Efficient rooting for establishment of papaya plantlets by micropropagation

A low cost micropropagation protocol to produce high quality root systems which are easy and economical to acclimatize is essential for large-scale micropropagation of papaya (Carica papaya L.). In this study, individual shoots (>0.5 cm) with 2_∼3 leaves from in vitro papaya multiple shoots were cultured on MS agar medium containing 2.5 μM IBA under dark conditions for 1 week for root induction. They were then transferred to agar or vermiculite media, containing half strength MS medium, under aerated or non-aerated conditions, for root development. Rooting percentage of shoots cultured for 2 weeks in aerated vermiculite was 94.5%, compared with 90.0% in non-aerated vermiculite, 71.1% in aerated agar, and 62.2% in non-aerated agar. Shoots with roots were acclimated in vermiculite under 100% RH for 1 week and then under ambient conditions for 2 weeks in a temperature-controlled growth chamber (28 °C). The survival rates of the plantlets were 94.5% from aerated vermiculite, 87.8% from non-aerated vermiculite, 42.2% from aerated agar, and 35.6% from non-aerated agar. Thus, root induction in low-concentration IBA agar medium followed by root development in vermiculite containing half strength MS medium under aerated conditions results in efficient rooting of in vitro papaya shoots. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effects of Humate, Agar-agar, and EDTA on the Development of Tomato Seedlings in Aerated and Non-aerated Water Cultures

The effect of sodium humate on the development of tomato seedlingsin aerated and non-aerated water cultures has been comparedwith the effect of colloidal solutions of agar-agar. Similarcomparisons have been made between the activities of humateand EDTA. The experiments have been conducted in fresh mediaas well as in nutrient solutions that had been formerly usedfor growing other tomato seedlings. Finally, a comparison betweeneffects of humate and EDTA on tomato seedlings growing in nutrientsolutions with various concentrations of calcium, magnesium,and iron salts in aerated and non-aerated water cultures hasbeen carried out. Media used for growth experiments have beentested microbiologically, and qualitative analyses of theirorganic compounds have been made. A distinct relationship between physiological activities ofhumate, EDTA, and the concentrations of calcium, magnesium,and iron cations in the medium has been found. It has also beenshown that in non-aerated media the plants mostly anifer thedeficiency of available iron; this is prevented by the additionof either humate or EDTA.     

Adaptation of barley roots to low oxygen supply and its relation to potassium and sodium uptake

The uptake of Na⁺ and K⁺ by barley seedlings grown on aerated or non-aerated solutions was studied. Plants growing in culture solution took up K⁺ with high selectivity whether the solution was aerated or not. Roots of plants grown on aerated CaSO₄ and transferred to a solution of KCl and NaCl had a lower preference for K⁺ than roots of plants grown on non-aerated CaSO₄. Both kinds of low-salt roots were much less able to discriminate between K⁺ and Na⁺ than high-salt roots grown on a culture solution. The different levels of K⁺ selectivity are suggested to be related to H⁺ release from the tissue.     

Seasonal Variations and Aeration Effects on Water Quality Improvements and Physiological Responses of Nymphaea Tetragona Georgi

Seasonal variations and aeration effects on water quality improvements and the physiological responses of Nymphaea tetragona Georgi were investigated with mesocosm experiments. Plants were hydroponically cultivated in six purifying tanks (aerated, non-aerated) and the characteristics of the plants were measured. Water quality improvements in purifying tanks were evaluated by comparing to the control tanks. The results showed that continuous aeration affected the plant morphology and physiology. The lengths of the roots, petioles and leaf limbs in aeration conditions were shorter than in non-aeration conditions. Chlorophyll and soluble protein contents of the leaf limbs in aerated tanks decreased, while peroxidase and catalase activities of roots tissues increased. In spring and summer, effects of aeration on the plants were less than in autumn. Total nitrogen (TN) and ammonia nitrogen (NH₄ ⁺-N) in aerated tanks were lower than in non-aerated tanks, while total phosphorus (TP) and dissolved phosphorus (DP) increased in spring and summer. In autumn, effects of aeration on the plants became more significant. TN, NH₄ ⁺-N, TP and DP became higher in aerated tanks than in non-aerated tanks in autumn. This work provided evidences for regulating aeration techniques based on seasonal variations of the plant physiology in restoring polluted stagnant water.     

