Sucrose application causes hormonal changes associated with potato tuber induction

Stems of potato plants (Solanum tuberosum L. cv. Dianella) were immersed in solutions containing water (control), sucrose, glucose, paclobutrazol, and gibberellic acid. The effects of these treatments on the ethylene release, levels of endogenous gibberellins, glucose, sucrose, and starch were correlated with tuberization of nodal cuttings, excised from potato stems. Paclobutrazol and sucrose improved the percent of tuberization and/or increased tuber weight. In contrast, GA₃ inhibited tuber formation compared with the control. The level of endogenous free GAs was negatively correlated with percent tuberization. However, the level of conjugated GAs was positively correlated with both percent tuberization and tuber weight. The effect of sucrose on potato tuber induction in relation to the possible role of sucrose in GA-conjugate formation is discussed.     

A Comparison of the Endogenous Hormone Levels in Stolons and Sprouts of the Potato Solanum tuberosum

Whereas the gibberellin and inhibitor levels of potato (Solanum tuberosum L. cv. BP-1) sprouts and stolons differ, their auxin and cytokinin levels apparently remain more or less unchanged. The levels of gibberellin in the sprouts is higher than that of the stolons; however, with regard to inhibitor levels the reverse is true. It would appear as if an internal balance of different hormones controls whether or not tuberization will occur. High gibberellin and low inhibitor levels (sprouts) may favour cell elongation and at the same time limit cell division. Decreased gibberellin and increased inhibitor levels (stolons) seems to favour cell division. As cell division is not retarded despite high inhibitor levels, it would appear as if the effect of the inhibitor is mainly directed at countering the gibberellins.     

Methionine metabolism and ethylene biosynthesis in senescing petals of Tradescantia

The endogenous content of methionine in isolated petals of Tradescantia was found to increase during petal senescence while the levels of S-methylmethionine and protein were found to decline. The increase in free methionine was, at least in part, the result of protein degradation. Methionine and homocysteine were shown to be intermediates in ethylene biosynthesis while S-methylmethionine was not involved. Application of 1-aminocyclopropane-1-carboxylic acid (ACC) to all floral tissues resulted in large stimulations of ethylene production. ACC was shown to be an endogenous amino acid the internal levels of which correlated positively with the rate of ethylene production. Application of l-methionine-[U-¹⁴C] led to a rapid appearance of radioactivity in both ethylene and ACC. The specific radioactivity of C-2 and C-3 of ACC and that of ethylene were found to be nearly identical which indicated that ACC was the immediate precursor of ethylene in senescing petals of Tradescantia.     

Development of Nitrogen Assimilation Enzymes during Photoautotrophic Growth of Chenopodium rubrum Suspension Cultures

We have shown previously that ethylene, which accumulates in the air spaces of submerged stem sections of rice (Oryza sativa L. cv “Habiganj Aman II”), is involved in regulating the growth response caused by submergence. The role of gibberellins in the submergence response was studied using tetcyclacis (TCY), a new plant growth retardant, which inhibits gibberellin biosynthesis. Stem sections excised from plants that had been watered with a solution of 1 micromolar TCY for 7 to 10 days did not elongate when submerged in the same solution or when exposed to 1 microliter per liter ethylene in air. Gibberellic acid (GA₃) at 0.3 micromolar overcame the effect of TCY and restored the rapid internodal elongation in submerged and ethylene-treated sections to the levels observed in control sections that had not been treated with TCY. The effect of 0.01 to 0.2 micromolar GA₃ on internodal elongation was enhanced two- to eight-fold when 1 microliter per liter ethylene was added to the air passing through the chamber in which the sections were incubated. GA₃ and ethylene caused a similar increase in cell division and cell elongation in rice internodes. Thus, ethylene may cause internodal elongation in rice by increasing the activity of endogenous GAs. In internodes from which the leaf sheath had been peeled off, growth in response to submergence, ethylene and GA₃ was severely inhibited by light.     

The Relationship between Exogenous Growth Inhibitors and Endogenous Levels of Ethylene, and Tuberization of Dahlias

Treatments with abscisic acid (ABA), succinic acid-2,2-dimethylhydrazide (SADH), or 2 chlorethyl phosphonic acid (ethephon) promoted the tuberization of dahlia plants in long-days. This effect was smaller, however, than the effect short days have on tuberization. In contrast, gibberellic acid (GA) treatments inhibited tuberization. SADH and ethephon treatments of budless leaf-cuttings inhibited tuberization whereas ABA treatments slightly enhanced it. Evolution of endogenous ethylene reached a peak between the second and third week after the start of short-day treatments, and then decreased to the low level found in plants growing under long days. The peak in ethylene evolution occurred one week before the onset of tuberization.     

