The Effects of Oxygen, Carbon Dioxide and Ethylene on Ethylene Biosynthesis in Relation to Shoot Extension in Seedlings of Rice (Oryza sativa) and Barnyard Grass (Echinochloa oryzoides)

Exposing dark-grown seedlings for 3 d to oxygen deficiency (0or 5 kPa) or to additions of carbon dioxide (10 kPa) or ethylene(0·1 Pa) slowed shoot extension in Echinochloa oryzoides,while in rice it was promoted by these treatments, except that5 kPa oxygen was without effect. In E. oryzoides this was dueto reduced growth of the mesocotyl, and in rice to enhancedgrowth of the coleoptile. These responses to carbon dioxideand oxygen deficiency were not consequences of increased ethyleneproduction, since this remained unchanged by carbon dioxideand depressed by oxygen shortage in both species. Furthermore,exogenous ethylene and the ethylene action inhibitor 2,5-norbornadieneeach failed to influence extension in anoxic seedlings, indicatingno regulatory role for ethylene in the absence of oxygen. However,concentrations of the ethylene precursor 1 -aminocyclopropane-1-carboxylic acid (ACC) were increased by carbon dioxide and0 kPa or 5 kPa oxygen, although after 72 h without oxygen totalACC production (i.e. changes in ethylene + ACC + MACC) was suppressedin both species. There was little effect on bound ACC [putativemalonyl-ACC (MACC)] formation. Transferring anaerobic (0 kPa)seedlings to oxygenated conditions (21 kPa) resulted in abnormallyfast rates of ethylene formation, possibly due to the accumulationof ACC under anoxia. This post-anoxic ethylene may have contributedto the faster extension by rice coleoptiles and slower extensionby mesocotyls of E. oryzoides compared with those of seedlingsmaintained continuously in air. Echinochloa oryzoides [Ard.] Fritsch, barnyard grass, Oryza sativa L, rice, oxygen shortage, carbon dioxide, ethylene biosynthesis, shoot extension, 1-aminocyclopropane-1-carboxylic acid (ACC), malonyl-ACC, GC-MS     

Injury to Rice Plants Caused by Complete Submergence; A Contribution by Ethylerie (Ethene)

Complete submergence of rice plants (Oryza sativa L. cv. ‘IR42’)in dilute nutrient solution for 3–6 d almost stopped theaccumulation of dry matter, depressed soluble carbohydrate concentrationby over 75% and promoted chlorosis in fully expanded leaves.Increase in fresh weight by the shoots was not impaired. Extensionby the youngest visible leaf was stimulated. Extension by thenext leaf to appear was retarded by submergence. These growthresponses to submergence were associated with a 1-5-fold increasein the partial pressure of endogenous ethylene (ethene). Applying ethylene (0.3–0.35 Pa) in the gas-phase to non-submergedplants reproduced some, but not all, of these effects of submergence.Thus, greater leaf extension and chlorosis of submerged plantscould be attributable to accumulated ethylene but neither theslow relative growth rate nor the decreased extension of leavesemerging after the start of submergence could be so attributed. Two cultivars (‘FR13A’ and ‘Kurkaruppan’)already known to tolerate submergence, differed little fromsubmergence-intolerant ‘IR42’ in their relativegrowth rate and soluble carbohydrate concentration during submergence.However, their underwater leaf extension was less than in ‘IR42’and chlorosis was much less prevalent, especially in ‘FR13A’.Similarly, ethylene supplied to non-submerged plants was a lesseffective promotor of leaf extension and chlorosis in the twosubmergence tolerant cultivars. Application of 1.0 kPa carbondioxide in the gas-phase prevented the chlorosis response toethylene. The results indicate that accumulated ethylene is a likely causeof fast leaf extension and chlorosis in submergence intolerantforms of rice, particularly when amounts of dissolved carbondioxide are minimal. Key words: Oryza sativa L., aeration, ethylene (ethene), stress-tolerance     

The Effect of Oxygen and Turbulence on Elongation of Coleoptiles of Submergence-Tolerant and -Intolerant Rice Cultivars

