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     

Epidermal Structure in the Ligule of Rice (Oryza sativa L.)

The structure of the abaxial epidermis of the ligule of rice(Oryza sativa L.) as seen in the light and scanning electronmicroscope is described. Long cells, silica cells, cork cells,prickle hairs, microhairs and stomata-like structures were found.Reasons for considering these latter structures to be true stomataare given. Results of an X-ray analysis for silicon are presentedand the distribution of silica discussed in relation to itsfunctional role in the ligule. Oryza sativa L, rice, Gramineae, ligule, epidermis, scanning electron microscopy, X-ray analysis, silica, stomata     

Cortical cell death, cell proliferation, macromolecular movements and rTip1 expression pattern in roots of rice (Oryza sativa L.) under NaCl stress

The mode of action of NaCl in terms of cell proliferation and cell death was examined in seminal roots of rice plants (Oryza sativa L.). Salt/sodium chloride was inhibitory to cell number increase and to cell death in cortical tissue, whereas final cortical cell size was the same as in control roots that were not exposed to NaCl. It seems that NaCl may stimulate the transition phase from cell division to cell elongation. Further analysis of the role of NaCl in the suppression of cortical cell death was confined to a delay in the early stage of cell collapse, which was caused by tonoplast disruption, and plasma-membrane destruction. Sodium chloride did not have any effect on the cell-to-cell movement of macromolecules in the root cortex. In-situ hybridization studies indicated that expression of the gene for tonoplast intrinsic protein (rTip1) was localized predominantly in the epidermal and exodermal cells as well as in metaxylem cells in seminal roots. Upon NaCl treatment, the intensity of rTip1 gene expression was raised in the cortical parenchyma, suggesting that salt plays a role in the rapid onset of cell elongation. Received: 2 April 1998 / Accepted: 18 September 1998     

Transverse Vein Differentiation Associated with Gas Space Formation--Fate of the Middle Cell Layer in Leaf Sheath Development of Rice

In monocotyledons, the leaf vascular network consists of a hierarchicalsequence of vertical vascular bundles and numerous transverseveins that interconnect adjacent vertical veins. In the leafsheath of these species, especially grasses, lysigenous gascavities (gas spaces) are developed into intervascular spacesand provide a gas conducting system to non-aerial parts underflooded conditions. The spatial relationship between gas spaceformation and transverse vein differentiation was investigatedusing the leaf sheath of rice (Oryza sativa L.). Histochemicalobservation showed that patterns of differentiation of the transversevein are distinct from those of vertical vascular bundles. Onthe other hand, gas spaces are formed through the processesof cell death (collapse). Both events are initiated at a specificcell position in the middle layers of the leaf sheath, fromwhich the vascular system of the leaf is derived; this indicatesthat differentiation of transverse veins is associated withgas space formation. The cell-to-cell movement of fluoresceinisothiocyanate-conjugated dextran injected into middle layercells coincided with the area where cell collapse occurred,indicating a close relationship between the middle and adaxialcell layers, but not abaxial cell layers. A uniform cell numberbetween each transverse vein in the leaf sheath suggested theinvolvement of spatial regulation in transverse vein formationregardless of clonal history at the later stage of leaf veincanalization. Copyright 2000 Annals of Botany Company Cell collapse, leaf development, middle cell layer, microinjection, Oryza sativa L., rice, programmed cell death.     

Formation of Nodular Structures on Rice Seedlings by Rhizobia

Nodular structures have been induced on rice roots by treatingthe roots of 2-d-old seedlings with a cell wall degrading enzymemixture consisting of 1.0% cellulase YC, 0.1% pectolyase Y23,and 8% mannitol, followed by inoculation with rhizobia in thepresence of polyethylene glycol. Rhizobia were located bothwithin the degenerating cytoplasm of cells and between the cellsof spherical and elongate nodular structures. Although nitrogenaseactivity in such structures was at the limit of sensitivityof the assay procedure, this first report of the induction ofnodular structures on rice by rhizobia opens up the possibilityof extending effective nodulation to non-legumes, includingcereals, by facilitating the entry of rhizobia through cellwall degradation coupled with polyethylene glycol treatment. Key words: Rice (Oryza sativa L.), Rhizobium, cell wall degrading enzymes, polyethylene glycol, nodular structures, acetylene reduction     