Ethylene production by suspension-cultured pear fruit cells as related to their senescence

Suspension-cultured pear fruit cells produce low levels of ethylene during growth and division in auxin containing medium. When deprived of auxin, division gradually ceases and ethylene production falls to barely discernible levels. However, notable ethylene production can now be induced by indoleacetic acid, CuCl₂, or 1-aminocyclopropane-1-carboxylic acid. If the auxin-deprived cells are transferred to `aging' medium that lacks auxin but contains 0.4 molar mannitol, inducible ethylene production increases several-fold reaching levels of 40 to 60 nanoliters/10⁶ cells per hour. Maximum inducible ethylene productivity is attained at varying times (1-6 days) after transfer to aging medium and appears to be temporally related to cell survival, i.e. the time of subsequent cell death. It is argued that auxin depletion initiates senescence which, in turn, leads to a transient increase in inducible ethylene production and eventual death. The limitations and potentials of the suspension-cultured pear cells as a system for the study of cellular senescence are discussed.     

Further Studies on the Relationship between Growth Regulators and Tuberization of Dahlias

Abscisic acid (ABA) and gibberellin (GA) treatments, applied to budless leaf cuttings of Dahlia under long day conditions, did not increase the total weight of the cuttings but did affect the region where photosynthates accumulate. GA treatment inhibited the growth of tubers and roots, and enhanced thickening at the petiole base. ABA did not affect the weight of roots or petiole, but promoted the growth of tuberous roots. Anatomical examination of the tuberization revealed that it is caused by inter-fascicular cambial activity producing parenchyma cells inward. No anatomical difference could be observed between the GA-induced thickening at the petiole base and the natural or ABA-induced thickening of the roots. In single node cuttings, taken from various sites along the branch, tuberization ability increased with proximity to the base of the branch, where axillary buds are more inhibited. Short-day conditions which promote tuberization, increased the endogenous level of ABA-like inhibitors in intact plants. External conditions and growth substances which inhibit growth also inhibited tuberization. Benzyladenine treatments did not affect tuberization. It is suggested that in addition to their respective indirect effects via the enhancement or suppression of top growth, GA and ABA directly control tuberization. ABA levels which increased under short-day conditions seem to produce a sink in the roots. GA probably inhibits tuberization by interfering with the trans-location of photosynthates to the root area.     

Cytokinin Profiles of AtCKX2-Overexpressing Potato Plants and the Impact of Altered Cytokinin Homeostasis on Tuberization In Vitro

Genes encoding cytokinin oxidase/dehydrogenase (CKX) enzymes have been used lately to study cytokinin homeostasis in a variety of plant species. In this study AtCKX2-overexpressing potato plants were engineered and grown in vitro as a model system to investigate the effects of altered cytokinin levels on tuber formation and tuber size. Protein extracts from shoots and roots of transformed potato plants exhibited higher CKX activity compared to control plants. Total endogenous cytokinin levels were generally not decreased in AtCKX2 overexpressors. However, levels of bioactive cytokinins were markedly lowered, which was accompanied by increased levels of O- and N-glucosides in some transgenic lines. The AtCKX2-overexpressing plants displayed reduced shoot growth but other symptoms of the ??cytokinin deficiency syndrome?? were not recorded. The transgenic plants were able to produce tubers in noninducing conditions. In inducing conditions they developed larger tubers than control. Tubers were also formed on a greater portion of the analyzed AtCKX2 plants, but with a lower number of tubers per plant compared to control. Taken together, our data suggest that cytokinins cannot be regarded simply as positive or negative regulators of tuberization, at least in vitro. Interactions with other plant hormones that play an important role in control of tuberization, such as gibberellins, should be further studied in detail.     

Cell division and cell enlargement during potato tuber formation

Cell division and cell enlargement were studied to reveal the developmental mechanism of potato tuberization using both in vivo in vitro culture systems. Distribution of cells in S-phase was visualized by immunolabelling of incorporated bromodeoxyuridine (BrdU). Mitosis was detected in DAPI (4,6-di-amidino-2-phenylindole) or toluidine blue-stained sections. Timing and frequency of cell division were determined by daily cell counting, and cell enlargement was deduced from measurements of cell diameters.Under in vivo conditions, lateral underground buds developed into stolons due to transverse cell divisions and cell elongation in the apical region of the buds. At the onset of tuber formation, the elongation of stolons stopped and cells in pith and cortex enlarged and divided longitudinally, resulting in the swelling of the stolon tip. When tubers had a diameter of 0.8 cm, longitudinal divisions had stopped but randomly oriented division and cell enlargement occurred in the perimedullary region and continued until tubers reached their final diameter.In vitro tubers were formed by axillary buds on single node cuttings cultured under tuber-including conditions. They stopped growing at a diameter of 0.8 cm. Pith and cortex were involved in tuberization such as that found during the early stage of in vivo tuberization (<0.8 cm in diameter). The larger size of in vivo tubers is, however, due to further development of the perimedullary region, which is lacking in vitro conditions.Keywords: Cell division, cell enlargement, DNA synthesis, in vitro culture, potato, tuber formation.      

