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PubMed1990

Relationship between canopy depth and other dimensions of coastal Pinus thunbergii Parlat. forests in Japan.

The relationship between canopy depth and other dimensions of coastal Pinus thunbergii Parlat. forests was analyzed from data for 29 stands using a mathematical model describing canopy depth. The model was derived from the relationship between mean diameter at breast height (D) and mean tree height (H). Maximum mean tree height (H(max)), which was calculated as H(max) = AD(B) where A and B are parameters, approximated the upper 95 percentile of H for a given D. Maximum canopy depth (L(cmax)) was calculated as L(cmax) = H(max) - H(B), where H(B) is mean clear length. Relative maximum canopy depth (R(cmax); defined as L(cmax)/H(max)) was approximated as 1/R(cmax) = 1/(aS(R) (b)) + 1, where a and b are parameters. The term S(R) was defined as N(-1/2)/H, where N is stand density (trees m(-2)). From these equations, the canopy depth (L(c))was derived as L(c) = H - H(max)/(aS(R) (b) + 1), because L(c) = H - H(B). From these relationships a diagram showing possible points of regulation of canopy depth was developed.

Journal Article↗

PubMed1990

Influence of male and female cones on assimilate production of Pinus contorta trees within a forest stand.

Studies on branches of field-grown Pinus contorta Dougl. trees showed that: (i) branches with fully developed male or female cones were heavier than vegetative branches; (ii) the production of female cones decreased neither the number of laterals nor the length of the terminal or lateral shoots on the cone-bearing branches; (iii) the production of male cones decreased both the number of laterals and the number of needles on the cone-bearing branches; (iv) needles on male cone-bearing branches had significantly higher photosynthetic rates and needles on female cone-bearing branches generally also fixed more CO(2) per quantum of light. In addition, both male and female cones refixed a significant proportion of respired CO(2) when illuminated. The simulation model, MAESTRO, was used to estimate light penetration through two stands (3690 and 1845 trees ha(-1)) of P. contorta trees and to calculate the resultant CO(2) assimilation of a vegetative, a male and a female cone-bearing tree on sunny, partly sunny and cloudy days in Scotland. Generally, the CO(2) efflux from female cones was less than 3% of the CO(2) assimilated by the whole tree. The presence of male cones resulted in an average 33% decrease in needle complement, but an increased quantum efficiency of associated needles in the autumn. It was estimated that, on a sunny day, male cone-bearing trees assimilated as much CO(2) as vegetative trees in the high density stand and only 6% less in the low density stand. On cloudy days, the smaller needle complement was beneficial to the carbon economy of the tree because of lower respiration losses compared with a vegetative tree.

Journal Article↗

PubMed1991

Influence of drought stress and low irradiance on plant water relations and structural constituents in needles of Pinus ponderosa seedlings.

The influence of low light on tolerance to prolonged drought was tested on unshaded and shaded seedlings of ponderosa pine (Pinus ponderosa var. scopulorum Dougl. ex Laws.). Unshaded seedlings of P. ponderosa var. ponderosa were also drought stressed to compare varietal responses to drought. The maximum irradiance received by shaded seedlings was 10% of full light. Seedlings were progressively drought stressed until predawn water potentials (Psi(x)) were -5.0 MPa. Relative water content (RWC) and the reciprocal of Psi(x) were analyzed by means of an unusual application of the pressure-volume relationship for determination of RWC of the apoplast (RWC(a)), osmotic potential at full turgor (Psi(oft)), and ratio of fully turgid weight to dry weight. Major varietal differences in drought response were in RWC(a) and needle cellulose content. The shaded seedlings showed tissue damage at relative water contents < 60%, and were killed by water deficits from which unshaded seedlings recovered. Correspondingly, shaded plants had significantly higher cell volume/cell mass ratio, Psi(oft), less cellulose in needle tissue, and lower RWC(a) than unshaded plants. These differences suggest that low irradiance restricts drought adaptation in ponderosa pine.

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PubMed1991

Distribution of (14)C-labeled photosynthate in loblolly pine (Pinus taeda) seedlings as affected by season and time after exposure.