Root porosity and oxygen movement in waterlogging-tolerant Trifolium tomentosum and -intolerant Trifolium glomeratum

Trifolium tomentosum and T. glomeratum are small (< 0·5 mg) seeded pasture legumes which are considered to be waterlogging tolerant and intolerant, respectively. The root porosity of the two species was compared for plants raised for 10 d in aerated nutrient solution and then transferred to either aerated (0·25 mol O₂ m^–3) or ‘hypoxic’ (0·031–0·069 mol O₂ m^–3) solutions for a further 7 and 21 d. After 21 d, T. tomentosum developed a root porosity of 11·2% in ‘hypoxic’ solution, which was significantly higher than the 6·1% developed by T. glomeratum. When grown in aerated solution, T. tomentosum also had a larger constitutive porosity (6·7%) than T. glomeratum (3·9%). Cylindrical root-sleeving O₂ electrodes were used to measure the rates of radial O₂ loss (ROL) from roots of the two species when in an O₂-free medium. In general, roots previously grown in ‘hypoxic’ solution had higher rates of ROL than roots grown in aerated solution. Moreover, the rates of ROL along the main root of T. tomentosum were ≈ 5-fold faster than from equivalent locations along roots of T. glomeratum. Manipulations of the shoot O₂ concentration resulted in rapid changes in ROL near the root tip of T. tomentosum plants raised in aerated or ‘hypoxic’ solutions, whereas for T. glomeratum ROL only increased for roots of plants raised in ‘hypoxic’ solution. Thus, the cortical air spaces in roots of both species raised in ‘hypoxic’ solution formed a continuous, low resistance pathway for O₂ diffusion from the shoots to the roots. ROL from the lateral roots was also evaluated and it was 3-fold faster from T. tomentosum than from T. glomeratum. Moreover, ROL from lateral roots of T. tomentosum was 10–20-fold higher than from a position on the primary root axis the same distance from the root/shoot junction. Relatively, high rates of ROL were also recorded for young (40 mm in length) lateral roots of T. glomeratum which were previously grown in ‘hypoxic’ solution, but the ROL was low for the older lateral roots of this species. The substantial amounts of ROL from the lateral roots may limit O₂ supply to the lower parts of the primary root axis, so that the laterals probably become the main functional root system in waterlogged soils.     

Comparative Physiology of Apple Rootstocks: II. The Effect of pH and Aeration on the Growth of Malling Apple Rootstocks in Water Cultures

Three Malling apple rootstocks, M.IX, M.II, and M.XVI, weregrown in aerated and non-aerated nutrient solutions at pH 3·6,4·0, and 5·5, and their growth and survival comparedwith rootstocks grown in sand and in soil. No plants survivedin the non-aerated cultures due to severe bacterial and fungalattack on the roots. In the aerated series a marked differencein susceptibility of rootstocks to root infection was found,M.XVI being considerably more resistant than M.II or M.IX. Morerootstocks survived at the lower pH's (4·0 and 3·6)but among surviving plants of M.XVI growth was much less thanat pH 5·5. It would appear therefore that factors makingfor survival do not of necessity make for optimum growth. Insand and soil cultures all plants survived.     

Effects of the pO2 on swimming activity and food utilization in ophiocephalus striatus

The obligatory air-breathing fish Ophiocephalus striatus, fed on goat-liver, surfaced 906 times, swimming 270 m/day in aerated water (mean pO₂: 151 mm Hg) and 883 times, travelling 325 m/day in non-aerated water (92 mm Hg); surfacing and swimming activities increased below the pO₂ of 74 mm Hg. Hanging duration was more (8.4 hr/day) in the aerated series than that in the non-aerated series (5.9 hr/day). Rates of feeding, absorption and metabolism of either series averaged to 131,126 and 93 (= 0.8 ml O₂/g/hr) g cal/g live fish/day. Conversion rate and efficiency slightly increased from 22 g cal/g/day and 17% in the aerated series. Culturing O. striatus in aerated waters may not be advantageous. Starving groups in the aerated and non-aerated aquaria surfaced 317 times, hung for 14.7 hr, and swam 61 m/day, expending 15 g cal/ g/day (= 0.1 ml O₂/g/hr).     

