THE MORPHOLOGICAL EFFECTS OF PROTEIN SYNTHESIS INHIBITION IN MARSILEA

Marsilea vestita and M. drummondii were grown in sterile cultures to which concentrations of the protein synthesis inhibitors, 2-thiouracil (10 mg/liter) and 5-fluorouracil (1 mg/liter) had been added. When young sporelings are grown in a solution of thiouracil at optimum concentration, there is an inhibition of the rate of leaf formation, a retardation of the leaf heteroblastic series, and all leaves develop as land forms. When thiouracil is added to plants which are already producing typical adult, quadrifid leaves, the effects depend on whether the treated plants are water or land forms. Plants which are typically water forms convert to land forms. After treatment successive leaves develop typical sunken stomata on both leaf surfaces. The tissues of the rhizome, root and petiole are more compact and, in general, the cells of the plant have thicker walls. Vascular patterns are not changed, though the size of the rhizome, root and petiole may be reduced. Plants which are typically land forms are less affected than the water forms, but they show a small reduction in apex volume and an apparent reversion of the leaflet number from the typical quadrifid leaf to a trifid, bifid, or single lamina condition. In both land and water forms apical dominance may be broken by treatment with 10 mg/liter thiouracil or 1 mg/liter fluorouracil and numerous lateral branches develop. Higher concentrations (15–25 mg/liter of thiouracil) may result in abnormal development of lateral axillary buds, petiole bases and leaflets. The meristems of the plant are differentially sensitive to thiouracil; leaflet meristems are most sensitive, the root meristems are the least sensitive. It appears that a true reversion to juvenile leaf development need not occur even though protein synthesis and the volume of the apex are reduced. The development of the land or water form in Marsilea appears to depend on rate of growth. Hence inhibition of the growth of typical water forms, through inhibition of protein synthesis, causes a shift in development toward the morphology typical of land forms.     

The Correlation of Physiological Differences and Varions Leaf Forms of an Aqnatic Plant

An analysis of the celluar constituents of the leaves of an aquatic fern, Marsilea vestita, was done after and during the formation of different leaf forms. Land leaves of Marsilea produced almost twice as much dry material, soluble carbohydrate, insoluble protein and soluble peptides compared to submerged leaves on a fresh weight basis, but on a dry weight basis there is tittle difference between the two types of leaves. the submerged leaf soluble nitrogen fraction consisted mostly of ammonia and free amino acids, while that of the land leaf probably con tained mostly peptides. Free arginine was present in large quantities in megaspores and as the plant matured, at the expense of the megaspore, the amount of arginine left in the pool diminished. A correlation was found between soluble sugar increase and leaf elongation, Just prior to the production of submerged leaves the developing leaves contained low levels of protein and soluble sugar, but high levels of soluble nitrogen and an increased amount of starch. A theory which involves metabolism was proposed to account for the leaf form changes in Marsilea.     

MORPHOLOGY OF MARSILEA VESTITA. III. MORPHOGENESIS OF THE LEAVES OF ETIOLATED PLANTS

The laminae of etiolated Marsilea vestita leaves develop by means of marginal meristems. Unlike light-grown plants, the form of the etiolated plant is not affected by growth on a solid medium. All of the young leaves isolated from light-grown submerged plants will elongate in darkness. The smallest, etiolated, uncoiled leaves develop into land leaves when they are placed in light, and this development occurs regardless of whether the leaves remain on the plant or are isolated on nutrient agar. Only these smallest leaves (2.3 mm average length) are actually capable of being converted from a submerged leaf form to a land leaf form by darkness.     

