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Seed Germination in Chenopodium album L: Relationships between Nitrate and the Effects of Plant Hormones.

Effects of ethylene, gibberellins, and kinetin on the germination of two lots of Chenopodium album L. seeds, collected from the field in 1982 and 1983, were studied in relation to the availability of nitrate. The experiments were conducted in darkness and at temperatures ranging from 12 to 32 degrees C. Ethylene induced over 75% germination in the 1983 seed but had little effect on the 1982 seed. Nitrate was only slightly promotive in either of the two seed lots. A combination of ethylene and nitrate, however, acted synergistically on 1982 seed, resulting in as much germination as that induced in 1983 seed by ethylene alone. In 1983 seed, a combination of ethylene and nitrate was only marginally more effective than ethylene. A similar relationship was observed in the effects of gibberellic acid(4+7) (GA(4+7)) and nitrate on seeds from the two lots. The 1982 seed, which responded synergistically to combinations of nitrate with ethylene or GA(4+7) was found to contain an extremely low endogenous level of nitrate as compared to 1983 seed. Thus, high levels of either endogenous or applied nitrate appeared to enhance the germination response to ethylene or GA(4+7).Kinetin had no effect on 1982 seed and only a small promotive effect on 1983 seed. There was no synergism between kinetin and nitrate in either of the seed lots.

Journal Article↗

Induction of Secondary Dormancy in Chenopodium bonus-henricus L. Seeds by Osmotic and High Temperature Treatments and Its Prevention by Light and Growth Regulators.

Factors controlling the establishment and removal of secondary dormancy in Chenopodium bonus-henricus L. seeds were investigated. Unchilled seeds required light for germination. A moist-chilling treatment at 4 C for 28 to 30 days removed this primary dormancy. Chilled seeds now germinated in the dark. When chilled seeds were held in the dark in -8.6 bars polyethylene glycol 6000 solution at 15 C or in water at 29 C a secondary dormancy was induced which increased progressively with time as determined by subsequent germination. These seeds now failed to germinate under the condition (darkness) which previously allowed their germination. Continuous light or daily brief red light irradiations during prolonged imbibition in polyethylene glycol solution at 15 C or in water at 29 C prevented the establishment of the secondary dormancy and caused an advancement of subsequent germination. Far red irradiations immediately following red irradiation reestablished the secondary dormancy indicating phytochrome participation in "pregerminative" processes. The growth regulator combination, kinetin + ethephon + gibberellin A(4)+A(7) (GA(4+7)), and to a relatively lesser extent GA(4+7), was effective in preventing the establishment of the secondary dormancy and in advancing the germination or emergence time. Following the establishment of the secondary dormancy by osmotic or high temperature treatments the regulator combination was relatively more active than light or GA(4+7) in removing the dormancy. Prolonged dark treatment at 29 C seemed to induce changes that were partially independent of light or GA(4+7) control. The data presented here indicate that changes during germination preventing dark treatment determine whether the seed will germinate, show an advancement effect, or will become secondarily dormant. These changes appear to be modulated by light and hormones.

Journal Article↗

Exogenous gibberellins inhibit coffee (Coffea arabica cv. Rubi) seed germination and cause cell death in the embryo.

The mechanism of inhibition of coffee (Coffea arabica cv. Rubi) seed germination by exogenous gibberellins (GAs) and the requirement of germination for endogenous GA were studied. Exogenous GA(4+7) inhibited coffee seed germination. The response to GA(4+7) showed two sensitivity thresholds: a lower one between 0 and 1 microM and a higher one between 10 and 100 microM. However, radicle protrusion in coffee seed depended on the de novo synthesis of GAs. Endogenous GAs were required for embryo cell elongation and endosperm cap weakening. Incubation of coffee seed in exogenous GA(4+7) led to loss of embryo viability and dead cells were observed by low temperature scanning microscopy only when the endosperm was surrounding the embryo. The results described here indicate that the inhibition of germination by exogenous GAs is caused by factors that are released from the endosperm during or after its weakening, causing cell death in the embryo and leading to inhibition of radicle protrusion.

Cell Death↗

Localization of Stem Elongation Control in Cucumis sativus L.