Coleoptiles of rice (Oryza saliva L. ) elongated more rapidlyin stagnant solution than in water-saturated air: elongationrates in aerated solution were intermediate. Elongation wasstimulated between 170% to 390% in stagnant solution in 17 cultivarsscreened. On this basis, a relatively submergence-intolerantcultivar and two submergence-tolerant cultivars were studiedfurther. When seedlings were grown in a range of oxygen concentrationsimposed by bubbling solutions rapidly with gas, germinationand early coleoptile growth (up to 5.0 mm) were inhibited atlow oxygen concentrations, particularly in the submergence-intolerantcultivar. Weight per unit length declined with time in thiscultivar, even in mild oxygen deficits. However, later stagesof coleoptile elongation were unaffected by low oxygen supply:elongation proceeded at control rates even in anoxia. It isconcluded that processes unique to germination and early stagesof coleoptile elongation (possibly cell division) have greateroxygen requirements than cell extension. Rates of elongation in stagnant solution were much faster thanthose in any other treatment, including bubbled solutions atsimilar oxygen concentrations (0.125 mmol O₂ I_–1). Therefore,elongation appears to be stimulated by accumulation of endogenouscompounds which are dispersed in bubbled solutions and in air.The principal volatile compound was almost certainly ethylene,since Ag+ (an antagonist of ethylene) reduced the rate of elongationin stagnant solution by 53%. Differences in turgor pressure were only partly responsiblefor rapid coleoptile elongation observed in stagnant solution:there must have also been differences in cell wall propertiesbetween coleoptiles grown in stagnant and bubbled solution.The rapid elongation in stagnant solution hastens coleoptileemergence from the solution, thus establishing an adequate oxygensupply for the seedling. Leaf and root growth then proceeds.     

Ethylene-Enhanced Transport of Uranine, a Fluorescent Dye, in Rice-Seedling Explants in Relation to Ethylene-Stimulated Coleoptile Growth

Ethylene enhanced the transport of uranine, a fluorescent dye,through the coleoptiles of rice (Oryza sativa L. cv. Sasanishiki)seedlings explants after a 2-h lag period. The uranine transportincreased with increasing concentration of ethylene, reachinga maximum at 1 µ1.liter^–1 ethylene. It decreasedwhen some metabolic inhibitors, 2,4-dinitrophenol, carbonylcyanide 3-chlorophenylhydrazone and N-ethylmaleimide, were appliedto the scutella. Mechanical rupture or heating at the vascular-bundlesides of the coleoptiles also interfered with the ethylene-enhancedtransport of uranine. The wounding and removal of the scutellasimilarly blocked uranine transport. Moreover, strong fluorescenceof uranine was observed on the phloem regions of two vascularbundles of a coleoptile under a fluorescence microscope. Itis suggested that the ethylene-enhanced transport of uraninein the coleoptile of rice-seedling explants occurs through thephloem. On the other hand, the removal of the endosperm or therupture of the vascular bundles inhibited the ethylene-stimulatedelongation of the rice coleoptiles. Moreover, ethylene increaseddry weight of the shoots. These results suggest that the ethylene-stimulatedelongation of rice coleoptiles was supported partly by the ethylene-enhancedtranslocation of materials from the endosperms to the coleoptiles. (Received September 6, 1984; Accepted November 15, 1984)     

Aerenchyma (Gas-space) Formation in Adventitious Roots of Rice (Oryza sativa L.) is not Controlled by Ethylene or Small Partial Pressures of Oxygen

Jackson, M. B., Fenning, T. M., and Jenkins, W. 1985 Aerenchyma(gas-space) formation in adventitious roots of rice (Oryza sativaL.) is not controlled by ethylene or small partial pressuresof oxygen.—J. exp. Bot. 36: 1566–1572. The extent of gas-filled voids (aerenchyma) within the cortexof adventitious roots of vegetative rice plants (Oryza sativaL. cv. RB3) was estimated microscopically from transverse sectionswith the aid of a computer-linked digitizer drawing board. Gas-spacewas detectable in 1-d-old tissue and increased in extent withage. After 7 d, approximately 70% of the cortex had degeneratedto form aerenchyma. The extent of the voids in 1-4-d-old tissuewas not increased by stagnant, poorly-aerated external environmentscharacterized by sub-ambient oxygen partial pressures and accumulationsof carbon dioxide and ethylene. Treatment with small oxygenpartial pressures, or with carbon dioxide or ethylene appliedin vigorously stirred nutrient solution also failed to promotethe formation of cortical gas-space. Furthermore, ethylene productionby rice roots was slowed by small oxygen partial pressures typicalof stagnant conditions. Silver nitrate, an inhibitor of ethylene action, did not retardgas-space formation; similarly when endogenous ethylene productionwas inhibited by the application of aminoethoxyvinylglycine(A VG), aerenchyma development continued unabated. Cobalt chloride,another presumed inhibitor of ethylene biosynthesis, did notimpair formation of the gas in rice roots nor did it decreasethe extent of aerenchyma even if A VG was supplied simultaneously.These results contrast with those obtained earlier using rootsof Zea mays L. We conclude that in rice, aerenchyma forms speedily even inwell-aerated environments as an integral part of ordinary rootdevelopment There seems to be little or no requirement for ethyleneas a stimulus in stagnant root-environments where aerenchymais likely to increase the probability of survival. Key words: Rice (Oryza sativa L.), ethylene, flooding, aeration, aerenchyma, environmental stress     

