THE ROLE OF LIGHT IN HISTOGENESIS AND DIFFERENTIATION IN THE SHOOT OF PISUM SATIVUM. II. THE LEAF

Thomson , Betty F., and Pauline Monz Miller . (Connecticut Coll., New London.) The role of light in histogenesis and differentiation in the shoot of Pisum sativum, II. The leaf. Amer. Jour. Bot. 49(4): 383–387. Illus. 1962.—Development of the form and anatomy of leaves was studied in plants of Pisum sativum grown in vermiculite under constant conditions and exposed daily to red or white light or kept in continuous darkness. The red light used had an intensity in the morphogenetically active red region of the spectrum of 70–75% that of the white light. Light had no effect on the manner of initiation or early development of leaf primordia. Quantitative data from older leaves showed that light has no effect on the pattern of later development but does affect the rate and extent of development. Under all light conditions, the length of the leaflet is closely correlated with the state of its internal anatomy. “Mature” etiolated leaves duplicate young stages of light-grown leaves. Mature leaves grown in red light duplicate not-quite-mature leaves grown in white light. The difference between white-light and red-light leaves is attributed here to light intensity and resembles that between sun and shade leaves.     

INTERACTION OF LIGHT QUALITY AND NUCLEASES IN THE GROWTH OF THE GAMETOPHYTES OF ASPLENIUM NIDUS

Appropriate concentrations of ribonuclease A and B selectively inhibited initiation of two-dimensional morphology in the gametophytes of Asplenium nidus, grown under a photoperiod of 5½ hr white light. Filamentous growth was promoted in such sporelings, the individual cells of which were significantly longer than corresponding cells of the control. Higher concentrations of enzymes were required to inhibit two-dimensional growth in the gametophytes grown in blue light. Concentrations of ribonuclease A or B which inhibited two-dimensional growth in white light promoted growth in length of the protonema in red light. Growth modifications in the sporelings induced by deoxyribonuclease in different light conditions were similar to those induced by the ribonucleases. The results lend further support to the postulated role of RNA in the regulation of two-dimensional growth in fern gametophytes.     

THE EFFECT OF AUXIN AND GUANINE ON CELL EXPANSION AND CELL DIVISION IN THE GAMETOPHYTE OF THE FERN,ONOCLEA SENSIBILIS

Miller , J. H. (Yale U., New Haven, Conn.) The effect of auxin and guanine on cell expansion and cell division in the gametophyte of the fern, Onoclea sensibilis. Amer. Jour. Bot. 48(9): 816–819. Illus. 1961.—Auxin and guanine promote cell expansion in 0. sensibilis gametophytes. The optimum concentration of auxin for total expansion is 10^−-5 M, but the optimum for elongation is 10^−-6 M. Above this concentration the cells expanded isodiametrically. Guanine is active at higher concentrations than auxin. Increasing concentrations of auxin progressively inhibit red light-induced cell division, while guanine has no effect on cell division. Neither kinetin nor adenine promotes cell expansion or cell division.     

The Role of Light in the Interrelated Processes of Morphogenesis and Photosynthesis in the Fern Gametophyte

The interrelationship of photosynthesis and photomorphogenesis in the gametophyte of the fern Pteridium aquilinum was examined in detail. High energy red light produces filamentous gametophytes composed of greatly elongated cells. Blue light of equal energy produces normal cordate gametophytes composed of isodiametric cells. The assimilative rates of the two forms are identical during the initial stages of development. However, the filamentous gametophytes eventually exhibit a decline in assimilation not matched by their normal counterparts. Although this decrease in assimilation of gametophytes exposed to red light may be traced to a decrease in the photo-synthetic rate due to a decrease in chlorophyll synthesis, these events occur subsequent to the establishment of the filamentous mode of growth and are therefore a result, not a cause, of filamentous development. Gametophyte assimilation is also impeded under low energy blue light, due to a reduction in the photosynthetic rate at this low energy; and the initial filamentous stage, in which the cells remain normally isodiametric, is not exceeded. The addition of extra exogenous sucrose restores the capacity for growth and normal cordate prothalli result. It may therefore be concluded that the ultimate form of the gametophyte is under the control of a photoreceptor that is sensitive to blue light, and activated at a relatively low energy level. Red light at any level is ineffective in promoting normal cordate development, but is effective in supporting growth through vigorous photosynthesis, yielding a filament composed of greatly elongated cells. The eventual photosynthetic decline of the filamentous form is due directly to this particular growth mode. Low energy blue light is sufficient to effect form (the cells remain isodiametric), but insufficient to provide the necessary energy for growth through vigorous photosynthesis. Therefore, in order for the gametophyte to reach maturity it must be supplied with light energy that not only satisfies the requirement for form but the requirement for growth as well. The implications of these requirements are discussed in the text.     