Structural Differentiation of the Nucellar Epidermis in the Caryopsis of Rice (Oryza sativa)

In the developing caryopsis of rice (Oryza sativa L.) the nucellarepidermis forms a uniseriate layer through which assimilatesare transported to the endosperm. An anatomical study demonstratedthat the nucellar epidermal cells are fusiform in shape andare hexagonally packed. The anticlinal walls of the nucellarepidermis are characterized by ribs of wall-thickening whichare orientated radially with respect to the caryopsis. The wall-thickeningsappear to be cellulosic primary walls, as indicated by theirstaining with Calcofluor and periodic acid-Schiff's reagent.It is proposed that the geometry of the nucellar cells and theribs of wall-thickening are structural adaptations to resistthe compressional force which is placed on the nucellar epidermisduring the latter stages of grain filling. Oryza sativa, rice, caryopsis, grain filling, nucellar epidermis, wall-thickening     

Biochemical Basis of Different Sensitivity to Methotrexate in Daucus carota and Oryza sativa Cell Cultures

The biochemical basis of the different sensitivity to methotrexateof Daucus carota and Oryza sativa cell cultures has been investigated.Carrot cells have a dihydrofolate reductase activity about tentimes higher than rice cells. In addition, they show a loweruptake rate of the inhibitor. No relevant differences have beenfound in the K_m value for the dihydrofolate of the two enzymesand in the degree of inhibition of their activity by methotrexate. Key words: Dihydrofolate reductase, Methotrexate resistance, Plant cell suspension cultures, Oryza sativa, Daucus carota     

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     

Photothermal Responses of Flowering in Rice (Oryza sativa)

Durations from sowing to panicle emergence in 16 diverse genotypesof rice (Oryza sativa L.) were recorded in 13 different photothermalregimes, comprising constant and diurnally alternating temperaturesbetween 16 and 32 °C and photoperiods between 10.5 and 15.0h d^–1—all provided by controlled-environment growthcabinets. In 11.5 h days and at sub-optimal temperatures, relationsbetween the rate of progress towards panicle emergence and meantemperature were linear in all genotypes, and amongst thesethe base temperature at that photoperiod varied between 6.6and 11.9 °C. In most cases progress was most rapid at 24–26°C, i.e. the optimum temperature was much cooler than expectedfrom previously published values of times to panicle emergencein a less extensive range of photothermal regimes. Only in threecultivars was it warmer than 28 °C, and in these there weresufficient data to establish that relations between rates ofprogress to panicle emergence and photoperiod in the diurnallyalternating temperature regime of 28–20 °C are alsolinear. Also, the responses of these three cultivars provideno evidence of any interaction between the effects of photoperiodand temperature. We conclude, then, that the model in whichrate of development is a linear function of both temperatureand photoperiod with no interaction, which has been shown tobe common to many other species, also applies to rice. Differencesamong genotypes in relative sensitivity of rate of progresstowards panicle emergence to both temperature and to photoperiodwere considerable; japonica cultivars tended to be more sensitiveto temperature and less sensitive to photoperiod than indicacultivars. Four indica cultivars bred and selected at The InternationalRice Research Institute (IRRI) in the Philippines did not differ(P > 0.10) in their relations between rate of progress towardspanicle emergence and sub-optimal temperatures in a daylengthof 11.5 h, but the optimum temperature for cv. IR 36 was appreciablywarmer than that for the cvs IR 5, IR 8 and IR 42. Oryza sativa, rice, flowering, temperature, photoperiod, photothermal responses     

Lysigenous aerenchyma formation involves non-apoptotic programmed cell death in rice (Oryza sativa L.) roots

