A possible regulatory role for ethylene in the pollen-pistil interaction in a sporophytic self-incompatible system

The sporophytic type of self-incompatibility exhibited by Ipomoea cairica Sweet (Convolvulaceae) was partially overcome in vitro by treating the pollen and/or stigma with 10^–6 to 10^–1 M methionine, a precursor of ethylene. The implications of these observations are discussed in relation to other experiments involving use of the ethylene antagonist AgNO₃, individually and in combination with methionine and an optimum level of indole-3-acetic acid (10^–2 M). The results suggest a role for ethylene (which could also be IAA-induced) in regulating pollen germination and further tube growth in sporophytic self-incompatible systems. A hypothesis on the action of hormones in pollen germination and tube growth in a sporophytic self-incompatible (SSI) system is presented.     

Is the ‘rejection reaction’ inducing ability in sporophytic self-incompatible systems restricted only to pollen and tapetum?

Summary The present investigation describes the stigmatic rejection response, induced by anther or pollen at developmental stages ranging from anther primordia to mature pollen, as well as in other floral and vegetative tissues. This has been studied in a sporophytic self-incompatible system, Brassica campestris (Brassicaceae). The implications of these observations have been discussed.     

Spectrophotometric method for determining gibberellic acid in fermentation broths

A novel method for the quantitative determination of gibberellic acid in fermentation broths has been developed. It is based on the kinetic of the reaction of conversion of gibberellic acid to gibberellenic acid. The method is simple, reliable, faster than most of methods known, and free of the interferences which commonly affect spectrophotometric methods currently in use. Its threshold sensitivity is 0.1 g and its accuracy is greater than 97% for concentrations of gibberellic acid ranging from 0.1 to 1 g l(-1).     

Possible role of soluble invertase in the gibberellic acid berry-sizing effect in Sultana grape

It is well known that post-bloom applications ofgibberellic acid (GA₃) increase seedless grapeberry size by enhancing cell division, or cellenlargement, or both. As a consequence, total waterand sugar per berry are increased. Soluble invertaseis considered to be one of the key enzymes in theaccumulation of sugar in grape berries. To study apossible role of invertase in the GA₃berry-sizing effect, different rates of post-bloomGA₃ were applied to seedless grape cv. Sultanaand hexose concentration and invertase activity weremeasured. GA₃ stimulated both parameters as earlyas 24 and 32 h after applications, respectively.Moreover, the increment in sugar content and enzymeactivity remained throughout the growing of the berries period and, at ripening, increases in hexosescontent (102%) and invertase activity (60%) weredetected when GA₃ was applied at a rate of 45 ppm.At the same GA₃ rate the pericarp cellsdoubled in size. Furthermore, positive correlationswere found between berry-size, invertase activity andhexose content, suggesting that GA₃ stimulationof invertase could be one of the factors involved in theberry sizing-effect of GA₃.     

Bioprocess strategies and recovery processes in gibberellic acid fermentation

Gibberellic acid (GA₃) is a commercially important plant growth hormone, which is gaining much more attention all over the world due to its effective use in agriculture and brewing industry. Industrially it is produced by submerged fermentation technique using Ascomycetous fungusGibberella fujikuroi. Solid state and immobilized cell fermentation techniques had also been developed as an alternative to obtain higher yield of GA₃. This review summarizes the problems of GA₃ fermentation such as production of co-secondary metabolites along with GA₃, substrate inhibition and degradation of GA₃ to biologically inert compound gibberellenic acid, which limits the yield of GA₃ in the fermentation medium. These problems can be overcome by various bioprocessing strategiese.g. two-stage and fed batch cultivation processes. Further research on bioreactor operation strategies such as continuous and/or extractive fermentation with or without cell recycle/retention system need to be investigated for improvement in yield and productivity. Down stream processing for GA₃ isolation is also a challenge and procedures available for the same have been critically evaluated.     

Nutritional and gibberellic acid requirements in kiwifruit vitroponic cultures

Summary Residual macronutrients, carbon source, and gibberellic acid (GA₃) in the culture medium were measured throughout the whole culture period of kiwi (Actinidia deliciosa Chev. cv. Hayward) explants cultured in liquid medium using cellulose plugs as explant support. The objective of this study was to adapt the composition of the culture medium to an automated culture system with the possibility of applying 6-benzyladenine pulses so as to improve the efficiency of kiwifruit micropropagation. Ammonium and phosphate were the most consumed ions at the end of the culture period (35 d). Murashige and Skoog medium might be too rich for the culture of kiwifruit in liquid medium except with respect to phosphate which decreased significantly toward the end of the culture period. Sucrose, and the glucose resulting from the hydrolysis of the former, were taken up by the kiwi explants throughout the whole culture period. GA₃ absorption by the kiwi explants occurred gradually throughout the subculture, although 67% of this initially added plant growth regulator remained in the culture medium at the end of the 35-d culture period.     

