A Physical Explanation for Solute Leakage from Dry Pea Embryos During Imbibition

The pattern of solute leakage from imbibing dead pea (Pisumsativum L.) embryos was the same as that from living embryos,with an initially high leakage declining to a low constant rateof leakage in the first 3 min of imbibition. The same patternof leakage occurred during each imbibition phase of repeatedimbibe/dry cycles of dead embryos. Living and dead seeds alsoshowed this pattern of leakage. These observations are usedto argue that leakage during imbibition of embryos and seedsis a physical diffusion phenomenon. Vital staining of livingembryos after imbibition revealed positive staining for dehydrogenaseenzymes in the cells on the outer surface of the cotyledonsonly when 0.5 mM sodium succinate solution was present duringimbibition and/or staining. This is discussed in relation tothe effect of rapid water uptake on these cells.     

Possible involvement of aquaporins in water uptake by pea seeds differing in quality

Using the method of room temperature phosphorescence (RTP), we divided air-dry pea (Pisum sativum L.) seeds subjected to accelerated ageing (40°C, 85% relative humidity) into three fractions: (I) high-quality seeds, (II) weakened seeds, and (III) dead seeds. In the process of ageing, seed germinability firstly decreased and then increased due to so-called “improved” seeds of fraction II, which returned to fraction I as judged from the RTP level; the germinability of these seeds became equal to that of fraction I seeds. Seeds capable of germination (fractions I and II) differed in the rates of imbibition, which depended on plasma membrane permeability (opened or closed water channels) but not on the presence of the seed coat. A low activation energy of seed imbibition in fraction II (less than 5 kcal/mol) indicates that water channels are open. A mercury-containing compound (5 μM p-chloromercuribenzoate (PCMB) reduced the rate of water uptake by these seeds, and dithiothreitol restored it. A high activation energy of fraction I seed imbibition (more than 12 kcal/mol) corresponded to the water uptake mainly across the lipid bilayer when water channels are closed. PCMB did not affect the rate of fraction I seed imbibition. We supposed that mature air-dry pea seeds had open water channels. During the first stages of fraction I seed imbibition, these channels were closed, limiting water uptake. NaF (100 μM), an inhibitor of phosphatase, prevented channel closing and accelerated the imbibition of fraction I seeds. It did not affect the imbibition rate of fraction II seeds, indicating their water channels to be opened. However, NaF did not affect the water uptake of “improved” fraction II seeds as well. It seems likely that their channels were closed during accelerated ageing but otherwise than via dephosphorylation. The results obtained indicate the possibility of water inflow regulation in the weakened seeds via the state of aquaporins, which form water channels in the membranes.     

Chilling Stress to Soybeans during Imhibition

Embryos, excised from seed coats of soybeans (Glycine max Merr. cv. `Wayne'), leak profusely during the first minutes of imbibition. A discontinuity of temperature/leakage patterns occurs between 10 and 15 C; as embryos imbibe at 10 C or lower, disproportionately more solutes leak out per unit of water imbibed. Short periods of imbibition at or below 12 to 14 C reduce embryo germination and axis elongation; injury results from imbibition at 2 C for as little as 5 minutes. Humidifying embryos to 35 to 50% moisture before imbibition reduced leakage during imbibition and imparted some resistance to imbibitional chilling injury.

The period of profuse leakage is interpreted as a time of membrane reorganization. Imposing a low temperature during this period prolongs the rapid leakage, suggesting delayed or faulty membrane reorganization. Reduced cold sensitivity of embryos with an initial 35 to 50% moisture content is presumed to be due to at least partial membrane reorganization in the embryo before imbibition. These data collectively are taken to indicate that low temperature interferes with normal membrane reorganization during imbibition, probably by modifying the physical state of membrane phospholipids, and that the consequent abnormal organization of membranes is a basic cause of low temperature injury.

     

Partial Sequence Analysis of the 5S to 18S rRNA Gene Region of the Maize Mitochondrial Genome

Measurements of the rate of imbibition by isolated cotyledons of soybean (Glycine max (L.) Merr cv Wayne) indicated that water uptake rates were slowed by low temperatures and by low initial moisture contents of the tissue. The role of water viscosity in the temperature effects on imbibition was examined, and a linear relation between imbibition rate and the reciprocal of viscosity was found only for seeds of very high initial moisture content. Adding solutes which lowered the surface tension, or increased the wetting ability of the water, yielded markedly increased rates of imbibition, especially for tissue of low initial moisture content. The data are interpreted as indicating a first component of water entry which is a wetting reaction influenced by the surface tension of the water, and a second component which resembles water flow through a porous matrix and is influenced by the water viscosity. We speculate that damage to soybeans during imbibition, such as in the case of chilling injury, may be particularly related to the initial wetting reaction rather than to the longer-term imbibitional rate.     