In waterlogged soil, deficiency of oxygen triggers development of aerenchyma in roots which facilitates gas diffusion between roots and the aerial environment. However, in contrast to other monocots, roots of rice (Oryza sativa L.) constitutively form aerenchyma even in aerobic conditions. The formation of cortical aerenchyma in roots is thought to occur by either lysigeny or schizogeny. Schizogenous aerenchyma is developed without cortical cell death. However, lysigenous gas-spaces are formed as a consequence of senescence of specific cells in primary cortex followed by their death due to autolysis. In the last stage of aerenchyma formation, a ‘spoked wheel’ arrangement is observed in the cortical region of root. Ultrastructural studies show that cell death is constitutive and no characteristic cell structural differentiation takes place in the dying cells with respect to surrounding cells. Cell collapse initiation occurs in the center of the cortical tissues which are characterized by shorter with radically enlarged diameter. Then, cell death proceeds by acidification of cytoplasm followed by rupturing of plasma membrane, loss of cellular contents and cell wall degradation, while cells nuclei remain intact. Dying cells releases a signal through symplast which initiates cell death in neighboring cells. During early stages, middle lamella-degenerating enzymes are synthesized in the rough endoplasmic reticulum which are transported through dictyosome and discharged through plasmalemma beneath the cell wall. In rice several features of root aerenchyma formation are analogous to a gene regulated developmental process called programmed cell death (PCD), for instance, specific cortical cell death, obligate production of aerenchyma under environmental stresses and early changes in nuclear structure which includes clumping of chromatin, fragmentation, disruption of nuclear membrane and apparent engulfment by the vacuole. These processes are followed by crenulation of plasma membrane, formation of electron-lucent regions in the cytoplasm, tonoplast disintegration, organellar swelling and disruption, loss of cytoplasmic contents, and collapse of cell. Many processes in lysing cells are structural features of apoptosis, but certain characteristics of apoptosis i.e., pycnosis of the nucleus, plasma membrane blebbing, and apoptotic bodies formation are still lacking and thus classified as non-apoptotic PCD. This review article, describes most recent observations alike to PCD involved in aerenchyma formation and their systematic distributions in rice roots.     

Time Lapse Analysis of Root Elongation Rates of Rice and Sorghum During the Day and Night

This paper describes a technique to monitor the root elongationrate (RER) per hour for several days, and variation in RER duringthe day and night. Rice (Oryza sativaL.) and sorghum (SorghumbicolorMoench) were grown in root boxes placed inside a growthchamber set at 25 °C with a 12 h photoperiod. Seminal rootaxes were sandwiched between a transparent acrylic board andfilter paper placed on a loamy sand soil. The roots were photographedunder dim green light using a CCD camera connected to a timelapse video recorder. The environment of the root, includingtemperature, light, nutrient, water and air supply, was controlledprecisely and maintained constant. RER fluctuated hourly insorghum and to a greater extent in rice. Maximum RERs were 1.4to 4.4 times faster than minimum rates. RERs during the dayand night did not differ statistically when temperatures werethe same.Copyright 1998 Annals of Botany Company Oryza sativaL., periodicity, root elongation,Sorghum bicolorMoench, time lapse.     

Short-term Salinity and High Temperature Stress-associated Ultrastructural Alterations in Young Leaf Cells ofOryza sativaL.

