[show abstract][hide abstract] ABSTRACT: Gibberellins (GAs) are the plant hormones that control many aspects of plant growth and development, including stem elongation. Genes encoding enzymes related to the GA biosynthetic and metabolic pathway have been isolated and characterized in many plant species. Gibberellin 2-oxidase (GA2ox) catalyzes bioactive GAs or their immediate precursors to inactive forms; therefore, playing a direct role in determining the levels of bioactive GAs. In the present study, we produced transgenic plants of the liliaceous monocotyledon Tricyrtis sp. overexpressing the GA2ox gene from the linderniaceous dicotyledon Torenia fournieri (TfGA2ox2). All six transgenic plants exhibited dwarf phenotypes, and they could be classified into two classes according to the degree of dwarfism: three plants were moderately dwarf and three were severely dwarf. All of the transgenic plants had small or no flowers, and smaller, rounder and darker green leaves. Quantitative real-time reverse transcription-polymerase chain reaction (PCR) analysis showed that the TfGA2ox2 expression level generally correlated with the degree of dwarfism. The endogenous levels of bioactive GAs, GA1 and GA4, largely decreased in transgenic plants as shown by liquid chromatography-mass spectrometry (LC-MS) analysis, and the level also correlated with the degree of dwarfism. Exogenous treatment of transgenic plants with gibberellic acid (GA3) resulted in an increased shoot length, indicating that the GA signaling pathway might normally function in transgenic plants. Thus, morphological changes in transgenic plants may result from a decrease in the endogenous levels of bioactive GAs. Finally, a possibility of molecular breeding for plant form alteration in liliaceous ornamental plants by genetically engineering the GA metabolic pathway is discussed.
Journal of plant physiology 06/2013; · 2.50 Impact Factor
[show abstract][hide abstract] ABSTRACT: Germination of Styrax japonicus seeds is promoted by warm stratification (WS) at 18–20°C followed by cold stratification (CS) at 4–5°. The objective of this
work was to analyze the state and mobility of water molecules measured by 1H-NMR and endogenous gibberellins (GAs) and abscisic acid (ABA) by ultra performance liquid chromatography/mass spectrometry/mass
spectrometry (UPLC-MS/MS) as influenced by WS and CS treatments had not previously been investigated. Styrax seeds that received 35 days of WS (35D WS) followed by 63 days of CS (63D CS) (35D WS + 63D CS) germinated. Seeds that received
only 35D WS failed to germinate. Endogenous GA1, GA8, GA19, GA20, and GA53 were identified as well as GA17, GA23, GA28, GA29, and GA97 by gas chromatography/MS (GC/MS) and UPLC-MS/MS in seeds that were treated with warm and cold stratification (WS + CS). This
suggests that the early C-13 hydroxylation pathway [-GA53-(GA44)-GA19-GA20-GA1-GA8] of GAs is a major biosynthetic pathway in the seeds. The concentration of GA53 and GA19 increased following WS and that of GA53 increased after WS + CS. The concentration of GA19 increased only slightly after WS + CS. The concentration of GA1 increased only after WS + CS. ABA concentration significantly decreased following the WS treatment. It is concluded that
the mobility of water molecules and water content in cotyledons and endosperm is increased following WS + CS treatments. The
occurrence of C-13 hydroxylated GAs suggests that the early C-13 hydroxylation pathway, → GA53 → GA44 → GA19 → GA20 → GA1 → GA8, is a major biosynthetic pathway in Styrax seeds.
Additional key wordsgibberellin metabolism–mass spectrometry–mass spectrometry (UPLC-MS/MS)–ultra performance liquid chromatography–water mobility
[show abstract][hide abstract] ABSTRACT: Three new 11β-hydroxy C20 gibberellins have been isolated from immature loquat fruit and their structures were established as 11β-hydroxy-GA12, 11β-hydroxy-GA15 and 11β-hydroxy-GA53, respectively, by direct GC–MS comparisons with authentic samples obtained from gibberellic acid by multistep syntheses. An advanced intermediate (30) was prepared in 20 steps from which 6 11β-hydroxy C20 gibberellins were prepared by parallel routes involving up to a further 5 steps for each sequence. The key steps involved a much improved synthesis of gibberellenic acid derivatives, a Lewis acid catalysed cyclisation of a diazoketone, a domino-hydroboration of a diene and oxidative cleavage of a ketone derived enolate.