Stimulation of ethylene production and gas-space (aerenchyma) formation in adventitious roots of Zea mays L. by small partial pressures of oxygen

Adventitious roots of two to four-weekold intact plants of Zea mays L. (cv. LG11) were shorter but less dense after extending into stagnant, non-aerated nutrient solution than into solution continuously aerated with air. Dissolved oxygen in the non-aerated solutions decreased from 21 kPa to 3–9 kPa within 24 h. When oxygen partial pressures similar to those found in non-aerated solutions (3, 5 and 12 kPa) were applied for 7 d to root systems growing in vigorously bubbled solutions, the volume of gas-space in the cortex (aerenchyma) was increased several fold. This stimulation of aerenchyma was associated with faster ethylene production by 45-mm-long apical root segments. When ethylene production by roots exposed to 5 kPa oxygen was inhibited by aminoethoxyvinylglycine (AVG) dissolved in the nutrient solution, aerenchyma formation was also retarded. The effect of AVG was reversible by concomitant applications of 1-aminocyclopropane-1-carboxylic acid, an immediate precursor of ethylene. Addition of silver nitrate, an inhibitor of ethylene action, to the nutrient solution also prevented the development of aerenchyma in roots given 5 kPa oxygen. Treating roots with only 1 kPa oxygen stimulated ethylene production but failed to promote gas-space formation. These severely oxygen-deficient roots seemed insensitive to the ethylene produced since a supplement of exogeneous ethylene that promoted aerenchyma development in nutrient solution aerated with air (21 kPa oxygen) failed to do so in nutrient solution supplied with 1 kPa oxygen. Both ethylene production and aerenchyma formation were almost completely halted when roots were exposed to nutrient solutions devoid of oxygen. Thus both processes require oxygen and are stimulated by oxygen-deficient surroundings in the 3-to 12-kPa range of oxygen partial pressures when compared with rates observed in air (21 kPa oxygen).Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AVG aminoethoxyvinylglycine     

Gas and Liquids in Intercellular Spaces of Maize Roots

Oils are spontaneously absorbed by gas-filled intercellularspaces (IS) in maize root cortex. The network of these spacesin living root sections was imaged by confocal laser scanningmicroscopy using a fluorescent solution of Nile red in oil.The gas volume fraction (GVF) of root segments was quantifiedby the increase in weight (differentiated zones) or tissue density(2–3 mm root tips) due to complete vacuum infiltration.Cooling to 6 °C or inhibition of oxidative phosphorylationdiminished the GVF of root tips but did not significantly affectthe GVF of differentiated root zones. The threshold pressuredifference for measurable infiltration of isolated root segmentsis lower (10 to 15  kPa) than the threshold for infiltrationof comparable zones of attached roots or of detached roots withthe cut surface sealed (>60 kPa). In the absence of an opencut, pressure-driven infiltration of the root cortex is acceleratedby microscopic fissures within the epidermal/hypodermal barrier.The GVF of the root cortex was reduced after transferring rootsfrom sugar solutions (0.1 to 0.3M ) to water. This points toefficient water transport from the medium to sugar-containingcortical cell walls through epidermal and hypodermal protoplasts.When 2-cm-long primary roots were vacuum infiltrated in situand then allowed to grow on aerated mineral medium for a further5 d, cortical IS of the originally infiltrated root bases remainedfilled with liquid but the subsequently grown apical root zoneshad a normal GVF. Copyright 1999 Annals of Botany Company Apoplastic and protoplasmic route, maize, infiltration, intercellular spaces, oil absorption, confocal laser scanning microscope, water transport, Zea mays L.     