Salinity and high temperature stresses adversely affect growthand development of rice plants. To investigate the responseof rice cells to these stresses, we have analysed short-termstress-induced subcellular alterations in undifferentiated leafcells of rice seedlings by transmission electron microscopy.Perturbations noted particularly with respect to plasma membrane,mitochondrial membranes, endoplasmic reticulum, polyribosomesand dictyosomes are highlighted. The subcellular changes evokedby both stresses after 4 h were lysis of the cytoplasm, accumulationof electron-dense granules in the cytoplasm, distension in theER membranes, enhanced association of ribosomes with the endoplasmicreticulum, reduction in the number of mitochondrial cristae,as well as disorganization of cell wall fibrillar material.Certain changes were found to be unique to either the salinityor high temperature stress. Plasmolysis and increased cytoplasmicvesiculation were seen only in response to salinity stress,while discontinuity in the plasma membrane with close associationof the osmiophilic granules were observed only in response tohigh temperature.Copyright 1997 Annals of Botany Company Electron dense granules; high temperature stress; leaf cells; Oryza sativaL.; rice; salinity; ultrastructure     

Contrasting dynamics of radial O2-loss barrier induction and aerenchyma formation in rice roots of two lengths

Background and Aims

Many wetland species form aerenchyma and a barrier to radial O₂ loss (ROL) in roots. These features enhance internal O₂ diffusion to the root apex. Barrier formation in rice is induced by growth in stagnant solution, but knowledge of the dynamics of barrier induction and early anatomical changes was lacking.

Methods

ROL barrier induction in short and long roots of rice (Oryza sativa L. ‘Nipponbare’) was assessed using cylindrical root-sleeving O₂ electrodes and methylene blue indicator dye for O₂ leakage. Aerenchyma formation was also monitored in root cross-sections. Microstructure of hypodermal/exodermal layers was observed by transmission electron microscopy (TEM).

Key Results

In stagnant medium, barrier to ROL formation commenced in long adventitious roots within a few hours and the barrier was well formed within 24 h. By contrast, barrier formation took longer than 48 h in short roots. The timing of enhancement of aerenchyma formation was the same in short and long roots. Comparison of ROL data and subsequent methylene blue staining determined the apparent ROL threshold for the dye method, and the dye method confirmed that barrier induction was faster for long roots than for short roots. Barrier formation might be related to deposition of new electron-dense materials in the cell walls at the peripheral side of the exodermis. Histochemical staining indicated suberin depositions were enhanced prior to increases in lignin.

Conclusions

As root length affected formation of the barrier to ROL, but not aerenchyma, these two acclimations are differentially regulated in roots of rice. Moreover, ROL barrier induction occurred before histochemically detectable changes in putative suberin and lignin deposits could be seen, whereas TEM showed deposition of new electron-dense materials in exodermal cell walls, so structural changes required for barrier functioning appear to be more subtle than previously described.     

Storage-protein Deposition in the Developing Rice Caryopsis in Relation to the Transport Tissues

The developing caryopsis of rice (Oryza sativa L.) was examinedhistologically at successive stages of grain-filling in orderto identify the factors which determine the distribution ofstorage protein in the endosperm, and which terminate the depositionof endosperm protein. The storage protein was deposited at theperiphery of the endosperm, and this distribution was apparentlycaused by the radial pattern of cell development in the endosperm,and by the proximity of the peripheral endosperm cells to thenucellar epidermis. The nucellar epidermis directly surroundsthe endosperm and functions as the pathway for amino acid transportto the endosperm. During the later stages of caryopsis developmentthe nucellar epidermis became compressed by being ‘sandwiched’between the expanding endosperm and the rigid hull (the tightlylocked palea and lemma) which encloses the caryopsis. It isproposed that this compression of the nucellar epidermis blocksthe supply of amino acids to the endosperm and thereby terminatesthe deposition of storage protein in the rice grain. Oryza sativa, rice, caryopsis (development), endosperm, grain filling, nucellar epidermis, storage protein     

Durations of the Photoperiod-sensitive and Photoperiod-insensitive Phases of Development to Flowering in Four Cultivars of Rice (Oryza sativa L.)