THE RELATIONSHIP BETWEEN THE PROMOTION OF ELONGATION OF FERN PROTONEMATA BY LIGHT AND GROWTH SUBSTANCES

Germinating spores of the fern Onoclea sensibilis L. were grown in darkness, so that they developed as filaments (protonemata). Brief daily exposure of the filaments to red, far-red or blue light increased the rate of filament elongation. Filament elongation was also promoted by indoleacetic acid. When filament elongation was promoted with both indoleacetic acid and exposure to light, the growth promotions caused by red and far-red light were additive to auxin-induced growth. Blue light promoted elongation only at sub-optimal concentrations of auxin. Elongation induced by guanine was additive to red- and far-red-induced elongation. Gibberellic acid had no effect on elongation under any condition. Blue-light-induced elongation resembled auxin-induced elongation in its requirement for exogenous sucrose and sensitivity to inhibition by parachlorophenoxyisobutyric acid. Red and far-red light were active regardless of the presence or absence of sucrose and promoted elongation at a concentration of parachlorophenoxyisobutyric acid which completely inhibited blue-light-induced elongation.     

THE ROLE OF LIGHT IN HISTOGENESIS AND DIFFERENTIATION IN THE SHOOT OF PISUM SATIVUM. I. THE APICAL REGION

Thomson , B. F., and P. M. Miller . (Connecticut Coll., New London.) The role of light in histogenesis and differentiation in the shoot of Pisum sativum. I. The apical region. Amer. Jour. Bot. 49(3): 303–310. Illus. 1962.—Seedlings of Pisum sativum grown under constant conditions and kept in total darkness or exposed daily to red or white light were harvested at the same plastochron age and examined histologically to determine what specific aspects of histogenesis and differentiation are affected by light. The tissue organization of the shoot apex is the same in all light conditions to a point below the 2 youngest leaf primordia. The first detectable difference is a slight thickening of the internode in light due to more and larger cells. The first effect on longitudinal growth appears below the fourth youngest primordium and consists of an increase of internode length in light-grown plants. This is associated with a greater distance between the apex and the first mature protoxylem. The distance from apex to the first pith, provascular strands, and protophloem and the distances between the 4 youngest leaf primordia are not affected by light.     

THE ROLE OF LIGHT IN HISTOGENESIS AND DIFFERENTIATION IN THE SHOOT OF PISUM SATIVUM. III. THE INTERNODE

Thomson, Betty F., and Pauline Monz Miller. (Connecticut Coll., New London.) The role of light in histogenesis and differentiation in the shoot of Pisum sativum. III. The internode. Amer. Jour. Bot. 50(3): 219–227. Illus. 1963.—Seedlings of Pisum sativum were grown under constant conditions and exposed daily to red or white fluorescent light or kept in total darkness. Counts and measurements of internodal cells in both transverse and longitudinal directions show that light does not alter the sequence or pattern of tissue differentiation, including the sequence of xylem maturation within the vascular bundle. Light does accelerate the rate of a constant course of differentiation. Light advances the time of division and enlargement of cortex, xylem, phloem, and pith cells in the longitudinal direction but reduces both the final number and the final length attained in all cases. It is concluded that light accelerates all phases of shoot growth and differentiation and that cell division and elongation in the later phases of internodal growth are reduced by light because of accelerated cell maturation.     

THE EFFECT OF LIGHT INTENSITY AND SUCROSE FEEDING ON THE FINE STRUCTURE IN CHLOROPLASTS AND ON THE CHLOROPHYLL CONTENT OF ETIOLATED LEAVES

Changes in the fine structure of proplastids of etiolated leaves exposed to various conditions of light and darkness for 24 and 48 hours were investigated, and the chlorophyll content of the leaves so treated was determined in vivo. The light treatments were given while the leaves were floated on tap water or on a 0.2 M sucrose solution. Leaves floated on water under low light intensity (2 foot-candles) were low in chlorophyll and contained plastids with concentric rows of vesicles. Transferring the leaves back to darkness resulted in the disappearance of the concentric rigs and re-formation of vesicular centers together with straight rows of vesicles and tubules, evenly spaced throughout the stroma. Chloroplasts of leaves floated on a sucrose solution under low light showed large vesicular centers together with stacks of rows of elongated tubules. The same chloroplast structure was found in leaves floated on a sucrose solution in the dark, after having been exposed to weak light for 24 hours. Chlorophyll content in these leaves was the same as in leaves floated on water under high light intensity, where the chloroplasts had normal grana and lamellae. The effect of the investigated factors on plastid development is discussed.     