[show abstract][hide abstract] ABSTRACT: Immature pumpkin (Cucurbita maxima) seeds contain gibberellin (GA) oxidases with unique catalytic properties resulting in GAs of unknown function for plant growth and development. Overexpression of pumpkin GA 7-oxidase (CmGA7ox) in Arabidopsis (Arabidopsis thaliana) resulted in seedlings with elongated roots, taller plants that flower earlier with only a little increase in bioactive GA4 levels compared to control plants. In the same way, overexpression of the pumpkin GA 3-oxidase1 (CmGA3ox1) resulted in a GA overdose phenotype with increased levels of endogenous GA4. This indicates that, in Arabidopsis, 7-oxidation and 3-oxidation are rate-limiting steps in GA plant hormone biosynthesis that control plant development. With an opposite effect, overexpression of pumpkin seed-specific GA 20-oxidase1 (CmGA20ox1) in Arabidopsis resulted in dwarfed plants that flower late with reduced levels of GA4 and increased levels of physiological inactive GA17 and GA25 and unexpected GA34 levels. Severe dwarfed plants were obtained by overexpression of the pumpkin GA 2-oxidase1 (CmGA2ox1) in Arabidopsis. This dramatic change in phenotype was accompanied by a considerable decrease in the levels of bioactive GA4 and an increase in the corresponding inactivation product GA34 in comparison to control plants. In this study, we demonstrate the potential of four pumpkin GA oxidase-encoding genes to modulate the GA plant hormone pool and alter plant stature and development.
[show abstract][hide abstract] ABSTRACT: Long day (LD) exposure of rosette plants causes rapid stem/petiole elongation, a more vertical growth habit, and flowering; all changes are suggestive of a role for the gibberellin (GA) plant growth regulators. For Arabidopsis (Arabidopsis thaliana) L. (Heynh), we show that enhancement of petiole elongation by a far-red (FR)-rich LD is mimicked by a brief (10 min) end-of-day (EOD) FR exposure in short day (SD). The EOD response shows red (R)/FR photoreversibility and is not affected in a phytochrome (PHY) A mutant so it is mediated by PHYB and related PHYs. FR photoconversion of PHYB to an inactive form activates a signaling pathway, leading to increased GA biosynthesis. Of 10 GA biosynthetic genes, expression of the 20-oxidase, AtGA20ox2, responded most to FR (up to a 40-fold increase within 3 h). AtGA20ox1 also responded but to a lesser extent. Stimulation of petiole elongation by EOD FR is reduced in a transgenic AtGA20ox2 hairpin gene silencing line. By contrast, it was only in SD that a T-DNA insertional mutant of AtGA20ox1 (ga5-3) showed reduced response. Circadian entrainment to a daytime pattern provides an explanation for the SD expression of AtGA20ox1. Conversely, the strong EOD/LD FR responses of AtGA20ox2 may reflect its independence of circadian regulation. While FR acting via PHYB increases expression of AtGA20ox2, other GA biosynthetic genes are known to respond to R rather than FR light and/or to other PHYs. Thus, there must be different signal transduction pathways, one at least showing a positive response to active PHYB and another showing a negative response.
[show abstract][hide abstract] ABSTRACT: Flower pigmentation is one of the most important traits for ornamental plants. To clarify the genetic basis for carotenoid pigmentation in flower tepals of Asiatic hybrid lily (Lilium sp.), we evaluated the segregation of a tepal-carotenoid content among F1 plants derived from a cross between ‘Montreux’ (having a small amount of carotenoids) and ‘Connecticut King’ (having a large amount of carotenoids), and mapped genetic loci for the carotenoid pigmentation onto the molecular linkage maps of ‘Montreux’ and ‘Connecticut King’ that we constructed previously. The tepal-carotenoid content among the F1 plants showed continuous segregation, indicating that several genes are associated with this trait. Quantitative trait loci (QTL) analysis identified one QTL, qCARmon6, on the sixth linkage group of the ‘Montreux’ map. qCARmon6 explained 58.2% of the total phenotypic variation, that is, this locus had a large effect on the carotenoid accumulation. The result that qCARmon6 was mapped on the linkage group of ‘Montreux’ which has a small amount of carotenoid pigments in tepals indicates that this locus has a dominant negative effect on carotenoid pigmentation.