Reciprocal grafts, and applications of gibberellin (GA) and indoleacetic acid (IAA) were used to localize the site of control for stem elongation in cucumber (Cucumis sativus L.). Dwarf and tall plants were reciprocally grafted to determine influence of stems and roots on stem elongation. At 21 days there were no significant differences in length between stems grafted to their own roots and those grafted to roots of the other type. GA(3), GA(4+7), and IAA were applied to seedlings with and without live apical buds. Seedlings with live apical buds responded to level of added GA, but not to added IAA. GA(4+7) was more effective than GA(3). Hypocotyls of tall plants responded more to both GA treatments than did those of the dwarves when both types had live apical buds. When either GA(4+7) or IAA was applied to seedlings with dead apical buds, elongation of the hypocotyl responded to level of the growth regulator, but there was no difference in response between the dwarf and tall plants.

Journal Article↗

Dual Effect of Light on the Gibberellin- and Nitrate-Stimulated Seed Germination of Sisymbrium officinale and Arabidopsis thaliana.

Red light (R) has a dual effect on the seed germination of the two related species Arabidopsis thaliana and Sisymbrium officinale. The two species provide different means to separate the light-effects. In S. officinale, stimulation of germination by R depends on the stimultaneous presence of nitrate (light-effect I). The effect of both factors is completely blocked by tetcyclacis, an inhibitor of gibberellin (GA)-biosynthesis. Addition of a mixture of gibberellins A(4) and A(7) (GA(4+7)) antagonizes the inhibition. In the absence of nitrate, R shifts germination to lower GA-requirement (light-effect II). In A. thaliana a similar second light-effect is seen on the GA-requirement of GA-deficient ga-1 mutant seeds. R stimulates germination of wild type seeds in water (light-effect I). For both species, light-effect I shows a fluence threshold value of approximately 10(-5) moles per square meter, which is independent of the nitrate concentration. Increasing nitrate concentrations narrow the fluence-range required for maximal germination whereby the product of nitrate concentration and fluence value determines the germination level, indicating a multiplicative interaction between R and nitrate. Fluence-response curves for light-effect II are similar for both species. Germination occurs in the range of 10(-6) to 10(-2) moles per square meter fluence. The maximal level of germination is determined by the level of dark-germination and light-effect II. Increasing GA(4+7) concentrations induce a shift to lower fluence values. It is shown that in the second effect the co-action of R and exogenous GA(4+7) is clearly additive. It is concluded that light-effect I induces a chain of events leading to GA biosynthesis. Light-effect II seems to enhance the sensitivity of the seeds to GAs.

Journal Article↗

Gibberellin and ethylene control endoreduplication levels in the Arabidopsis thaliana hypocotyl.

We have previously shown that endoreduplication levels in hypocotyls of Arabidopsis thaliana (L.) Heynh. are under negative control of phytochromes. In this study, the hormonal regulation of this process was analysed using a collection of A. thaliana mutants. The results show that two hormones in particular, gibberellin (GA) and ethylene, play distinct roles. Hypocotyl cells of the GA-deficient mutant ga1-11 grown in the dark did not elongate and showed a greatly reduced endoreduplication. Normal endoreduplication could be restored by supplying 10(-9) M of the gibberellin GA(4+7), whereas the restoration of normal cell growth required 100-fold higher concentrations. The GA-insensitive mutant gai showed reduced cell elongation but normal ploidy levels. We conclude that (i) GA(4+7) has a global positive effect on endoreduplication and (ii) that endoreduplication is more sensitive to GA(4+7) than cell elongation. Ethylene had a completely different effect. It induced an extra round of endoreduplication both in light- and dark-grown seedlings and acted mainly on discrete steps rather than having a global effect on endoreduplication. The genes EIN2 and CTR1, components of the ethylene signal transduction pathway were both involved in this process.

Arabidopsis↗

Functional expression of the cotton gibberellic acid oxidase homologous gene GhGA20ox1 in tobacco.