Effect of gibberellic acid on photosynthesis and glycollate dehydrogenase in Anacystis nidulans

Gibberellic acid at 10^-4 Mxxx was optimal for enhancement of growth, O₂ evolution, photosystem II and I and the activity of glycollate dehydrogenase of Anacystis nidulans. A stimulatory effect was observed on photosystem II. Other concentrations of gibberellic acid were inhibitory to O₂ evolution and photosystem I. Syntheses of phycocyanin, phycoerythrin and -carotene were significantly enhanced after 48 h incubation with gibberellic acid at 10^-3 Mxxx but the chlorophyll content began to increase 3 h after adding 10^-4 Mxxx gibberellic acid.The author is with the Department of Biological Sciences, Faculty of Science, University of Science and Technology, Irbid, Jordan.     

Stimulation of the gibberellic acid synthesis by Aspergillus niger in submerged culture using a precursor

Stimulation of Aspergillus niger in submerged culture using a commonly known precursor, mevalonic acid (MVA), was investigated in terms of growth and gibberellic acid production. Increasing concentrations of MVA up to 60 M enhanced product and growth yields. Above this amount, gibberellic acid yields and growth were gradually decreased.     

The effects of gibberellic acid,fluridone, abscisic acid and paclobutrazol on anther culture of Brussels sprouts

Both gibberellic acid (GA₃) and fluridone, a non-specific inhibitor of ABA biosynthesis, promoted embryo production in anther cultures of Brussels sprouts cv. Hal, but not in cv. Gower. Abscisic acid (ABA) and the gibberellin-biosynthesis inhibitor paclobutrazol inhibited embryo production in both cultivars.     

Capture of volatile metabolites for tracking the evolution of gibberellic acid in a solid-state culture of Gibberella fujikuroi

The evolution of volatile compounds produced during solid substrate cultivation (SSC) of Gibberella fujikuroi on wheat bran was tracked looking for volatile metabolites related with GA₃ production. Ethyl acetate and isoamyl acetate gave identical profiles and sharp increases that abated as the production of GA₃ began, while ent-kaurene displayed a profile matching that of the development of GA₃. ent-Kaurene is a precursor in the synthesis of gibberellins and was the most abundant compound found.     

Effects of abscisic acid,gibberellic acid and fusicoccin on the transmembrane potential during the early phases of germination in radish (Raphanus sativus L.) seeds

During germination, the transmembrane electric potential (PD) of cortical cells of the embryonal axis of radish seeds (Raphanus sativus L.) rises from-120 mV initially to a maximum of-150 mV after 5 h incubation, then falls again to stable values of around-120 mV. Treatments inhibiting germination block the transitory PD increase. Administration of uncoupling agents or low temperatures, during the process of germination, produces a marked fall of the PD transitory increase. Abscisic Acid has a parallel inhibitory effect on PD and germination, while fusicoccin produces a rise in both; administration of abscisic acid with fusicoccin inhibits germination, while the PD remains at the high levels given by fusicoccin. These results are discussed in relation to ion exchange at membrane level.Abbreviations ABA abscisic acid - FC fusicoccin - GA₃ gibberellic acid - PD electric potential difference (between the vacuole and the external medium) - CH cycloheximide - DNP dinitrophenol - FCCP (p-trifluormethoxy)-carbonylcyanide-phenylhydrazone - DCCD N,N-dicyclohexylcarbodiimide     

Factors affecting gibberellic acid production by Fusarium moniliforme in solid-state cultivation on starch