Arginase activity in the cotyledons of soybean seedlings

The influence of naphthylacetic acid, abscisic acid, gibberellic acid and kinetin on the formation of aerenchyma in seedling roots of Zea mays L. cv. Capella has been studied in relation to reported changes of their concentration in poorly aerated roots, which readily form aerenchyma, and to the effects of these hormones on the production of ethylene, a major factor promoting aerenchyma formation. Because the absence of nitrate accelerates aerenchyma formation in aerated roots, their influence on these roots was compared. The growth regulators were added to roots growing in non-aerated and aerated nutrient solutions, and aerenchyma formation and the production and endogenous concentration of ethylene were measured. Naphthylacetic acid prevented aerenchyma formation in both aerated roots without nitrate and in non-aerated roots although it enhanced the ethylene concentration of the roots. Abscisic acid also prevented aerenchyma formation, but without affecting the ethylene concentration. Gibberellic acid promoted aerenchyma formation in aerated roots only, but ethylene production in both aerated and non-aerated roots. Kinetin promoted aerenchyma formation in both aerated and non-aerated roots. It stimulated ethylene production in aerated roots, but slightly inhibited it in non-aerated roots. Co²⁺ and Ag⁺, which suppress ethylene production and action, respectively, reduced the promoting effects of gibberellic acid, but not those of kinetin. It is concluded that the effects of the plant growth regulators on aerenchyma formation in maize roots were, with a possible exception for gibberellic acid, not the result of altered ethylene concentrations in the roots. Their influence on aerenchyma formation is discussed in relation to their reported actions on cell membranes.     

Oxygen distribution and movement,respiration and nutrient loading in banana roots (Musa spp. L.) subjected to aerated and oxygen-depleted environments

Excessive soil wetness is a common feature where bananas (Musa spp.) evolved. Under O₂ deficiency, a property of wet soils, root growth and functions will be influenced by the respiratory demand for O₂ in root tissues, the transport of O₂ from the shoot to root and the supply of O₂ from the medium. In laboratory experiments with nodal roots of banana, we examined how these features influenced the longitudinal and radial distributions of O₂ within roots, radial O₂ loss, solute accumulation in the xylem, root hydraulic conductivity, root elongation and root tip survival. In aerated roots, the stele respired about 6 times faster than the cortex on a volume basis. Respiratory O₂ consumption decreased substantially with distance from the root apex and at 300–500 mm it was 80% lower than at the apex. Respiration of lateral roots constituted a sink for O₂ supplied via aerenchyma, and reduced O₂ flow towards the tip of the supporting root. Stelar anoxia could be induced either by lowering the O₂ partial pressure in the bathing medium from 21 to 4 kPa (excised roots) or, in the case of intact roots, by reducing the O₂ concentration around the shoot. The root hair zone sometimes extended to 1.0 mm from the root surface and contributed up to a 60% drop in O₂ concentration from a free-flowing aerated solution to the root surface. There was a steep decline in O₂ concentration across the epidermal-hypodermal cylinder and some evidence of a decline in the O₂ permeability of the epidermal-hypodermal cylinder with increasing distance from the root apex. The differences in O₂ concentration between cortex and stele were smaller than reported for maize and possibly indicated a substantial transfer rate of dissolved O₂ from cortex to stele in banana, mediated by a convective water flow component. An O₂ partial pressure of 4 kPa in the medium reduced net nutrient transfer into the vascular tissue in the stele within 1 or 2 h. Hypoxia also caused a temporary decrease in radial root hydraulic conductivity by an order of magnitude. In O₂ deficient environments, the stele would be among the first tissues to suffer anoxia and O₂ consumption within the root hair zone might be a major contributor to root anoxia/hypoxia in banana growing in temporarily flooded soils.     

Colonisation patterns of root tissues byPhytophthora nicotianae var.parasitica related to reduced disease in mycorrhizal tomato

Tomato plants pre-colonised by the arbuscular mycorrhizal fungusGlomus mosseae showed decreased root damage by the pathogenPhytophthora nicotianae var.parasitica. In analyses of the cellular bases of their bioprotective effect, a prerequisite for cytological investigations of tissue interactions betweenG. mosseae andP. nicotianae v.parasitica was to discriminate between the hyphae of the two fungi within root tissues. We report the use of antibodies as useful tools, in the absence of an appropriate stain for distinguishing hyphae ofP. nicotianae v.parasitica from those ofG. mosseae inside roots, and present observations on the colonisation patterns by the pathogenic fungus alone or during interactions in mycorrhizal roots. Infection intensity of the pathogen, estimated using an immunoenzyme labelling technique on whole root fragments, was lower in mycorrhizal roots. Immunogold labelling ofP. nicotianae v.parasitica on cross-sections of infected tomato roots showed that inter or intracellular hyphae developed mainly in the cortex, and their presence induced necrosis of host cells, the wall and contents of which showed a strong autofluorescence in reaction to the pathogen. In dual fungal infections of tomato root systems, hyphae of the symbiont and the pathogen were in most cases in different root regions, but they could also be observed in the same root tissues. The number ofP. nicotianae v.parasitica hyphae growing in the root cortex was greatly reduced in mycorrhizal root systems, and in mycorrhizal tissues infected by the pathogen, arbuscule-containing cells surrounded by intercellularP. nicotianae v.parasitica hyphae did not necrose and only a weak autofluorescence was associated with the host cells. Results are discussed in relation to possible processes involved in the phenomenon of bioprotection in arbuscular mycorrhizal plants.     

