Effects of exogenous nitrogen-compounds on the concentrations of allantoin and various constituents in several organs of soybean plants

The effects of nitrogen compounds supplied to culture solutionson the concentrations of allantoin and various constituentsin several organs of soybean plants A62-1 (nodulating variety)were studied to elucidate the symbiotic relation. A62-1 plantsbearing well developed nodules accumulated a large quantityof allantoin in the upper stems, roots, developing leaves, podsand maturing seeds in the reproductive stage. However, the additionof N lowered the allantoin accumulation without changing thesoluble Kjeldahl-N concentration in any organs of the host plants.Also addition of N increased the amino-N concentration in upperstems and roots in contrast with the lack of change in developingleaves, pods and maturing seeds. The decrease of allantoin accumulationwas parallel with the weakened formation of nodules. The additionof N also scarcely affected the concentrations of reducing sugarand sugar in the upper stems. There were few differences inthe concentrations of allantoin, amino-N and soluble Kjeldahl-Namong nodules attached to the A62-1 plants grown in variousamounts of nitrate. Statistical calculations showed that the allantoin concentrationin A62-1 plants was correlated negatively with N applicationand positively with nodule weights. Significant levels of bothcorrelation coefficients were attained in the reproductive stage.Thevariation in allantoin concentration in A62-1 plants was notcorrelated with that in the sugar/soluble Kjeldahl-N ratio andthe reducing sugar/soluble Kjeldahl-N ratio. A large quantity of nitrate added to the nutrient solutionsof the A62-2 (non-nodulating variety) plants elicited only asmall amount of allantoin accumulation in the upper stems, witha consequent increase in the concentrations of amino-N and solubleKjeldahl-N and a decrease in the concentrations of sugar andreducing sugar. (Received August 25, 1976; )     

Distribution and change in the contents of allantoin and allantoic acid in developing nodulating and non-nodulatng soybean plants

Distribution and change in contents of allantoin¹ in each organof nodulating variety, A62-1, and non-nodulating variety, A62-2,of soybean plants were measured over the growth period, andthe physiological significance of allantoin in soybean plantsis discussed. Allantoin in the cotyledons of both varieties increased andthen decreased in the germination stage. The allantoin levelin stems, roots and nodules of A62-1 was raised with the growthand attained a maximum at the green pod stage and then decreased.On the other hand, those organs of A62-2 accumulated littleallantoin over the growth period. The allantoin level in thestems of A62-1 was the highest compared with other organs. Inthe leaves of A62-1, the level was higher in the developingleaves than lower mature leaves. The level decreased just beforethe end of leaf development and became trace in the lower fullydeveloped leaves. The allantoin level in the pods of A62-1 duringthe young stage was fairly high; whereas that of A62-2 was lowbut significant, and then decreased with maturing. The dry seedsin both varieties showed low levels. Allantoin was concluded to be accumulated in roots and stemsof developing soybean plants bearing nodules and then decreasedin the stage of seed formation. ¹ In this article the sum of allantoin and allantoic acid ismeasured. Therefore, the expression "allantoin" in the textand abstract includes allantoic acid. (Received August 19, 1976; )     

Comparative Response of Nodulated and Nitrogen-Supplied Cowpea (Vigna unguiculata (L.) Walp. var. Tuy) Plants to Infection by Cowpea Mosaic Virus