A CORRELATION BETWEEN THE APICAL CELL AND THE HETEROBLASTIC LEAF SERIES IN MARSILEA

Plants of three species of Marsilea (M. vestita, M. villosa, M. drummondii) were grown in sterile culture under controlled conditions, and stem apices were sampled at one of the three heteroblastic leaf forms typical of this plant: spatulate, bifid, or quadrifid leaves. Statistical analyses were made of the relationship between the area of the apical cell and the leaf form which the plant produces under varied growth conditions. For all three species there is a statistically significant correlation (1% level) between apical cell area and leaf form. The analysis indicates that 83% of the variation in apical cell area in Marsilea vestita, 52% in M. villosa, and 54% in M. drummondii can be related to the change in leaf form. An increase in the glucose concentration of the culture medium increases the average apical cell area, and the addition of the protein synthesis inhibitor 2-thiouracil at concentrations of 10 mg/liter and 25 mg/liter decreases the average apical cell area. A certain average apical cell area is necessary for the production of a particular leaf form in the series. The area is a relative rather than an absolute size under any one particular growth condition. Any growth condition which inhibits or reverses the increase in apical cell size (which is typical of the normal growth pattern) will inhibit or reverse the normal heteroblastic series. Under constant conditions, the average size at which particular leaf forms develop appears to be species specific. These results confirm a generally held idea that the apex size is related to the heteroblastic leaf series, and they indicate that the area of the apical cell is the key factor rather than the volume or the number of cells of the apex. Undoubtedly the area of the apical cell is only a reflection of the physiological or morphological characteristics of the apical meristem that underlie the heteroblastic leaf series and which currently do not appear to lend themselves to more direct quantitative analysis.     

CAUSAL MECHANISMS OF LEAF DIMORPHISM IN THE AQUATIC ANGIOSPERM CALLITRICHE HETEROPHYLLA

This report investigates the physiological basis for the production of dimorphic leaves on the aquatic angiosperm Callitriche heterophylla. In nature, the leaf morphology of this plant depends on whether the shoot apex is submerged in or emergent from water. The water-form leaves that develop on submerged apices assume a long, linear shape in contrast to the short, obovate appearance of land forms on emergent apices. The parameters of length/width ratio and stomatal density were used as developmental indices to characterize how natural conditions, fluctuating water levels and other experimental treatments affect leaf shape. Transferring submerged and emergent shoots to the alternative culture conditions caused immature leaves to assume the characteristics appropriate to their new environment. Moreover, the treatments of 0.24 mol mannitol, high temperature (30 C) and 10^−-5 m abscisic acid induced submerged shoots to produce land-form leaves whereas 10^−-5 m gibberellic acid mediated the development of water-form leaves on emergent shoots. Water, osmotic and pressure potentials of immature leaves in the control and experimental treatments were determined by thermocouple psychrometry. Under natural conditions, growing water forms exhibited high turgors (3–5 bars) while developing land forms showed much lower turgors (0–1 bar). Similar correlations between turgor pressure and leaf morphology were observed in the case of the gibberellic acid and mannitol treatments. However, abscisic acid and high temperature caused the developing land-form leaves to exhibit high turgors without a concomitant change to the water-form morphology. Microscopic measurements of epidermal cells established that irrespective of the experimental conditions, water-form leaves had longer and narrower epidermal cells with less convoluted anticlinal walls than land forms. Cell counts indicated that the numbers of epidermal cells did not account for the observed differences in leaf morphology. The results are interpreted in terms of how cell expansion might regulate leaf morphology in aquatic angiosperms.     

Blue Light and Abscisic Acid Independently Induce Heterophyllous Switch in Marsilea quadrifolia

In natural habitats Marsilea quadrifolia L. produces different types of leaves above and below the water level. In aseptic cultures growth conditions can be manipulated so that leaves of the submerged type are produced continuously. Under such conditions the application of either blue light or an optimal concentration of abscisic acid (ABA) induced the development of aerial-type leaves. When fluridone, an inhibitor of ABA biosynthesis, was added to the culture medium it did not prevent blue light induction of aerial leaf development. During blue light treatment the endogenous ABA level in M. quadrifolia leaves remained unchanged. However, after the plants were transferred to an enriched medium, the ABA level gradually increased, corresponding to a transition in development from the submerged type of leaves to aerial leaves. These results indicate that the blue light signal is not mediated by ABA. Therefore, in the regulation of heterophyllous determination, discrete pathways exist in response to environmental signals.     

ABSCISIC ACID INDUCES LAND FORM CHARACTERISTICS IN MARSILEA QUADRIFOLIA L.

In aseptically cultured rhizome segments of the aquatic fern Marsilea quadrifolia, the addition of abscisic acid (ABA) to the liquid medium induced development of morphological characteristics distinctive to the land form of the plant. The land-form characteristics induced by ABA included a change in leaf morphology, an increase in the surface area of leaflets, differentiation of stomata and trichomes, elongation of petioles and roots, development of lateral roots, shortening of rhizome, and reduction in the number of leaves and roots formed on each plant.     