GROWTH PATTERNS OF PEA SEEDLINGS IN DARKNESS AND IN RED AND WHITE LIGHT

Thomson , Betty F., and Pauline Monz Miller . (Connecticut Coll., New London.) Growth patterns of pea seedlings in darkness and in red and white light. Amer. Jour. Bot. 48(3): 256–261. Illus. 1961.—Seedlings of peas were grown in vermiculite at 22°C. and exposed 16 hr. daily to red or white light or kept in darkness. Others were grown in soil in the greenhouse. Samples harvested daily to 16 days were dissected, the length of each internode and leaf measured and the total number of leaves and leaf primordia counted. The form of the stem apex and youngest primordia and interrodes is the same in light as in darkness. Leaf production is accelerated very slightly and the growth of leaves and internodes is decidedly accelerated by light. Leaf-leaf, leaf-internode and internode-internode correlations indicate that the morphogenetic effect of light is limited to later stages of organ growth. Dry weight is consumed more rapidly in light than in darkness, probably because of more rapid growth and slightly greater amounts of respiring tissue in light-grown plants.     

PHYLOGENETIC AND BIOGEOGRAPHIC AFFINITIES OF THE HALYMENIALES AND RHODYMENIALES (RHODOPHYTA) FROM THE GULF OF MEXICO

A kelp/red algal symbiosis is described from nature based on extensive collections from the San Juan Islands, Washington. Kelp gametophytes were found as endophytes in the cell walls of seventeen species of red algae in three different kelp communities. Host red algae were mostly filamentous (e.g., Pleonosporium vancouverianum ) or polysiphonous (e.g. Polysiphonia paniculata ). The kelp gametophytes completed vegetative and reproductive development in the hosts with gametangia formed at the host surface and with sporophytes up to several mm in height being produced while still attached to the host. To date, none of the kelp gametophytes from nature have been identified to genus or species, although the gametophyte of Nereocystis luetkeana is a potential candidate for the symbiosis. Preliminary observations from Nova Scotia and the Isle of Man have not found the association in the Atlantic Ocean. Laboratory studies in Korea successfully reconstructed the symbiosis in the red alga Aglaothamnion oosumiense using zoospores of Undaria pinnatifida but not Laminaria religiosa. Here we outline the development of the symbiosis and discuss the potential adaptive significance of the kelp/red algal interaction.     

THE TRANSITION FROM FILAMENTOUS TO TWO-DIMENSIONAL GROWTH IN FERN GAMETOPHYTES. III. INTERACTION OF CELL ELONGATION AND CELL DIVISION

Periodic cell divisions were induced in gametophytes of Pteridium aquilinum by daily irradiation with white light. In white-dark cycles, the rate of cell division was promoted by increased time in white light; cell elongation was not affected. The time of transition to two-dimensional growth (days to 5% 2-D) was closely associated with the mitotic rate. For white-red cycles, the rate of elongation was controlled by the intensity of red light (wavelengths over 550 nm). This increased elongation delayed the initiation of 2-D development. In both cases the rate of transition to 2-D growth was correlated with the amount of elongation per division.     

THE EFFECTS OF TEMPERATURE ON THE GERMINATION AND RADICLE GROWTH OF PHOTOSENSITIVE LETTUCE SEED

1. The time course of germination of Grand Rapids lettuce seedshas heen followed with different combinations of temperature(3°–35°) and irradiation (red or far-red light).For each set of conditions the following three parameters weredetermined: (i) the time required for half maximum germination,(ii) the rate of germination during the actively germinatingphase, and (iii) the maximum germination attained. In general,as the temperature was lowered, with dark-imbibed seeds, (i)became longer, (ii) became lower, but (iii) became progressivelyhigher. The effect of red light at any temperature was to shorten(i) and increase (ii) and (iii) over the values dark controls.Far-red light exerted an effect opposite to that of red light.Temperatures higher than 25° inhibited (ii) and (iii) underany light conditions. The optimum temperature to the actionof red and far-red light is 25°, at which the stimulatoryeffect of red light and the inhibition of this effect by far-redlight are both maximal. 2. The growth of the radicles of de-coated seeds of Grand Rapidslettuce shows two phases at all temperatures studied. PhaseI is characterized by slow but linear growth which continuesuntil shortly after visible differentiation of the radicle intothe hypocotyl and the root. Phase II is a phase of active growthin which the total length reflects mainly the length of theroot. The optimum temperature for Phase I is 25°-35°,and that, for Phase II is 25°. In neither phase, and atnone of the temperatures studied, is there any effect of redor far-red radiation on the growth of the radicle. The firstvisible sign of radicle elongation in red light induced seeds,however, takes place at exactly the same time as that of germination. 3. Similarities and dissimilarities between the germinationand the growth are pointed out, and it is concluded that thetwo phenomena are different, but proceed at sites closely associatedin the embryo. ¹Present address: Johnson Foundation for Medical Physics, Universityof Pennsylvania, Philadelphia, Pa., U.S.A.     

