Maturation in larch : I. Effect of age on shoot growth, foliar characteristics, and DNA methylation

The time course of maturation in eastern larch (Larix laricina [Du Roi] K. Koch) was examined by grafting scions from trees of different ages onto 2-year-old root stock and following scion development for several years. Height, diameter, foliar chlorophyll content, and rooting ability of scion-derived cuttings all varied linearly as a function of log₁₀ age. Chlorophyll content (milligrams per gram of dry weight) increased while height, diameter, and ability to root decreased with age (P < 0.01). The tendency toward orthotropic growth and branch formation per centimeter of main stem decreased abruptly between age 1 and 5 years (P < 0.01). Total chlorophyll content of both long and short shoot foliage increased by 30 to 50% with increasing age, but the chlorophyll a/b ratio did not change. Also, juvenile long shoot needles were significantly longer than mature (P < 0.01). Surprisingly, the juvenile scions produced more total strobili over two successive years, but the mature scions produced a significantly higher proportion of male strobili (P < 0.001 year 1; P < 0.02 year 2). The age-related changes in foliar traits were not associated with changes in DNA methylation between juvenile and mature scions. Using HPLC, we found that 20% of foliar DNA cytosine residues were methylated in both scion types.     

Expression of cab Genes in Douglas-Fir Is Not Strongly Regulated by Light

Dark-grown Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco) seedlings had approximately 30% of the major polypeptide of the light-harvesting chlorophyll a/b binding protein, 30% of cab mRNA, 54% of psbA mRNA, and 14% of total chlorophyll, in comparison with amounts in light-grown seedlings. Seedlings entrained under a 24-hour photoperiod of light and dark showed small diurnal fluctuations in cab and psbA mRNA levels and, when transferred to continuous conditions, no circadian rhythms in mRNA levels were apparent. These results suggest that regulation of cab gene expression in Douglas-fir differs from regulation in angiosperms, because in the latter, both light and circadian factors strongly influence the expression of cab genes.     

Influence of Cell Age on Chlorophyll Formation in Light-grown and Etiolated Wheat Seedlings

A method is described for relating the age of a cereal leaf cell to its distance from the leaf base. The rates of chlorophyll synthesis per plastid in the first leaf of light-grown and of greening etiolated seedlings of wheat (Triticum aestivum, var. Maris Dove) increase with cell age. Normally developing plastids of light-grown wheat take over 24 hours to reach the chlorophyll a/b ratio characteristic of mature wheat chloroplasts (4.5), but mature etioplasts need only 8 hours light to achieve this a/b ratio. Plastid greening potential depends only on cell age, whereas the chlorophyll a/b ratio is influenced both by cell age and by light.     

A review of light interception in plant stands from leaf to canopy in different plant functional types and in species with varying shade tolerance

Changes in the efficiency of light interception and in the costs for light harvesting along the light gradients from the top of the plant canopy to the bottom are the major means by which efficient light harvesting is achieved in ecosystems. In the current review analysis, leaf, shoot and canopy level determinants of plant light harvesting, the light-driven plasticity in key traits altering light harvesting, and variations among different plant functional types and between species of different shade tolerance are analyzed. In addition, plant age- and size-dependent alterations in light harvesting efficiency are also examined. At the leaf level, the variations in light harvesting are driven by alterations in leaf chlorophyll content modifies the fraction of incident light harvested by given leaf area, and in leaf dry mass per unit area (M _A) that determines the amount of leaf area formed with certain fraction of plant biomass in the leaves. In needle-leaved species with complex foliage cross-section, the degree of foliage surface exposure also depends on the leaf total-to-projected surface area ratio. At the shoot scale, foliage inclination angle distribution and foliage spatial aggregation are the major determinants of light harvesting, while at the canopy scale, branching frequency, foliage distribution and biomass allocation to leaves (F _L) modify light harvesting significantly. F _L decreases with increasing plant size from herbs to shrubs to trees due to progressively larger support costs in plant functional types with greater stature. Among trees, F _L and stand leaf area index scale positively with foliage longevity. Plant traits altering light harvesting have a large potential to adjust to light availability. Chlorophyll per mass increases, while M _A, foliage inclination from the horizontal and degree of spatial aggregation decrease with decreasing light availability. In addition, branching frequency decreases and canopies become flatter in lower light. All these plastic modifications greatly enhance light harvesting in low light. Species with greater shade tolerance typically form a more extensive canopy by having lower M _A in deciduous species and enhanced leaf longevity in evergreens. In addition, young plants of shade tolerators commonly have less strongly aggregated foliage and flatter canopies, while in adult plants partly exposed to high light, higher shade tolerance of foliage allows the shade tolerators to maintain more leaf layers, resulting in extended crowns. Within a given plant functional type, increases in plant age and size result in increases in M _A, reductions in F _L and increases in foliage aggregation, thereby reducing plant leaf area index and the efficiency of light harvesting. Such dynamic modifications in plant light harvesting play a key role in stand development and productivity. Overall, the current review analysis demonstrates that a suite of chemical and architectural traits at various scales and their plasticity drive plant light harvesting efficiency. Enhanced light harvesting can be achieved by various combinations of traits, and these suites of traits vary during plant ontogeny.     