The production of gibberellic acid (GA₃) by Fusarium moniliforme M-7121 in solid-state culture was evaluated in flask cultures as well as in 3-I horizontal rotary reactors. The highest production rate of GA₃ was with 80% (w/v) maize flour mixed with wheat bran. The optimum initial moisture content was inversely dependent on the ambient relative humidity. The initial water activity range for optimal growth and GA₃ accumulation was about 0.98 to 0.99, which is unusually high for a filamentous fungus. A low O₂ concentration resulted in a much decreased GA₃ yield and the appearance of a yellow to reddish pigmentation in the mycelium. The lag phase was short and rapid growth continued for up to 2 days in the rotary reactor, with a maximum specific growth rate of 0.12 h^–1. The maximum rate of GA₃ production occurred during the subsequent 3 to 10 days of incubation and the final GA₃ concentration reached was 18.7 mg to 19.3 mg/g dry culture. The point of maximum GA₃ accumulation after 10 to 12 days of incubation was usually marked by a sharp increase in pH.The authors are with the Department of Microbiology and Biochemistry, University of the Orange Free State, P.O. Box 339, 9300 Bioemfontein, Republic of South Africa     

Production of steroidal alkaloids by hairy roots of Solanum aviculare and the effect of gibberellic acid

Cultures of Solanum aviculare hairy roots were established after transformation with Agrobacterium rhizogenes A4. High levels of steroidal alkaloids measured as solasodine equivalents were produced in shake-flasks and bioreactor, even though relatively low concentrations are found in roots in vivo. In shake flasks the maximum alkaloid yield was 32 mg g^-1 dry weight; in a 3-1 air-driven bioreactor the yield was 29 mg g^-1. These yields represent a 5-fold increase over previous reports for in vitro production, and are comparable with levels found in the aerial parts of intact S. aviculare plants. Production of steroidal alkaloids was growth-associated. High sugar levels at stationary phase and insensitivity to increased levels of medium components suggest that root cultures were limited by oxygen mass-transfer. In Petri-dish culture with and without exogenous gibberellic acid, root length and number of root tips increased exponentially; growth proceeded with a constant length per root tip of about 35 mm. Addition of gibberellic acid enhanced growth but reduced the specific steroidal-alkaloid level. Taking into account both growth and alkaloid yield, accumulation of steroidal alkaloids was improved by about 40% at gibberellic-acid concentrations of 10 and 100 g l^-1.     

Effects of foliar applied gibberellic acid and benzyladenine upon yield components in sunflower (Helianthus annuus L.)

Sunflower yield is determined by seed number/m^–2 and by achene weight. Frequently, a high percentage of empty achenes in the inner portion of the capitulum, probably due to a reduced vascularization of that section of the flower head, decreases final yield. The objective of the present research is to determine if foliarly applied gibberellic acid (GA) and benzyladenine (BA) can enhance the vascularization in the inner portion of the capitulum, improving photoassimilate translocation. Field experiments were conducted during 1989/90 with hybrid SPS 894 and during 1990/91 with hybrid ACA 882. GA (150 mg/l^–1), BA (150 and 250 mg/l^–1) and GA 150+BA 150 mg/l^–1 each were foliarly applied 20, 40, or 60 days after emergence. For both seasons and hybrids plant growth regulator (PGRs) applications significantly reduced the percentage of empty achenes, increased achene weight, achene weight (× 1000) and achene number in the inner portion of the capitulum and in the middle and outer portion during 1990/91. A 25% increase in seed yield was achieved due to PGR application and the capitulum partition index (achene weight/receptacle weight^–1, CPI) was significantly increased due to an improvement in photoassimilate distribution. A distribution model was derived showing that preferential allocation of photoassimilates in the outer portion of the capitulum can be modified by PGR application, demonstrating that photoassimilate distribution is under hormonal control.Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET)     

Biosynthesis of glycoproteins involved in the pollen-stigma interaction of incompatibility in developing flowers of Brassica oleracea L.

De-novo synthesis of the S-allele-specific glycoproteins of Brassica oleracea is demonstrated in stigmas at different developmental stages. Excised stigmas incorporate ¹⁴C-labeled amino acids into their S-glycoproteins early in development and before the self-incompatibility response is acquired, but the rate of synthesis accelerates prior to anthesis, resulting in the accumulation of high levels of the S-glycoproteins in the stigma and coinciding with the acquisition of the pollen-stigma incompatibility response. Since the self-compatible and self-incompatible zones of developing inflorescences are very sharply delineated, a threshold quantity of S-glycoproteins appears to be critical for the onset of self-incompatibility. Incorporation experiments in which [^35Smethionine was applied to intact stigma surfaces indicate that the papillae are the main sites of synthesis of the S-specific glycoproteins.Abbreviations IEF isoelectric focusing - SC self-compatibility - SDS sodium dodecyl sulfate - SI self-incompatibility     