Metabolic evidence for stelar anoxia in maize roots exposed to low o(2) concentrations

This investigation presents metabolic evidence to show that in 4- to 5-day-old roots of maize (Zea mays hybrid GH 5010) exposed to low external O₂ concentrations, the stele receives inadequate O₂ for oxidative phosphorylation, while the cortex continues to respire even when the external solution is at zero O₂ and the roots rely solely on aerenchyma for O₂ transport. Oxygen uptake rates (micromoles per cubic centimeter per hour) declined at higher external O₂ concentrations in excised segments from whole roots than from the isolated cortex; critical O₂ pressures for respiration were greater than 0.26 moles per cubic meter O₂ (aerated solution) for the whole root and only 0.075 moles per cubic meter O₂ for the cortex. For plants with their shoots excised and the cut stem in air, ethanol concentrations (moles per cubic meter) in roots exposed to 0.06 moles per cubic meter O₂ were 3.3 times higher in the stele than in the cortex, whereas this ethanol gradient across the root was not evident in roots exposed to 0 moles per cubic meter O₂. Alanine concentrations (moles per cubic meter) in the stele of roots exposed to 0.13 and 0.09 moles per cubic meter O₂ increased by 26 and 44%, respectively, above the levels found for aerated roots, whereas alanine in the cortex was unchanged; the increase in stelar alanine concentration was not accompanied by changes in the concentration of free amino acids other than alanine. For plants with their shoots intact, alcohol dehydrogenase and pyruvate decarboxylase activities (micromoles per gram protein per minute) in roots exposed to 0.13 moles per cubic meter O₂ increased in the stele by 40 to 50% over the activity in aerated roots, whereas there was no appreciable increase in alcohol dehydrogenase and pyruvate decarboxylase activity in the cortex of these roots. More convincingly, for roots receiving O₂ solely from the shoots via the aerenchyma, pyruvate decarboxylase in the cortex was in an “inactive” state, whereas pyruvate decarboxylase in the stele was in an “active” state. These results suggest that for roots in O₂-free solutions, the aerenchyma provides adequate O₂ for respiration in the cortex but not in the stele, and this was supported by a change in pyruvate decarboxylase in the cortex to an active state when the O₂ supply to the roots via the aerenchyma was blocked.     

Morphological and physiological responses of rice (Oryza sativa) to limited phosphorus supply in aerated and stagnant solution culture

BACKGROUND AND AIMS: Rain-fed lowland rice commonly encounters stresses from fluctuating water regimes and nutrient deficiency. Roots have to acquire both oxygen and nutrients under adverse conditions while also acclimating to changes in soil-water regime. This study assessed responses of rice roots to low phosphorus supply in aerated and stagnant nutrient solution. METHODS: Rice (Oryza sativa 'Amaroo') was grown in aerated solution with high P (200 micro m) for 14 d, then transferred to high or low (1.6 micro m) P supply in aerated or stagnant solution for up to 8 d. KEY RESULTS: After only 1 d in stagnant conditions, root radial oxygen loss (ROL) had decreased by 90 % in subapical zones, whereas near the tip ROL was maintained. After 4 d in stagnant conditions, maximum root length was 11 % less, and after 8 d, shoot growth was 25 % less, compared with plants in aerated solution. The plants in stagnant solution had up to 19 % more adventitious roots, 24 % greater root porosity and 26 % higher root/shoot ratio. Rice in low P supply had fewer tillers in both stagnant and aerated conditions. After 1-2 d in stagnant solution, relative P uptake declined, especially at low P supply. Aerated roots at low P supply maintained relative P uptake for 4 d, after which uptake decreased to the same levels as in stagnant solution. CONCLUSIONS: Roots responded rapidly to oxygen deficiency with decreased ROL in subapical zones within 1-2 d, indicating induction of a barrier to ROL, and these changes in ROL occurred at least 2 d before any changes in root morphology, porosity or anatomy were evident. Relative P uptake also decreased under oxygen deficiency, showing that a sudden decline in root-zone oxygen adversely affects P nutrition of rice.     