Growth from two transient apical initials in the meristem of Selaginella kraussiana

A major transition in land plant evolution was from growth in water to growth on land. This transition necessitated major morphological innovations that were accompanied by the development of three-dimensional apical growth. In extant land plants, shoot growth occurs from groups of cells at the apex known as meristems. In different land plant lineages, meristems function in different ways to produce distinct plant morphologies, yet our understanding of the developmental basis of meristem function is limited to the most recently diverged angiosperms. To redress this balance, we have examined meristem function in the lycophyte Selaginella kraussiana. Using a clonal analysis, we show that S. kraussiana shoots are derived from the activity of two short-lived apical initials that facilitate the formation of four axes of symmetry in the shoot. Leaves are initiated from just two epidermal cells, and the mediolateral leaf axis is the first to be established. This pattern of development differs from that seen in flowering plants. These differences are discussed in the context of the development and evolution of diverse land plant forms.     

Genotypic Differences in Leaf Water Potential, Abscisic Acid and Proline Concentrations in Spring Wheat during Drought Stress

Recent work with spring wheat has revealed significant genotypicvariation in changes of water potential and abscisic acid (ABA)concentration in response to drought Two experiments with eightspring wheat genotypes have been carried out to check the earlierwork on relationships between water potential and ABA concentrationand to examine causes of genotypic variation in the rate ofdecline of water potential during drought Changes in prolineconcentration were also studied Plants were grown in controlled environment cabinets with nutrientsolution culture and were stressed by withholding water as thefifth or sixth leaf on the main stem emerged. Plants were harvested4, 5 and 6 days after the treatment commenced and measurementsof leaf water potential, stomatal conductance, ABA and prolineconcentrations, and tissue d wts were taken. Significant genotypic variation was found in the decrease ofwater potential with time and in the slopes of linear regressionsof ABA concentration on water potential, confirming earlierresults When differences between leaf areas at the start of the treatmentwere minimised by varying the genotype sowing date significantgenotypic variation in water potentials at harvest was stillobtained. The change in water potential was significantly positivelycorrelated with shoot root d wt ratios at harvest and pre-treatmentstomatal conductances. Proline concentrations were significantly correlated with waterpotential for every genotype, although there was no clear evidenceof genotypic variation in proline concentrations at a givenwater potential The possible role of ABA concentration in drought resistanceof cereals is discussed Triticum aestivum L, spring wheat, water potential, abscisic acid, proline, drought stress     

Co-existence of three species of amphibious plants in relation to spatial and temporal variation: Investigation of plant responses

Plant responses in Ludwigia peploides (Kunth) Raven ssp. montevidensis (Spreng.) Raven and Marsilea mutica Mett., and the introduced Myriophyllum aquaticum (Vellozo) Verdc. were observed and related to co-existence of the species in a freshwater lagoon in the Sydney basin. Water levels vary in the lagoon, and plants of all three species grow readily from stem fragments on freely draining, waterlogged and submerged soils. In Marsilea, sporelings develop rapidly from ruptured sporocarps when wet. In Ludwigia, the seeds germinate in white and in red light, but not in darkness or far-red light. They germinate readily on soil and in water, floating or when held submerged. At 30°C, germination is rapid, at 40°C slower and less, and at 10°C nil, but seeds imbibed at 10°C and then exposed at 15°C have greater and more rapid germination than seeds maintained at 15°C. In relation to seasonal variation, plants of all three species grew at 10, 20, 30 and 40°C on freely draining, waterlogged and submerged soils, with the optimum temperature for growth apparently lower in Myriophyllum than in the other two species. In relation to water depth, plants of all three species grew when submerged to I m; the stems of Ludwigia and Myriophyllum elongating and reaching the water surface, and the petioles in Marsilea elongating bringing the leaf blades on to the surface, each within a week during summer. The floating leaf blades in Marsilea increase in size with depth of submergence of the subtending shoots. Mean relative growth rates (R?) under the most favourable experimental conditions were least in plants of Marsilea, the species occurring along more exposed parts of the shoreline in the lagoon. It was concluded that, while water levels in the lagoon were stable, differences in growth form between the species were probably more critical in their co-existence than quantitative differences in their growth: the floating leaves of Marsilea, being more resistant to wind and wave action than the floating shoots of Ludwigia and Myriophyllum, occupy the more exposed parts of the lagoon 'S edge. The most sheltered parts are occupied by the shoots of Myriophyllum which ride high on the water and overtop shoots of Ludwigia. Shoots of Ludwigia appear to be more resistant than those of Myriophyllum to removal and damage by wind and wave action, and achieve their greatest abundance on the water surface in situations of intermediate exposure to wind and wave action. Using Grubb's (1977) classification of niches, in Bushells Lagoon the three species appear to be primarily differentiated in habitat niche, less in regeneration niche and even less in phenological niche. In the lagoon, their habitat niches apparently overlap in water depth but are differentiated along a gradient of exposure.     