[show abstract][hide abstract] ABSTRACT: Gibberellin (GA) 20-oxidase (GA20ox) is a key enzyme that normally catalyzes the penultimate steps in GA biosynthesis. One of the GA20ox genes in rice (Oryza sativa L.), OsGA20ox2 ( SD1 ), is well known as the "Green Revolution gene", and loss-of function mutation in this locus causes semi-dwarfism. Another GA20ox gene, OsGA20ox1, has also been identified, but its contribution to plant stature has remained unclear because no suitable mutants have been available. We isolated a mutant, B142, tagged with a T-DNA containing three CaMV 35S promoters, which showed a tall, GA-overproduction phenotype. The final stature of the B142 mutant reflects internode overgrowth and is approximately twice that of its wild-type parent. This mutant responds to application of both GA3 and a GA biosynthesis inhibitor, indicating that it is a novel tall mutant of rice distinct from GA signaling mutants such as slr1 . The integrated T-DNAs, which contain three CaMV 35S promoters, are located upstream of the OsGA20ox1 open reading frame (ORF) in the B142 mutant genome. Analysis of mRNA and the endogenous GAs reveal that biologically active GA level is increased by up-regulation of the OsGA20ox1 gene in B142. Introduction of OsGA20ox1 cDNA driven by 35S promoter into the wild type phenocopies the morphological characteristics of B142. These results indicate that the elongated phenotype of the B142 mutant is caused by up-regulation of the OsGA20ox1 gene. Moreover, the final stature of rice was reduced by specific suppression of the OsGA20ox1 gene expression. This result indicates that not only OsGA20ox2 but also OsGA20ox1 affects plant stature.
[show abstract][hide abstract] ABSTRACT: In this study, we present data showing that two members of the GRAS family of genes from rice, CIGR1 and CIGR2 (chitin-inducible gibberellin-responsive), inducible by the potent elicitor N -acetylchitooligosaccharide (GN), are rapidly induced by exogenous gibberellins. The pattern of mRNA accumulation was dependent on the dose and biological activity of the gibberellins, suggesting that the induction of the genes by gibberellin is mediated by a biological receptor capable of specific recognition and signal transduction upon perception of the phytoactive compounds. Further pharmacological analysis revealed that the CIGR1 and CIGR2 mRNA accumulation by treatment with gibberellin is dependent upon protein phosphorylation/dephosphorylation events. In rice calli derived from slender rice 1, a constitutive gibberellin-responsive mutant, or d1, a mutant deficient in the alpha -subunit of the heterotrimeric G-protein, CIGR1 and CIGR2 were induced by a GN elicitor, yet not by gibberellin. Neither gibberellin nor GN showed related activities in defense or development, respectively. These results strongly suggested that the signal transduction cascade from gibberellin is independent of that from GN, and further implied that CIGR1 and CIGR2 have dual, distinct roles in defense and development.
[show abstract][hide abstract] ABSTRACT: Correlation of coloration and anthocyanin profile in tulip flowers was investigated. Pelargonidin 3-rutinoside, cyanidin 3-rutinoside, delphinidin 3-rutinoside and their acetyl derivatives were major anthocyanins. Anthocyanins in purple, red, orange and pink perianthes of tulip were analyzed by HPLC. The occurrence of delphinidin 3-rutinoside and its acetyl derivatives is responsible for purple coloration. Pelargonidin 3-rutinoside, cyanidin 3-rutinoside and their 2'''-acetyl esters were major anthocyanins in perianthes of the red, orange and pink cultivars with their variant composition ratios. Tissue specific coloration and distribution of anthocyanins among the perianth, perianth-bottom, anther and pollen of the tulip flower were analyzed by using the cultivars 'Ben van Zanten' and 'Ile de France' in detail. Pelargonidin 3-rutinoside, cyanidin 3-rutinoside and their acetyl esters were major anthocyanins in the red perianthes while the cyanidin 3-rutinoside and delphinidin 3-rutinoside were major anthocyanins in the dark purple perianth-bottoms. Delphinidin 3-rutinoside and its acetyl esters were present in the dark purple anthers and pollens. Anthocyanins seem to be tissue-specifically bio-synthesized, resulting in formation of a color pattern between inner and outer parts of the tulip flower.
