Abscisic Acid Alters the Metabolism of Indole-3-Acetic Acid in Senescing Flowers of Cucumis melo L

Experiments were conducted to investigate indole-3-acetic acid (IAA) and abscisic acid (ABA) metabolism associated with postanthesis senescence of ovaries from nonpollinated muskmelon (Cucumis melo L.) flowers. Flowers attached to the vine were allowed to senesce for 4 days after anthesis or were harvested at full anthesis and aged for the same time interval with or without access to water. The IAA ester, amide-linked forms of IAA, free ABA, and ABA esters increased in senescent ovaries from flowers left attached to the vine. Detaching flowers from the vine resulted in an accumulation of free and amide-linked IAA in the senescing ovary but suppressed accumulation of ester IAA. Free ABA failed to increase in ovaries detached from the vine. Subjecting detached flowers to water stress had no effect on the endogenous level of free ABA but resulted in the accumulation of ABA ester and suppression of any increase in free IAA. However, detached flowers treated with 0.1 millimolar ABA accumulated 75% less free IAA and initiated the synthesis of ester IAA. Detached flowers treated with ABA also accumulated high levels of ester ABA. These results suggest that the metabolism of free IAA in muskmelon ovary tissue is regulated in situ and not the consequence of external synthesis and importation. ABA appears to be transported into the senescing ovary from an external source and alters the IAA metabolism in such a manner as to suppress the level of free IAA while stimulating accumulation of the ester IAA.     

Hormonal Status in Wheat and Rice Seedlings under Anoxia

A comparative analysis of the effects of anoxia on growth, fresh weight gain, and phytohormones in wheat (Triticum aestivum L.) and rice (Oryza sativa L.) seedlings was performed. In both plant species, a total cessation of root growth occurred during the initial hours of anoxia. In an anaerobic environment, the fresh weight of wheat seedlings decreased. An increase in the shoot length and weight under the stress conditions was found only in rice seedlings. During the initial hours of anoxia, the level of free ABA in wheat and rice tissues increased manifold, and the accumulation of a free ABA form occurred at the expense of the hydrolysis of its bound forms. The IAA content in plant tissues also increased. In wheat, the accumulation of IAA was short, but in rice, a high hormone level was retained during the entire experiment, and, as a result, its concentration exceeded that of ABA. A level of cytokinins in the tissues of both plant species was affected by anoxia to a lesser extent than that of other phytohormones. This level somewhat decreased under anoxia similarly to the level in darkness under aeration. It is suggested that IAA accumulation in hypoxia-tolerant rice seedlings under anoxia favors maintenance of shoot growth and simultaneous inhibition of root growth. At the same time, in the hypoxia-sensitive wheat, an increase in the ABA level resulted in growth cessation.     

Hormonal changes associated with fruit set and development in mandarins differing in their parthenocarpic ability

Satsuma [Citrus unshiu (Mak) Marc.] and Clementine [Citrus reticulata (Hort.) Ex. Tanaka, cv. Oroval] are two related species of seedless mandarins which differ in their tendency to set parthenocarpic fruits. Satsuma fruits naturally set parthenocarpically whereas Clementine mandarins show very low ability to set fruit in the absence of cross-pollination. The endogenous levels of gibberellins (GAs) and free and conjugated indole-acetic acid (IAA) and abscisic acid (ABA) throughout early stages of fruit development were investigated in seedless cultivars of both species. Analyses performed by full-scan combined gas chromatography-mass spectrometry (GC-MS) of extracts from ovaries at anthesis demonstrated the presence of GA₁₉, GA₂₀, GA₂₉, GA₁, GA₈, GA₃ and iso-GA₃ in Satsuma mandarin, whereas only GA₂₉, GA₃ and trace levels of GA₈ were detected in Clementine. At this developmental stage GA-like substances, as estimated by bioassay, reached their highest levels in Satsuma, while Clementine mandarins contained relatively lower levels. In both species the highest levels of free IAA were found at petal-fall stage at which time free ABA levels also peaked. Developing fruits of Clementine had higher amounts of both free IAA and ABA. In Satsuma, levels of conjugated IAA remained low throughout reproductive development whereas in Clementine they increased as the free form declined. In contrast, conjugated ABA was at low levels in Clementine but reached higher concentrations in Satsuma. These results suggest that in these mandarins the potential for setting parthenocarpic fruits is mainly influenced by the hormonal status of the fruit during the later stages of cell division and early stages of cell enlargement. Thus, the condition of low ability to set parthenocarpic fruits appears to be associated with lower levels of active GAs, lower capability to catabolize ABA to conjugated ABA and higher ability to conjugate IAA during this period.     