Effect of Localized Anoxia on Phaseolus vulgaris L. Root Growth

Heterogeneity within the root environment results in differentialgrowth within root systems. The response of five Phaseolus vulgarisL. cultivars to non-uniform root aeration was evaluated. Threetreatments were applied to a split root system for a periodof 72 h. Treatments consisted of an aerated control, a non-aeratedcontrol (both halves non-aerated, using N₂). and localized anoxia(one-half the root system aerated and the remaining half subjectedto N₂). Shoot and root growth were reduced in the anoxic controlbut not in the aerated control or localized anoxia treatment. Root growth was greatest in the aerated portion of the localizedanoxia treatment for all genotypes. Contributions of the rootcomponents to the compensatory responses differed dependingon the plant cultivar examined. The growth of branched and lateralroots present before the treatment period increased by 65% inline 31908. A 50% increase in the growth of lateral roots whichemerged during the treatment period occurred in another line(Swan Valley). Other genotypes responded in an intermediatemanner. These observations indicate differences in cultivarresponses to localized soil stress. Key words: Phaseolus vulgaris L., Anoxia, Root growth     

Aerenchyma develops by cell lysis in roots AND CELL SEPARATION IN LEAF PETIOLES IN Sagittaria lancifolia (Alismataceae)

Aerenchyma gas spaces are important for plants that survive flooding because these spaces provide an internal pathway for oxygen transport to the root zone. The objective of this study was to characterize the development of aerenchyma gas spaces in Sagittaria lancifolia L., a dominant species in freshwater wetlands adjacent to the Gulf of Mexico. Tissue at different developmental stages was collected from hydroponically grown plants, embedded in plastic, and sections were observed with a light microscope. In S. lancifolia roots, lysigeny (cell lysis) produced gas spaces that increased in volume from the root meristem to the most mature root tissue. Shoot aerenchyma occurred in the large petioles of S. lancifolia and through the blade midrib, but not in the laminar portion of the blade. In contrast to the roots, gas spaces in the petiole were formed by schizogeny (cell separation during development). Shoot initials produced cells that formed interlocking cylinders in the cortex and diaphragm cells that bridged the central portion of the cylinders. Division and expansion of both these cell types increased the diameter of the cylinders and created schizogenous gaps between diaphragm layers that produced large gas spaces in mature tissue. Therefore, aerenchyma development occurs by two different processes in S. lancifolia.     

Influence of Tops and Roots on Net Assimilation Rate of Sugar-beet and Spinach Beet and Grafts between them

Sugar-beet has a larger storage root and greater net assimilationrate (E) than spinach beet. To determine whether the greaterroot was a result or cause of the greater E, grafts were madebetween tops and roots of sugar-beet and spinach-beet in allfour possible combinations. Grafted plants with sugar-beet roots had greater E and rootdry weight, less leaf area and top dry weight and lower concentrationof sugar in the leaf lamina, than those with spinach-beet roots,irrespective of the type of top. Grafted plants with sugar-beettops had greater E, total and root dry weight, but less leafarea, than those with spinach-beet tops, irrespective of thetype of root. The difference in E between grafted plants withsugar-beet tops and spinach-beet tops was similar to that betweengrafted plants with sugar-beet roots and spinach-beet roots.It increased with time to 60 per cent. Increases in E probably represent increases in rate of photosynthesis.Sugar-beet roots probably increased photosynthesis by providinga better sink for assimilates than spinach-beet roots.     

Sensitivity of chickpea and faba bean to root‐zone hypoxia,elevated ethylene,and carbon dioxide

During soil waterlogging, plants experience O₂ deficits, elevated ethylene, and high CO₂ in the root‐zone. The effects on chickpea (Cicer arietinum L.) and faba bean (Vicia faba L.) of ethylene (2 μL L^?1), CO₂ (2–20% v/v) or deoxygenated stagnant solution were evaluated. Ethylene and high CO₂ reduced root growth of both species, but O₂ deficiency had the most damaging effect and especially so for chickpea. Chickpea suffered root tip death when in deoxygenated stagnant solution. High CO₂ inhibited root respiration and reduced growth, whereas sugars accumulated in root tips, of both species. Gas‐filled porosity of the basal portion of the primary root of faba bean (23%, v/v) was greater than for chickpea (10%), and internal O₂ movement was more prominent in faba bean when in an O₂‐free medium. Ethylene treatment increased the porosity of roots. The damaging effects of low O₂, such as death of root tips, resulted in poor recovery of root growth upon reaeration. In conclusion, ethylene and high CO₂ partially inhibited root extension in both species, but low O₂ in deoxygenated stagnant solution had the most damaging effect, even causing death of root tips in chickpea, which was more sensitive to the low O₂ condition than faba bean.     