Light Intensity-Induced Changes in cab mRNA and Light Harvesting Complex II Apoprotein Levels in the Unicellular Chlorophyte Dunaliella tertiolecta

During a transition from high growth irradiance (700 micromoles quanta per square meter per second) to low growth irradiance (70 micromoles quanta per square meter per second), the unicellular marine chlorophyte Dunaliella tertiolecta Butcher increases the cellular pool size of the light-harvesting complex of photosystem II (LHC II). We showed that the increase in LHC II apoproteins and in chlorophyll content per cell is preceded by an approximately fourfold increase in cab mRNA. The increase in cab mRNA is detectable within 1.5 hours following a shift from high to low light intensity. An increase in the relative abundance of cab mRNA was also found following a shift from high light to darkness and from high light to low light in the presence of gabaculine, a chlorophyll synthesis inhibitor. However, the LHC II apoproteins did not accumulate in the latter experiments, suggesting that LHC II apoprotein synthesis is coupled to chlorophyll synthesis at or beyond translation. We propose that changes in energy balance brought about by a change in light intensity may control a regulatory factor acting to repress cab mRNA expression in high light.     

Effect of Morpholine Systemic Fungicides on the Chloroplast Pigments of Wheat and Maize

Abstract

The growth, chlorophyll and carotenoid contents of wheat and maize seedlings as influenced by a soil drench of 250 μM of tridemorph or fenpropimorph, were studied. It was found that the length of the shoot, primary leaf, and root, as well as the leaf area were significantly reduced in the fungicide treated seedlings. Although the dry matter percentage was higher in the treated seedlings as compared to the control, the fresh or dry weight per shoot remained lower in the fungicide treated seedlings. The chloroplast pigments in the treated seedlings of both crops were unaffected, suggesting that the fungicides had no significant effect on the chlorophyll or carotenoid biosynthesis.     

Carbon dioxide enrichment reduces shoot growth in sweet chestnut seedlings (Castanea sativa Mill.)*

Abstract. Two-year-old potted sweet chestnut seedlings were grown at 350 ppm CO₂ and 700 ppm, day and night in constantly ventilated tunnels during two full growing seasons, near Paris, France (48° N, 2° E). Enrichment with CO₂ caused an unusual shoot growth response. After the end of July, stem elongation ceased in 62% of the CO₂ enriched plants as compared with 37% in the control. The leaves of CO₂-enriched seedlings showed early senescence, indicated by premature yellowing and a decrease in chlorophyll content. This was associated with nutrient dilution brought about by the rapid growth of these trees. The increase in total dry weight of the CO₂-enriched seedlings was essentially the result of increase in the root dry weight (69%). Shoot weight decreased by 22% relative to the control. Total leaf area per enriched plant was 25% smaller than the control. This unusual pattern of growth and carbon allocation of the CO₂ treated Chestnut trees emphasizes the concept of a response specificity within trees to an increase of atmospheric CO₂.     