Temperature effects on oxidative metabolism of dormant sugar pine seeds

When dormant sugar pine (Pinus lambertiana L.) seeds were imbibed at 5°C, they showed a rapid increase in O₂ uptake, ATP level, and moisture content during the first 4 days. This was followed by a plateau phase until 60 days, after which a second significant increase in all three features occurred as dormancy was broken. During the plateau phase, conventional CN-sensitive respiration accounted for 74 to 79% of the total O₂ uptake. When dormant sugar pine seeds were imbibed at and maintained at 25°C, a different pattern occurred. Water uptake was much more rapid during the first 4 days and no second increase occurred after 60 days because the seeds did not break dormancy. There was an initial burst of O₂ uptake and ATP formation, but these both declined abruptly after 24 to 48 hours. Levels about half those of seeds at 5°C were maintained through the rest of a 90-day period. CN-sensitive respiration declined during imbibition at 25°C, and accounted for only 55 to 61% of the total O₂ uptake. The inability of dormant sugar pine seeds to germinate at temperatures above about 17°C may therefore result from initial temperature effects on membrane properties, leading to reduced O₂ uptake, reduced cytochrome oxidase electron transport activity, and lowered ATP levels.     

Effect of temperature on glycerol metabolism in membranes and on phospholipases C and D of germinating pea embryos

The effect of temperature of imbibition on the synthesis and turnover of membrane phosphatidyl choline was studied. Pea seeds (Pisum sativum cv. Alaska) were imbibed in [U-¹⁴C]glycerol and then germinated. Seeds were kept constantly either at 5° or 25°, or were imbibed at one temperature and then germinated at the other one. Glycerol incorporation into phosphatidyl choline in the ER and the plasma membrane, obtained from the embryonic axes after germination, and the glycerol pool were measured. Embryos from seeds kept constantly at 25° showed a rapid incorporation of glycerol into membranes followed by a loss of label; in embryos from seeds kept at 5° incorporation was much lower. Embryos from seeds transferred from 25° to 5° behaved as if continuously kept at 25°, while the behaviour of the embryos from seeds transferred from 5° to 25° resembled embryos from seeds maintained at 5°. The glycerol content of the axes rose during imbibition and fell thereafter. The activities of phospholipases C and D also responded to the initial temperature of imbibition, but the two activities changed differently. The results are discussed in relation to the effect of transient exposure to temperature changes in the seed membranes and the possible way in which such changes are sensed.     

The Importance of Genetically Determined Seed Coat Characteristics to Seed Quality in Grain Legumes

Comparisons of five pairs of isogenk lines of peas, differingonly in the A gene for seed coat colour showed that white seeds(genotype aa) imbibed more rapidly than coloured seeds (AA),suffered greater imbibition damage revealed by dead tissue onthe cotyledons, and higher solute leakage. Seed-coat pigmentationwas closely associated with slow water uptake, since when expressionof the A gene was suppressed by the recessive pollens gene,the resulting white seeds {palpal AA) imbibed rapidly. The slowwater uptake by coloured seeds was not due to the restrictionof water entry by the seed coat since the differences in imbibitionrate were maintained when a portion of the seed coat was removedand seeds were imbibed with the exposed cotyledon in contactwith moist filter paper. Imbibition of similarly treated seedsby immersion in polyethylene glycol solutions (1–4%) whichincreased the seed/solution wettability, had little effect onthe water uptake of coloured seeds compared to imbibition inwater whereas that of white seeds increased in the first 10mins imbibition. Poor wettability of the inner surface of colouredseed coats did not therefore explain the slow imbibition ofthese seeds. The white seed coats loosened rapidly during imbibitionwhilst the coloured seed coats remained closely associated withthe cotyledons suggesting that the adherence of the seed coatto the cotyledons and therefore the ease of access of waterbetween the testa and cotyledons determines the rate of imbibition.The rapid water uptake by white-coated seeds and the subsequentimbibition damage may explain the high incidence of infectionof these seeds by the soil-bome fungus Pythhan after 2 d insoil. Improved seed quality and emergence may therefore be achievedby breeding for seed coat characteristics leading to reducedrates of imbibition Pisum sativum, isogenic lines, A gene, seed coat colour, imbibition, imbibition damage, wettability, pollens gene, seed quality, grain legumes     