Distribution of (14)C-labeled photosynthate was determined in field-grown loblolly pine (Pinus taeda L.) seedlings on August 9 and October 15, 1984 and January 15 and March 12, 1985. Leaves on a lateral branch fixed (14)C photosynthetically and amounts of (14)C in seven biochemical fractions in each of six plant parts were determined 8, 24, and 72 h later. In all treatments, (14)C uptake was approximately 96% of that originally presented. Respiratory loss of (14)C ranged from 22 to 87% of uptake and increased sharply with increasing time after exposure and as the seedlings grew larger later in the study. Most (14)C was found in exposed leaves and very little occurred above the exposed branch. Amounts of (14)C decreased in the exposed leaves and increased in the roots with time after exposure and date. Sugars were generally the most heavily labeled fraction. Labeled sugar content of exposed leaves decreased by more than half between 8 and 72 h as sugars were metabolized and translocated to other parts, primarily the roots. In roots, the labeling of starch and residue (structural compounds) increased greatly with transport time and season. In all plant parts, proteins and amino acids contained very little (14)C regardless of date or time.

Journal Article↗

PubMed1991

Seasonal variation in ethylene concentration in the wood of Pinus sylvestris L.

Ethylene concentrations were determined in gas samples extracted from sealed holes made in the sapwood and heartwood of stems of 70-100-year-old Scots pine (Pinus sylvestris L). Gas could be collected from the heartwood holes by lowering the pressure by means of a large syringe. However, attempts to extract gas from air spaces in the sapwood with the same technique failed, presumably because of lack of an interconnected system of gas-filled canals. High ethylene concentrations, usually in excess of 1 ppm, accumulated in the sapwood holes within one day after sealing. Ethylene concentrations in the sapwood rose to 3-7 ppm during the growing season, and decreased to 0.1-0.3 ppm during the winter. In response to extreme drought, sapwood ethylene concentration increased to 30 ppm, followed by a rapid decrease after the onset of rain. Ethylene concentrations in gas samples from the heartwood were consistently lower than 1 ppm. The lowest values, about 0.1 ppm, were found during the autumn and early winter, whereas values around 0.5 ppm were typical from February to August.

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PubMedDate not supplied

Modeling the water balance of a small Pinus radiata catchment.

An hourly biophysical model was used to calculate the water balance over a period of one year for an 8.7-ha catchment with a closed-canopy, 13-year-old Pinus radiata D. Don forest in the central North Island, New Zealand. Components of the model are transpiration from the dry tree canopy, evaporation from the partially wet tree canopy and stems, evaporation from the understory and soil, and drainage from a single-layer root zone. The model requires input of hourly weather data (net radiation, air and wet bulb temperatures, windspeed, and rainfall), tree stand characteristics (average height, tree number, leaf area index), physical characteristics of the site (root zone depth, relationship between root zone matric potential and volumetric water content, the relationship between the rate of drainage from the root zone and volumetric water content, and the area of open-stream channels). A submodel of the response of stomatal conductance to air saturation deficit and root zone matric potential is also required. Tree transpiration (704 mm year(-1) or 50% of annual rainfall) was a dominant component of the catchment water balance. Estimated evaporation from the wet tree canopy was 203 mm year(-1) (15%). Evaporation from the understory was much less, amounting to 94 mm year(-1) (7%) and an increase in water storage for the 3.5 m root zone depth was estimated to be 53 mm year(-1) (4%). Estimated daily rates of drainage generally agreed well with measurements of streamflow, although estimated annual drainage (349 mm year(-1), 24%) exceeded measured streamflow (234 mm year(-1)). The significance of the results is discussed in relation to closure of the hydrologic balance.

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PubMedDate not supplied

Interrelation between shoot characteristics and solar irradiance in the crown of Pinus contorta ssp. latifolia.

In Pinus contorta Doug. ssp. latifolia (Engelm.), annual shoot length increment, annual foliage production per shoot, and the daily integral of photosynthetically active radiation (DPAR) at shoot tips increased with height in the crown, whereas leaf longevity decreased. There was a decrease in DPAR along the shoot from the tip to beyond the oldest leaves, and the value reached at the point just proximal to the oldest leaf cohort was the same for all shoots from all crown locations, regardless of the age of the oldest leaves. The results suggest that shoot elongation, leaf production and leaf retention are controlled by the light environment within the crown. Because high irradiance promotes leaf production and low irradiance promotes leaf shedding, foliage is not randomly distributed in the crown.