Acclimation responses of mature Abies amabilis sun foliage to shading

This paper addresses two main questions. First, can evergreen foliage that has been structurally determined as sun foliage acclimate physiologically when it is shaded? Second, is this acclimation independent of the foliage ageing process and source-sink relations? To investigate these questions, a shading and debudding experiment was established using paired branches on opengrown Abies amabilis trees. For each tree, one branch was either shaded, debudded, or both, from before budbreak until the end of summer, while the other branch functioned as a control. Foliage samples were measured both prior to and during treatment for photosynthesis at light saturation (A _max), dark respiration, nitrogen content, chlorophyll content, chlorophyll-to-nitrogen ratio and chlorophyll a:b ratio. All age classes of foliage responded similarly during the treatment, although pre-treatment values differed between age classes. Within 1 month after the treatment began, A _max was lower in shaded foliage and remained lower throughout the treatment period. For debudded branches, A _max was lower than the controls only during active shoot elongation. At the end of the treatments in September, A _max in shade-treated sun foliage matched the rates in the true shade-formed foliage, but nitrogen remained significantly higher. By 1.5 months after treatment, chlorophyll content in shaded foliage was higher than in controls, and the chlorophyll a:b ratio was lower for the shaded foliage. On debudded branches, chlorophyll content and chlorophyll a:b ratio were similar to the values in control samples. Shading lowered the rate of nitrogen accumulation within a branch, while removing debudding decreased the amount of sequestered N that was exported from the older foliage to supply new growth. By September, chlorophyll content in shade-treated foliage was higher than that in the control sun foliage or in true shade foliage. The chlorophyll increase as a result of shading was unexpected. However, the chlorophyll-to-nitrogen ratio was identical for the shade-treated sun foliage and the true shade foliage while being significantly lower than the control sun foliage. It appears that acclimation to shading in mature foliage involves a reallocation of nitrogen within the leaf into thylakoid proteins. A redistribution of resources (nitrogen) among leaves is secondary and appears to function on a slower time scale than reallocation within the leaf. Thus, A. amabilis foliage that is structurally determined as sun foliage can acclimate to shade within a few months; this process is most likely independent of ageing and is only slightly affected by source-sink relations within a branch.     

Growth of regenerated tree seedlings associated with microclimatic change in a mature larch plantation after harvesting

In order to further develop methods of sustainable forest management, we evaluated the effects of logging practices during the winter on microclimatic factors and growth of four seral deciduous broad-leaved tree seedlings regenerated in a larch plantation in northern Japan. We found that winter logging practices drastically changed microclimatic factors, especially in terms of light intensity and the vertical distribution pattern of CO₂ concentration. Harvesting overstory larch trees increased photosynthetic photon flux (PPF), which may enhance the photosynthesis of understory plants. We examined the undergrowth for tree seedlings of the following species: two late successional species, Prunus ssiori and Carpinus cordata; one gap phase species, Magnolia hyporeuca; and one mid-late successional species, Quercus mongolica var. crispula. All of the four studied tree species responded well to the practices of winter logging after the second year of harvesting overstoried larch trees. The current shoot diameter and current shoot length increased significantly. Therefore, we conclude that winter logging practices should improve shoot growth of deciduous broad-leaved tree seedlings regenerated in a larch plantation through the change in environmental factors that accompanies larch harvest.     

Characteristics of Photosynthate Partitioning during Chloroplast Development in Avena Leaves

The first leaves (40 millimeters long) of 4-day-old light-grown Avena sativa L. cv Victory I seedlings contained a complete age sequence of cells from the base to the tip, and within these tissues all stages of chloroplast development could be observed. Although chloroplasts underwent progressive development, a marked increase in number of thylakoids per granum, in chloroplast volume, and in chlorophyll content occurred in the region between 20 and 30 millimeters from the base. Photosynthetic CO₂ fixation (per unit chlorophyll) increased markedly during chloroplast development and closely followed structural changes in chloroplasts. It was also found that the partitioning of photosynthates differed greatly in the segment from 30 to 40 millimeters (at the tip of the leaf) compared with the segment nearer to the leaf base, although both total ¹⁴CO₂ fixation and chlorophyll content per segment did not change significantly along the length of the leaves. As the thylakoid system reached full maturation, partitioning of photosynthates into sucrose increased but partitioning decreased into starch, lipids, and phosphorylated intermediates.     

The influence of micrografting in vitro on tissue culture behavior and vegetative propagation of old European larch trees

The influence of a new micrografting method in vitro was tested by using long shoot meristems of adult 140 year old European larch trees grafted onto larch seedling rootstocks in sterile peat pellets. Five months after grafting and transfer of grafts to ex vitro conditions, new shoots from sprouting scions were re-established into tissue culture. The propagation behavior of shoots derived from these explants was compared with shoots established via bud culture from the same donor trees. Shoot explants from micrografts multiplied more rapidly than shoots explants from the original adult donor trees. Rooting experiments with cuttings from six micrografted clones resulted in 49.9 ± 11.9% rooting. Cuttings from donor trees invariably showed no root formation. The results confirm a rejuvenating influence of micrografting in larch.     