MORPHOLOGY AND REPRODUCTION OF THE RED ALGA ACROCHAETIUM PECTINATUM IN CULTURE12

The life history of the marine red alga Acrochaetium pectinatum (Kylin) Hamel was studied in unialgal culture using supplemented natural seawater media. The tetrasporophytes are larger than the gametophytes, have a compact filamentous basal system, and produce monosporangia and tetrasporangia. Mono-spores give rise to tetrasporophytes. Tetraspores develop into small gametophytes with unicellular bases. The gametophytes are heterothallic when small (usually less than 500 μ) but as some females become larger (2-3 mm) they produce spermatangia as well as carpogonia. Gametophytes may bear mono-sporangia in addition to carpogonia or spermatangia. These monospores give rise to gametophytes. Fertilization of the carpogonia has not been observed. The tetrasporophytes produce only monosporangia in day-lengths of 12-16 hr, but both tetrasporangia and monosporangia are formed in daylengths of 6–10 hr. Tetrasporangial production is reduced at 15 C compared with 10 C. Light intensity in the range of 5-200 ft-c (cool white fluorescent lighting) has no apparent influence on induction of tetrasporangia. Induction of tetrasporangia is not a photoperiodic response because their development is not inhibited by a brief light break in the middle of the dark period in short daylengths. Plastid morphology, origin, and frequency of sporangia and vegetative branching are variable during the ontogeny, and consequently are somewhat unreliable as taxonomic criteria. Differences in basal systems between gametophytes and tetrasporophytes also indicate that this feature, which is used to distinguish major subgeneric groupings in Acrochaetium, may not be as useful as previously thought.     

PHOTOMORPHOGENESIS OF THE GAMETOPHYTES OF LYGODIUM JAPONICUM

Protonemata of Lygodium japonicum turn biplanar in both red and blue light regimes and remain filamentous in far-red light. Biplanar gametophytes formed in red light are longer than broad with long, rectangular cells, whereas in blue light they appear broader than long with short, isodiametric cells. Transfer of protonemata of all ages from far-red regime to red or blue light induces a morphological type of growth characteristic of the new light regime. However, only relatively young biplanar forms transferred from red or blue light are able to resume filamentous type of growth in a subsequent regime of far-red light.     

Growth Promotion of Vegetative Gametophytes of <Emphasis Type="Italic">Undaria pinnatifida</Emphasis> by Blue Light

Through an acclimation period of 10 days, compared to white light, the maximal net photosynthetic rates were significantly higher for gametophytes of Undaria pinnatifida cultivated under blue light (400–500 nm), and were lower under red light (600–700 nm). Chlorophyll c and the carotenoid content of gametophytes were similar under blue light and red light but were much lower under white light. The growth rate of female gametophytes under blue light was higher than that under other lights, and the growth rate of male gametophytes showed little variation with respect to blue and white light. Male and female gametophytes were mixed together to form sporophytes under white, blue and red light. After approximately 5 days, 50% gametophytes became fertile under blue and white light, but remained vegetative under red light after 10 days.     