The effect of infection by the Cowpea Mosaic Virus (CpMV) onseveral parameters relevant to symbiotic nitrogen fixation wasdetermined in cowpea (Vigna unguiculata (L.) Walp. var. Tuy)plants nodulated with two strains of Rhizobium cowpea: IVIC–124and IVIC–38. Plants were virus-infected at the seedlingstage before Rhizobium inoculation. The effect of CpMV infectionon plant growth was analysed in nodulated and nitrogen-suppliedplants at 18, 25 and 35 d after germination. At all developmentalstages of nodulated plants CpMV infection caused a reductionof leaf chlorophyll content, leaf area, dry weight of shootsand roots, total nodule weight and nodule number. Most of thenodules from 18- and 25-d-old CpMV-infected plants did not exhibitleghaemoglobin pigmentation. CpMV infection delayed the onsetof nitrogenase activity in nodules of the rhizobial strain IVIC–124and the enzyme activity measured on a per plant basis was reducedin both strains at the first and second harvests. Significantnitrogenase activity was detected in 35-d-old infected plants.Some of the nodules of the rhizobial strain IVIC-124 and mostof the nodules from plants nodulated with the strain IVIC-38developed leghaemoglobin; however, the nodule-specific nitrogenaseactivity, estimated on a milligram nodule dry weight basis,was always higher in virus-infected plants, particularly in18-d-old CpMV-infected plants harbouring the IVIC–124strain. CpMV-infected nodules had a larger peribacteroidal space,a reduced number of peribacteroid units, a greater number ofbacteroids per unit, a lower number of vesicles and 88% lowertotal reducing sugar content. Starch accumulation was detectedin infected leaves of nodulated plants during the first harvest,while high levels of leaf reducing sugars and protein were presentat the second harvest. In healthy nodulated plants the rhizobialstrain IVIC–124 was shown to be more efficient than IVIC–38in promoting plant growth. However, the results indicate thatnodulation by rhizobial strain IVIC–124 and growth ofplants harbouring this strain were affected to a greater extentby virus infection. The effect of CpMV infection on leaf chlorophyllcontent, leaf area, carbohydrate level, leaf proteins and growthof nitrogen-supplied plants, as well as the symptoms inducedin the leaves, were less conspicuous than in nodulated plants. Key words: Cowpea, Rhizobium, virus infection, nodule untrastructure     

Probable site of allantoin formation in nodulating soybean plants

Nodulated soybean plants contain high concentration of allantoin in all parts. Excision of nodules from the roots brought about a marked decrease in allantoin. To examine the function of nodules in allantoin production, nodulated and nodule-detached soybeans were fed with ¹⁵NH₃ for 1 week. High abundance of ¹⁵N was found in the amino acid-N fraction of both plants. In the root and stem of the nodulated plants, ca 80% of the nitrogen in this fraction was derived from the NH₃ added in the medium. Excess ¹⁵N was detected also in allantoin-N fraction, but the ¹⁵N content was very low in contrast to that in amino acid-N fraction. The site involved in the allantoin formation and the possible significance of its synthesis are discussed in relation to symbiotic nitrogen fixation.     

Effects of combined nitrogen on anapleurotic carbon assimilation and bleeding sap composition in Phaseolus vulgaris L.

Dwarf french beans, Phaseolus vulgaris L., were grown with or without inoculation with rhizobia (strain 3644), and with or without a combined nitrogen source (nitrate or ammonium ions). The distribution of radioactivity into products of dark ¹⁴CO₂ assimilation was studied in roots or nodules from these plants. A detailed study was also made of the distribution and rates of excretion of nitrogen in xylem bleeding sap in 28 day old plants grown on the various sources of nitrogen. Whereas detached nodules accumulated radioactive glycine, serine and glutamate when incubated with ¹⁴CO₂, bleeding sap from plants root fed ¹⁴CO₂ contained low levels of radioactivity in these compounds but higher levels in allantoin. Chemical analysis showed allantoin to be the major compound transported in the xylem of nodulated plants, whether or not they were fed on combined nitrogen. In contrast uninoculated plants accumulated mainly amino acids in the bleeding sap, the amount and chemical composition of which depended on the combined nitrogen source.Abbreviations PEP phosphoenol pyruvate - OAA oxaloacetate     

Increase in the Formation and Nitrogen Fixation of Soybean Nodules by Vesicular-Arbuscular Mycorrhiza

Symbiotic interactions of the tripartite association of soybeanplant, vesiculararbuscular (VA) mycorrhizal fungus and Rhizobiumjaponicum were shown. Mycorrhizal plants absorbed more P, Ca and Mg and had higherP, Ca and Mg contents in their stems or leaves than non-mycorrhizalplants. Phosphorus concentration was also higher in the nodulesof mycorrhizal plants. VA mycorrhizae increased nodule number, nodule weight and acetylenereduction activity of nodules. Concomitantly seed productionand N content of leaves were enhanced. Both nodulating (A62-1) and non-nodulating (A62-2) cultivarsof soybean plants [Glycine max (L.) Merr.] were colonized byVA mycorrhizal fungi, identified as belonging to the genus Glomus. (Received August 12, 1985; Accepted January 14, 1986)     