Plantlet regeneration from somatic hybrids of rice (Oryza sativa L.) and barnyard grass (Echinochloa oryzicola Vasing)

Summary Somatic hybridization of rice (Oryza sativa L.) and barnyard grass (Echinochloa oryzicola), a close relative of barnyard millet, was attempted using electrofusion and a new culture method developed for rice protoplasts (Kyozuka et al. 1987) to incorporate some of the agronomically important characters of the latter species into rice. Selection of hybrids was based on inactivation of rice protoplasts by iodoacetamide and the inability of barnyard grass protoplasts to divide. A total of 166 calli were identified as hybrids by isozyme and chromosome analyses. Hybrid calli were highly morphogenic, and 44 shoots were obtained. Most of them, however, were abnormal, and nine grew to plantlets whose morphology was distinct from that of either parent. Our study clearly demonstrates the totipotency of protoplasts in graminaceous monocots.     

Auxin and Ethylene Control of Growth in Seedlings of Zea mays L. and Avena sativa L

The effects of applied ethylene on the growth of coleoptilesand mesocotyls of etiolated monocot seedlings (oat and maize)have been compared with those on the epicotyl of a dicot seedling(the etiolated pea). Significant inhibition of elongation by ethylene (10 µll^–1for 24 h) was found in intact seedlings of all three species,but lateral expansion growth was observed only in the pea internodeand oat mesocotyl tissue. The sensitivity of the growth of seedlingparts to ethylene is in the decreasing order pea internode,oat coleoptile and oat mesocotyl, with maize exhibiting theleast growth response. Although excised segments of mesocotyland coleoptile or pea internode all exhibit enhanced elongationgrowth in IAA solutions (10^–6–2 ? 10^–5 moll^–1), no consistent effects were found in ethylene. Ethyleneproduction in segments was significantly enhanced by applicationof auxin (IAA, 10^–5 mol l^–6 or less) in all tissuesexcept those of the eat mesocotyl. Segments of maize show a slow rate of metabolism of applied[2-¹⁴C]IAA (30 per cent converted to other metabolites within9 h) and a high capacity for polar auxin transport. Ethylene(10 µl l^–1 for 24 h) has little effect on eitherof these processes. The oat has a smaller capacity for polartransport than maize and the rate ef metabolism of auxin isas fast as in the pea (90 per cent metabolized in 6 h). Althoughethylene pretreatment does not change the rate of auxin metabolismin oat, there is a marked reduction in auxin transport. It is proposed that the insensitivity of maize seedlings toethylene is related to the supply and persistence of auxin whichcould protect the seedling against the effects of applied orendogenously produced ethylene. Although the mesocotyl of oatis sensitive to applied ethylene it may be in part protectedagainst ethylene in vivo by the absence of an auxin-enhancedethylene production system. The results are discussed in relationto a model for the auxin and ethylene control of cell growthin the pea.     

