GROWTH REGULATION BY ETHYLENE IN FERN GAMETOPHYTES. IV. INVOLVEMENT OF PHOTOSYNTHESIS IN OVERCOMING ETHYLENE INHIBITION OF SPORE GERMINATION

The inhibitory effects of ethylene on spore germination were investigated. In darkness spore germination was completely inhibited by 10 μ1 · 1⁻¹ ethylene. Light partially overcame this inhibition, and the effect of continuous irradiation with white fluorescent light saturated at about 450 μW · cm⁻². Monochromatic red, blue and far-red light were effective in overcoming ethylene inhibition, whereas green was not. Short periodic exposures to red or far-red light were not sufficient to overcome ethylene inhibition. This suggested that phytochrome was not involved. The photosynthetic inhibitor DCMU blocked the effect of light. Infrared gas analysis showed that photosynthesis saturated at about 450 μW · cm⁻² in white light. Red, blue and far-red light were more efficient photosynthetically than green light; DCMU blocked photosynthesis. Normalized curves of photosynthesis and germination vs. light intensity showed a similar dependence on light energy. It was concluded that light appears to overcome the inhibitory effects of ethylene through some process dependent on photosynthesis.     

Effects of Anaerobiosis and Respiratory Inhibitors on Blue-Light Inhibition of Spore Germination in Pteris vittata

In a fern, Pteris vittata, inhibition by low-energy blue lightof phytochrome-dependent spore germination was counteractedby anerobiosis and respiratory inhibitors, such as KCN and NaN₃.A 50% inhibition of spore germination in a medium containing0.3 mM NaN₃ required about 8 times longer duration of blue lightirradiation compared with the control. The counteracting effectof NaN₃ continued for about 32 hr after withdrawal of the inhibitor.However, NaN₃ neither induced dark germination nor counteractedthe far-red light inhibition of spore germination. Reducingagents and uncouplers were tested and dithionite and arsenateslightly reversed the blue light inhibition of spore germination. (Received December 17, 1981; Accepted July 8, 1982)     

PHOTOMORPHOGENESIS IN PTERIS VITTATA: I. PHYTOCHROME-MEDIATED SPORE GERMINATION AND BLUE LIGHT INTERACTION

1. Spores of the fern Pteris vittata did not germinate under totaldark conditions, while an exposure of the spores to continuouswhite light brought about germination. The germination was mosteffectively induced by red light and somewhat by green and far-red,but not at all by blue light. The sensitivity of spores to redlight increased and leveled off about 4 days after sowing at27–28. The promoting effect of red light could be broughtabout by a single exposure of low intensity. Far-red light givenimmediately after red light almost completely reversed the redlight effect, and the photoresponse to red and far-red lightwas repeatedly reversible. The photoreversibility was lost duringan intervening darkness between red and far-red irradiations,and 50% of the initial reversibility was lost after about 6hr of darkness at 27–28. These observations suggest thatthe phytochrome system controls the germination of the fernspore.
2. When the imbibed spores were briefly exposed to a low-energyblue light immediately before or after red irradiation, theirgermination was completely inhibited. The blue light-inducedinhibition was never reversed by brief red irradiation givenimmediately after the blue light. The escape reaction of redlight-induced germination as indicated by blue light given aftervarious periods of intervening darkness was also observed, andits rate was very similar to that determined by using far-redlight. Spores exposed to blue light required 3 days' incubationin darkness at 27–28 to recover their sensitivity tored light. The recovery in darkness of this red sensitivitywas temperature-dependent. It is thus suggested that an unknownbluelight absorbing pigment may be involved in the inhibitionof phytochrome-mediated spore germination.

(Received August 21, 1967; )     

