Endogenous gibberellins in immature seeds of Prunus persica L.: identification of GA118, GA119, GA120, GA121, GA122 and GA126

Nakayama, Masayoshi; Koshioka, Masaji; Matsui, Hiroyuki; Ohara, Hitoshi; Mander, Lewis N.; Leitch, Sarah K.; Twitchin, B.; Kraft-Klaunzer, Petra; Pharis, Richard P.; Yokota, Takao

doi:10.1016/s0031-9422(01)00154-6

Cited by 9 publications

(8 citation statements)

References 26 publications

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“…Operating conditions of GC–MS were as previously described (Nakayama et al . ). The carrier gas was helium at a flow rate of 1 ml min −1 , and the interface temperature was 250 °C.…”

Section: Methodsmentioning

confidence: 97%

“…Running conditions for HPLC were similar to those described in Nakayama et al . (), retention times of IAA in HPLC were of 23–24 min. The HPLC fractions were completely dried with N 2 and derivatised by adding ethereal diazomethane then dried with N 2 .…”

Section: Methodsmentioning

confidence: 99%

“…The pellet was re-dissolved in 30% aqueous MeOH containing 0.1% acetic acid (HOAc) and injected into a Beckman System Gold HPLC system with a LiChrosphere RP-18 column (250 9 4 mm diameter, 5-lM particle size; Merck, Darmstadt, Germany). Running conditions for HPLC were similar to those described in Nakayama et al (2001), retention times of IAA in HPLC were of 23-24 min. The HPLC fractions were completely dried with N 2 and derivatised by adding ethereal diazomethane then dried with N 2 .…”

Section: Indole-3-acetic Acid Extraction and Quantificationmentioning

confidence: 99%

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Arabidopsis thaliana transgenics overexpressing IBR3 show enhanced susceptibility to the bacterium Pseudomonas syringae

et al. 2012

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The gene, indole-3-butyric acid (IBA)-RESPONSE (IBR) 3, is thought to participate in peroxisomal β-oxidation of IBA to indole-3-acetic acid. Here we show that IBR3 may also play a role in Arabidopsis thaliana defence response to microbial pathogens. IBR3 is up-regulated during infection by virulent Pseudomonas syringae pv. tomato (Pst) DC3000 bacteria. Although mutant ibr3-4 did not show a pathogen phenotype, lines overexpressing IBR3 demonstrated enhanced susceptibility to Pst DC3000. Increased susceptibility phenotypes of IBR3 overexpressors were correlated with defective SA defence signalling and impairment of pattern-triggered immunity (PTI) activation. Notably, reactive oxygen species production was reduced in IBR3 overexpressors after treatment with the microbe-associated molecular patterns flg22 and efl26. Later PTI responses, such as accumulation of FRK1 transcripts and callose deposition were also reduced in transgenics overexpressing IBR3 after inoculation with the Type III secretion system deficient bacterial mutant Pst DC3000 hrcC or treatment with flg22 or elf26. Importantly, overexpression of IBR3 did not affect indole-3-acetic acid content or auxin-responsive gene expression. These results suggest a novel role for IBR3 in A. thaliana defence response against bacterial pathogens.

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“…Operating conditions of GC–MS were as previously described (Nakayama et al . ). The carrier gas was helium at a flow rate of 1 ml min −1 , and the interface temperature was 250 °C.…”

Section: Methodsmentioning

confidence: 97%

Section: Methodsmentioning

confidence: 99%

Section: Indole-3-acetic Acid Extraction and Quantificationmentioning

confidence: 99%

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Arabidopsis thaliana transgenics overexpressing IBR3 show enhanced susceptibility to the bacterium Pseudomonas syringae

et al. 2012

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“…In contrast, in G. fujikuroi direct 1,2-desaturation by loss of the 1␣, 2␣-H atoms (33) is the major reaction and is accompanied by 2␣-hydroxylation. It is of interest that immature seeds of Prunus persica contain a number of 3-deoxy 1,2-didehydro GAs including GA 120 that are probably formed by direct 1,2-desaturation (34). The seeds also contain 1␣-hydroxy GAs, which could be formed as byproducts of the desaturation.…”

Section: Figmentioning

confidence: 99%

Characterization of the Final Two Genes of the Gibberellin Biosynthesis Gene Cluster of Gibberella fujikuroi