Leakage during Seed Imbibition

When pea embryos are placed in water solutes leak from themat a rate which declines rapidly at first and then more slowly.Embryos can be dried down over calcium chloride and will thenleak as before when returned to water. Similar results wereobtained with Ricinus seeds after removal of the testa. Pea embryos that have first been allowed to imbibe some waterthrough a small part of their surface (by placing them on dampfilter-paper) leak relatively slowly when subsequently immersedin water; the greater the initial imbibition the slower thesubsequent leakage. Likewise, embryos taken from peas that wereharvested when succulent and tender show only slow leakage It is proposed that as seeds dry out in the course of developmentcell membranes lose their integrity. When such dry seeds areallowed to imbibe water there is a short period, before membraneintegrity is re-established, during which solutes can leak outof the cells.     

Role of the testa in preventing cellular rupture during imbibition of legume seeds

Studies with the seeds of soybean, navy bean, pea, and peanut were made to determine the extent of leakage of intracellular enzymes during imbition. Embryos with intact testae from all four species were found to leak detectable activities of either intracellular enzymes of the cytosol (glucose-6-phosphate dehydrogenase) or enzymes found in both the cytosol and organelles (malate dehydrogenase, glutamate dehydrogenase, glutamate oxaloacetate transaminase, and NADP-isocitrate dehydrogenase) after 6 hours imbition at 25 C. Pea and peanut embryos with testae leaked considerably lower levels of activity for these enzymes than did those of soybean and bean. Leakage of mitochondrial marker enzymes (fumarase, cytochrome c oxidase, and adenylate kinase) was not detected from embryos with testae, suggesting that a differential diffusion of intracellular components out of cells occurred. Soybean and bean embryos without testae leaked high, and proportionally (per cent dry seed basis) similar, levels of all cytosol, cytosol-organelle, and mitochondrial marker enzymes and protein during imbibition, indicating that cell membranes were not differential to leakage and that they had ruptured. Pea and peanut embryos without testae leaked detectable activities of all cytosol and cytosol-organelle enzymes, although fumarase was the only detectable mitochondrial marker enzyme leaked, suggesting that some degree of differential leakage may have occurred in these species. The outermost layers of embryo cells of seeds without testae of all four species absorbed and sequestered the nonpermeating pigment Evan's blue after 5 to 15 minutes imbibition, indicating that membranes had ruptured. This occurred to a much lesser extent in seeds with intact testae. Both soybean and bean embryos without testae were observed to disintegrate during imbibition, whereas those of pea and peanut did not. These data indicate that seeds of certain legumes are susceptible to cellular rupture during imbibition when seed coats are damaged or missing.     

The Role of Imbibition Damage in Determining the Vigour of White and Coloured Seed Lots of Dwarf French Beans (Phaseolus vulgaris)

Seeds of dwarf French beans showed evidence of damage alterimbibition in water in the form of dead tissue and high soluteleakage. Damage was more extensive in seeds of a cultivar withwhite testae (Tenderette) both at 20?C and with the initial6 h imbibition at 4?C where damage was more severe, whereasa cultivar with brown testae (Provider) was damaged only alterthe 4?C treatment. Tenderette imbibed more rapidly than Providerin both imbibition treatments. The significant correlationswithin each cultivar of both solute leakage (positive) and theextent of living tissue (negative) with the rate of water uptake,and the reduced damage when seeds imbibed slowly in PEG indicatedthat this was imbibition damage due to rapid water uptake. Differencesin the rates of water uptake were also observed in sand at 15%and 20% mc where rapid imbibition was associated with reducedgermination and the production of more abnormal seedlings. Atemperature of 4?C during the first 6 h germination in sandat 15% mc also reduced germination, particularly in Tcnderette.It is suggested that the more rapid water uptake by white seededcultivars of dwarf French beans and their greater susceptibilityto imbibition damage are responsible for their low vigour. Key words: Imbibition damage, vigour, dwarf French beans     