Metabolism and translocation of allantoin in ureide-producing grain legumes

Transfer of the nitrogen and carbon of allantoin to amino acids and protein of leaflets, stems and petioles, apices, peduncles, pods, and seeds of detached shoots of nodulated cowpea (Vigna unguiculata L. Walp. cv. Caloona) plants was demonstrated following supply of [2-¹⁴C], [1,3-¹⁵N]allantoin in the transpiration stream. Throughout vegetative and reproductive growth all plant organs showed significant ureolytic activity and readily metabolized [2-¹⁴C]allantoin to ¹⁴CO₂. A metabolic pathway for ureide nitrogen utilization via allantoic acid, urea, and ammonia was indicated. Levels of ureolytic activity in extracts from leaves and roots of nodulated cowpea were consistently maintained at higher levels than in non-nodulated, NO₃⁻ grown plants.

[¹⁴C]Ureides were recovered in extracts of aphids (Aphis craccivora and Macrosiphum euphorbieae) feeding at different sites on cowpea plants supplied with [2-¹⁴C]allantoin through the transpiration stream or to the upper surface of single leaflets. The data indicated that the ureides were effectively transferred from xylem or leaf mesophyll to phloem, and then translocated in phloem to fruits, apices, and roots.

     

Effect of Salinity on Growth, Nodulation and Nitrogen Yield of Chickpea (Cicer arietinum L.)

Chickpea cultivar ILC 482 was inoculated with salt-tolerantRhizobium strain Ch191 in solution culture with different saltconcentrations added either immediately with inoculation or5 d later. The inhibitory effect of salinity on nodulation ofchickpea occurred at 40 dS m^–1 (34.2 mol m^–3 NaCl)and nodulation was completely inhibited at 7 dS m^–1 (61.6mol m^–3 NaCl); the plants died at 8 dS m^–1 (71.8mol m^–3 NaCl). Chickpea cultivar ILC 482 inoculated with Rhizobium strain Ch191^spcstrwas grown in two pot experiments and irrigated with saline water.Salinity (NaCl equivalent to 1–4 dS m^–1) significantlydecreased shoot and root dry weight, total nodule number perplant, nodule weight and average nodule weight. The resultsindicate that Rhizobium strain Ch191 forms an infective andeffective symbiosis with chickpea under saline and non-salineconditions; this legume was more salt-sensitive compared tothe rhizobia, the roots were more sensitive than the shoots,and N₂ fixation was more sensitive to salinity than plant growth. Key words: Cicer arietinum, nodulation, N₂ fixation, Rhizobium, salinity     

PvUPS1, an allantoin transporter in nodulated roots of French bean

Nodulated legumes receive their nitrogen via nitrogen-fixing rhizobia, which exist in a symbiotic relationship with the root system. In tropical legumes like French bean (Phaseolus vulgaris) or soybean (Glycine max), most of the fixed nitrogen is used for synthesis of the ureides allantoin and allantoic acid, the major long-distance transport forms of organic nitrogen in these species. The purpose of this investigation was to identify a ureide transporter that would allow us to further characterize the mechanisms regulating ureide partitioning in legume roots. A putative allantoin transporter (PvUPS1) was isolated from nodulated roots of French bean and was functionally characterized in an allantoin transport-deficient yeast mutant showing that PvUPS1 transports allantoin but also binds its precursors xanthine and uric acid. In beans, PvUPS1 was expressed throughout the plant body, with strongest expression in nodulated roots, source leaves, pods, and seed coats. In roots, PvUPS1 expression was dependent on the status of nodulation, with highest expression in nodules and roots of nodulated plants compared with non-nodulated roots supplied with ammonium nitrate or allantoin. In situ RNA hybridization localized PvUPS1 to the nodule endodermis and the endodermis and phloem of the nodule vasculature. These results strengthen our prediction that in bean nodules, PvUPS1 is involved in delivery of allantoin to the vascular bundle and loading into the nodule phloem.     