Involvement of Ethylene in Potato Microtuber Dormancy

Potato (Solanum tuberosum L.) single-node explants undergoing in vitro tuberization produced detectable amounts of ethylene throughout tuber development, and the resulting microtubers were completely dormant (endodormant) for at least 12 to 15 weeks. The rate of ethylene production by tuberizing explants was highest during the initial 2 weeks of in vitro culture and declined thereafter. Continuous exposure of developing microtubers to the noncompetitive ethylene antagonist AgNO₃ via the culture medium resulted in a dose-dependent increase in precocious sprouting. The effect of AgNO₃ on the premature loss of microtuber endodormancy was observed after 3 weeks of culture. Similarly, continuous exposure of developing microtubers to the competitive ethylene antagonist 2,5-norbornadiene (NBD) at concentrations of 2 mL/L (gas phase) or greater also resulted in a dose-dependent increase in premature sprouting. Exogenous ethylene reversed this response and inhibited the precocious sprouting of NBD-treated microtubers. NBD treatment was effective only when it was begun within 7 d of the start of in vitro explant culture. These results indicate that endogenous ethylene is essential for the full expression of potato microtuber endodormancy, and that its involvement may be restricted to the initial period of endodormancy development.     

Ethylene, a regulator of young fruit abscission

In an earlier study we reported that detached cotton flowers produced sufficient ethylene before the period of natural abscission to suggest that ethylene might be a natural regulator of young fruit abscission. The present report explores this probability further. Intact cotton (Gossypium hirsutum L.) fruits produced ethylene at rates as high as 36 μl ethylene/kg fresh wt·hr during the 2 days before they abscised. Direct measurements of ethylene in gas samples withdrawn from fruits indicated that production of 1 μl ethylene/kg fresh wt·hr is equivalent to an internal concentration of approximately 0.1 μl/l. Fumigation of fruiting cotton plants with only 0.5 μl/l caused 100% abscission of young fruits and floral buds within 2 days. This correlated with the estimated endogenous levels of ethylene. Reduced pressure, which reduced the internal levels of ethylene, delayed abscission of young fruits and leaves, a result which supports our conclusion from this study— that ethylene is one of the regulators of young fruit abscission in cotton.     

Fusicoccin-ethylene Interaction in the Growth of Etiotated Pea Seedlings

Intact etiolated seedlings of pea (Pisum sativum L. cv. guangzhou honqhua) were used to investigate the changes in the physiological activities by the treatment with fusicoccin (FC), ethylene, and FC + ethylene respectively with the aim of understanding the fusicoccin-ethylene interaction. After the combined application of FC and ethylene, FC may partly removes the inhibition of elongation and fresh weight caused by ethylene. However, that the reduction of DNA synthesis and cell division resulted from ethylene can not eliminated by FC. Ethylene induces the microfibril of cell wall to orientate longitudinally while FC alters microfibrillar orientation by random deposition. FC is also capable of decreasing the inhibition of K+ uptake, H+ secretion and respiratory rate caused by ethylene. The authors consider that the cause of FC partly antagonize ethylene on reversing ethylene-inhibited growth appears not to be reduced the content of endogenous ethylene, but may be caused by raising the respiratory rate, stimulating the H+ secretion and K+ uptake. Eventually, cells uptake much water and result in increase of their elongation and swelling.     

Effect of Ethylene on Cell Division and Deoxyribonucleic Acid Synthesis in Pisum sativum

Ethylene and supraoptimal levels of 2,4-dichlorophenoxyacetic acid inhibit the growth of the apical hook region of etiolated Pisum sativum (var. Alaska) seedlings by stopping almost all cell divisions. Cells are prevented from entering prophase. The hormones also retard cell division in intact root tips and completely stop the process in lateral buds. The latter inhibition is reversed partially by benzyl adenine. In root tips and the stem plumular and subhook regions, ethylene inhibits DNA synthesis. The magnitude of this inhibition is correlated with the degree of repression of cell division in meristematic tissue, suggesting that the effect on cell division results from a lack of DNA synthesis. Ethylene inhibits cell division within a few hours with a dose-response curve similar to that for most other actions of the gas. Experiments with seedlings grown under hypobaric conditions suggest that the gas naturally controls plumular expansion and cell division in the apical region.     