To determine the physiological function of GhGA20ox1, a homologous gene of GA 20-oxidase from elongating cotton fibers, we expressed this gene ectopically in Nicotiana benthamiana. Reverse transcription-PCR analysis showed that the GhGA20ox1 gene was expressed in the transgenic plants at various levels. It was demonstrated that overexpression of GhGA20ox1 enhanced preferentially the GA(4+7) biosynthesis in N. benthamiana and conferred GA-overproduction characters to transformants. The extent of phenotypic alteration in the transgenic plants was found to correlate with the transcriptional levels of GhGA20ox1 and GA contents. Results indicated that the GhGA20ox1 gene promoted the biosynthesis of the active GAs (GA(4+7)) in transgenic tobacco plants therefore represents a useful gene for manipulating GA levels.

Gene Expression Regulation, Enzymologic↗

Response of seedlings of a dwarf and a normal strain of watermelon to gibberellins.

Hypocotyl and root elongation in a dwarf and a normal strain of watermelon (Citrullus lanatus [Thunb.] Matsu.) in the absence or presence of different gibberellins was investigated in seedlings grown under gold fluorescent light or in darkness. The normal strain, "Sugar Baby," responded only slightly to the gibberellic acids employed. At appropriate concentrations all of the gibberellic acids were capable of normalizing growth in the monorecessive dwarf strain, WB-2, in darkness or in light. Gibberellins A(4+7) and A(7) were effective in stimulating hypocotyl elongation at concentrations 10 to 15 times lower than that needed for a response to GA(1) or GA(3). Dark-grown dwarfs responded to about a 3-fold lower concentration of GA(4+7) than those grown in light.In contrast to hypocotyl elongation, root elongation was greater in the dwarf than in the normal strain. Concentrations of gibberellic acids which enhanced hypocotyl elongation of WB-2 plants, inhibited root growth proportionately.Anatomically, the response of the dwarf strain to GA(4+7) was primarily in terms of increased cell division. Dark-enhanced elongation of both SB and WB-2 was due almost solely to increased cell elongation. The results suggest that, at least in some species, a major component of dark-enhanced growth (or light-inhibited growth) is physiologically distinct from gibberellic acid-stimulated elongation.

Journal Article↗

Gibberellins and antheridiogen on sex in Blechnum spicant L.

The role of gibberellins (GAs) in determining sex in the gametophyte of the fern Blechnum spicant L. was studied through (a) the effect of exogenous GA(4+7) and GA3 (b) quantitation of the endogenous levels of GA1, GA3, GA4, GA7, GA9, and GA20 in male and female gametophytes, and (c) the effect of flurprimidol, a GAs biosynthesis inhibitor of the steps of oxidation of ent-kaureno to ent-kaurenoic acid. Our results show that GA(4+7) had a slight effect of inducing either male or female sexual organs, antheridia and archegonia, respectively. The endogenous GAs content was not significantly different between sexes, but the GA4, GA7, and GA20 levels were raised above those of the other GAs in both sexes. Neither antheridiogen biosynthesis nor antheridia formation was inhibited by flurprimidol. Gametophytes regenerated from homogenized mature gametophytes (HG) show a different physiological behavior than spore-derived gametophytes. In the first case, gametophytes are males and synthesize antheridiogen before they attain maturity, in contrast to what occurs in spore-derived gametophytes which are females and synthesize antheridiogen when mature.

Abscisic Acid↗

Vacuolar H(+)-ATPase is expressed in response to gibberellin during tomato seed germination.

Completion of germination (radicle emergence) by gibberellin (GA)-deficient (gib-1) mutant tomato (Lycopersicon esculentum Mill.) seeds is dependent upon exogenous GA, because weakening of the endosperm tissue enclosing the radicle tip requires GA. To investigate genes that may be involved in endosperm weakening or embryo growth, differential cDNA display was used to identify mRNAs differentially expressed in gib-1 seeds imbibed in the presence or absence of GA(4+7). Among these was a GA-responsive mRNA encoding the 16-kD hydrophobic subunit c of the V(0) membrane sector of vacuolar H(+)-translocating ATPases (V-ATPase), which we termed LVA-P1. LVA-P1 mRNA expression in gib-1 seeds was dependent on GA and was particularly abundant in the micropylar region prior to radicle emergence. Both GA dependence and tissue localization of LVA-P1 mRNA expression were confirmed directly in individual gib-1 seeds using tissue printing. LVA-P1 mRNA was also expressed in wild-type seeds during development and germination, independent of exogenous GA. Specific antisera detected protein subunits A and B of the cytoplasmic V(1) sector of the V-ATPase holoenzyme complex in gib-1 seeds only in the presence of GA, and expression was localized to the micropylar region. The results suggest that V-ATPase plays a role in GA-regulated germination of tomato seeds.