The durations of the photoperiod-sensitive and photoperiod-insensitivephases of development to panicle emergence were estimated infour contrasting indica cultivars of rice (Oryza sativa L.)in a reciprocal-transfer experiment. Plants were grown in potsin glasshouses maintained at warmer (32/26 C) or cooler (28/20C) day/night temperatures, and the durations from sowing topanicle emergence were determined for plants moved from relativelyshort (11 h) to relatively long (13.5 h) days and vice versaat various times after sowing. Panicle emergence was delayedby long days in all cultivars, but the traditional cvs Carreonand Peta were much more sensitive to photoperiod than the moderncvs IR8 and IR36 The durations of the photoperiod-insensitivepre-inductive phase (equivalent to some definitions of the basicvegetative phase) varied from 14.4 d in cv. Carreon at 32/26C to 42.0 d in cv. IR8 at 28/20 C. In all cultivars this initialphase was of a longer duration in the cool than in the warmregime. The duration of the photoperiod-insensitive post-inductivephase was also consistently greater, but usually only slightso, at cool than relatively warm temperatures; it varied from6.8 d in cv. IR8 at 32/26 C to 272 d in cv. Carreon at 28/20C. As expected, the length of the intervening photoperiod-sensitiveinductive phase was greater in long days, but the effect oftemperature on these durations was not consistent; for example,these durations were longer in warm than in cool temperaturesin cv. 1R8 but, if anything, they were slightly longer in coolthan in warm temperatures in cv. IR36. This difference is compatiblewith previous findings that cv. IR36 has a warmer optimum temperaturefor rate of progress towards panicle emergence than cv. IR8.A subsequent reciprocal-transfer experiment with cv. Peta providedestimates of the durations of the photoperiod-insensitive andphotoperiod-sensitive phases of pre-flowering development whichwere compatible with our earlier estimates. Furthermore, panicleinitiation was found to occur after about 80% of the photoperiod-sensitiveinductive phase had elapsed. We conclude that although the durationof the photoperiod-insensitive pre-inductive phase in rice isgreater than in many other annual crops, genotypic variationin this duration may well be less than was previously deduced.We also conclude that, despite common assumptions to the contrary,photoperiod-sensitivity during rice plant development does notend at panicle initiation. Oryza sativa L., rice, panicle initiation, panicle emergence, photoperiodism, temperature     

Transformation of Indica Rice (Oryza sativa L.) Mediated by Agrobacterium tumefaciens

A system has been developed for the transfer of genes by Agrobacteriumtumefaciens to indica rice (Oryza sativa L. cv. Taichung Native1), and optimal conditions for such Agrobacterium-mediated transformationhave been determined. Excised stems, leaves, and roots from3- to 4-day-old seedlings were infected with strains of Agrobacteriumthat carried plasmids with chimeric derivatives of genes forß-glucuronidase (GUS) and neomycin phosphotransferase(NPTII). After selection with the antibiotic G418, only rootssurvived and formed calli. The integration of chimeric genesin the rice genome was demonstrated by Southern analysis andassays of enzymatic activity. We obtained a transformation frequencyof 37.5% for transgenic callus. In addition, the absence ofhybridization with vir probes indicated that transformants werenot contaminated with Agrobacterium. The complete failure oftransformation in the absence of medium from potato suspensioncultures (PSC) indicates that PSC is essential for Ti-mediatedtransformation in indica rice. A high efficiency of transformationwas observed upon infection with the cointegrate vector pGV2260::NG1. (Received August 13, 1991; Accepted April 28, 1992)     

Comparison of Growth Responses of Barnyard Grass (Echinochloa oryzoides) and Rice (Oryza sativa) to Submergence, Ethylene, Carbon Dioxide and Oxygen Shortage