Chlorophyll accumulation and breakdown in light-grown barley transferred to darkness: effect of seedling age

Diurnally grown barley (Hordeum vulgare L. cv. Clipper) seedlings of various ages (3–4, 5–6 and 10–11-days-old) were transferred to darkness for 17 h and changes in leaf fresh weight, chlorophyll a, chlorophyll b and protochlorophyllide measured. The results were consistent with previous evidence of a light-independent chlorophyll biosynthetic pathway in light-grown barley. There was a net gain in chlorophyll (μg leaf^-1) in 5–6- and 10–11-day-old plants after 17 h dark treatment. The amounts of chlorophyll that accumulated were similar (5.9 and 4.3 μg Chl leaf^-1), despite a twofold difference in leaf size at T₀. The rate of leaf expansion in 5–6-day-old plants greatly exceeded the rate of chlorophyll accumulation and leaves were noticeably paler after dark treatment i.e. there was a reduction in chlorophyll concentration (μg g fresh weight^-1) in spite of an increase in chlorophyll content (μg leaf^-1). The ability of light-grown barley to accumulate chlorophyll in darkness was a function of seedling age. Very young seedlings (3–4-day-old) generally lost chlorophyll in darkness. The decrease in chlorophyll per leaf resulted mainly from loss of chlorophyll b. Preferential loss of chlorophyll b resulted in dramatic increases in the chlorophyll a:b ratio. Since 3–4-day-old seedlings (1) accumulated 5-aminolevulinic acid in the presence of levulinic acid at a rate comparable to older seedlings, and (2) converted exogenous 5-aminolevulinic acid to chlorophyll in the absence of light, it is unlikely that failure of the youngest plants to accumulate chlorophyll in darkness was due to blocks at these steps in the pathway. Net loss of chlorophyll (μg leaf^-1) in 3–4-day-old seedlings in darkness was eliminated by the addition of chloramphenicol, which occasionally produced a small, but significant, gain in total chlorophyll. These results imply that chlorophyll degradation in young barley leaves is strongly influenced by the chloroplast genome, and is a major factor influencing changes in chlorophyll levels in darkness. The present findings are consistent with the suggestion that the failure of 3–4-day-old barley seedlings to accumulate chlorophyll in darkness may be due to chlorophyll turnover in which the rate of degradation exceeds the rate of synthesis.     

Action Spectra for the Inhibition of Hypocotyl Growth by Continuous Irradiation in Light and Dark-Grown Sinapis alba L. Seedlings

Action spectra for the inhibition by continuous (24-hour) irradiation of hypocotyl growth in 54-hour-old Sinapis alba L. seedlings were measured using seedlings which had had four different pretreatments. These seedlings were either (a) dark-grown with a high total phytochrome level, (b) dark-grown with a low total phytochrome level, (c) light-grown with chlorophyll, or (d) light-grown with no chlorophyll [treated with 4-chloro-5-(methylamino)-2-(α,α,α-trifluoro-m-tolyl)-3(2H) -pyridazinone (San 9789)].     

The response of maize seedlings to cadmium stress under hydroponic conditions

The effects of cadmium (Cd) supply level in nutrient solution (0, 12.5, 25, 50, 100, 200, 400, and 800 μM) on growth, Cd accumulation ability, and the related physiological indices of maize (Zea mays L.) seedlings were studied under hydroponic conditions. The results showed that the increments in the shoot height and biomass were stimulated at relatively low external Cd supply levels (<100 μM), while they were inhibited at Cd supply levels over 200 μM. Cd accumulation ability of the maize seedlings also showed the similar stimulation/inhibition pattern as shoot growth, and the Cd contents in the shoots and roots reached the peaks (389.5 and 505.5 mg/kg dry wt, respectively) at 50 μM Cd. The contents of chlorophyll a, chlorophyll b, and carotenoids in the maize leaf blades decreased with increasing external Cd supply level. At the highest Cd supply level (800 μM), the contents of chlorophyll a, chlorophyll b, and carotenoids in the leaf blade were only 38.9, 46.0, and 29.7% of the control plants, respectively. Moreover, chlorophyll b was more sensitive to the Cd stress than chlorophyll a. The increased proline content in the leaf blade of maize seedlings resulted from external Cd stress indicates that maize can adapt to the adversity menace via changing the content of proline.     