UV‐A/BLUE LIGHT–INDUCED REACTIVATION OF SPORE GERMINATION IN UV‐B IRRADIATED ULVA PERTUSA (CHLOROPHYTA)1

Recent reduction in the ozone shield due to manufactured chlorofluorocarbons raised considerable interest in the ecological and physiological consequences of UV‐B radiation (λ=280–315 nm) in macroalgae. However, early life stages of macroalgae have received little attention in regard to their UV‐B sensitivity and UV‐B defensive mechanisms. Germination of UV‐B irradiated spores of the intertidal green alga Ulva pertusa Kjellman was significantly lower than in unexposed controls, and the degree of reduction correlated with the UV doses. After exposure to moderate levels of UV‐B irradiation, subsequent exposure to visible light caused differential germination in an irradiance‐ and wavelength‐dependent manner. Significantly higher germination was found at higher photon irradiances and in blue light compared with white and red light. The action spectrum for photoreactivation of germination in UV‐B irradiated U. pertusa spores shows a major peak at 435 nm with a smaller but significant peak at 385 nm. When exposed to December sunlight, the germination percentage of U. pertusa spores exposed to 1 h of solar radiation reached 100% regardless of the irradiation treatment conditions. After a 2‐h exposure to sunlight, however, there was complete inhibition of germination in PAR+UV‐A+UV‐B in contrast to 100% germination in PAR or PAR+UV‐A. In addition to mat‐forming characteristics that would act as a selective UV‐B filter for settled spores under the parental canopy, light‐driven repair of germination after UV‐B exposure could explain successful continuation of U. pertusa spore germination in intertidal settings possibly affected by intense solar UV‐B radiation.     

Control of Mitosis by Phytochrome and a Blue-Light Receptor in Fern Spores

The first mitosis in spores of the fern A. capillus-veneris was observed under a microscope equipped with Nomarski optics with irradiation from a safelight at 900 nm, and under a fluorescent microscope after staining with 4[prime],6-diamidino-2-phenylindole. During imbibition the nucleus remained near one corner of each tetrahedron-shaped dormant spore, and asymmetric cell division occurred upon brief irradiation with red light. This red light-induced mitosis was photoreversibly prevented by subsequent brief exposure to far-red light and was photo-irreversibly prevented by brief irradiation with blue light. However, neither far-red nor blue light affected the germination rate when spores were irradiated after the first mitosis. Therefore, the first mitosis in the spores appears to be the crucial step for photoinduction of spore germination. Furthermore, experiments using a microbeam of red or blue light demonstrated that blue light was effective only when exposed to the nucleus, and no specific intracellular photoreceptive site for red light was found in the spores. Therefore, phytochrome in the far-red absorbing form induces the first mitosis in germinating spores but prevents the subsequent mitosis in protonemata, whereas a blue-light receptor prevents the former but induces the latter.     

Photocontrol of spore germination in the fern Thelypteris kunthii

The light requirement for germination in spores of the fern Thelypteris kunthii (Desv.) Morton was fully satisfied by a long period of continuous red light or partially by intermittent, short periods of red light. Red light-potentiated spore germination was inhibited by brief far-red light irradiation, indicating phytochrome involvement. Repeated exposure of spores to prolonged red and short far-red irradiations, or exposure of red-potentiated spores to far-red light after an extended period in darkness, led to their escape from inhibition of germination by far-red light. Prolonged irradiation of spores with blue light before or after red light treatment partially antagonized the effect of red light.     

Light dependent spore germination in Woodwardia radicans (L.) Sm

Abstract

The spores of Woodwardia radicans can germinate indifferently either in water or in culture media containing mineral salts at temperatures (15-24°C) falling within a range believed optimal for many other ferns (15-30 C).

The spores are photosensitive, will not germinate in the dark and the addition of gibberellic acid is ineffective in substituting a light requirement. Spore germination was induced by white and red light and phytochrome seems to be implicated in the control of germination since far-red light (and not the blue irradiation) can reverse the stimulating effect of the red light.

Spore morphology and spore germination pattern was studied using light and scanning electron microscopes.

It was concluded that the progressive disappearance of W. radicans from the Italian localities is not due to difficulties in spore germination but is related to problems that arise during the subsequent stages.     

Photomorphogenesis in Pteris vittata IV. Action spectra for inhibition of phytochrome-dependent spore germination

Action spectra between 350 and 500 nm for the inhibition ofphytochrome-dependent spore germination in the fern Pteris vittatawere obtained. Both action spectra obtained before and afterred light irradiation have peaks at about 440 nm and 380 nmand shoulders from 440 to 480 nm. These results suggest thatthe phytochrome system is not involved in the inhibitory processof spore germination induced by short irradiation with bluelight. (Received October 8, 1970; )     