Phyllody Malformation in Flowers of Rosaxhybrida cv. Motrea: Effects of Rootstocks, Flower Position, Growth Regulators and Season

Phyllody, a common malformation of Rosaxhybrida cv. Motrea,is characterized by the formation of vegetative structures inplace of stamens and carpels. The incidence of phyllody in thiscultivar is dependent on the type of the root system. The highestproportion of malformed flowers was found in ungrafted plants,this decreased in plants grafted on R. indica major and waslowest on R. canina inermis. Flowers formed on shoots, whichwere located closer to the roots, were more prone to phyllodythan those grown on the upper part of the plants. The incidenceof phyllody was lower in winter than in summer, while shadingof the plants reduced the proportion of malformed flowers regardlessof their root system and the season. Application of gibberellicacid (GA₃) or benzylaminopurine (BAP) greatly promoted the incidenceof phyllody, while paclobutrazol (PBZ) reduced it. The activityof endogenous cytokinins in the xylem exudates from ungraftedplants was higher than in exudates from those grafted on R.indica major, and the lowest activity was found in exudatesfrom plants grafted on R. canina inermis. The involvement ofplant hormones in the development of floral organs in rose flowersis discussed. Key words: Cytokinins, flower malformation, gibberellic acid, homeosis, phyllody, roses     

Effect of Removal of Flower Buds, Open Flowers, Young Pods and Shoot Apex on Growth and Pod Set in Soybean

The complete removal of the reproductive structures once andshoot apices of soybeans (Glycine max L. Merrill) during earlyanthesis but before the rapid seed development stage significantlyincreased flowering and pod set in greenhouse and field grownplants. The treated plants had darker green leaves, shorterstems and petioles and retained their chlorophyll content longerthan control plants. Pod maturation was also delayed. Althoughdecapitation and the removal of reproductive structures increasedthe number of 3- or more-seeded pods in all varieties tested,seed weight per plant was not consistently increased. The possibleinvolvement of endogenous hormones in pod set and multi-loculepod production in soybeans is discussed. Key words: Glycine max (L.) Merr, Reproductive structures, Shoot apex, Growth, Flowering, Pod set, Multi-locule pods     

Effects of Allopurinol [4-Hydroxypyrazolo(3,4-d)Pyrimidine] on the Metabolism of Allantoin in Soybean Plants

Some studies on the effects of xanthine oxidase inhibitor allopurinol [4-hydroxypyrazolo(3,4-d)pyrimidine] on allantoin metabolism of soybean plants (Glycine max cv. Tamanishiki) are reported. Soybean seedlings, aseptically germinated for 96 hours on agar containing 1 millimolar allopurinol, contained only slight amounts of allantoin, allantoic acid, and urea as compared with controls. Analysis of purines and pyrimidines of the allopurinol-treated seedlings showed marked accumulation of xanthine both in the cotyledons and seedling axes. No hypoxanthine accumulation was found. Xanthine accumulation due to allopurinol treatment was relatively low after the cotyledons had fallen. For nodulated plants, allopurinol caused a significant drop in allantoin (+allantoic acid) in the stems and nodules, accompanied by a striking accumulation of xanthine in the nodules. The xanthine concentration in the nodules far exceeded that in the germinated seedlings. Allopurinol at a concentration of 50 micromolar strongly inhibited xanthine oxidase prepared from soybean nodules.

The results suggested that the main pathway of allantoin formation in soybean plants was through purine decomposition, via xanthine-uric acid. It was specially noted that a very active purine-decomposing system existed in soybean nodules.

     

Symbiotic nitrogen fixation and physiological performance of bean (Phaseolus vulgaris L.) plants as affected by Rhizobium inoculum position and bean rugose mosaic virus infection