Correlations of Growth Rate and De-etiolation with Rate of Ent-Kaurene Biosynthesis in Pea (Pisum sativum L.)

Biosynthesis of the gibberellin precursor ent-kaurene-¹⁴C from mevalonic acid-2-¹⁴C was assayed in cell-free extracts of shoot tips of etiolated and light-grown Alaska (normal) and Progress No. 9 (dwarf) peas (Pisum sativum L.). During ontogeny of light-grown Alaska peas, kaurene-synthesizing activity increased from an undectectable level in 3-day-old epicotyls to a maximum in shoot tips of 9-day-old plants and remained relatively constant thereafter until postanthesis. The capacity for kaurene synthesis in extracts from shoot tips of 10-day-old etiolated Alaska seedlings increased approximately exponentially during the first 12 hr of de-etiolation in continuous high intensity white light and remained relatively constant during the succeeding 24 hr of irradiation. Extracts from light-grown Alaska (normal) shoot tips possessed greater capacity for kaurene synthesis than did extracts from light-grown Progress No. 9 (dwarf) shoot tips. Extracts from shoot tips of either light-grown cultivar displayed greater kaurene-synthesizing capacity than was observed in extracts from their dark-grown counterparts. It is concluded that gibberellin biosynthesis in pea shoot tips is subject to partial regulation by factors controlling the rate of biosynthesis of kaurene.     

Proanthocyanidins and potential precursors in needles of douglas fir and in cell suspension cultures derived from seedling shoot tissues

Proanthocyanidins and their potential precursors have been analyzed by paper chromatography and C₁₈ reversed phase columns with high performance liquid chromatography. Total proanthocyanidins on a dry weight basis in cell suspension cultures derived from seedlings of Douglas fir (Pseudotsuga menziesii) were equal to or greater than those found in mature needles of randomly selected outdoor-grown trees. The major monomer and dimer were catechin and epicatechin-catechin, respectively. Although only procyanidins were detectable in cell suspension cultures, mature needles of outdoor-grown trees contained prodelphinidins as well. Immature needles (flush growth) of the same trees contained only trace amounts of prodelphinidins. Eriodictyol-7-glucoside and dihydroquercetin-3′-glucoside were present in all tissues examined. The amount of eriodictyol-7-glucoside was strongly correlated with total proanthocyanidins in immature needles of flush growth (r = 0.89, p = 0.001). The most complex pattern of flavonoids was found in flush growth needles, which contained in addition to the above, naringenin-7-glucoside and five to six flavone glycosides. Chlorogenic acid was detected only in seedlings and in flush growth needles.     

Light Intensity Alters the Extent of Arsenic Toxicity in Helianthus annuus L. Seedlings

The present study is aimed at assessing the extent of arsenic (As) toxicity under three different light intensities—optimum (400 μmole photon m^?2 s^?1), sub-optimum (225 μmole photon m^?2 s^?1), and low (75 μmole photon m^?2 s^?1)—exposed to Helianthus annuus L. var. DRSF-113 seedlings by examining various physiological and biochemical parameters. Irrespective of the light intensities under which H. annuus L. seedlings were grown, there was an As dose (low, i.e., 6 mg kg^?1 soil, As₁; and high, i.e., 12 mg kg^?1 soil, As₂)-dependent decrease in all the growth parameters, viz., fresh mass, shoot length, and root length. Optimum light-grown seedlings exhibited better growth performance than the sub-optimum and low light-grown seedlings; however, low light-grown plants had maximum root and shoot lengths. Accumulation of As in the plant tissues depended upon its concentration used, proximity of the plant tissue, and intensity of the light. Greater intensity of light allowed greater assimilation of photosynthates accompanied by more uptake of nutrients along with As from the medium. The levels of chlorophyll a, b, and carotenoids declined with increasing concentrations of As. Seedlings acquired maximum Chl a and b under optimum light which were more compatible to face As₁ and As₂ doses of As, also evident from the overall status of enzymatic (SOD, POD, CAT, and GST) and non-enzymatic antioxidant (Pro).