Factors influencing the production of hardened glaucous carnation plantlets in vitro

Medium type, its water status and the relative humidity in the culture vessel modified carnation leaf development in vitro. Carnation shoot apices cultured on liquid or on 0.8% agar solidified media developed into plantlets having succulent and translucent leaves which are not transplantable to non-aseptic conditions. Increasing the agar and/or sucrose concentration in the medium as well as decreasing the relative humidity in the culture vessel by a desiccant promoted glaucous leaf production. Increased water status (H₂O and relative humidity) increased shoot proliferation and translucency of leaves. Decreased water status reduced shoot proliferation but induced the formation of glaucous leaves. The culture of apices for 5–6 days on liquid medium prior to their sub-culture to 1.5% agar medium improved shoot proliferation and normal leaf development. An agar slant prevented the submergence of apices in water accumulating on the medium and thus reduced leaf translucency. Survival was further increased by the transfer of plantlets in uncapped culture vessels to a desiccator for 1–2 weeks prior to transplanting to soil.     

THE INFLUENCE OF WATER STRESS AND TEMPERATURE ON LEAF PUBESCENCE DEVELOPMENT IN ENCELIA FARINOSA

The degree of leaf pubescence development in the arid land shrub Encelia farinosa Gray is affected by air temperature, leaf water potential, and previous history of the apical meristem during the current growing season. Changes in leaf pubescence levels change leaf spectral characteristics and affect both leaf temperature and photosynthesis. Decreasing leaf water potentials and increasing air temperatures both independently increase pubescence development as measured by decreased leaf absorptances. During any one growing season leaf absorptance may change reversibly coincident with air temperature changes, but with respect to water stress leaf absorptance only decreases as the season progresses. The ecological significance of regulation of the leaf spectral characteristics is discussed.     

Differential expression of parental genomes during formation of embryonic organs in the early development of the fernMarsilea quadrifolia

This paper reports some experimental results related to the expression of parental genomes during the development of the water fernMarsilea quadrifolia L., a plant with graduated heteroblastic development. We found a non-random segregation of the original chromatids of the paternal genome, as shown by autoradiography of embryo sections after fertilization of eggs with [³H]thymidin-tagged sperm. From the 16-cell stage on, label is mainly concentrated in the sectors where apical cells will differentiate, and later in or near the apical areas of the young organs. Similar results had been found previously inMarsilea vestita. We also observed aberrations in primary organ development after fertilization with sperm treated with actinomycin, ethidium bromide, hydroxyurea, bromodeoxyuridin, or chlorambucil. The growth of the first organs was quite normal, indicating that they develop under the control of the maternal genome. The subsequent organs displayed abnormal development, indicating that they are under the control of both parental genomes.     

Altitudinal variation in stomatal conductance,nitrogen content and leaf anatomy in different plant life forms in New Zealand