Journal Article↗

PubMed1993

Purification and Characterization of Catalase from Loblolly Pine (Pinus taeda L.) Megagametophytes.

Catalase (EC 1.11.1.6) was purified to near homogeneity from isolated megagametophytes of germinated loblolly pine (Pinus taeda L.) seeds, and monospecific antibodies were elicited in rabbits. Following a procedure that involved acetone extraction, (NH4)2SO4 fractionation, and four chromatographic steps (i.e. DE-52 cellulose, Superdex-200, hydroxylapatite, and phenyl-Sepharose CL-4B), catalase was purified about 140-fold to a final specific activity of 2215 mmol min-1 mg-1 of protein. Cotton isocitrate lyase antibodies were used, and protein immunoblots revealed that the resolution on hydroxylapatite and phenyl-Sepharose allowed for the complete separation of catalase from contaminating isocitrate lyase. The molecular masses of the native enzyme and its subunit are 235 and 59 kD, respectively, indicating that the pine holoenzyme is a homotetramer. Loblolly pine catalase exists as multiple isoforms. When megagametophytes taken 7 d after imbibition at 30[deg]C were extracted, subjected to nondenaturing isoelectric focusing, and stained for catalase activity, at least four catalase isoforms were observed, including one dominant form with an isoelectric point of 6.87. Purified pine catalase is not a glycoprotein and has a ratio of absorbance at 208 nm to absorbance at 405 nm of 1.5. When probed with loblolly pine catalase antibodies, protein blots of cell-free extracts from megagametophytes of mature, stratified, and germinated loblolly pine seeds, the megagametophyte glyoxysomal fraction, and purified loblolly pine catalase all revealed one immunoreactive 59-kD polypeptide. This indicates that no detectable change in the enzyme's monomeric molecular mass occurs during seed stratification and germination, early seedling growth, and purification.

Journal Article↗

PubMed1993

Molecular Responses to Photooxidative Stress in Pinus sylvestris (L.) (II. Differential Expression of CuZn-Superoxide Dismutases and Glutathione Reductase.

The influence of photooxidative stress on genes expressing superoxide dismutase (Sod) and glutathione reductase (Gor) was analyzed in needles of top and side shoots of 3-year-old Pinus sylvestris (L.) seedlings. The study was carried out in the field during spring recovery. From mid-April the top shoots of seedlings protruded above the snow and thus were exposed to sunlight, whereas the side shoots were covered with snow until May 4. Needles were sampled from top and side shoots on five different occasions. At the beginning of May the mRNA levels for cytosolic CuZn-Sod were significantly higher in top-shoot needles than in side-shoot needles. Similar results were obtained for chloroplastic CuZn-Sod mRNA. After May 6 we could not detect any significant differences between top- and side-shoot needles for either CuZn-Sod mRNA level. Transcript accumulation for the chloroplastic CuZn-Sod was up to 4-fold higher than for cytosolic CuZn-Sod in both types of shoots. On June 1 minimum transcript levels were observed for both CuZn-SOD isoforms. Protein activity analysis for CuZn-SOD isozymes did not reveal any significant differences between top- and side-shoot needles during the whole period of measurements. The mRNA level for chloroplastic Gor was similar in both types of shoots. However, the total GR activity was significantly higher in top-shoot needles than in side-shoot needles at the beginning of May. The analysis of mRNA accumulation for chloroplastic CuZn-Sod and Gor indicates that transcript levels were at least 5- to 20-fold higher for CuZn-Sod than for chloroplastic Gor. The differential expressions of Sod and Gor genes are discussed in relation to regulation of the enzymic scavenging system during photooxidative stress conditions.

Journal Article↗

PubMed1994

Effects of the Indole-3-Acetic Acid (IAA) Transport Inhibitors N-1-Naphthylphthalamic Acid and Morphactin on Endogenous IAA Dynamics in Relation to Compression Wood Formation in 1-Year-Old Pinus sylvestris (L.) Shoots.