Light germination ofAnthoceros miyabeanus spores

The effects of light on the spore germination of a hornwort species,Anthoceros miyabeanus Steph., were investigated. Spores of this species were photoblastic, but their sensitivities to light quality were different. Under either continuous white, red or diffused daylight, more than 80% of the spores germinated, but under blue light none or a few of them germinated. Under continuous far-red light or in total darkness, the spores did not germinate at all.Anthoceros spores required red light irradiation for a very long duration, i.e., over 12–24 hr of red light for saturated germination. However, the spore germination showed clear photo-reversibility by repeated irradiation of red and far-red light. The germination pattern clearly varied with the light quality. There were two fundamental patterns; (1) cell mass type in white or blue light: spores divide before germination, and the sporelings divide frequently and form 1–2 rhizoids soon after germination, and (2) germ tube type in red light: spores germinate without cell division, and the single-cell sporelings elongate without cell division and rhizoid formation.     

Photo-Inhibition of Red-Light-Induced Spore Germination in Pteris vittata: Cyanide, Azide and Ethanol Counteracts Restorable Inhibitory Action of Near UV and Blue-light but not That of Far UV

In Pteris vittata, red-light-induction of spore germinationwas completely inhibited by subsequent irradiation with farUV (260 nm), near UV (380 nm) or blue (440 nm) monochromaticlight produced at the Okazaki Large Spectrograph. Germinationbut recovered from these photo-inhibition after less than 48h of darkness. Near UV- and blue-light-induced inhibition werestrongly counteracted by addition of 1 mM KCN, 1 RIM NaN₃ or100 mM ethanol. Far UV- and far red light-induced inhibition,however, was not influenced by these chemicals. Consequentlythe heights of peaks of action spectrum for this photo-inhibitionof spore germination was changed by addition of these chemicalsin the blue and near UV region but not at 260 nm. The resultssuggest that either or both of the photoreceptor system andthe signal transduction chain of the photo-inhibition are qualitativelydifferent between the shorter (i.e. far UV) and the longer (i.e.near UV and blue) wavelength regions. (Received August 31, 1989; Accepted February 19, 1990)     

GROWTH REGULATION BY ETHYLENE IN FERN GAMETOPHYTES. V. ETHYLENE AND THE EARLY EVENTS OF SPORE GERMINATION

Early events during the germination of spores of the fern Onoclea sensibilis were studied to determine the time during germination when ethylene had its greatest inhibiting effect. Water imbibition by dry spores was rapid and did not appear to be inhibited by ethylene. During normal germination DNA synthesis occurred about four hours before the nucleus moved from a central position to the spore periphery. Following nuclear movement, mitosis and cell division occurred, partitioning the spore into a small rhizoid cell and a large protonemal cell. Cell division was complete approximately six hours after nuclear movement. Ethylene treatment of the spores blocked DNA synthesis, nuclear movement, and cell division. The earliest DNA replication in uninhibited spores was observed after 14 hours of germination, and the maximal rate of spore labeling with ³H-thymidine was between 16 and 20 hours. Spores were most sensitive to ethylene, however, during the stages of germination prior to DNA synthesis, and it was concluded that ethylene did not directly inhibit DNA replication but blocked germination at some earlier fundamental step. The effects of ethylene were reversible. since complete recovery from inhibition of germination was possible if ethylene was released and the spores were kept in light. Recovery was much slower in darkness. It was hypothesized that light acted photosynthetically to overcome the ethylene inhibition of germination. Consistent with this, it was shown that spores exhibit net photosynthesis after only two hours of germination.     