Summary This study is part of a series of investigations on the influence of altitude on structure and function of plant leaves. Unlike most other mountain areas, the Southern Alps of New Zealand provide localities where physiologically effective moisture stress occurs neither at high nor at low elevation, but the changes in temperature and radiation with elevation are similar or even steeper than in most other regions. In comparison with results from other mountains, where moisture may impair plant functioning at low elevation, this study allows an estimation of the relative role of water for the expression of various leaf features typically associated with alpine plants. Maximum leaf diffusive conductance (g), leaf nitrogen content (LN), stomatal density (n) and distribution, as well as area (A), thickness (d) and specific area (SLA) of leaves were studied. Three different plant life forms were investigated over their full altitudinal range (m): trees, represented by Nothofagus menziesii (1,200 m), ericaceous dwarf shrubs (1,700 m), and herbaceous plants of the genus Ranunculus (2,500 m). In all three life forms g, LN, and n increased, while SLA and A decreased with elevation. Recent investigations have found similar trends in other mountains from the temperate zone, but the changes are larger in New Zealand than elsewhere. Herbs show the greatest differences, followed by shrubs and then trees.It is concluded that g is dependent upon light climate rather than water supply, whereas SLA and related structural features appear to be controlled by the temperature regime, as they show similar altitudinal changes under different light and moisture gradients. The higher leaf nitrogen content found at high elevations in all three life forms, suggests that metabolic activity of mature leaves is not restricted by low nitrogen supply at high altitude. In general, the leaves of herbaceous plants show more pronounced structural and functional changes with altitude than the leaves of shrubs and trees.     

Localization of Ca++-containing antimonate precipitates during mitosis

Intracellular bound Ca++ has been localized throughout mitosis and cytokinesis in two plant species by means of in situ precipitation with potassium antimonate and electron microscope visualization. Identification of Ca++ as the major cation precipitated was made by comparing solubility properties in water, EDTA, and EGTA of the intracellular deposits with respect to those of K+-, Mg++-, and Ca++- antimonate standards. In spermatogenous cells of the water fern, Marsilea vestita, and stomatal complex cells of barley, Hordeum vulgare, antimonate deposits have been found associated with the endoplasmic reticulum (ER), vacuoles, euchromatin/nucleoplasm, and mitochondria. The last contain a much higher density of precipitates in Marsilea than in Hordeum. Dictyosomes and the nuclear envelope of Marsilea also contain antimonate deposits, as do the plasmalemma, cell wall, and phragmoplast vesicles of Hordeum. Microtubule-organizing centers such as kinetochores and the blepharoplast of Marsilea do not stain. In spite of differences in associated antimonate between certain organelles of the two species, the presence of antimonate aong the ER throughout the cell cycle is common to both. Of particular interest are those precipitates seen along the tubules and cisternae of the extensive smooth ER that surrounds and invades the mitotic spindle in both species. The ability to bind divalent cations makes the mitotic apparatus (MA)-associated ER a likely candidate for regulation of free Ca++ levels in the immediate vicinity of structural components and processes that are Ca++-sensitive and proposed to be Ca++-regulated.     

Water sources and water-use efficiency of desert plants in different habitats in Dunhuang,NW China

To understand habitat associated differences in desert plant water-use patterns, water stable oxygen isotope composition was used to determine water source and leaf carbon isotope composition (δ ¹³C) was used to estimate long-term water-use efficiency in three typical habitats (saline land, sandy land and Gobi) in Dunhuang. The primary findings are: (1) in the three habitats, plant species used mainly deep soil water (>120 cm), except for Kalidium foliatum in the saline land, which relied primarily on 0–40 cm soil water; (2) in the saline land and Gobi habitat, Alhagi sparsifolia had the most negative foliar δ ¹³C; in the sandy land, Elaeagnus angustifolia leaf was enriched in ¹³C than the other three species in 2011, but no species differences in foliar δ ¹³C was observed among the four species in 2012; (3) common species (Tamarix ramosissima and A. sparsifolia) may alter their water sources to cope with habitat differences associated changes in soil water availability with deeper water sources were used in the Gobi habitat with lower soil water content (SWC) compared to in the saline land and sandy land; (4) we detected significant habitat differences in foliar δ ¹³C in A. sparsifolia which may have resulted from differences in SWC and soil electrical conductivity. However, no habitat differences in foliar δ ¹³C were observed in T. ramosissima, which may attribute to the strong genetic control in T. ramosissima or the ability to access stable deep soil water. Overall, the results suggest that extremely arid climate, root distribution and soil properties worked together to determine plant water uptake in Dunhuang area.     