Relationship between Coleoptile Elongation and Alcoholic Fermentation in Rice Exposed to Anoxia. II. Cultivar Differences

The relationship between coleoptile elongation and survivalvs. alcoholic fermentation of rice under anoxia is examinedusing eight cultivars differing in submergence tolerance. Anoxiawas imposed on either 1 or 4 d aerated seeds either by N₂ flushingsubmerged tissues or by incubating tissues in stagnant deoxygenatedagar at 0·1% w/v; the latter simulated the stagnant conditionsof waterlogged soil. Two cultivars that were most tolerant tosubmergence also had the greatest tolerance to anoxia, whilea submergence intolerant cultivar was also intolerant to anoxia. Coleoptile growth under anoxia was related to rates of ethanolsynthesis (R_E), however differences between growth during anoxiaand survival after anoxia indicated that post-anoxic injurymay also be important in rice seeds exposed to anoxia. The correlationbetween coleoptile growth and R_E measured on a tissue basisusing intact seeds was r² = 0·67 among six varietiesover 0-3 d anoxia. This correlation improved to about r² = 0·85using R_E of (embryos plus coleoptiles) over 0-3 d, or coleoptilesat 3 d after anoxia. Coleoptile growth of individual seeds wasusually poorly correlated to R_E in these cultivars at 2-3 dafter anoxia. When coleoptiles of similar lengths were obtainedfrom different cultivars using 4 d aerated seeds, there weredifferences in R_E and coleoptile growth which were related tocoleoptile growth during 3 or 5 d anoxia, either on a tissue(r² = 0·85) or a fresh weight basis (r² = 0·70-0·97respectively). Results are discussed in relation to factorswhich may limit ethanol synthesis in rice exposed to anoxiaand the importance of growth to the survival of seeds and matureplants during submergence in the natural environment.Copyright1994, 1999 Academic Press Anoxia, ethanol, alcoholic fermentation, Oryza sativa L., rice, submergence     

Changes in cell wall composition of rice mesocotyls in relation to growth and treatment with 4-ethoxy-1-(p-tolyl)-s-triazine-2,6(1H, 3H)-dione

A synthetic substance, 4-ethoxy-l-(p-tolyl)-s-triazine-2,6(1H,3H)-dione [TA] dramatically promoted mesocotyl growth in dark-grownrice (Oryza sativa L. cv. Nato) seedlings, the optimal concentrationbeing 0.1–0.2 mM. Changes in the cell wall compositionof the rice mesocotyls were examined in relation to growth andtreatment with 0.1 mM TA. The amount of the cell wall increasedduring the elongation in control and treated mesocotyls. Particularly,TA caused a large increase in the amount of the cell wall permesocotyl but a decrease per unit length of mesocotyl. Hydrolysisof the cell wall with trifluoroacetic acid liberated xylose,glucose, arabinose, galactose, and trace amounts of rhamnose,fucose and mannose. An increase in the relative amount of xyloseand a decrease in that of glucose in the noncellulosic fractionduring growth were found in control and treated mesocotyl walls.On the 2nd day after planting when the mesocotyl emerged, TAsignificantly affected the cell wall composition; TA decreasedthe relative amount of -cellulose in the wall, and caused anincrease in the relative amount of glucose and decreases inthose of xylose and arabinose in the noncellulosic fraction. ¹This paper is Part 7 in the series "Plant Growth-regulatingActivities of Isourea Derivatives and Related Compounds." (Received March 18, 1980; )     

Mesocotyl elongation in japonica rice: Effect of high temperature pre-treatment and ethylene

Ethylene only slightly increased mesocotyl length in a japonicatype of rice, but when seeds were pre-treated with high temperature,added ethylene dramatically stimulated mesocotyl elongation.Removal of endogenously evolved ethylene apparently reducedmesocotyl length in seedlings. Elongation of cotyledonary noderoots was inhibited by ethylene. (Received November 25, 1971; )     

Coleoptile Senescence in Rice (Oryza sativa L.)

We investigated the cellular events associated with cell deathin the coleoptile of rice plants (Oryza sativa L.). Seeds germinatedunder submergence produced coleoptiles that were more elongatedthan those grown under aerobic conditions. Transfer of seedlingsto aerobic conditions was associated with coleoptile opening(i.e. splitting) due to death of specific cells in the sideof the organ. Another type of cell death occurred in the formationof lysigenous aerenchyma. Senescence of the coleoptile was alsonoted, during which discolouration of the chlorophyll and tissuebrowning were apparent. DNA fragmentation was observed by deoxynucleotidyltransferase-mediateddUTP nick end labelling (TUNEL) assay, and further confirmedby the appearance of oligonucleosomal DNA ladders in senescentcoleoptile cells. Two nucleases (Nuc-a and Nuc-b) were detectedby in-gel-assay from proteins isolated from coleoptiles. Nuc-a,commonly observed in three cell death phases required eitherCa²⁺or Mg²⁺, whereas Nuc-b which appeared during senescencerequired both Ca²⁺and Mg²⁺. Both nucleases were strongly inhibitedby Zn²⁺. Copyright 2000 Annals of Botany Company Aerenchyma, rice, cell death, coleoptile, fragmentation, nuclease, Oryza sativa, senescence, split, submergence, TUNEL     

