Ethylene and in vitro rooting of hazelnut (Corylus avellana) cotyledons

Ethylene may be one of the many factors that play a role in rooting. However, in some studies ethylene promoted rooting, while in others it was inhibitory or had no effect. Using cotyledons of hazelnut ( Corylus avellana L. cv. Casina) observations were made of the effect of ethylene precursors on adventitious root formation. l-methionine (Met) or 1-aminocyclopropane-1-carboxylic acid (ACC) added to a standard indole-3-butyric acid (IBA)-kinetin-containing medium did not enhance rooting, while 2-chloroethylphosphonic acid (CEPA) did. The ethylene inhibitor, aminoethoxyvinylglycine (AVG), inhibited root formation, but its effect was reversed by ACC when cotyledonary segments were transferred to rhizogenic medium plus ACC at day 10. Ethylene production by cotyledons cultured on rhizogenic medium or rhizogenic medium plus CEPA was high at the beginning of rooting. Thus, the wound-induced ethylene is a key stimulatory factor in the formation of root primordia. The data support the hypothesis that ethylene plays a positive role in root formation.     

Kinetic properties of IAA oxidase from mung bean cotyledons

A crude enzyme preparation from mung bean cotyledons was separated into peroxidative and non-peroxidative IAA oxidase on a DEAE-cellulose column. Both fractions differed in their pH optima, K_m and V_max. The K_m and V_max of non-peroxidative IAA oxidase were higher than those of peroxidative IAA oxidase. Peroxidative IAA oxidase showed a linear increase in absorption at 247 and 254 nm after a short lag of 2–3 min. The addition of catalytic amounts of hydrogen peroxide eliminated the lag period and also enhanced the rate of IAA degradation. The non-peroxidative IAA oxidase fraction, however, did not exhibit any significant increase in absorption at 247 and 254 nm and showed a lag period of 5 min which was not affected by hydrogen peroxide. Instead, addition of the same catalytic amount of hydrogen peroxide inhibited the rate of IAA degradation. The peroxidative IAA oxidase fraction exhibited the reaction kinetics characteristic of peroxidase-catalysed IAA degradation. The rate of IAA oxidation by purified non-peroxidative IAA oxidase was very low. The slow rate of catalysis shown by non-peroxidative IAA oxidase appears to be due to the presence of inhibitor(s).     

Hormonal changes associated with leaf senescence in cotton (Gossypium barbadense)

To study the hormonal regulation of foliar senescence in cotton ( Gossypium barbadense L. cv. Giza 68, long staple), the sequential changes in gibberellins (GAs), indoleacetic acid (IAA) and abscisic acid (ABA) were examined in the cotyledons from the completion of expansion through senescence (days 12-24 after sowing). The onset of senescence could be detected on day 20, the stage of maximum accumulation of leaf metabolites. At this stage, free GAs quickly lost more than 40% of their initial activity. Further decrease of free GAs was then characteristically observed in the senescent leaves. A remarkable increase in free IAA and free ABA between days 18 and 20 immediately followed by nutrient depletion, suggests the contribution by both hormones to the senescence system. The definite drop in free IAA below its initial level occurred on day 24, when most of the leaf protein and chlorophyll were already broken down.     

Fatty acids of callus tissues of six species of cucurbitaceae

A comparative study was made of the fatty acid composition of the total lipids extracted from the cotyledons and the callus cultures derived from cotyledon segments of six species of Cucurbitaceae. Conditions for callus induction and growth of cultures were identical. The difference between the two systems was in the reversal of the ratio of total unsaturated to saturated acids in all callus cultures. In callus cultures, instead of linoleic, linolenic was the major unsaturated fatty acid. In Momordica charantia, α-elaeostearic acid present in the cotyledon was not detected in callus and oleic acid was the major unsaturated fatty acid.     

绿豆子叶衰老期间核酸代谢的变化

萌发绿豆的子叶自然衰老期间,核酸含量降低,RNA降低的幅度比DNA大。电泳分析结果表明,子叶衰老期间细胞核主带DNA明显降低;而迁移慢的卫星带DNA变化不大。在RNA各组分中,18S rRNA从衰老前期就开始降低;25S rRNA和4～5S小分子RNA到衰老后期才缓慢下降。DNase和RNase活性在子叶整个衰老期间都明显升高,是导致核酸含量下降的主要原因。~3H-核苷掺入试验表明,核酸的合成速率在子叶衰老前期有所上升,到衰老后期又降低。poly(A)~ -mRNA含量在子叶开始衰老时明显上升。     

Mitochondrial malate dehydrogenase from watermelon: sequence of cDNA clones and primary structure of the higher-plant precursor protein

The isolation and sequence of a cDNA clone encoding the complete mitochondrial malate dehydrogenase (mMDH) of watermelon cotyledons is presented. Taking advantage of the polymerase chain reaction technology partial cDNA clones from the central part, the 3 part and the 5 part of the mRNA were obtained with oligonucleotides based on directly determined amino acid sequences. Subsequently, two complete cDNA clones for mMDH were synthesized with a sense primer corresponding to the nucleotide sequence of the amino terminal end of pre-mMDH and two antisense primers corresponding to the major alternative adenylation sites found in the mRNA.The amino acid residues for substrate and cofactor binding identified by X-ray crystallography for pig heart cytoplasmic MDH are conserved in the 320 amino acid long mature higher-plant mMDH. A presequence of 27 amino acids is present at the amino terminal end of the precursor protein.     