Ethylene as a natural agent inducing plumular hook formation in pea seedlings

Summary Removing endogenous ethylene by hypobaric treatment, or displacing it with carbon dioxide inhibits hook development in etiolated pea seedlings. When seedlings are returned to a normal atmosphere, hook formation occurs in darkness. Addition of ethylene accelerates this process. When ethylene induces hook formation, cell division in the hook tissue is rather inhibited by the gas. These data suggest that endogenous ethylene causes formation of the hook by inducing expansion of certain cells.     

THE ROLES OF PLANT HORMONES IN STYLE AND STIGMA GROWTH IN GAILLARDIA GRANDIFLORA (ASTERACEAE)

Style and stigma elongation and stigma unfolding, and the roles of plant hormones in these processes in Gaillardia grandiflora Van Houtte were investigated. Style and stigma elongation in vivo began just after anthesis, and style elongation was accompanied by epidermal cell elongation (greatest near the stigma) and a fresh weight increase, but not by cell division or a dry weight increase. The stigma unfolded after the style and stigma elongated. Style-stigma units excised from young disc flowers of this composite were measured as they responded to plant growth regulators applied singly, as well as in sequential and simultaneous combinations, in vitro. Style elongation was promoted by auxin, was inhibited by gibberellins and ethylene, and was unaffected by other growth regulators. Stigma elongation followed a similar pattern of response. Endogenous auxin levels and ethylene production showed parallel variation and endogenous gibberellin levels showed inverse variation with style and stigma elongation. Stigma unfolding was more sensitive to auxin applications and was promoted by applied ethylene. Ethylene production showed parallel variation and endogenous auxin levels showed inverse variation with stigma unfolding. AVG and Co²⁺ applications decreased auxin-induced style elongation and fusicoccin promoted all of the growth responses of style-stigma units in vitro. A gibberellin-auxin-ethylene-acid growth interaction mode of control is proposed for these three growth processes.     

Ethylene directs auxin to control root cell expansion

Root morphogenesis is controlled by the regulation of cell division and expansion. We isolated an allele of the eto1 ethylene overproducer as a suppressor of the auxin-resistant mutant ibr5, prompting an examination of crosstalk between the phytohormones auxin and ethylene in control of root epidermal cell elongation and root hair elongation. We examined the interaction of eto1 with mutants that have reduced auxin response or transport and found that ethylene overproduction partially restored auxin responsiveness to these mutants. In addition, we found that the effects of endogenous ethylene on root cell expansion in eto1 seedlings were partially impeded by dampening auxin signaling, and were fully suppressed by blocking auxin influx. These data provide insight into the interaction between these two key plant hormones, and suggest that endogenous ethylene directs auxin to control root cell expansion.     

Comparison of Propylene-induced Responses of Immature Fruit of Normal and rin Mutant Tomatoes

Continuous application of propylene to 40 to 80% mature fruits of normal tomato strains (Lycopersicon esculentum Mill.) advanced ripening in fruits of all ages by at least 50%. Although preclimacteric respiration was stimulated by propylene treatment, there was no concomitant increase in ethylene production. Once ripening commenced, the rates of endogenous ethylene production were similar in both propylene-treated and untreated fruits. Continuous exposure to propylene also stimulated respiration in immature fruits of rin, a nonripening mutant. Although respiration reached rates similar to those during the climacteric of comparable normal fruits there was no change in endogenous ethylene production which remained at a low level. Internal ethylene concentrations in attached 45 to 75% mature fruits of rin and a normal strain were similar. It is suggested that the onset of ripening in normal tomato fruit is not controlled by endogenous ethylene, although increased ethylene production is probably an integral part of the ripening processes.     

Role of Ethylene Metabolism in Amaranthus retroflexus

¹⁴C-Ethylene was metabolized by etiolated pigweed seedlings (Amaranthus retroflexus L.) in the manner similar to that observed in other plants. The hormone was oxidized to ¹⁴CO₂ and incorporated into ¹⁴C-tissue components. Selected cyclic olefins with differing abilities to block ethylene action were used to determine if ethylene metabolism in pigweed is necessary for ethylene action. 2,5-Norbornadiene and 1,3-cyclohexadiene were effective inhibitors of ethylene action at 800 and 6400 microliters per liter, respectively, in the gas phase, while 1,4-cyclohexadiene and cyclohexene were not. However, all four cyclic olefins inhibited the incorporation and conversion of ¹⁴C-ethylene to ¹⁴CO₂ by 95% with I₅₀ values below 100 microliters per liter. The results indicate that total ethylene metabolism does not directly correlate with changes in ethylene action. Additionally, the fact that inhibition of ethylene metabolism by the cyclic olefins did not result in a corresponding increase in ethylene evolution indicates that ethylene metabolism does not serve to significantly reduce endogenous ethylene levels.