Cooperation of Ethylene and Auxin in the Growth Regulation of Rice Coleoptile Segments

Elongation of coleoptile segments, having or not having a tip,excised from rice (Oryza sativa L. cv. Sasanishiki) seedlingswas promoted by exogenous ethylene above 0.3 µl l^–1as well as by IAA above 0.1 µM. Ethylene production ofdecapitated segments was stimulated by IAA above 1.0µM,and this was strongly inhibited by 1.0 µM AVG. AVG inhibitedthe IAA-stimulated elongation of the decapitated segment witha 4 h lag period, and this was completely recovered by ethyleneapplied at the concentration of 0.03 µl l^–1, whichhad no effect on elongation without exogenous IAA. The effectsof IAA and ethylene on elongation were additive. These factsshow that ethylene produced in response to IAA promotes ricecoleoptile elongation in concert with IAA, probably by prolongingthe possible duration of the IAA-stimulated elongation, butthat they act independently of each other. Moreover, AVG stronglyinhibited the endogenous growth of coleoptile segments withtips and this effect was nullified by the exogenous applicationof 0.03 µl l^–1 ethylene. These data imply that theelongation of intact rice coleoptiles may be regulated cooperativelyby endogenous ethylene and auxin in the same manner as foundin the IAA-stimulated elongation of the decapitated coleoptilesegments. Key words: oryza sativa, Ethylene, Auxin, Coleoptile growth     

Relationship between Coleoptile Elongation and Alcoholic Fermentation in Rice Exposed to Anoxia. I. Importance of Treatment Conditions and Different Tissues

The relationship between coleoptile elongation and alcoholicfermentation of rice under anoxia is examined using seeds either:(a) N₂ flushed during submergence, (b) incubated in stagnantdeoxygenated agar at 0·1% w/v to simulate the stagnantconditions of waterlogged soil, or (c) incubated in waterloggedsoil. Coleoptile elongation growth was greater for N₂ flushing> stagnant agar > soil; seed survival was also greatestin this order over 1-5 d. Ethanol concentrations in coleoptiles and intact seeds (cv.IR42) were approximately 300 and 100 mol m^-3 respectively whenseeds were grown 3 d in stagnant agar, however 92% of the ethanolin seeds diffused into the external medium when solutions weremixed for 5-10 s. Coleoptile growth under anoxia was relatedto rates of ethanol synthesis (R_E) in different treatments;there was greater coleoptile growth and R_E for seeds in N₂ flushedsolutions than in stagnant deoxygenated agar. Coleoptile growthof individual seeds was also related to the R_E of each seedat 2-3 d after anoxia (r² = 0·46). Analysis of different tissues was important in evaluating growthand metabolism of coleoptiles. Although the coleoptile onlyaccounted for 0·7% of seed dry weight at 3 d after anoxia,it contained 21% of the ethanol produced by rice seeds. Therewere also three-fold higher rates of R_E on a fresh weight basisin expanding tissues in the base of the coleoptile relativeto the elongated tissues at the apex. Results are discussedin terms of the importance of environmental conditions usedto impose anoxia, quantification of R_E in specific tissues andthe possibility that under stagnant conditions high ethanolconcentrations in tissues may limit R_E and coleoptile growth.Copyright1994, 1999 Academic Press Anoxia, ethanol, alcoholic fermentation, Oryza sativa L., rice, submergence     