Concentration of Indole-3-acetic Acid and Its Derivatives in Plants

Seeds of oat, coconut, soybean, sunflower, rice, millet, kidney bean, buckwheat, wheat, and corn and vegetative tissue of oat, pea, and corn were assayed for free indole-3-acetic acid (IAA), esterified IAA, and peptidyl IAA. Three conclusions were drawn: (a) all plant tissues examined contained most of their IAA as derivatives, either esterified or as a peptide; (b) the cereal grains examined contained mainly ester IAA; (c) the legume seeds examined contained mainly peptidyl IAA. Errors in analysis of free and bound IAA are discussed.     

Activity of trypsin inhibitors in wheat seedlings exposed to pathogenic fungus Tilletia caries and phytohormones

We compared the effect of common bunt (caused by fungus Tilletia caries Tul.) and treatment with phytohormones IAA, ABA, and cytokinins (CK) on the activity of trypsin inhibitors (TI) in wheat seedlings. The experiments were conducted with pathogen-susceptible species of wheat Triticum aestivum L. (cv. Zhnitsa) and resistant species T. timopheevii Zhuk. (accession k-58666 from the collection of All-Russian Institute of Plant Industry). In the resistant wheat, the fungus elevated the activity of TI and the content of CK, whereas in the susceptible wheat, it induced accumulation of IAA. In the seedlings of wheat T. aestivum, TI activity increased under the effect of CK, same as upon the action of pathogen. ABA briefly increased the activity of TI, whereas IAA did not considerably affect it. It was concluded that among the investigated hormones, CK play a leading role in the regulation of defense responses of wheat plants involving TI.     

Seasonal Changes in the Levels of Free and Bound Abscisic Acid in Blackcurrant (Ribes nigrum) Buds and Beech (Fagus sylvatica) Buds

Free and bound abscisic acid (ABA) levels in blackcurrant andbeech buds have been determined by an extraction procedure involvingseveral thin-layer chromatography purification stages and finalbioassay in the wheat coleoptile straight-growth test. In bothspecies the highest level of free ABA occurs in the autumn atabout the time of onset of winter dormancy. The free ABA contentthen declines throughout the winter months reaching its lowestvalue just before bud burst. These results strengthen the viewthat free ABA plays an important role in the induction and maintenanceof winter dormancy. The bound ABA level increases in both species throughout theautumn and winter until halfway through the bud swelling phaseand then declines. It is proposed that the bound ABA accumulatesin the buds because it cannot be translocated or further metabolizedduring the winter months. The ratios of free to bound ABA were plotted and the resultingcurves show a distinctive annual cycle with the highest freeABA/ bound ABA ratio (F/B) occurring in the autumn and the highestbound ABA/ free ABA ratio (B/F) coinciding with bud burst. Agradual increase in the B/F ratio during bud swelling may indicatea feed-back reaction between free and bound ABA which maintainsthe free ABA at a sufficiently high level to put a gentle brakeon growth. At bud burst the B/F ratio falls very sharply anda period of very rapid growth ensues.     

Overexpression of wheat <Emphasis Type="Italic">TaNCED</Emphasis> gene in <Emphasis Type="Italic">Arabidopsis</Emphasis> enhances tolerance to drought stress and delays seed germination

Abscisic acid (ABA) regulates various plant physiological processes, especially participates in the plant responses to harsh environments. The 9-cis-epoxycarotenoid dioxygenase (NCED) is a key enzyme in ABA biosynthesis pathway. Here, a TaNCED with an 1 887-bp open reading frame was cloned from wheat, which encodes a peptide of 628 amino acids. A chloroplast transit peptide sequence was found at the N-terminus of the TaNCED protein. Multiple sequence alignments indicate that the TaNCED protein shared high similarities with other NCEDs from different species. Real-time quantitative PCR analysis shows that expression of TaNCED was strongly up-regulated by treatments with ABA, polyethylene glycol, and drought stress, and it was down-regulated during germination of the wheat seeds. Ectopic overexpression of the TaNCED gene in Arabidopsis resulted in an increase of endogenous ABA and free proline content. A lower water loss rate and stomatal conductance of leaves were found in the transgenic plants in comparison with the wild type. Subsequently, the transgenic plants displayed an enhanced tolerance to drought stress but delayed seed germination. These data provide evidence that the TaNCED might play a primary role in regulation of ABA content during water stress and seed dormancy.     

Comparative analysis of the spatiotemporal distribution of endogenous hormones in the ovaries of wheat and dandelion in the period of ovule activity

New data are reported on the spatiotemporal distribution of free and bound forms of phytohormones (cytokinins, auxins, abscizins) in the ovaries of the amphimictic species Triticum aestivum L. and the apomictic species Taraxacum officinale Web. The data are attributed to four domains of the ovary and three intervals composing the period of ovule activity (from the moment of the ovule at relative rest to the moment of the ovule at the first division). The data were obtained by modifications of the ELISA method. The modifications were elaborated for each phytohormone. Comparative analysis of the data implies that basic events of the reproduction process in the ovary in the period of the ovule activity are under the endogenous hormonal control system. The distributions of free and bound forms of phytohormones in the ovaries of two species show both common regularities, probably characteristic of amphimicts and apomicts, and special features. The latter may be related to the specificity of the reproduction type.     