Both N-1-naphthylphthalamic acid (NPA) and methyl-2-chloro-9-hydroxyfluorene-9-carboxylic acid (CF) inhibit the polar transport of indole-3-acetic acid (IAA) and, therefore, are attractive tools for investigating IAA's role in the regulation of plant growth. Ringing an intact conifer shoot with lanolin containing NPA or CF induces the formation of compression wood above the ring. This induction has been attributed to a postulated accumulation of IAA above the application site of the IAA transport inhibitor, but the validity of this postulation has never been confirmed. Using gas chromatography-selected ion monitoring-mass spectroscopy with [13C6]IAA as an internal standard, we measured the levels of endogenous free and conjugated IAA in 1-year-old Pinus sylvestris (L.) shoots ringed with NPA or CF. The level of free IAA was dramatically decreased below the ring, indicating that the polar transport of endogenous IAA was inhibited by the treatment. However, the free IAA level above the ring, where compression wood was formed, was also slightly lower than in control shoots. The lack of IAA accumulation above the site of the IAA transport inhibitor could not be explained by an increase in IAA conjugation. Furthermore, the turnover of [2-14C]IAA, measured using high-performance liquid chromatography with on-line radioactivity monitoring, was the same in NPA-treated and control shoots. The decrease in IAA level above a NPA or CF ring is attributed to these substances being transported acropetally and interfering with polar IAA transport along the shoot. It is concluded that compression wood formation above a NPA or CF ring is not associated with an overall increase in cambial region IAA level or increased IAA turnover. Instead, we suggest that acropetally transported NPA and CF induce compression wood formation by interacting with the NPA receptor in differentiating tracheids, thereby locally increasing IAA in these cells.

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PubMed1996

Biochemistry of Short-Chain Alkanes (Tissue-Specific Biosynthesis of n-Heptane in Pinus jeffreyi).

Short-chain (C7-C11) alkanes accumulate as the volatile component of oleoresin (pitch) in several pine species native to western North America. To establish the tissue most amenable for use in detailed studies of short-chain alkane biosynthesis, we examined the tissue specificity of alkane accumulation and biosynthesis in Pinus jeffreyi Grev. & Balf. Short-chain alkane accumulation was highly tissue specific in both 2-year-old saplings and mature trees; heart-wood xylem accumulated alkanes up to 7.1 mg g-1 dry weight, whereas needles and other young green tissue contained oleoresin with monoterpenoid, rather than paraffinic, volatiles. These tissue-specific differences in oleoresin composition appear to be a result of tissue-specific rates of alkane and monoterpene biosynthesis; incubation of xylem tissue with [14C]sucrose resulted in accumulation of radiolabel in alkanes but not monoterpenes, whereas incubation of foliar tissue with 14CO2 resulted in the accumulation of radiolabel in monoterpenes but not alkanes. Furthermore, incubation of xylem sections with [14C]acetate resulted in incorporation of radiolabel into alkanes at rates up to 1.7 nmol h-1 g-1 fresh weight, a rate that exceeds most biosynthetic rates reported with other plant systems for the incorporation of this basic precursor into natural products. This suggests that P. jeffreyi may provide a suitable model for elucidating the enzymology and molecular biology of short-chain alkane biosynthesis.

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PubMed1996

Developmental and light-regulated expression of individual members of the light-harvesting complex b gene family in Pinus palustris.

Angiosperms requires light for multiple aspects of chloroplast development, including chlorophyll synthesis and induction of expression of the mRNAs encoding the major polypeptides of the light-harvesting complex of photosystem II (Lhcb genes). In contrast, many conifers, including pines, firs, and spruces, can accumulate chlorophyll and the light-harvesting chlorophyll a/b-binding proteins of photosystem II in complete darkness. To understand the factors responsible for the regulation of expression of individual Lhcb mRNAs in the pine Pinus palustris, we have prepared sequence-specific cDNA probes for each of three family members, Lhcb1*Pp1, Lhcb2*Pp1, and Lhcb2*Pp2, and have studied the expression of two of these, Lhcb1*Pp1 and Lhcb2*Pp2, in detail. The levels of expression of each sequence were disparate, and Lhcb1*Pp1-encoded transcripts were the most abundant in the light. Both Lhcb1*Pp1 and Lhcb2*Pp2 mRNAs were expressed in stems and cotyledons, but Lhcb1*Pp1 mRNA was present at about 10-fold lower levels in stems than in cotyledons, in contrast to Lhcb2*Pp2 mRNA, which was expressed at higher levels in stems than in cotyledons. Both Lhcb1*Pp1 and Lhcb2*Pp2 mRNAs were absent in embryos but were expressed during seedling development. The levels increased with age in both the light and the dark and in both cases were about 2-fold higher in the light than in the dark. Despite the expression of Lhcb1*Pp1 and Lhcb2*Pp2 mRNAs during development in darkness, the levels of both mRNAs increased in dark-grown seedlings given red light in the low fluence range within 2 h of treatment.