Hypoxia and Imbibition Injuries to Aging Seeds

The development of hypoxia and primary injuries were examined during the imbibition of aging pea seeds (Pisum sativum L., cv. Nemchinovskii). The distribution of air-dry pea seeds by their room-temperature phosphorescence revealed the presence of two fractions (I and II) in a seed lot with 72% germinability and three fractions (I, II, and III) in a seed lot with 50% germinability. The water uptake during imbibition was slower in the fraction I seeds than in the fraction-II seeds. The fraction-I seeds produced normal seedlings, whereas the fraction-II seeds either produced seedlings with morphological defects (abnormal) or did not germinate at all. The fraction-III seeds were all dead. The phosphorescence of endogenous porphyrins, emitted only at low O₂ content, was measured after 20-h seed imbibition. The fraction-I seeds emitted no discernible phosphorescence. The fraction-II comprised highly phosphorescent seeds incapable of radicle protrusion and moderately phosphorescent seeds producing abnormal seedlings. The fraction-II seeds experienced hypoxia during the imbibition because of rapid oxygen consumption by the embryo and restrictions to O₂ diffusion imposed by the seed coat. In the fraction-I seeds, the rate of oxygen consumption by the embryo was slower and the seed coat resistance to oxygen diffusion was lower than in the fraction-II seeds. Therefore, hypoxia did not arise in the fraction-I seeds. The submergence of seeds in water caused lethal injuries. The imbibition of seeds without any contact with water caused no lethal damages but did not reduce the percentage of seeds dying of hypoxia. A slow imbibition of seeds in the media containing either an osmoticum (PEG) or an inhibitor of aquaporin channels (p-chloromercuribenzoate) prevented the lethal injuries at early stages of seed hydration and retarded the appearance of oxygen deficiency in fraction-II seeds. Different rates of water uptake by fraction-I and fraction-II seeds were controlled by permeability of cell membranes rather than by permeability of seed coat. It is proposed that low permeability of plasma membranes to water in fraction-I seeds results from the predominantly closed aquaporin channels, whereas a higher permeability of weak seeds (fraction II) is due to open channels.     

Water Relations of Seed Development and Germination in Muskmelon (Cucumis melo L.) : IV. Characteristics of the Perisperm during Seed Development

We previously reported that an apparent water potential disequilibrium is maintained late in muskmelon (Cucumis melo L.) seed development between the embryo and the surrounding fruit tissue (mesocarp). To further investigate the basis of this phenomenon, the permeability characteristics of the tissues surrounding muskmelon embryos (the mucilaginous endocarp, the testa, a 2- to 4-cell-layered perisperm and a single cell layer of endosperm) were examined from 20 to 65 days after anthesis (DAA). Water passes readily through the perisperm envelope (endosperm + perisperm), testa, and endocarp at all stages of development. Electrolyte leakage (conductivity of imbibition solutions) of individual intact seeds, decoated seeds (testa removed), and embryos (testa and perisperm envelope removed) was measured during imbibition of freshly harvested seeds. The testa accounted for up to 80% of the total electrolyte leakage. Leakage from decoated seeds fell by 8- to 10-fold between 25 and 45 DAA. Presence of the perisperm envelope prior to 40 DAA had little effect on leakage, while in more mature seeds, it reduced leakage by 2- to 3-fold. In mature seeds, freezing, soaking in methanol, autoclaving, accelerated aging, and other treatments which killed the embryos had little effect on leakage of intact or decoated seeds, but caused osmotic swelling of the perisperm envelope due to the leakage of solutes from the embryo into the space between the embryo and perisperm. The semipermeability of the perisperm envelope of mature seeds did not depend upon cellular viability or lipid membrane integrity. After maximum seed dry weight is attained (35-40 DAA), the perisperm envelope prevents the diffusion of solutes, but not of water, between the embryo and the surrounding testa, endocarp, and mesocarp tissue.     

Protein Synthesis in Embryos of Dormant and Germinating Agrostemma githago L. seeds

The time course of protein synthesis in embryos of dormant and afterripened Agrostemma githago seeds was studied. In embryos of afterripened geminating seeds, protein synthesis increased in three successive stages: (a) concurrent with swelling; (b) during the lag phase between the completion of water uptake and the onset of growth; and (c) immediately after protrusion through the seed coat. Embryos of dormant seeds showed the first increase but not the second unless dormancy was broken by imbibition at 4°C. This indicates that dormancy affects processes prior to the onset of growth. The third increase was largely due to higher oxygen availability after the rupture of the seed coat and not to actual growth. It could also be elicited in dormant embryos by isolating them from the seeds.