Amino Acid Sequence↗

A physical, enzymatic, and genetic characterization of perturbations in the seeds of the brownseed tomato mutants.

The brownseed mutants (bs(1), bs(2), and bs(4)) of tomato all possess dark testae and deleteriously affect seed germination speed and/or final percentage. Poor germination performance of the bs(1) but not the bs(4) mutant, was due to greater impediment to radicle egress. Testa toughening (bs(1)) was prevented by drying in N(2). However, poor germination speed was hardly affected by drying. GA(4+7) did not ameliorate germination percentage or speed (bs(1), bs(2)), whereas bs(4) seeds commenced radicle protrusion sooner and had a greater germination percentage. bs(1) mutant seeds have two times more catalase activity while those of bs(4) contained six times more peroxidase and almost two times more catalase activity than WTs. bs(4) release only half of the reactive oxygen species into the media than WT during imbibition. EPR detected the presence of free radicals in bs(1) and its WT. bs mutants were epistatic to 12 anthocyaninless mutations, at least some of which produce seeds of lighter than usual testa colour. Macro-arrays of subtractive, suppressive PCR products identified differentially regulated transcripts between seeds of bs(4) and WT. EST identity suggests bs(4) does not exit the developmental programme upon attaining maturity.

Anthocyanins↗

GA4+7 alleviates pear fruit semi-russeting partly by suppressing PRX-mediated lignin deposition.

Pear fruit semi-russeting is a surface disorder that frequently occurs during fruit development and significantly diminishes fruit appearance quality and commercial value. Although Gibberellin 4 + 7 (GA4+7) has been used to reduce fruit surface defects in horticultural crops, the physiological and molecular mechanisms underlying its inhibitory effect on pear fruit semi-russeting remain poorly understood. In this study, preharvest GA4+7 treatment of 'Cuiguan' pear significantly reduced russet coverage and lignin accumulation in mature fruit skin without adversely affecting fruit size, fruit shape index, or total soluble solids content. Integrated metabolomic and transcriptomic analyses revealed that GA4+7 treatment was associated with the repression of phenylpropanoid and lignin biosynthesis at both metabolic and transcriptional levels. Among the lignin-related differentially expressed genes, two class III peroxidase genes, PpyPRX22 and PpyPRX65, were strongly downregulated by both GA4+7 and bagging treatments. Both proteins localized to the cell wall, and transient expression assays in pear fruit skin supported positive roles for PpyPRX22 and PpyPRX65 in lignin deposition. Furthermore, dual-luciferase reporter assays combined with transient overexpression experiments suggested that several PpyMYB transcription factors may regulate PpyPRX expression and lignin accumulation, with PpyMYB138 and PpyMYB139 significantly activating PpyPRX22 and/or PpyPRX65 promoter activity. Taken together, these results suggest that GA4+7 alleviates pear fruit semi-russeting at least partly by reducing lignin deposition in the fruit skin, with PpyPRX22 and PpyPRX65 potentially contributing to this process.

Class III peroxidase↗

Contents and recovery of gibberellins in monoecious and gynoecious cucumber plants.

Diffusates from seedlings and root exudates from 6-week-old plants of a monoecious line of cucumber, Cucumis sativus L., contained considerably higher levels of gibberellin-(GA-) like substances than did those from plants of an isogenic gynoecious line. Most of the GA-like activity was found in a chromatogram region typical of GA(1) and GA(3); some activity, particularly in root exudates, appeared also at an R(F) similar to that of GA(4) and GA(7).When seedlings were treated with (3)H-labeled GA(1), more radioactivity was found in the diffusates from monoecious seedlings than from gynoecious ones. The same was true of biological activity in root diffusates from older plants which had been treated with gibberellin A(4+7).In conjunction with evidence present in literature, these results support the idea that endogenous GAs play a part in the regulation of sex expression in cucumber, relatively high levels favoring the formation of staminate flowers.

Journal Article↗