Regulation of growth in rice seedlings

Etiolated rice seedlings (Oryza sativa L.) exhibited marked morphological differences when grown in sealed containers or in containers through which air was passed continuously. Enhancement of coleoptile and mesocotyl growth and inhibition of leaf and root growth in the sealed containers (enclosure syndrome) were accompanied by accumulation of CO₂ and C₂H₄ in and depletion of O₂ from the atmosphere. Ethylene (1 l 1^–1), high levels of CO₂, and reduced levels of O₂ contributed equally to the increase in coleoptile and mesocotyl growth. The effect of enclosure could be mimicked by passing a gas mixture of 3% O₂, 82% N₂, 15% CO₂ (all v/v), and 1 l l^–1) C₂H₄ through the vials containing the etiolated seedlings. The effects of high CO₂ and low O₂ concentrations were not mediated through increased C₂H₄ production. The enclosure syndrome was also observed in rice seedlings grown under water either in darkness or in light. The length of the rice coleoptile was positively correlated with the depth of planting in water-saturated vermiculite. The length of coleoptiles of wheat, barley, and oats was not affected by the depth of planting. In rice, the length of coleoptile was determined by the levels of O₂, CO₂, and ethylene, rather than by light. This regulatory mechanism allows rice seedlings to grow out of shallow water in which the concentration of O₂ is limiting.     

Effects of abscisic acid and its related compounds on rice seedling growth

Rice seedlings with the mesocotyl and coleoptile (the undeveloped leaves enclosed in the coleoptile) are here referred to as MC type seedlings and are considered to be suitable for deep sowing. We investigated the effects of abscisic acid (ABA) and several of its related compounds on the occurrence of MC type seedlings and on rice mesocotyl growth. Rice (Oryza sativa L. cv. JC 91) seedlings were grown on 0.8% agar medium in the presence or absence of various kinds of ABAs under aseptic conditions at 30 °C in the dark for 14 days. The activity of the R isomer of ABA (R-ABA) was slightly less than that of the naturally occurring S form (S-ABA) concerning the occurrence of MC type rice seedlings and the growth of the rice mesocotyl. In addition, the racemate of R-and S-ABA (RS-ABA) is less effective than R-ABA and S-ABA alone.Trans-ABA had no activity in relation to both percent occurrence of MC type seedlings and mesocotyl growth. The results of the present study suggest that the occurrence of MC type rice seedlings and the growth of rice mesocotyls were closely related to structure-activity relationships with analogs of ABA.     

Anaerobic conditions strongly promote extension by stems of overwintering tubers of Potamogeton pectinatus L

Elongation by dark-grown shoots of Potamogeton pectinatus tuberswas enhanced by the absence of oxygen. This promoting effectwas located in the stem and was stronger under water than ina gas phase and also stronger in unsparged water compared tosparged water. Anaerobic shoots elongated under water by almost13 cm in 5 d. This was the outcome of longer cells of the steminternodes and of some cell division coupled with leaf extensionwhich continued in the absence of oxygen, but at a slower rate.Continued attachment to a starch-filled tuber was required forsignificant anaerobic elongation, which could be sustained forat least 14 d. Switching intact, growing tubers from aerobicto anaerobic conditions stimulated stem extension within 24h. Conversely, stem elongation was slowed when tubers were transferredfrom an anaerobic to an aerobic environment. A marked gravitropicresponse occurred in anaerobic conditions, which involved bothstem and leaf tissue, and indicated that active internal growth-regulatingmechanisms continued to operate without oxygen. Shoot extension by tubers was also stimulated by hypo-aerobicconditions (5–8 kPa O₂) compared with fully aerated solutions(20.8 kPa O₂). This acceleration was smaller than that obtainedby the complete removal of oxygen, but still involved stem ratherthan leaf growth. Unlike elongation by apical shoots of tubers,that by shoot tips of rhizomes taken from mature light-grownplants was strongly inhibited by lack of oxygen, and in somecases shoot tips died within 5 d. All shoots and leaves werehighly aerenchymatous and the gas-filled lacunae were connectedby side-pores. Key words: Water plants, environmental stress, Potamogeton pectinatus, elongation, anaerobiosis, gravitropism     