ANALYSIS OF STARCH ACCUMULATION AND GERMINATION IN ONOCLEA SPORES

Previous studies have shown that low fluences of light accelerate starch accumulation and enhance germination of Onoclea spores. Fluence response curves for induction of starch accumulation were compared with fluence response curves for enhancement of germination in order to determine if the two photoresponses in Onoclea spores have a common photoreceptor. Fluence response curves indicate that both responses were proportional to the log of the fluence and that the relative fluence efficiencies of the four wavelength regions tested were similar for induction of both germination and starch accumulation. Red (600–720 nm) irradiation was the most efficient, while green (500–600 nm), blue (400–520 nm), and far-red (720–900 nm) irradiations showed a decreasing order of efficiency for induction of the responses. A correlation coefficient between the amount of starch accumulated as a result of red irradiation and the final percent germination was calculated to be 0.964. These results support the hypothesis that a common photoreceptor mediates both photoinduced germination and starch accumulation. 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) inhibits photosynthesis by blocking the flow of electrons from Photosystem II to Photosystem I. At 0.1 mM DCMU failed to inhibit both photoinduced starch acumulation and germination. This result and the greater efficiency of red than blue light, the low fluence functional for induction, and the fluence dependency argue against the participation of photosynthesis in photoinduced starch accumulation. A similar conclusion has been previously drawn for photoenhancement of Onoclea spore germination. Additionally, the effects 0.01–1.0 mm cycloheximide and 100 μl/l ethylene on photoinduced starch accumulation were investigated. Neither agent inhibited starch accumulation, whereas both substances inhibited germination 70–90% when applied at a time coincidental with the period of rapid starch accumulation. These results indicate that the photoinduction of starch accumulation does not have an ethylene sensitive stage nor does it require protein synthesis as does photoenhancement of germination of Onoclea spores.     

Hydrolytic enzyme activities in germinating spores ofAdiantum capillus-veneris L.

Changes in hydrolytic enzyme activities were investigated during spore germination ofAdiantum capillus-veneris L. The spores were incubated for 3 days in the dark at 25 C for imbibition, and then germination of the spores was induced by continuous irradiation with red light. At day 2 after onset of the red light irradiation, rhizoids appeared out of spore coats and protonemal cells became visible on the following day. Lipase occurred in dry spores and its activity decreased during 3 days of dark incubation. The activity started to increase when the spore germination was induced by red light irradiation. On the other hand, amylolytic and aminopeptidase activities which were also detected in dry spores decreased continuously during the dark incubation and following the germination process. RNase activity also decreased during 3 days of dark incubation but the activity was retained thereafter at a constant level with or without red light irradiation. Developmental patterns of these hydrolytic enzymes were classified into two groups: One decreased during imbibition and dark incubation but increased after red light irradiation and the other continuously decreased during dark incubation and germination. These results are discussed in relation to compositional changes of cell constitutions such as lipid, sugars, proteins and amino acids during spore germination.     

A NEW METHOD FOR EXAMINING INDUCTION OF MOSS BUDS BY CYTOKININ

Protonemata of Polytrichum juniperinum were grown in the dark, in sucrose- and cytokinin-supplemented media. Under these conditions gametophore buds form shortly after spore germination. Of the cytokinins tested, DMAAP was most effective in bud induction. This system holds much promise for study of cytokinin action.     

INDUCTION OF SPORE GERMINATION IN SCHIZAEA PUSILLA (SCHIZAEACEAE)

Requirements for spore germination in the rare and native New Jersey fern, Schizaea pusilla Pursh., were examined. Spores did not germinate in darkness and gibberellins (GA) did not induce germination in the dark. However, a dark pretreatment promoted germination in a subsequent light treatment and low temperatures during the dark pretreatment greatly enhanced germination in culture. Three wks of dark pretreatment were required for maximum germination. GA₃ promoted germination in red light more effectively than GA₄₊₇. Greater than ten days of continuous illumination was necessary for germination. Spores given red light reached half-maximum germination six days earlier than spores under white light. Red light promoted germination while blue light did not. Far-red light alone could stimulate germination and enhanced the promotive effect of red light; typical phytochrome photoreversibility was not observed. Blue light reduced the effect of red light.     

Photomorphogenesis in Pteris vittata II. Recovery from blue-light-induced inhibition of spore germination

1. 1. In the fern Pteris vittata, low-energy blue-light-inducedinhibition of phytochrome-dependent spore germination and darkrecovery from this inhibition were repeatedly observed severaltimes at intervals of 3 days at 26. The same amount of incidentenergy of blue light was required for inhibition in each successivetreatment.
2. 2. The recovery from blue-light-induced inhibitionof germinationwas markedly accelerated by continuous illuminationwith redlight, and this red light effect was not affected bythe presenceof CMU.
3. 3. The recovery process was not influencedby a single exposureto redlight, but was definitely promotedby brief red irradiationsgiven intermittently, at least 2 times,at equal intervals duringthe first 8 hr after blue light treatment.The effect of intermittentlygiven red light was annulled wheneach red exposure was followedby brief far-red irradiation.These facts suggest that phytochromemay be involved in therestoration of the ability of sporesto germinate (in responseto red light) which had been lostby blue irradiation.

