to 0 5 1 capacity During plant (6 weeks), characterized by a high abscission-rate of flower Downloaded from https://academic.oup.com/aob/article/70/1/11/129022 by guest on 29 September 2021 growth pots were spread to minimize canopy interference buds, flowers and immature pods Tubenzation appeared to The pots were arranged in a completely randomized be initiated during flowering The active tuber growth was design Eight replicate pots of the treatments ( + N —I and delayed till after pod dryness, due to the 'strong sink' of the — N + I) were sampled on five occasions during the first 71 d pods During pod development disintegration of nodules after emergence (DAE), and four replications of the treatments ( + N-I and —N + I) were sampled from 85 (A) DAE to 185 DAE on six occasions, le a total of 11 70 samplings during the experiment The reference — N —I were sampled on seven occasions till 99 DAE The leaf area 60 of the plants was determined in all 11 samplings The leaf area ot the leaflets was measured by an area meter (Model 50 Li-3000) Plants were separated into root, nodules,
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