CRITICAL LEVELS OF LIGHT AND TEMPERATURE REGULATING THE GAMETOGENESIS OF THREE LAMINARIA SPECIES (PHAEOPHYCEAE)1

Gametophytes of three Laminaria species occurring near Helgoland, North Sea, were cultivated 4 wk in a 12:12 LD regime at different temperatures in artificial light fields, and in the sea at different water depths. In the artificial light fields underwater spectral distribution was simulated according to Jerlov water Types 5, 7, 9. Blue light in the simulated light fields amounted to 17, 12 or 4% of total quanta. The rate of vegetative growth did not depend on spectral distribution, was light-saturated at 4–6 W · m^?2, and increased with temperature up to 15 C. L. saccharina (L.) Lamour. exhibited the highest tolerance towards temperature, light and UV. Gametophytes survived 1 wk at 21 C ± 0.1, but not 22 C ± 0.1. Gametophytes of L. hyperborea (Gunn.) Fosl. and L. digitata (Huds.) Lamour. survived 1 wk at 20 C ± 0.1, but not at 21 C ± 0.1. In sunlight, and in the light field of a xenon lamp, 50% of L. saccharina gametophytes were killed by a quantum dose of 50 μEin · cm^?2, and 100% of the plants by 90 μEin · cm^?2. Approximately half of these quantum doses killed the corresponding percent of the other species gametophytes. Appreciably higher quantum doses were survived in visible light, with red being the most detrimental. Fertility depended on a critical quantum dose of blue light which decreased almost exponentially with decreasing temperature. The quantum dose (400–512 nm) required for induction of fertilization of 50% of the female gametophytes (males react similarly) was 90 μEin · cm^?2 at 5 C, 110 μEin · cm^?2 at 10 C, 230 (560 in L. digitata)μEin · cm^?2 at 15 C, and 560 (L. hyperborea) or about 850 (other 2 species) μEin · cm^?2 at 18 C. In the sea the gametophytes survived the dark winter months in the unicellular stage, with almost no vegetative growth of the primary cell, due to lack of light. In early spring the female gametophytes matured in the unicellular, and the males in a few-celled stage at the depth of 2 m, as did the laboratory cultures under conditions inducing maximal fertility.     

THE EFFECT OF GROWTH CONDITIONS AND THE STAGE OF LEAF DEVELOPMENT ON THE HILL REACTION IN HOMOGENATES OF PISUM SATIVUM LEAVES

Miller, John H. (Yale U., New Haven, Conn.) The effect of growth conditions and the stage of leaf development on the Hill reaction in homogenates of Pisum sativum leaves. Amer. Jour. Bot. 47(7): 532–540. Illus. 1960.—With plants grown under short-day conditions (8 hr. light and 16 hr. dark), crude chloroplast suspensions from young leaves have a higher Hill-reaction activity between 1 and 3 hr. after the beginning of illumination than suspensions from older leaves, while after 5–7 hr. of illumination, this activity difference is not found. These differences result from a marked diurnal rise and fall in the Hill reaction. The magnitude of the rise depends on the age of the leaf from which the chloroplast suspension is prepared. Peak activity occurs after the plants have received between 3 and 4 hr. of light and is highest in suspensions prepared from young leaves. Suspensions from the oldest leaves show no diurnal change in activity. No diurnal changes in activity are found in chloroplast suspensions from plants which are grown under continuous light, and the diurnal rise and fall is dependent on the plant receiving an alternation of light-dark-light.     

Physiological and morphogenetic studies of fern gametophytes and sporophytes in aseptic culture

Summary Prolonged irradiation with red light and addition of sucrose to the basal medium induced two-dimensional growth in gametophytes of Pteris vittata. The results suggest that two-dimensional growth is controlled by the following three factors: 1. a photoreactive system which depends on blue light; 2. a photoreactive system which can utilize red light; and 3. the photosynthetic system which the level of carbohydrate. Media containing 2.4-dichlorophenoxyacetic acid and sucrose, and complex media containing peptone caused formation of tumor-like calli.     

BIOGEOGRAPHY AND ECOLOGY OF THE KELP/RED ALGAL SYMBIOSIS

A kelp/red algal symbiosis is described from nature based on extensive collections from the San Juan Islands, Washington. Kelp gametophytes were found as endophytes in the cell walls of seventeen species of red algae in three different kelp communities. Host red algae were mostly filamentous (e.g., Pleonosporium vancouverianum) or polysiphonous (e.g. Polysiphonia paniculata). The kelp gametophytes completed vegetative and reproductive development in the hosts with gametangia formed at the host surface and with sporophytes up to several mm in height being produced while still attached to the host. To date, none of the kelp gametophytes from nature have been identified to genus or species, although the gametophyte of Nereocystis luetkeana is a potential candidate for the symbiosis. Preliminary observations from Nova Scotia and the Isle of Man have not found the association in the Atlantic Ocean. Laboratory studies in Korea successfully reconstructed the symbiosis in the red alga Aglaothamnion oosumiense using zoospores of Undaria pinnatifida but not Laminaria religiosa. Here we outline the development of the symbiosis and discuss the potential adaptive significance of the kelp/red algal interaction.