¹Present address: Botany Department, Faculty of Science, Universityof Tokyo, Hongo, Tokyo 113.     

The Photocontrol of Spore Germination in the Fern Ceratopteris richardii

This paper describes how different wavelengths of light regulatespore germination in the fern Ceratopteris richardii. This speciesdoes not exhibit any dark germination. Maximum photosensitivityof the spores is reached 7 to 10 d after imbibition. An increasein the red light fluence above the threshold fluence of 10¹⁶quanta.m^–2 leads to a corresponding increase in germination.In sequential irradiation experiments, farred light can reversethis red light-mediated germination to the level observed withthe far-red light control. Blue light fluences above 10²⁰ quanta.m^–2can also block the germination response to red light. Moreover,this antagonistic effect of blue light is not reversed by subsequentirradiation with red light. It is therefore concluded that phytochromeand a distinct blue light photoreceptor control C. richardiispore germination. These interpretations are entirely consistentwith the published literature on other fern genera. (Received November 28, 1986; Accepted April 6, 1987)     

Ethylene Inhibition of Phytochrome-Induced Germination in Potentilla norvegica L. Seeds

Germination of Potentilla norvegica L. (rough cinquefoil) seeds stimulated by fluorescent irradiations of nearly 24 hours was inhibited by ethylene at <1 microliter per liter. Sensitivity to ethylene inhibition was highest during and immediately after the irradiation. By delaying ethylene treatment until about a day after the light potentiation, seeds escaped the inhibition. Ethylene inhibition may be readily reversed upon release of the gas and reirradiation of the seeds. Imbibition of seeds at 10 or 15°C, or at high temperatures of 35 and 40°C, partially prevented subsequent inhibition by ethylene. Alternating temperatures during germination nearly overcame the inhibition from 1 microliter per liter ethylene, but not higher doses. With brief red-irradiation and alternating temperatures, 0.1 microliter per liter ethylene promoted germination about 2-fold. These data suggest that ethylene may loosely associate on a site required for phytochrome action. The effect of temperature that opposed the inhibition may be to deny the association of ethylene with the site. Loose association is supported by the reversal of inhibition by gas release and increased temperature during germination. A blocking effect was shown by the failure of phytochrome to act when ethylene was present.     

EFFECTS OF VISIBLE AND ULTRAVIOLET RADIATION ON THE GERMINATION OF PHACELIA TANACETIFOLIA

Schulz , Sister M. Richardis , O.P., and Richard M. Klein . (N. Y. Bot. Gard., N. Y., N. Y.) Effects of visible and ultraviolet radiation on the germination of Phacelia tanacetifolia. Amer. Jour. Bot. 50(5): 430–434. Illus. 1963.—Germination of Phacelia tanacetifolia was suppressed by exposure to white light increasing with intensity and length of illumination. The light effect decayed during 24 hr of darkness. Seeds were most sensitive to the suppressive effects of light 13–17 hr after the beginning of imbibition. Light suppression was caused by a photocatalytic reaction. Wavelengths causing the suppression lie in the far-red, red, blue, near-ultraviolet and far-ultraviolet regions of the spectrum. At equal energies, blue light was less effective than far-red, red or ultraviolet radiation. There was no evidence for the existence of the phytochrome system. Simultaneous irradiation with red and blue light or simultaneous irradiation with red and far-red induced a synergistic repression of germination. The presentation of different wavelengths in various sequential patterns markedly altered the germination response. An interaction between elevated temperatures and visible radiation affecting germination response was also noted.     

粗茎鳞毛蕨孢子萌发研究

以经过3年低温储藏的粗茎鳞毛蕨孢子为实验材料,从孢子离心、孢子消毒、培养基种类、光质等4方面对孢子萌发进行研究,结果表明:在离心转数≤14 000 r.min-1、离心时间≤30 min条件下,离心处理对孢子萌发基本无影响;对孢子进行1%NaClO水溶液浸泡处理20～30 min为最佳消毒条件;改良Knop’s培养基为最佳孢子萌发培养基;黑暗条件下孢子不能萌发,但是黑暗处理能够明显提高孢子萌发整齐性;红光比白光能促进孢子提早萌发1 d左右,但对提高萌发率效果不显著。