Amino Acid Sequence↗

PubMed1996

Changes in Dehydrodiferulic Acids and Peroxidase Activity against Ferulic Acid Associated with Cell Walls during Growth of Pinus pinaster Hypocotyl.

Hydroxycinnamic acids associated with hypocotyl cell walls of dark-grown seedlings of Pinus pinaster Aiton were extracted with 1 N NaOH and identified by gas chromatography-mass spectrometry. The main hydroxycinnamic acid found was ferulic acid. Diferulic acid dehydrodimers were also found, with the 8,8-coupled isomer (compound 11) being the dehydrodiferulate present in the highest amount. However, the 5,5-coupled isomer, commonly referred to referred to as diferulic acid, was not detected. Two truxillic acids, 4-4[prime]-dihydroxy-3-3[prime]-dimethoxy-[alpha]-truxillic acids I and II, were tentatively identified. The 8,8-coupled dehydrodiferulic acid (compound 11) was the phenolic acid that showed the most conspicuous changes with hypocotyl age as well as along the hypocotyl axis. Peroxidase activity against ferulic acid was found in the apoplastic fluid as well as being ionically and covalently bound to the cell walls. The peroxidase activity increased with hypocotyl age as well as from the subapical toward the basal region of the hypocotyls. A key role in the cell-wall stiffening of 8,8 but not 5,5 dimerization of ferulic acid catalyzed by cell-wall peroxidases is proposed.

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PubMed1999

Genes expressed in Pinus radiata male cones include homologs to anther-specific and pathogenesis response genes.

We describe the isolation and characterization of 13 cDNA clones that are differentially expressed in male cones of Pinus radiata (D. Don). The transcripts of the 13 genes are expressed at different times between meiosis and microspore mitosis, timing that corresponds to a burst in tapetal activity in the developing anthers. In situ hybridization showed that four of the genes are expressed in the tapetum, while a fifth is expressed in tetrads during a brief developmental window. Six of the seven cDNAs identified in database searches have striking similarity to genes expressed in angiosperm anthers. Seven cDNAs are homologs of defense and pathogen response genes. The cDNAs identified are predicted to encode a chalcone-synthase-like protein, a thaumatin-like protein, a serine hydrolase thought to be a putative regulator of programmed cell death, two lipid-transfer proteins, and two homologs of the anther-specific A9 genes from Brassica napus and Arabidopsis. Overall, our results support the hypothesis that many of the reproductive processes in the angiosperms and gymnosperms were inherited from a common ancestor.

Acyltransferases↗

PubMed2000

Compression wood-responsive proteins in developing xylem of maritime pine (Pinus pinaster ait.).

When a conifer shoot is displaced from its vertical position, compression wood (CW) is formed on the under side and can eventually return the shoot to its original position. Changes in cell wall structure and chemistry associated with CW are likely to result from differential gene/protein expression. Two-dimensional polyacrylamide gel electrophoresis of differentiating xylem proteins was combined with the physical characterization of wooden samples to identify and characterize CW-responsive proteins. Differentiating xylem was harvested from a 22-year-old crooked maritime pine (Pinus pinaster Ait.) tree. Protein extracted from different samples were revealed by high-resolution silver stained two-dimensional polyacrylamide gel electrophoresis and analyzed with a computer-assisted system for single spot quantification. Growth strain (GS) measurements allowed xylem samples to be classified quantitatively from normal wood to CW. Regression of lignin and cellulose content on GS showed that an increase in the percentage of lignin and a decrease of the percentage of cellulose corresponded to increasing GS values, i.e. CW. Of the 137 studied spots, 19% were significantly associated with GS effect. Up-regulated proteins included 1-aminocyclopropane-1-carboxylate oxidase (an ethylene forming enzyme), a putative transcription factor, two lignification genes (caffeic O-methyltransferase and caffeoyl CoA-O-methyltransferase), members of the S-adenosyl-L-methionine-synthase gene family, and enzymes involved in nitrogen and carbon assimilation (glutamine synthetase and fructokinase). A clustered correlation analysis was performed to study simultaneously protein expression along a gradient of gravistimulated stressed xylem tissue. Proteins were found to form "expression clusters" that could identify: (a) Gene product under similar control mechanisms, (b) partner proteins, or (c) functional groups corresponding to specialized pathways. The possibility of obtaining regulatory correlations and anticorrelations between proteins provide us with a new category of homology (regulatory homology) in tracing functional relationships.