毛乌素沙地油蒿叶建成成本及相关叶性状沿降水梯度的变化

干旱区植物在如何提高水分利用效率与降低叶建成成本之间可能存在一种权衡。我们假设: 与湿润区植物相比, 干旱区植物能通过调节叶功能性状之间的关系(如通过提高单位面积叶氮含量(N_area)), 实现在相同的叶建成成本下具有更高的水分利用效率。为了验证这一假设, 该研究以毛乌素沙地的广布物种油蒿(Artemisia ordosica)为研究对象, 分析了油蒿叶建成成本沿降水梯度的变化规律及其与比叶面积(SLA)、单位质量叶氮含量(N_mass)、N_area和叶碳稳定同位素比率(δ ¹³C)的关系。结果表明: 油蒿单位质量叶建成成本(CC_m)在不同降水条件下差异不显著, 而单位面积叶建成成本(CC_a)在不同降水条件下虽有显著差异, 但并未随降水减少而明显增加。油蒿CC_m与SLA无显著相关性, 与叶δ ¹³C值呈显著正相关关系。油蒿叶建成成本与N_area呈显著正相关关系, 但这种关系格局在低降水量(264 mm)区与高降水量(310-370 mm)区之间存在策略位移现象——即在相同叶建成成本下, 低降水量区植物比高降水量区植物具有更高的N_area。以上结果表明, 尽管高水分利用效率与高叶建成成本相关, 但与高降水量区植物相比, 低降水量区植物具有较高的N_area并没有导致其叶建成成本增加。     

六盘山林区几种土地利用方式对土壤中可溶性有机碳浓度影响的初步研究

 测定分析了六盘山林区典型天然次生林（杂灌林、山杨（Populus davidiana）和辽东栎（Quercus liaotungiensis）林）、人工林（13、18和25年华北落叶松（Larix principis_rupprechti））、农田和草地中穿透雨、凋落物淋溶液、土壤溶液和渗漏水溶液及地下水和降雨中可溶性有机碳 (DOC)的浓度。结果显示：5～10月,雨水中DOC浓度为0.80～1.60mg•L－1,地下水中在2.43～7.66 mg•L－1。9～10月,穿透雨中DOC浓度为1.78～15.20 mg•L－1,其中天然次生林和人工林比农田和草地中高,这些DOC与地表凋落物碳年形成量正相关显著。凋落物浸泡24 h后淋溶产生的DOC浓度为12.30～64.79 mg•L－1,占凋落物碳贮量比例不足1%;浓度方面,天然次生林比农田和草地分别高400%和153%,人工林比农田和草地分别高194%和50%;比例方面,农田和草地比天然次生林分别高79%和98%、比人工林分别高180%和210%,这些DOC浓度与落叶、小枝、碎小物和腐解物碳贮量的正相关显著。9～10月,0～20 cm土层溶液中DOC浓度为7.88～88.44 mg•L－1,占土壤有机碳密度的比例不足0.1%,它们随土层加深而下降,其中天然次生林和人工林中下降幅度比农田和草地中大;浓度方面,天然次生林、人工林比农田和草地中高,差异主要在0～10 cm土层;比例方面,天然次生林DOC比例比农田和草地中低,人工林比它们高,差异主要在0～10 cm土层;这些DOC浓度与土壤湿度及凋落物层碳贮量正相关显著。5～10月,0～40 cm土层渗漏水中DOC浓度为5.76～58.84 mg•L－1,天然次生林、人工林比农田和草地高,差异主要在0～10 cm土层;它们随土层加深而下降,其中天然次生林和人工林下降幅度比农田和草地中大。这些差异可能由土地利用引起的植被和土壤性质改变及其对水文过程的影响所致,说明陆地生态系统中DOC浓度受土地利用变化的影响较大。     

The Effects of Changes in the Inorganic Nitrogen Supply on the Growth and Development of Marsilea in Aseptic Culture

The effects of changes in the inorganic nitrogen source of themedium on the growth and development of Marsilea in asepticculture are described. A change in nitrate concentration to0·2 or 5 x that of the normal medium had initially noeffect on growth or heteroblastic development, but in the 0·2nitrogen medium there was finally a reversion to juvenile stagesfollowed by early cessation of growth. Growth and heteroblastic development were stimulated by thesubstitution of the nitrate of the medium by ammonium saltsor urea. Nitrite was little inferior to nitrate as a nitrogensource, but hydroxylamine was toxic even at low concentrations.Addition of sodium chloride led to improved growth and developmentin nitrate media, an effect referred to beneficial effects ofthe chloride ion. Symptoms of toxicity and eventual death of the cultures whichoccurred after prolonged growth on ammonium media were due toan acid drift of the media rather than to a specific effectof the ammonium ion.