[show abstract][hide abstract] ABSTRACT: Shoot branching and plant height are among the key factors that define the overall architecture of plants. We found that overexpression of a cDNA for a zinc-finger protein of petunia, designated Lateral shoot-Inducing Factor (LIF), in transgenic petunia plants resulted in a dramatic increase in lateral shoots and reduced plant height. LIF overexpression also caused a decrease in the number of cells in the stem, leaf, and flower, accompanied by enlargement of cells. trans-Zeatin was decreased while N6-(Delta2-isopentenyl)adenine was increased in the leaves of LIF-overexpressed petunia. Most of the riboside, ribotide, and glucoside forms were also increased. Expression analysis using a LIF::GUS fusion gene and RT-PCR suggested that LIF is specifically expressed around the bases of axillary buds and weakly in basal part of flowers in wild-type petunia. GFP-LIF-GUS fusion proteins were translocated into the nucleus when transiently expressed in onion epidermal cells. LIF overexpression resulted in enhanced branching also in tobacco and Arabidopsis, indicating the conservation of the response to LIF overexpression among dicotyledonous plants. On the basis of these results we discuss about possible functions of LIF.
The Plant Journal 01/2004; 41(4):512-523. · 6.58 Impact Factor
[show abstract][hide abstract] ABSTRACT: To understand the genetic background of two floral anthocyanin pigmentation traits, anthocyanin pigmentation in the flower tepals and spot formation, in the Asiatic hybrid lily (2n = 24), segregation of the two traits among 96 F(1) plants derived from a cross between commercial cultivars 'Montreux' and 'Connecticut King' were investigated. 'Montreux' has anthocyanin pigmentation in the tepals with many spots, and 'Connecticut King' has flowers with carotenoid pigmentation without spots. The F(1) plants with or without anthocyanin pigment in the tepals segregated with a 1:1 segregation ratio, indicating that a single gene controls anthocyanin pigmentation in the tepals. The number of spots per square centimeter of all tepals showed continuous distribution in the F(1) plants. To map the loci for the two anthocyanin pigmentation traits, molecular linkage maps in the Asiatic hybrid lily were constructed using a double pseudo-testcross strategy, with the same F(1) plants used for phenotypic evaluation, and 212 PCR-based DNA markers. The trait for anthocyanin pigmentation in tepals was used as a trait marker. The map of 'Montreux' comprised 95 markers in 26 linkage groups, and the map of 'Connecticut King' used 119 markers in 24 linkage groups. The total map lengths were 867.5 and 1,114.8 cM, respectively. The trait locus for anthocyanin pigmentation in the tepals was between markers ASR35-180 and P506-40 in linkage group 1 of the 'Montreux' map with a map distance of 1.2 cM and 2.6 cM, respectively. A single-point analysis of quantitative trait loci (QTLs) for tepal spot number identified two putative QTLs in linkage groups 1 and 19 of the 'Connecticut King' map. One putative QTL in linkage group 19 explained 64% of the total phenotypic variation. Because both putative QTLs were mapped on the linkage map of 'Connecticut King' that has no spots, dominant alleles of them might suppress spot formation.
Theoretical and Applied Genetics 01/2003; 105(8):1175-1182. · 3.66 Impact Factor
[show abstract][hide abstract] ABSTRACT: Larvae of the diamondback moth, Plutella xylostella, a crucifer specialist, refuse to feed on a crucifer, Barbarea vulgaris, because of the presence of a feeding deterrent, which is extractable with chloroform. We isolated a feeding deterrent from B. vulgaris leaves, by successive fractionations with silica-gel, ODS, i.e., C18 reversed phase, and Sephadex LH-20 column chromatographies, and ODS-HPLC, guided by a bioassay for feeding deterrent activity. The structure of the compound was determined to be a monodesmosidic triterpenoid saponin, 3-O-[O-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranosyl]-hederagenin, based on FAB-MS, 1H- and 13C-NMR spectra, and hydrolysis experiments. When the compound was applied to cabbage leaf disks at greater than 0.18 microg/mm2, consumption of the disks by third instars was less than 11% of control disks treated with the solvent alone. Furthermore, all first instars died on the disks treated with the same concentrations. Because the concentration of the compound in the fresh leaves of B. vulgaris was comparable to the effective dose in the cabbage leaf disk tested, we conclude that the unacceptability of B. vulgaris to P. xylostella larvae is primarily due to this saponin.
Journal of Chemical Ecology 03/2002; 28(3):587-99. · 2.46 Impact Factor