Tobacco-mosaic-virus-induced increase in abscisic-acid concentration in tobacco leaves:

The concentrations of free and bound abscisic acid (ABA and the presumed ABA glucose ester) increased three- to fourfold in leaves of White Burley tobacco (Nicotiana tabacum L.) systemically infected with tobacco mosaic virus. Infected leaves developed a distinct mosaic of light-green and dark-green areas. The largest increases in both free and bound ABA occurred in dark-green areas. In contrast, virus accumulated to a much higher concentration in light-green tissue. Free ABA in healthy leaves was contained predominantly within the chloroplasts while the majority of bound ABA was present in non-chloroplastic fractions. Chloroplasts from light-green or dark-green tissues were able to increase stromal pH on illumination by an amount similar to chloroplasts from healthy leaf. It is unlikely therefore that any virus-induced diminution of pH gradient is responsible for increased ABA accumulation. Tobacco mosaic virus infection had little effect on free ABA concentration in chloroplasts; the virus-induced increase in free ABA occurred predominantly out-side the chloroplast. The proportional distribution of bound ABA in the cell was not changed by infection. Treatment of healthy plants with ABA or water stress increased chlorophyll concentration by an amount similar to that induced by infection in dark-green areas of leaf. A role for increased ABA concentration in the development of mosaic symptoms is suggested.Abbreviations ABA abscisic acid - TMV tobacco mosaic virus     

Changes in abscisic acid and phenols during flower development in two diverse species of rose

Changes in endogenous abscisic acid (ABA) and phenols were determined in petals of two diverse species of rose, viz., Rosa damascena Mill and Rosa bourboniana Desport during flower development. A progressive increase in free ABA was observed during flower development till full bloom in both the species with higher content of free ABA in Rosa damascena. While bound ABA level increased in Rosa damascena till stage 6, in Rosa bourboniana it continued to increase till full bloom. Acidic phenols increased slowly in both the species till stage 4, but sharply afterwards and no significant differences were noticed during full bloom period. Bound phenols content was higher in Rosa damascena during full bloom period. The significance of these changes in relation to flowering in the two diverse species of rose is discussed.     

Cloning of 9-cis-epoxycarotenoid dioxygenase gene (TaNCED1) from wheat and its heterologous expression in tobacco

Abscisic acid (ABA) regulates plant responses to various environmental stresses. Oxidative cleavage of cis-epoxycarotenoids catalyzed by 9-cis-epoxycarotenoid dioxygenase (NCED) is the critical step in the biosynthesis of ABA in higher plants. Using a homologous cloning approach, a NCED-like gene (designated as TaNCED1) was isolated from wheat (Triticum aestivum). It contained an open reading frame of 1 848 bp and encodes a peptide of 615 amino acids. Multiple sequence alignments showed that TaNCED1 shared high identity with NCEDs from other plants. Phylogenetic analysis revealed that TaNCED1 was most closely related to a barley HvNCED1 gene. The predicted 3D structure of TaNCED1 showed high similarity with other homologues. Southern blot analysis indicated that TaNCED1 was a single copy in the genome of wheat. TaNCED1 was differentially expressed in various organs and the expression was up-regulated by low temperature, drought, NaCl, and ABA. Heterologous expression of TaNCED1 in tobacco (Nicotiana tabacum) significantly improved its drought tolerance. Under drought treatment, TaNCED1-overexpressing transgenic tobacco plants exhibited higher germination rate, higher relative water content, content of soluble sugars and of ABA when compared with the wild type plants.     

Abscisic Acid Accumulation in Developing Seeds of Phaseolus vulgaris L

Free and bound abscisic acid (ABA) in the pod, seed coat, and embryo were determined separately throughout seed development of Phaseolus vulgaris L. cv. `Taylor's Horticultural.' An internal standard method of gas-liquid chromatography was used for ABA quantification. In the embryo, two peaks of free ABA occurred at days 22 (1.18 micrograms per gram or 5.5 micromolar) and 28 (1.74 micrograms per gram or 12 micromolar); and a single peak of bound ABA at day 30. In the seed coat, there was one peak of free ABA at day 22 and only small amounts of bound ABA. Very small amounts of ABA were detected in the pod at any stage of development. In cv. PI 226895, in which seed development is more rapid than in `Taylor's Horticultural,' the embryo ABA peaks occur on days 20 and 26. The timing of the ABA peak in the embryo, and the concentration attained, are consistent with previous reports on the natural pattern of RNA synthesis and with ABA inhibition of RNA synthesis in developing bean fruit.     