Synergistic interaction of gibberellic acid and triazinone derivatives in the promotion of rice shoot growth

Forty-five 1,3,5-triazine-2,6-dione derivatives were synthesized and their plant growth-promoting activities examined by the rice (Oryza sativa) seedling test in the presence or absence of gibberellic acid (GA). For high activity in promoting the growth of rice seedlings and acting as active GA-synergists, a para-substituted or a 2,4-disubstituted phenyl group, a hydrogen atom and an alkoxy group were required in the 1-, 3- and 4-positions of the 1,3,5-triazine-2,6-dione molecule. 4-Ethoxy-1-(p-tolyl)-s-triazine-2,6(1H, 3H)-dione [TA], one of the most potent triazinones, synergized the effect of GA on the shoot elongation of different varieties of rice including normal type, dwarf mutants and chlorophyll-mutants. TA synergistically increased the growth-promoting activity of GA by both a simultaneous treatment at the same sites and separate treatments at separate sites of rice seedlings.     

Increased photosynthetic rates following gibberellic acid treatments to the roots of tomato plants

This study was conducted to evaluate the effects of root applications of gibberellic acid (GA₃) on photosynthesis in tomato plants grown hydroponically. Photosynthetic rates (mg CO₂/dm²/hr) determined using an open infrared CO₂ gas exchange system showed a 40–50% increase within 5 hr after treatment with a 1.4 µM gibberellic acid (GA₃) to their roots. The effect was shown to persist for the duration of the experiment (9 days). Plants receiving pulses of 1.4 µM GA₃ to the roots for 1, 4, 8 or 12 hr exhibited significantly higher photosynthetic rates than the control for 6 days following treatment. By day 9 however, there was no significant difference. Continual treatments with 1.4 µM GA₃ to the roots maintained the photosynthetic rate significantly higher than the control for the duration of the experiment. Interestingly, at the lower light levels the percent stimulation was more dramatic. There was approximately a 90% increase in the photosynthetic rate at 80 µE m^-2 s^-1 while at saturating light conditions (560 µE m^-2 s^-1) there was approximately a 40% increase over the control rate. The light saturation point for both treated and control plants was 240 µE m^-2 s^-1. Applications of physiologically relevant concentrations of GA₃ to the roots of tomato plants stimulates photosynthesis more consistently than that achieved by previous studies involving foliar absorption.Approved for publication on May 28, 1981 as paper number 6242 in the Journal series of the Pennsylvania Agricultural Experiment Station.     

Numbers of sporophytic self-incompatibility alleles in populations of wild radish

Summary To estimate the numbers of sporophytic S-alleles in two adjacent populations of wild radish, we performed 701 reciprocal crosses among 50 individuals. Each cross was replicated five times in each direction. Sixteen plants were fully intercompatible, indicating the presence of at least 32 S-alleles in the two populations. A minimum of 22 S-alleles occur in a single population. The frequency of incompatibility was significantly higher for within-population crosses (14.5%) than for between-population crosses (7.8%). This suggests that the two populations differ in the composition and frequency of alleles at the S-locus.     

Responsiveness of Amaranthus retroflexus seeds to ethephon, 1-aminocyclopropane 1-carboxylic acid and gibberellic acid in relation to temperature and dormancy

Dormant Amaranthus retroflexus seeds do not germinate in the dark at temperatures below 35°C. Fully dormant seeds germinate only at 35–40°C whereas non-dormant ones germinate within a wider range of temperatures (15 to 40°C). Germination of non-dormant seeds requires at least 10% oxygen, but the sensitivity of seeds to oxygen deprivation increases with increasing depth of dormancy. 10^–6 to 10^–4 M ethephon, 10^–3 M 1-aminocyclopropane 1-carboxylic acid (ACC) and 10^–3 M gibberellic acid (GA₃) break this dormancy. In the presence of 10^–3 M GA₃ dormant seeds are able to germinate in the same range of temperatures as non-dormant seeds. The stimulatory effect of GA₃ is less dependent on temperature than that of ethephon, while ACC stimulates germination only at relatively high temperatures (25–30°C). The results obtained are discussed in relation to the possible involvement of endogenous ethylene in the regulation of germination of A. retroflexus seeds.Abbreviations ACC 1-aminocyclopropane 1-carboxylic acid - GA₃ gibberellic acid - SD standard deviation