Bean (Phaseolus vulgaris L. var. Tacarigua) plants were grownin sterilized Leonard jars under controlled conditions. Beforesowing, 1 g of gamma irradiated peat containing the Rhizobiumtropici strain CIAT899 was placed at either 2 or 10 cm belowthe sand surface. Mechanical infection of bean rugose mosaicvirus (BRMV) was carried out in 3-d-old seedlings. Thus, theearly events of nodulation occurred before the arrival of virusparticles to roots. Rhizobium inoculation at 2 cm deep resultedin the formation of nodule clusters close to the crown, in contrastto the homogeneous nodulation along the roots observed in plantsinoculated with Rhizobium at a depth of 10 cm. The uniform arrangementof nodules on the roots enhanced the plant shoot biomass, althoughthe total nodule mass per plant did not differ between Rhizobiuminoculation treatments. Nodules located on deeper roots resultedin higher ureide concentrations in shoots and leaves and inreduced carbohydrate concentrations in leaves. In healthy plants,nodules formed on deeper roots had higher allantoinase activityand a greater carbohydrate concentration when compared to thatof nodules located close to the crown. Deeper nodules had ureideconcentrations similar to those of upper nodules, probably asa consequence of increased translocation of N-compounds to aerialorgans. A similar pattern of nodule formation and response toinoculum position was observed in BRMV-infected plants at allharvests. However, virus infection resulted in reduced totalnodule mass, shoot biomass, total leaf area and induced transitoryalterations in the ureide, -amino-N and carbohydrate concentrationin the different plant compartments. The effect of BRMV infectionon plant parameters was more evident during the vegetative stagesof growth. Nevertheless, the magnitude of the effect was alwaysmore pronounced in plants inoculated with Rhizobium at a depthof 2 cm compared to those Inoculated at 10 cm due to a greateractivity of deeper nodules despite virus infection. Deeper nodulesin BRMV-infected plants showed higher carbohydrate concentrationas well as higher allantoinase and uricase activity than thosedeveloped close to the crown, at all harvests. This observationwas further supported by ultrastructural analysis of virus-infectednodules, since virus replication took place in cells containingbacteroids of upper and lower nodules, but only in the interstitialcells of the latter. BRMV infection did not hinder the allantoinaseactivity and the chlorophyll content of uppermost mature leavesregardless of inoculum position. At the flowering and fruitingstages, healthy and BRMV-infected plants did not differ withregard to any of the tested parameters. Only inoculum positionhad an effect. The nearly normal functioning of the symbioticprocess at these stages of growth was attributed to the formationof a new generation of nodules in BRMV-infected plants subjectedto each of the Rhlzobium inoculation treatments. Key words: Bean rugose mosaic virus, symbiotic nitrogen fixation, bean, Rhizobium inoculum position, nodule ultrastructure     

Transport of nitrogen in the xylem of soybean plants

Experiments were conducted to characterize the distribution of N compounds in the xylem sap of nodulated and nonnodulated soybean plants through development and to determine the effects of exogenous N on the distribution of N compounds in the xylem. Xylem sap was collected from nodulated and nonnodulated greenhouse-grown soybean plants (Glycine max [L.] Merr. “Ransom”) from the vegetative phase to the pod-filling phase. The sum of the nitrogen in the amino acid, nitrate, ureide (allantoic acid and allantoin), and ammonium fractions of the sap from both types of plants agreed closely with total N as assayed by a Kjeldahl technique. Sap from nodulated plants supplied with N-free nutrient solution contained seasonal averages of 78 and 20% of the total N as ureide-N and amino acid-N, respectively. Sap from nonnodulated plants supplied with a 20 millimolar KNO₃ nutrient solution contained seasonal averages of 6, 36, and 58% of total N as ureide-N, amino acid-N, and nitrate-N, respectively. Allantoic acid was the predominant ureide in the xylem sap and asparagine was the predominant amino acid. When well nodulated plants were supplied with 20 millimolar KNO₃, beginning at 65 days, C₂H₂ reduction (N₂ fixation) decreased relative to nontreated plants and there was a concomitant decrease in the ureide content of the sap. A positive correlation (r = 0.89) was found between the ureide levels in xylem sap and nodule dry weights when either exogenous nitrate-N or urea-N was supplied at 10 and 20 millimolar concentrations to inoculated plants. The results demonstrate that ureides play a dominant role in N transport in nodulated soybeans and that the synthesis of ureides is largely dependent upon nodulation and N₂ fixation.     