Benzyladenine induces the appearance of LHCP-mRNA and of the relevant protein in dark-grown excised watermelon cotyledons

Cotyledons were excised from imbibed watermelon seeds, grown for 4 days in darkness on water or 10 M benzyladenine (BA) and then tested for the presence of the light-harvesting chlorophyll a/b protein (LHCP) and its mRNA. LHCP was assayed immunologically by western blotting of SDS gels: the protein was present in plastids, but it was not recovered with the thylakoid fraction. Antibodies directed against LHCP precipitated a 32 kDa polypeptide from translation products of poly(A) RNA of cotyledons only if these had been grown on BA. Taken together the data suggest that in absence of light cytokinins are necessary for the maintenance of a detectable level of LHCP-mRNA as well as for synthesis of the protein.     

Non-morphogenic metabolism during shoot induction in radiata pine cotyledon explants

Summary The effects of excision, light and cytokinin (N⁶-benzyladenine) on¹⁴C-acetate metabolism in cotyledons ofPinus radiata (D. Don) were determined.¹⁴CO₂ was released and the distribution of radioactivity into lipids, sugars, organic acids and amino acids was determined. While light and cytokinin generally caused some increase in metabolism, the effect of excision, i.e., wounding, was most pronounced. Specific metabolites examined (citrate, malate, succinate, alanine, aspartate, glutamate and glutamine) were at least 50% greater in¹⁴C-labeling in excised cotyledons as compared to intact seedlings. This enhancement of wound metabolism would mask possible morphogenically-related changes occurring at that time. This research was supported by the Natural Sciences and Engineering Research Council of Canada Grant A-6467 to T.A. Thorpe.     

Assays for the effects of volatile compounds from artificially damaged cotyledons of subterranean clover on the redlegged earth mite, Halotydeus destructor

The effects on the redlegged earth mite (Halotydeus destructor) (Acarina: Penthaleidae) of volatile compounds released from artificially damaged cotyledons of subterranean clover (Trifolium subterraneum L.), a host plant, were investigated with different assays. Mites were repelled by the volatile compounds in flask tube and in trimmed cotyledon disc tests. No differences could be shown between different tissue amounts and between resistant and susceptible varieties. When a membrane sachet was used containing either 2-(E)-hexenal, a compound produced by damaged subclover cotyledons, or the total volatile compounds collected from damaged cotyledons of Dalkeith (susceptible) admixed with 1% glucose, mites gathered to low but were deterred by high concentrations of the compounds. Volatile compounds collected from the resistant variety DGI007 were more deterrent than those from the susceptible Dalkeith. Membrane sachets containing 30 p.p.m. of 2-(E)-hexenal and 1% glucose were less preferred than cotyledons of Dalkeith (susceptible) but were not different from DGI007 (resistant). By increasing the glucose concentration to 5%, the membrane sachets with 30 p.p.m. of 2-(E)-hexenal were preferred to cotyledons of either variety. The results indicate that the membrane feeding technique provides a sensitive way of assaying volatile compounds from damaged subclover cotyledons against the mite.     

Turgor-dependent unloading of photosynthates from coats of developing seed of Phaseolus vulgaris and Vicia faba. Turgor homeostasis and set points

Key physiological characteristics of turgor-dependent efflux of photosynthates were examined using excised coats and cotyledons of developing Phaseolus vulgaris (cv. Redland Poineer) and Vicia faba (cv. Coles Prolific) seed during the linear phase of seed fill. Exposure to solutions of high osmotic potential inhibited net uptake of [¹⁴C]sucrose by cotyledons at developmental stages less than 60% of their final dry weight. The effect could not be fully reversed by transferring cotyledons to solutions set at lower osmotic potentials. The inhibition became apparent at osmotic potentials that were higher than those that caused stimulation of efflux from seed coats. Net [¹⁴C]sucrose uptake by cotyledons at more advanced stages of development was unaffected by external osmotic potential. Specified tissue layers were removed from seed coats by pretreatment with pectinase. Efflux studies with the pectinase-modified coats of Phaseolus and Vicia seed demonstrated that the cellular site of turgordependent efflux was the ground parenchyma and thin-wall parenchyma transfer cells, respectively. Coats subjected to long-term (hours) incubations, under hypo-osmotic conditions, exhibited the capacity for turgor regulation. This was mediated by turgor-dependent efflux of solutes. The solutes exchanged were of nutritional significance to the developing embryo. The relationship between efflux and coat turgor was characterised by a turgor-independent phase at low turgors. Once turgor exceeded a minimal value (set point), efflux increased in proportion to the magnitude of the turgor deviation (error signal) from the set point. For coats of sink-limited seed of Vicia and Phaseolus, efflux exhibited apparent saturation at turgors above 0.25 and 0.5 MPa respectively. The putative turgor set point and slope of the turgor-dependent component of efflux varied with seed development, the prevailing source/sink ratio and genetic differences in seed growth rate. The nature of these kinetic variations was compatible with the competitive ability of the seed. A turgor homeostat model is proposed that incorporates the observed functional attributes of turgor-dependent efflux. Operationally, the model provides a mechanistic basis for the integration of assimilate demand by the cotyledons with assimilate import into and unloading from the seed coat.