Electrophoretic analysis of the newly synthesized proteins demonstrated that the patterns of dormant and afterripened embryos became significantly different in both axes and cotyledons only just prior to the onset of axis elongation. Thereafter, the differences became larger.

When afterripened or dormant seeds were transferred from a low, germination-permitting to a high, germination-inhibiting temperature, the seeds germinated at the high temperature if they had completed the lag phase to a sufficient extent at the low temperature. This shows that the processes during the lag phase were inhibited by the high temperature while the onset of growth was not affected.

     

Characterization of Solute Efflux from Dehydration Injured Soybean (Glycine max L. Merr) Seeds

Soybean (Glycine max L. Merr) seeds lose their tolerance of dehydration between 6 and 36 hours of imbibition. Soybean axes and cotyledons were excised 6 hours (tolerant of dehydration) and 36 hours (susceptible) after commencing imbibition and subsequently dehydrated to 10% moisture. Kinetics of the efflux of potassium, phosphate, amino acid, sugar, protein, and total electrolytes were compared in the four treatments during rehydration. Only slight differences were observed in the kinetics of solute efflux between the two cotyledon treatments dehydrated at 6 and 36 hours suggesting that the cotyledons may retain their tolerance of dehydration at this stage of germination. Several symptoms of injury were observed in the axes dehydrated at 36 hours. An increase in the initial leakage of solutes during rehydration, as quantified by the y-intercept of the linear regression line for solute efflux between 2 and 8 hours suggests an increased incidence of cell rupture. An increase in the rate of solute efflux (slope of regression line between 2 and 8 hours) from fully rehydrated axes was observed in comparison to axes dehydrated at 6 hours. The Arrhenius activation energy for potassium, phosphate, and amino acid efflux decreased and for protein remained unchanged. Both observations indicate an increase in membrane permeability in dehydration-injured tissue. Increasing the H⁺ concentration of the external solution increased K⁺ efflux from both control and dehydrated/rehydrated samples, increased sugar efflux from axes at 6 hours imbibition but decreased sugar efflux from axes at 36 hours imbibition, indicating changes in membrane properties during germination. The dehydration treatment did not alter the pattern of the pH response of axes dehydrated at 6 or 36 hours but did increase the quantity of potassium and sugar efflux from dehydration injured axes. These results are interpreted as indicating that dehydration of soybean axes at 36 hours of imbibition increased both the incidence of cell rupture during rehydration and altered membrane permeability of the rehydrated tissue.     

The effect of sugar solution type,sugar concentration and viscosity on the imbibition and energy intake rate of bumblebees

Nectar is an essential resource for bumblebees and many other flower-visiting insects. The main constituents of nectar are sugars, which vary in both composition and concentration between plant species. We assessed the influence of sugar concentration, sugar solution viscosity and sugar solution composition on the imbibition and energy intake rate of bumblebees, Bombus impatiens Cresson (Hymenoptera: Apidae). To do this, we measured their rate of solution intake for 49 different sugar solution treatments, which varied in both sugar composition and concentration. In general, the imbibition rates of bumblebees were found to increase with increasing sugar concentration, probably due to their preference for high sugar concentrations, up to a concentration of 27% (w/w), at which point solutions reached a threshold viscosity of approximately 1.5–1.6 mPa.s. Above this threshold, the increasing viscosity of the solutions physically inhibited the imbibition rates of bees, and imbibition rate began to decrease as the concentration increased. Nevertheless, bumblebee energy intake rate increased with increasing concentration up to about 42–56%. Although we found that sugar solution composition had an impact on both imbibition and energy intake rate, its effect was not as straightforward as that of sugar concentration and viscosity.     