Cellulose↗

PubMed2000

Multisite inhibition of Pinus pinea isocitrate lyase by phosphate.

Our results show that the phosphate ion is a nonlinear competitive inhibitor of Pinus pinea isocitrate lyase. In addition, this compound induces a sigmoidal response of the enzyme, which usually exhibits standard Michaelis-Menten kinetics. This peculiar behavior of P. pinea isocitrate lyase could be explained by a dimer (two-site) model, in which phosphate binds cooperatively, but the affinity of the vacant site for substrate (the magnesium-isocitrate complex) remains the same. As a result, the interaction of phosphate with free enzyme produces an inhibitor-enzyme-inhibitor species that is of significant importance in determining reaction rate; a possible regulatory role of the glyoxylate cycle by inorganic phosphate is suggested. The mode of phosphate inhibition is consistent with both the mechanism for magnesium ion activation of P. pinea isocitrate lyase and its site heterogeneity. Our results explain the cooperative effects observed by some authors in kinetic studies of isocitrate lyase carried out in phosphate buffers and also account for the higher K(m) values determined by using such assay systems. Phosphate buffer should be avoided in performing isocitrate lyase kinetics.

Buffers↗

PubMed1968

Seasonal Course of Photosynthesis, Respiration, and Distribution of C in Young Pinus resinosa Trees as Related to Wood Formation.

Rates of net photosynthesis and dark respiration, and distribution of (14)C were determined for new (current season's) and old (previous season's) needles at 10 times during the seasonal development of young Pinus resinosa Ait. trees. The seasonal changes in these factors associated with the development of the new shoot were related to known seasonal patterns of wood formation.Net photosynthesis per gram of needle dry weight (photosynthetic efficiency) was maximum in the old needles at the time of first new needle elongation; at the same time translocation of (14)C from old to new needles was greatest. Photosynthetic efficiency of new needles was maximum at the end of the period of rapid new needle elongation, when the new needles also began exporting much greater quantities of (14)C to other plant parts. In particular, the amount translocated from the new needles to the stem was greatly increased. At this time thick-walled xylem cells were first observed in the stem.These results, together with those of previous studies, indicate that the production of thick-walled xylem tracheids normally associated with latewood is physiologically correlated with maturation of the current season's needles. Because there is a lesser demand for photosynthate in the new shoot and a high rate of photosynthesis in the whole plant at the time of new needle maturity, a sharply increased amount of photosynthate becomes available for wall synthesis by cambial derivatives.

Journal Article↗

PubMed1969

Changes in Nucleic Acid Fractions of Seed Components of Red Pine (Pinus resinosa Ait.) during Germination.

Changes in nucleic acid fractions of Pinus resinosa during seed germination were studied. At various stages of seed germination, embryos and megagametophytes were surgically separated and nucleic acids were extracted separately by a phenol-method. Total nucleic acids were fractionated on single-layer methylated albumin kieselguhr (MAK) columns. Total nucleic acids in embryos increased significantly 2 days after seeds were moistened, whereas, in megagametophytes, total nucleic acids stayed almost at a constant level until they degenerated at the time of shedding. In embryos, ribosomal RNAs (rRNA) increased 2 days after seeds were sown, whereas soluble RNA (sRNA) increased at 3 days. By comparison, nucleic acid fractions of megagametophytes did not show any quantitative changes during germination, except that rRNA fractions decreased shortly before shedding of seed coats. In dormant embryos the proportion of DNA was high and the proportions of sRNA and rRNA were low, whereas in megagametophytes at dormancy the proportions were completely reversed. As seed germination progressed, proportions of nucleic acid fractions in embryos changed significantly. In megagametophytes, although proportions of individual fractions remained almost constant throughout the experimental period, incorporation of (32)P into sRNA and rRNA of megagametophytes indicated turnover of these fractions.

Journal Article↗