Tudzynski

Mihlan

Rojas

et al. 2003

Journal of Biological Chemistry

106

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Recently, six genes of the gibberellin (GA) biosynthesis gene cluster in Gibberella fujikuroi were cloned and the functions of five of these genes were determined. Here we describe the function of the sixth gene, P450 -3, and the cloning and functional analysis of a seventh gene, orf3, located at the left border of the gene cluster. We have thereby defined the complete GA biosynthesis gene cluster in this fungus. The predicted amino acid sequence of orf3 revealed no close homology to known proteins. High performance liquid chromatography and gas chromatography-mass spectrometry analyses of the culture fluid of knock-out mutants identified GA 1 and GA 4 , rather than GA 3 and GA 7 , as the major C 19 -GA products, suggesting that orf3 encodes the GA 4 1,2-desaturase. This was confirmed by transformation of the SG139 mutant, which lacks the GA biosynthesis gene cluster, with the desaturase gene renamed des. The transformants converted GA 4 to GA 7 , and also metabolized GA 9 (3-deoxyGA 4 ) to GA 120 (1,2-didehydroGA 9 ), but the 2␣-hydroxylated compound GA 40 was the major product in this case. We demonstrate also by gene disruption that P450-3, one of the four cytochrome P450 monooxygenase genes in the GA gene cluster, encodes the 13-hydroxylase, which catalyzes the conversion of GA 7 to GA 3 , in the last step of the pathway. This enzyme also catalyzes the 13-hydroxylation of GA 4 to GA 1 . Disruption of the des gene in an UV-induced P450 -3 mutant produced a double mutant lacking both desaturase and 13-hydroxylase activities that accumulated high amounts of the commercially important GA 4 . The des and P450 -3 genes differ in their regulation by nitrogen metabolite repression. In common with the other five GA biosynthesis genes, expression of the desaturase gene is repressed by high amounts of nitrogen in the culture medium, whereas P450-3 is the only gene in the cluster not repressed by nitrogen.

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“…MS coupled with advanced separation techniques, such as gas chromatography (Taylor et al ., ; Nakayama et al ., ; Jager et al ., ), liquid chromatography (LC) (Cao et al ., ; Gong et al ., ; Simura et al ., ) and capillary electrophoresis (Ge et al ., ; Chen et al ., ) is extensively used for GA qualitative and quantitative analysis. Among these, LC‐electrospray ionization‐tandem mass spectrometry (LC‐ESI‐MS/MS) in the multiple reaction monitoring (MRM) mode is becoming increasingly popular because of its excellent separation capability and high detection selectivity, specificity and sensitivity (Pan and Wang, ).…”

Section: Introductionmentioning

confidence: 99%

One‐pot sample preparation approach for profiling spatial distribution of gibberellins in a single shoot of germinating cereal seeds

Liu

et al. 2019

The Plant Journal

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Summary Sample preparation remains a bottleneck in the rapid and reliable quantification of gibberellins (GAs) for obtaining an insight into the physiological processes mediated by GAs. The challenges arise from not only the extremely low content of GAs in complex plant matrices, but the poor detectability of GAs by mass spectrometry (MS) in negative ion mode. In an effort to solve these urgent difficulties, we present a spatial‐resolved analysis method to investigate the distribution of GAs in tiny plant tissues based on a simplified one‐pot sample preparation approach coupled with ultrahigh‐performance liquid chromatography‐tandem MS. By integrating extraction and derivatization into one step, target GAs were effectively extracted from plant materials and simultaneously reacted with N‐(3‐dimethylaminopropyl)‐N′‐ethylcarbodiimide, the sample preparation time was largely shortened, the probability of sample loss was minimized and the detection sensitivity of MS was also greatly improved compared with underivatized GAs. Under optimal conditions, the method was validated from the quantification linearity, limits of detection and limits of quantification in the presence of plant matrices, recoveries, and precision. With the proposed method, 15 endogenous GAs were detected and, among these, 11 GAs could be quantified in 0.50 mg fresh weight (FW) wheat shoot samples, and five GAs were quantified in only 0.15 mg FW developing seed samples of Arabidopsis thaliana. The distribution patterns of GAs along both the non‐13‐hydroxylation pathway and the early 13‐hydroxylation pathway in a single shoot of germinating wheat, rice and maize seeds were finally profiled with a spatial resolution down to approximately 1 mm2.

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Endogenous gibberellins in immature seeds of Prunus persica L.: identification of GA118, GA119, GA120, GA121, GA122 and GA126

Cited by 9 publications

References 26 publications

Arabidopsis thaliana transgenics overexpressing IBR3 show enhanced susceptibility to the bacterium Pseudomonas syringae

Arabidopsis thaliana transgenics overexpressing IBR3 show enhanced susceptibility to the bacterium Pseudomonas syringae

Characterization of the Final Two Genes of the Gibberellin Biosynthesis Gene Cluster of Gibberella fujikuroi

One‐pot sample preparation approach for profiling spatial distribution of gibberellins in a single shoot of germinating cereal seeds

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