The Influence of Testa Condition on the Imbibition and Vigour of Pea Seeds

Seed lots of peas with high rates of water uptake which containeda large proportion of seeds with at least one crack in the testaand were low in vigour, as indicated by electrolyte leakageand poor field emergence. Comparisons of the tetrazolium chloridestaining of the cotyledons after imbibition in water and polyethyleneglycol of two seed lots both with and without scarificationshowed that cracks in the testa could cause cell death by increasingthe rate of water uptake. Although the major cause of poor seedvigour appeared to be the incidence of cracks in the testa whichresulted in rapid water uptake, cell death, and solute leakage,the condition of the embryo also influenced the response towater uptake. The practical significance of these observationsto the problems of field emergence and seed production in peasis discussed.     

The Damaging Effect of Water on Dry Pea Embryos During Imbibition

When pea seeds were imbibed in water without their seed coats,vital staining revealed that cells on the abaxial surface ofthe cotyledons were dead. No damage occurred on the surfaceof cotyledons when the seeds were imbibed intact, or beneaththe testa when only half of the testa was removed. Cell deathoccurred as a result of rapid water uptake within the first2 min of imbibition, since reducing the rate of imbibition insolutions of Carbowax 4000 lessened the damage. Cell death wasrestricted to the outer layers of the cotyledons; inner tissuesremained alive. These observations supported the hypothesisthat rapid early leakage during imbibition of dry embryos resultedfrom the death of cells caused by the physical disruption ofmembranes. Imbibition damage resulted in reduced respirationand germination, a decline in the rate of food reserve transferfrom the cotyledons to the growing axis, and a lower growthrate in the seedlings produced. Greater sensitivity of embryosto imbibition damage at low temperature, and similarities betweenfeatures of imbibition damage and chilling injury led to thesuggestion that so-called chilling injury is the result of imbibitiondamage rather than the effects of low temperature.     

Osmoregulation, growth and sucrose accumulation in germinated Trigonella foenum-graecum (fenugreek) seeds treated with polyethylene glycol

Germinated seeds of Trigonella foenum-graecum L. (fenugreek) were grown in water or in polyethylene glycol (PEG) solutions. After endosperm removal, the water relations, growth, dry weight, sucrose and reducing sugar content of the embryo were determined. Under water sstress conditions, water content and osmotic potential (π₀) at saturation, growth and dry weight were lower than in non-stressed controls. The reduction in dry weight indicated a lower uptake of solutes from the endosperm and the decrease in π₀ was not accompanied by an increase in the amount of the accumulated solutes. It is suggested that embryos of stressed fenugreek seeds control osmotic potential by reduction of water uptake and that this results in reduction of growth. Embryos isolated from germinated seeds ("naked" embryos) were grown in water or in PEG solutions, with or without galactose (as an external solute source substituting for the endosperm). The results indicate that a decrease in the external solute did not account for growth reduction under conditions of water stress, and that decreased solute transport to the embryo may be important. The sucrose contents of "naked" embryos and of embryos from whole seeds were higher after PEG treatment, while reducing sugar contents were lower compared to non-stressed controls. The increased sucrose accumulation may be due to decreased sucrose hydrolysis.     

Effect of laser irradiation on microviscosity of aqueous medium in imbibing maize seeds as studied with a spin probe method

A spin probe method was used to study microviscosity of aqueous medium in embryos and endosperm of maize (Zea mays L.) seeds after their irradiation with a Lvov-1 Elektronika laser device. Based on parameters of electron spin resonance (ESR) spectra of nitroxyl radicals (probes) absorbed by imbibing seeds during water uptake, correlation times τ_C were determined for the rotational diffusion of probes in embryos and endosperm of seeds. The τ_C values were found smaller in embryos of irradiated seeds than in untreated seeds; the dependence of τ_C on the duration of seed imbibition was determined. It is concluded that laser irradiation of seeds decreases the microviscosity of aqueous medium in embryo cells and elevates the mobility of the probes. Effect of laser irradiation on τ_C in seed endosperm was less pronounced but also led to the increase in probe mobility.     

Initiation of active metabolism in the early imbibitional phase of mungbean (phaseolus aureus) seeds

Metabolism of dry mungbean seeds(Phaseolus aureus) was compared with that of 3 h imbibed ones. Respiration rates sharply increased upon imbibition which was accompanied by large water uptake. Storage protein was degraded resulting in the rise in soluble nitrogen. RNA and water soluble protein content increased but DNA declined. It is suggested that some of the soluble nitrogen pools are diverted to RNA formation which in turn produce enzyme protein. In mungbean seeds imbibition of water thus triggers a series of metabolic changes within 3 h with the respiratory energy as the driving force.