The effect of some oligo-amines and -guanidines on membrane permeability in higher plants

The effect of a series of oligo-amines and -guanidines on the membranes of higher plants has been tested by measuring the efflux of betacyanin from beet root discs, and the loss of total ions from beet root and swede discs, beet and spinach leaf discs and apple cells in suspension culture. All of the naturally occurring di- and polyamines tested showed relatively little toxicity. Betacyanin efflux from beet root discs was reduced by diamines [NH₂(CH₂)_xNH₂] up to x = 10 or less. Ion efflux was minimal at x = 7. Within the triamine series [NH₂(CH₂)_xNH(CH₂)₃NH₂] for x = 8 or less, betacyanin efflux was reduced or unaffected, although total ion loss was increased by the triamines when x = 4 or more and especially by the longer chain amines (to x = 10). Similar behaviour was found in the tetra-amine series [NH₂(CH₂)₃NH(CH₂)_xNH(CH₂)₃NH₂] with betacyanin efflux reduced for x = 2–4 (spermine). Although spermine potentiated the toxicity effects of Guazatine {[NH₂C(NH)NH(CH₂)₈]₂NH} and Dodine [NH₂C(NH)NH(CH₂)₁₁,Me] in beet root discs, spermine and calcium ions reduced the ion efflux caused by these toxic guanidines and also by Synthalin B [NH₂C(NH)NH(CH₂)₁₂NHC(NH)NH₂] in swede discs, spinach leaves and apple cells. No significant reversal of ion loss was detected with putrescine, cadaverine or spermidine in swede discs. In the homologous series of monoguanidines [NH₂C(NH)NH(CH₂)_x?1Me] for x up to 18, greatest toxicity was shown for x = 10 and 11 in both betacyanin loss and total ion efflux in beet root discs. Greatest ion efflux from the apple cell suspension was found with x = 11 and 12. In the homologous series of diguanidines [NH₂C(NH)NH(CH₂),NHC(NH)NH₂] for x = 2–12 greatest toxicity was shown for x = 12 (the longest chain tested) in beet root and in the efflux of ions from apple cell suspension. Technical Guazatine was considerably more phytotoxic than pure Guazatine in all systems tested. p-Chloromercuribenzoate (p-CMB) potentiates the loss of betacyanin and total ions caused by Guazatine, Synthalin B, and Dodine in beet root discs. This effect of p-CMB is reversed by subsequent incubation in cysteine or mercaptoethanol, prior to treatment with the guanidines.     

Differential ultrastructural changes in tomato hormonal mutants exposed to cadmium

Cadmium (Cd) is a toxic heavy metal, which can cause severe damage to plant development. The aim of this work was to characterize ultrastructural changes induced by Cd in miniature tomato cultivar Micro-Tom (MT) mutants and their wild-type counterpart. Leaves of diageotropica (dgt) and Never ripe (Nr) tomato hormonal mutants and wild-type MT were analysed by light, scanning and transmission electron microscopy in order to characterize the structural changes caused by the exposure to 1 mM CdCl₂. The effect of Cd on leaf ultrastructure was observed most noticeably in the chloroplasts, which exhibited changes in organelle shape and internal organization, of the thylakoid membranes and stroma. Cd caused an increase in the intercellular spaces in Nr leaves, but a decrease in the intercellular spaces in dgt leaves, as well as a decrease in the size of mesophyll cells in the mutants. Roots of the tomato hormonal mutants, when analysed by light microscopy, exhibited alterations in root diameter and disintegration of the epidermis and the external layers of the cortex. A comparative analysis has allowed the identification of specific Cd-induced ultrastructural changes in wild-type tomato, the pattern of which was not always exhibited by the mutants.