Unlike germination of wheat (Triticum aestivum), millet (Eleucinecoracana), and sorghum (Sorghum caudatum), that of Echinochloaoryzoides (barnyard grass) and Oryza satwa (rice) was not inhibitedby poorly aerated solutions with 11 k Pa oxygen (equilibriumpartial pressure) or less In the dark, seedling shoots of riceincluded a coleoptile, and in Echinochloa, a mesocotyl alsoGrowth in fresh and dry weight of shoots was strongly depressedby poorly aerated solutions in both rice and Echinochloa butthe effects on extension differed in the two species in rice,coleoptile extension was promoted by solutions partly depletedof oxygen, and also by the absence of oxygen The stimulationin partly de-oxygenated solutions resulted from the combinedpromoting effects of small oxygen partial pressures, carbondioxide, ethylene and buoyant tension in contrast, these treatmentsneither promoted nor inhibited elongation by the Echinochloacoleoptile while severely inhibiting extension of the mesocotyl Overall, poorly aerated solutions lengthened the shoot of riceand shortened it in Echinochloa when compared with those submergedin well-aerated solutions These opposite effects were broughtabout by the same gaseous changes, i e oxygen shortage, elevatedethylene and carbon dioxide The effect on Echinochloa was almostentirely restricted to the mesocotyl, coleoptile extension beingremarkably insensitive to large increases in ethylene and carbondioxide, or to extreme oxygen shortage Seedlings of the twospecies thus have contrasting strategies for survival Stress, submergence, ethylene, oxygen shortage, carbon dioxide, adaptation, anaerobiosis, rice (Oryza sativa L), barnyard grass [Echinochloa oryzoides (Ard) Fritseh]     

The involvement of wheat and common bean lectins in the control of cell division in the root apical meristems of various plant species

The effects of wheat germ agglutinin (WGA) and phytohemagglutinin (PHA) at the concentration of 1 mg/l on the rate of cell division in the root apical meristem of wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), rice (Oryza sativa L.), and common bean (Phaseolus vulgaris L.) seedlings were compared. WGA enhanced cell division in the roots of barley and rice approximately similarly as in wheat roots but did not affect division of meristematic cells in the roots of common bean seedlings. In contrast PGA enhanced mitotic activity in the root apical meristem of common bean seedlings but did not affect division in the wheat and barley roots. Seedling treatment with lectins shifted the hormonal balance in them toward accumulation of growth activators (IAA and cytokinins). The relationship between lectin and hormonal systems in the control of cell division is discussed.     

Structural Aspects of Cluster Root Development and their Possible Significance for Nutrient Acquisition in Grevillea robusta (Proteaceae)

Light microscopy (LM), scanning (SEM) and transmission electronmicroscopy (TEM) were used to study structure and function ofcluster roots inGrevillea robusta . These roots were developedduring growth ofG. robustaseedlings in modified Hoagland's solutionlacking phosphate. Cluster rootlets formed root hairs, basipetally,only after completing their determinate development. The rootlethairs branched in two ways and some had apical swelling. Rootletswith hairs produced two different forms of exudate, one fibrousand the other globular in nature. The fibrous material appearedto be synthesised in the cortical cells. It is released by exocytosisfrom the epidermis. Rootlet hairs produced only fibrous exudate.They attached firmly to pieces of vermiculite. The significanceof cluster roots is discussed within the context of patchy soilresources. Grevillea robusta ; Silky oak; Proteaceae; cluster roots; morphology; mucilage; nutrient acquisition; root exudates     

Factors influencing plant regeneration from protoplasts isolated from long-term cell suspension culture of recalcitrant Indica rice cultivar IR36

Summary Factors influencing successful establishment of embryogenic cell-suspension cultures and plant regeneration from longterm cell suspension-derived protoplasts of the recalcitrant Indica rice cultivar IR36 were studied. The factors included cell and protoplast culture medium, protoplast culture procedure, the source of nurse cells, and the regeneration procedure. Embryogenic cell suspension cultures could only be established from mature seed-derived callus of IR36 in AA-based medium (Müller and Grafe, 1978). Protoplast-derived colonies could be obtained only using the filter-membrane nurse-culture procedure when Lolium multiflorum suspension cells served as nurse, rather than wild rice (Oryza ridleyi) and Japonica rice (Oryza sativa cv. Taipei 309) cells. The utilization of a two-step regeneration procedure led to regeneration of fertile plants from protoplasts isolated from 2-yr-old cell suspensions of IR36, one of the most important but recalcitrant rice cultivars.