Effect of Growing Conditions on Wheat Hormonal Status and Productivity in Experimental Ecological System

The levels of free and bound forms of IAA, ABA, cytokinins (CK), and gibberellins, as well as growth characteristics and productivity were investigated in two wheat lines. The plants were grown under controlled conditions in an artificial ecosystem that allowed the irradiance, CO₂ concentration, and rhizosphere temperature to be changed. The main difference in the hormonal status of leaves of tall spring wheat, line 232, and dwarf wheat, line 95-3, was the absence of GA₉ gibberellins in the latter. It was found that the light intensity and temperature of rhizosphere insignificantly affected the balance of endogenous phytohormones and HI in wheat. The elevation of CO₂ concentration resulted in a considerable increase in the content of free IAA, an appearance of free GA₉, and a rise in the productivity of wheat, line 232. The concentration of CO₂ was shown to be a major parameter that determined HI in the experimental ecological system.     

Distribution of free and bound forms of cis-trans and trans-trans abscisic acid in broad-bean plants in relation to apical dominance

Levels of endogenous abscisic acid (ABA; free and bound forms) have been determined by gas chromatography in stems and buds of broad-bean plants ( Vicia faba L. cv. Aguadulce) in relation to apical dominance. A downward gradient of free cis-trans ABA occurred along the stem, from the apical bud to the roots. Except for the actively growing apical bud the levels of free cis-trans ABA were higher in the buds than in the corresponding nodes. An inverse correlation can be set up between levels of free cis-trans ABA and growth of buds, except for the cotyledonary ones. High levels of bound ABA ( cis-trans form) are correlated with the growth of the apical bud and that of the axillary bud ax1. The hormonal regulation of the growth of the cotyledonary buds, which contained high levels of trans-trans ABA in bound forms, is apparently different from that of the other buds.     

Ethylene-induced Formation of ABA in Citrus Peel as Related to Chloroplast Transformations

When mature green harvested Shamouti oranges (Citrus sinensis L. Osbeck) were exposed to 35 μl/liter of ethylene, a 3-fold increase in free abscisic acid (ABA) of the flavedo could be detected after 12 hours and a 10-fold increase after 24 hours, while chlorophyll destruction did not exceed 20%. The increase in free ABA continued up to 24 hours and leveled off. Bound ABA accumulated strongly after 24 hours suggesting that excess of free ABA was being converted into the bound form. Similar increases in ABA upon exposure to ethylene occurred also in fully mature orange fruits which had already lost all of their chlorophyll, in white and green portions of the flavedo of variegated lemons, and in the colorless albedo of Eureka lemons.     

Identification of candidate SNPs for drug induced toxicity from differentially expressed genes in associated tissues

The full-length cDNA sequence (1158 bp) encoding a ribosomal L5 protein, designated as TaL5, was firstly isolated from common wheat (Triticum aestivum L.) using the rapid amplification of cDNA ends method (RACE). The open reading frame (ORF) of TaL5 gene was 906 bp, and its deduced amino acid sequence (301 residues) shared high similarity to those of other higher plant L5 proteins. TaL5 protein contained a putative 5S binding region (74 amino acids). TaL5 DNA sequence was further cloned, and sequence analysis showed that it contained 7 introns and 8 exons. Predicated using TargetP software, TaL5 protein was putatively located in mitochondria and contains a transit peptide of 12 amino acids. During grain filling period, temporal expression pattern of TaL5 gene was approximately consistent with the rates of starch accumulation in grains. Additionally, TaL5 gene was dramatically induced by salt, drought and freezing stresses, exogenous abscisic acid (ABA) and salicylic acid (SA) in wheat seedlings. These implied that TaL5 gene could function in growth, development and abiotic stresses in wheat plants.     

Optically pure abscisic Acid analogs-tools for relating germination inhibition and gene expression in wheat embryos

We report an examination of the structural requirements of the abscisic acid (ABA) recognition response in wheat dormant seed embryos using optically pure isomers of ABA analogs. These compounds include permutations to the ABA structure with either an acetylene or a trans bond at C-4 C-5, and either a single or double bond at the C-2′ C-3′ double bond. (R)-ABA and the three isomers with the same configuration at C-1′ as natural ABA were found to be effective germination inhibitors. The biologically active ABA analogs exhibited differential effects on ABA-responsive gene expression. All the ABA analogs that inhibited germination induced two ABA-responsive genes, wheat group 3 lea and dhn (rab). However, (R)-ABA and (S)-dihydroABA were less effective in inducing the ABA-responsive gene Em within the time that embryonic germination was inhibited.