Regulation of growth in rice seedlings

Etiolated rice seedlings (Oryza sativa L.) exhibited marked morphological differences when grown in sealed containers or in containers through which air was passed continuously. Enhancement of coleoptile and mesocotyl growth and inhibition of leaf and root growth in the sealed containers (“enclosure syndrome”) were accompanied by accumulation of CO₂ and C₂H₄ in and depletion of O₂ from the atmosphere. Ethylene (1 μl 1^?1), high levels of CO₂, and reduced levels of O₂ contributed equally to the increase in coleoptile and mesocotyl growth. The effect of enclosure could be mimicked by passing a gas mixture of 3% O₂, 82% N₂, 15% CO₂ (all v/v), and 1 μl l^?1) C₂H₄ through the vials containing the etiolated seedlings. The effects of high CO₂ and low O₂ concentrations were not mediated through increased C₂H₄ production. The enclosure syndrome was also observed in rice seedlings grown under water either in darkness or in light. The length of the rice coleoptile was positively correlated with the depth of planting in water-saturated vermiculite. The length of coleoptiles of wheat, barley, and oats was not affected by the depth of planting. In rice, the length of coleoptile was determined by the levels of O₂, CO₂, and ethylene, rather than by light. This regulatory mechanism allows rice seedlings to grow out of shallow water in which the concentration of O₂ is limiting.     

Mapping quantitative trait loci for seedling vigor in rice using RFLPs

Improving seedling vigor is an important objective of modern rice (Oryza saliva L.) breeding programs. The purpose of this study was to identify and map quantitative trait loci (QTL) underlying seedling vigor-related traits using restriction fragment length polymorphisms (RFLPs). An F₂ population of 204 plants was developed from a cross between a low-vigor japonica cultivar Labelle (LBL) and a high-vigor indica cultivar Black Gora (BG). A linkage map was constructed of 117 markers spanning 1496 Haldane cM and encompassing the 12 rice chromosomes with an average marker spacing of 14 cM. The length of the shoots, roots, coleoptile and mesocotyl were measured on F₃ families in slantboard tests conducted at two temperatures (18° and 25°C). By means of interval analysis, 13 QTLs, each accounting for 7% to 38% of the phenotypic variance, were identified and mapped in the two temperature regimes at a log-likelihood (LOD) threshold of 2.5. Four QTLs controlled shoot length, 2 each controlled root and coleoptile lengths and 5 influenced mesocotyl length. Single-point analysis confirmed the presence of these QTLs and detected additional loci for shoot, root and coleoptile lengths, these latter usually accounting for less than 5% of the phenotypic variation. Only 3 QTLs detected both by interval and singlepoint analyses were expressed under both temperature regimes. Additive, dominant and overdominant modes of gene action were observed. Contrary to what was predicted from parental phenotype, the low-vigor LBL contributed 46% of the positive alleles for shoot, root and coleoptile lengths. Positive alleles from the high-vigor parent BG were identified for increased root, coleoptile and mesocotyl lengths. However, BG contributed alleles with only minor effects for shoot length, the most important determinant of seedling vigor in water-seeded rice, suggesting that it would not be an ideal donor parent for introducing faster shoot growth alleles into temperate japonica cultivars.     

Effect of Ethylene and Culture Environment on Rice Callus Proliferation

Modifications to the gaseous envelope by callus during culturein Petri dishes were shown to reduce growth and promote necrosisof several rice (Oryza sativa L.) cultivars. Incubatingcallusunder a continuous flow of gas mixtures of known compositionsuggested that the inhibition of growth was caused by the accumulationof ethylene, the depletion of oxygen and, to a lesser extent,the accumulation of carbon dioxide. In order to evaluate theimportance of ethylene accumulation aminoethoxyvinylglycine(AVG), 1-aminocyclopropane-l-carboxylic acid (ACC and silvernitrate (AgNO₃), were added to the nutrient medium and ethylenemeasurements performed during callus culture. Ethylene restrictedcallus growth particularly under high (35 °C) as comparedto moderate (25 °C) temperatures and under illuminated ascompared to darkened incubation. Under illuminated incubationat 25 °C AVG (5 mmol m^–3) and AgNO°(50 mmol m^–3)significantly improvedcallus growth (100 and 60% respectively)while ACC (200 mmol m^–3) significantly decreased growth(40%). AVG and AgNO₃ were less effective under dark incubationat 25 °C where ethylene production was lower. Furthermore,callus growth was significantly better in large as comparedto small culture vessels since the ethylene concentration wasdiluted and more oxygen was available for respiration. Bettercontrol of ethylene and increased oxygen availability couldbe a way ofproducing healthy callus for the formation of embryogenictissues of otherwise recalcitrant cultivars of rice (e.g. IndicaIR42) and may be a way of improving manipulation of other cerealspecies. Key words: 1-Aminocyclopropane-1-carboxylic acid, aminoethoxyvinylglycine, callus, ethylene, Oryza sativa, silver nitrate