Carbon Flow from Nodulated Roots to the Shoots of Soybean (Glycine max L. Merr.) Plants: An Estimation of the Contribution of Current Photosynthate to Ureides in the Xylem Stream

Kouchi, H. and Higuchi, T. 1988. Carbon flow from nodulatedroots to the shoots of soybean {Glycine max L. Merr.) plants:An estimation of the contribution of current photosynthate toureides in the xylem stream.–J. exp. Bot. 39: 1015–1023. Well-nodulated, water-cultured soybean plants were allowed toassimilate ¹³CO₂ at a constant specific activity for 10 h andthe ¹³C-labelling of total carbon and ureides in xylem sap wasinvestigated. Labelled carbon appeared very rapidly in the xylem stream. Percentageof labelled carbon (relative specific activity, RSA) in xylemsap was 18% at 2 h after the start of ¹³CO₂ assimilation andreached 53% at the end of the 10 h assimilation. The amountof labelled carbon exported from nodulated roots to the shootsvia the xylem during the 10 h labelling period accounted for33% of total labelled carbon imported into the nodulated roots.Ureides (allantoin and allantoic acid) in xylem sap were stronglydependent on currently assimilated carbon. The RSA of ureidesin xylem sap had reached 83% at the end of the assimilationperiod. Labelled carbon in ureides accounted for 51% of totallabelled carbon returned from nodulated roots to the shootsvia the xylem during the 10 h assimilation period. A treatmentwith 20 mol m^–3 nitrate in the culture medium for 2 ddecreased the ureide concentration in the xylem sap slightly,but greatly decreased the RSA of ureides. By comparing the data with the results of analysis of the xylemsap of nodule-detached plants, it was concluded that the majorityof labelled carbon exported to the xylem stream from noduleswas in ureide form. A considerable amount of carbon was alsoreturned from roots to shoots via the xylem stream but it wasmore dependent on (non-labelled) carbon reserved in the roottissues. Key words: Soybean(Glycine max L.), root nodule, carbon partitoning, ¹³CO₂ assimilation, xylem     

A possible role of allantion and the influence of nodulation on its production in soybean plants

Summary To examine the influence of nodulation on the production of nitrogenous compounds, soybean plants (Glycine max var. Tamanishiki) were grown with or without N-fertilizer in the field, and the changes in amino-N and allantoin-N content in the different organs were determined throughout the growth period. In the stem allantoin-N markedly increased after the flowering period and then decreased during seed formation. Allantoin accumulated in the pod (up to 70 per cent of total alcohol soluble-N) during pod formation, while in the seed the main N-compounds were amino acids, the allantoin concentration being very low. In well-nodulated soybeans grown without N-fertilizer, allantoin content in every plant organ was always high compared to beans grown with N-fertilizer, but amino-N content was comparatively low.Another experiment, in which soybean plants were allowed to form nodules by growing on a N-free medium, and in which a fixed-N supply was then controlled by the addition of various levels of ammonium, was made in a greenhouse. When nodules were formed, the subsequent addition of high concentrations of ammonium caused the accumulation of allantoin as well as the accumulation of amides and arginine. A possible role for allantoin and some aspects of its production in soybeans are discussed. re]19760421     

Plant growth, nutrient acquisition and mycorrhizal symbioses of a waterlogging tolerant legume (Lotus glaber Mill.) in a saline-sodic soil

Seedlings of Lotus glaberMill., were grown in a native saline-sodic soil in a greenhouse for 50 days and then subjected to waterlogging for an additional period of 40 days. The effect of soil waterlogging was evaluated by measuring plant growth allocation, mineral nutrition and soil chemical properties. Rhizobiumnodules and mycorrhizal colonisation in L. glaberroots were measured before and after waterlogging. Compared to control plants, waterlogged plants had decreased root/shoot ratio, lower number of stems per plant, lower specific root length and less allocation of P and N to roots. Waterlogged plants showed increased N and P concentrations in plant tissues, larger root crown diameter and longer internodes. Available N and P and organic P, pH and amorphous iron increased in waterlogged soil, but total N, EC and exchangeable sodium were not changed. Soil waterlogging decreased root length colonised by arbuscular mycorrhizal (AM) fungi, arbuscular colonisation and number of entry points per unit of root length colonised. Waterlogging also increased vesicle colonisation and Rhizobium nodules on roots. AM fungal spore density was lower at the end of the experiment in non-waterlogged soil but was not reduced under waterlogging. The results indicate that L. glaber can grow, become nodulated by Rhizobium and colonised by mycorrhizas under waterlogged condition. The responses of L. glaber may be related its ability to form aerenchyma.     

Grafting of cucumber as a means to minimize copper toxicity

The aim of the current work was to determine whether grafting could improve copper (Cu) tolerance of cucumber, and to study the changes induced by the rootstock in the shoot growth at agronomical and physiological levels. A greenhouse experiment was carried out to determine yield, growth, fruit quality, leaf gas exchange, chlorophyll and carotenoids contents, electrolyte leakage, mineral composition and assimilate partitioning of cucumber plants (Cucumis sativus L. cv. ‘Akito’), either ungrafted or grafted onto the commercial rootstock ‘Shintoza’ (Cucurbita maxima Duchesne × Cucurbita moschata Duchesne) and cultured in nutrient film technique (NFT). Plants were supplied with nutrient solutions having three levels of Cu concentration [0.3 (control), 47, or 94 μM]. Significant depression of yield, shoot and root biomass production, and fruit quality (low fruit pH, and high Cu content) in Cu treated cucumber plants was observed, and this effect varied as a function of Cu concentration in NFT solution. The relative yield of cucumber decreased by ≈3.4% for each unit of increase in leaf tissue Cu concentrations above the threshold value (7.8 μg g⁻¹). At the two higher Cu concentrations (47 and 94 μM Cu), the percentages of yield, shoot and root biomass weight reductions were significantly lower in grafted plants in comparison to those of the ungrafted plants. Excessive Cu, especially at 94 μM Cu, inhibited photosynthesis, pigment synthesis, and membrane integrity. The Cu-related reductions in net assimilation, stomatal conductance, chlorophyll and carotenoid content were more severe in ungrafted plants in comparison with those grafted on 'Shintoza’. The percentage of electrolyte leakage was significantly higher in ungrafted plants especially those with severe Cu toxicity (94 μM Cu). The accumulation of Cu in leaf tissue at 47 and 94 μM Cu, with respect to control, were significantly lower in grafted plants (138 and 181%, respectively) in comparison to that of ungrafted plants (about 235 and 392%, respectively). Significant reduction of macro- (N, K, Ca and Mg) and micro-elements (Fe, Mn and Zn) in cucumber leaf tissue was found under moderate and severe Cu stresses especially on ungrafted plants. The improved crop performance of grafted plants was attributed to their strong capacity to inhibit Cu accumulation in the aerial parts and to maintain a better plant nutritional status.     

Evidence for a graft-transmissible substance which delays apical senescence in Pisum sativum L.

Apical senescence in an early flowering line of pea, G2, is greatly delayed by short days. This behavior is controlled by two dominant genes. Apical senescence of ungrafted, insensitive (I) lines is unaffected by photoperiod. When I-type scions with one of the two required genes were grafted onto G2, apical senescence of the I-type was delayed in short days, but not in long days. Flowering of the I-type was unaffected. The apex of the G2 stock was unaffected as well. Apical senescence of an I-type line lacking both photoperiod genes was not delayed when grafted on G2 in short days. It is concluded that G2 plants grown in short days produce a graft-transmissible factor which delays apical senescence of photoperiodically insensitive lines.     

Growth of Trifolium subterraneum L. Selected for Sparse and Abundant Nodulation as Affected by Root Temperature and Rhizobium Strain

Root temperature greatly affected plant growth whether or notplants depended on symbiotic nitrogen fixation. The two plantselections responded differently to the three strains of Rhizobiumand this response was differentially affected by root temperature. Plant yield was significantly decreased by each fall of 4 °Cin temperature from 19 to 7 °C by amounts that dependedboth on the host and Rhizobium strain. Symbiosis with strainTA1, originally isolated from a cold environment, was most tolerantof a root temperature of 11 °C; TA1 produced as much ormore plant material of the abundantly nodulating host in 40days growth at 7 and 11 °C as did the uninoculated plantsgiven KNO³. Root temperature affected the number, rate of formation, anddistribution of nodules on the root system. At 7 °C fewernodules formed than between 11 and 19 °C. At 7 °C nodulesdid not form on secondary roots by 40 days but at 11 °Cthe secondary roots nodulated rapidly between 30 and 40 days.Nodule formation at 19 °C was almost completed at 20 days,when secondary root nodules accounted for 60 per cent of thetotal. Within the range 15 to 19 °C, at which the originalselections for sparse and abundant nodulation were made, plantsnodulated true to selection, but not at 11 °C. At 7 and11 °C plants nodulated with TA1 yielded more with increasingnumber of nodules.