Isolation of gibberellin metabolic pathway genes from barley and comparative mapping in barley, wheat and rice

Spielmeyer, Wolfgang; Ellis, Marc H.; Robertson, Masumi; Ali, Shahjahan; Lenton, John R.; Chandler, Peter M.

doi:10.1007/s00122-004-1689-6

Cited by 82 publications

(86 citation statements)

References 20 publications

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“…The protein of Lp2ox was capable of 2-oxidation of GA 20 as a substrate when transiently expressed in a reticulocyte assay as described by Spielmeyer et al (2004). GA 1 as a substrate was not metabolized, so additional 2-and probably 20-oxidases are likely to be present in Lolium as in other monocots and dicots (Hedden and Phillips, 2000;Spielmeyer et al, 2004); however, to date, we have not succeeded in cloning these genes from a Lolium cDNA library. There can be complete amino acid identity between GAs, FLOWERING LOCUS T, genes from L. perenne and L. temulentum.…”

Section: Ga Analysismentioning

confidence: 99%

Regulation of Flowering in the Long-Day Grass Lolium temulentum by Gibberellins and the FLOWERING LOCUS T Gene

et al. 2006

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Seasonal control of flowering often involves leaf sensing of daylength coupled to time measurement and generation and transport of florigenic signals to the shoot apex. We show that transmitted signals in the grass Lolium temulentum may include gibberellins (GAs) and the FLOWERING LOCUS T (FT) gene. Within 2 h of starting a florally inductive long day (LD), expression of a 20-oxidase GA biosynthetic gene increases in the leaf; its product, GA20, then increases 5.7-fold versus short day; its substrate, GA19, decreases equivalently; and a bioactive product, GA5, increases 4-fold. A link between flowering, LD, GAs, and GA biosynthesis is shown in three ways: (1) applied GA19 became florigenic on exposure to LD; (2) expression of LtGA20ox1, an important GA biosynthetic gene, increased in a florally effective LD involving incandescent lamps, but not with noninductive fluorescent lamps; and (3) paclobutrazol, an inhibitor of an early step of GA biosynthesis, blocked flowering, but only if applied before the LD. Expression studies of a 2-oxidase catabolic gene showed no changes favoring a GA increase. Thus, the early LD increase in leaf GA5 biosynthesis, coupled with subsequent doubling in GA5 content at the shoot apex, provides a substantial trail of evidence for GA5 as a LD florigen. LD signaling may also involve transport of FT mRNA or protein because expression of LtFT and LtCONSTANS increased rapidly, substantially (>80-fold for FT), and independently of GA. However, because a LD from fluorescent lamps induced LtFT expression but not flowering, the nature of the light response of FT requires clarification.

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Section: Ga Analysismentioning

confidence: 99%

Regulation of Flowering in the Long-Day Grass Lolium temulentum by Gibberellins and the FLOWERING LOCUS T Gene

et al. 2006

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“…The productivity of semidwarf plant also increased because assimilates translocation is more directed to develop seeds rather than to develop new leaves or buds (Evans 1993). Dwarfisme can be due to either deficiency in gibberellin biosynthesis or inhibition in gibberellin signalling (Spielmeyer et al 2004). Application of paclobutrazol to create semidwarf plants was reported in rice subspecies "Keng", in which 22.5 g/mL paclobutrazol reduced stem height and increased tiller numbers (Minzi 1993).…”

Section: Introductionmentioning

confidence: 99%

EFFECTS OF BLUE LIGHT AND PACLOBUTRAZOL ON SEED GERMINATION, VEGETATIVE GROWTH AND YIELD OF BLACK RICE (Oryza sativa L. 'Cempo Ireng')

Dewi

Agustina²,

Nurmalika³

2017

BIOTROPIA

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Black rice (Oryza sativa L. "Cempo Ireng") is one of local rice varieties in Sleman Regency, Yogyakarta. The black color is caused by high anthocyanin content which is important source of antioxidant. The cultivation of black rice is still limited due to its tall phenotype, long vegetative stage and low productivity compared to white rice. Paclobutrazol is a growth retardant causing dwarfing in several crop plants and reducing lodging. Blue light can improve plant quality. This research was aimed to evaluate the effect of blue light and paclobutrazol on seed germination, vegetative growth and yield of black rice. The results showed that the average of seed germination as well as the activity of α-amylase of seeds subjected to blue light were lower compared to those subjected to sunlight; however, paclobutrazol concentrations did not affect seed germination percentage. The height of rice plants treated with paclobutrazol decreased in accordance with the increase of paclobutrazol concentration. Chlorophyll content and tiller numbers increased by paclobutrazol treatment of 12.5 ppm. Nitrate reductase activity was higher in rice seedlings subjected to blue light compared to those subjected to sunlight. Iron (Fe) content of rice plants treated with 25 or 50 ppm paclobutrazol increased compared to control. It was concluded that paclobutrazol application of 12.5 ppm already reduced plant height. The higher concentration of paclobutrazol applied the greater reduction on plant height was observed. Blue light treatment applied during black rice seed germination slightly reduced germination percentage and α-amylase activity in the germinated seeds. However, blue light treatment combined with paclobutrazol application during black rice seed germination increased chlorophyll content, tiller numbers and Fe content in black rice grain.

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“…Typical feedback regulation of HvGA3ox expression was not observed in the presence of active GA, GA 3 , and the GA biosynthesis inhibitor, uniconazol ( Figure 3B), a result similar to AtGA3ox2 regulation (Yamaguchi et al 2001), and in contrast to the regulation seen with OsGA3ox2 (Itoh et al 2001). These results indicate that there might be some functional differences between HvGA3ox2 and OsGA3ox2 in spite of amino acid sequence similarity and chromosomal synteny (Spielmeyer et al 2004).Base on the present results, it must be concluded that HvGA3ox2 regulates and stimulates germination through the production of active GAs that induce hydrolytic enzymes including a-amylase in barley seeds. The expression pattern of HvGA3ox2 is similar with AtGA3ox2, the Arabidopsis GA 3-oxidase gene that is expressed specifically during germination (Yamaguchi et al , 2001.…”

mentioning

confidence: 53%

Section: Discussionmentioning

confidence: 85%

Specific expression of the gibberellin 3.BETA.-hydroxylase gene, HvGA3ox2, in the epithelium is important for Amy1 expression in germinating barley seeds

Suzuki

Ishiyama

Kobayashi

et al. 2005

Plant Biotechnology

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We have cloned gibberellin (GA) 3b-hydroxylase cDNA, (HvGA3ox), a homolog of the rice gene OsGA3ox from germinating barley seeds. HvGA3ox cDNA was expressed in Escherichia coli as a fusion protein capable of oxidizing GA 9 to GA 4 and GA 20 to GA 1 . Deduced amino acid sequence analysis showed that HvGA3ox was identical to HvGA3ox2, which shares a high homology with the OsGA3ox2 isoform that plays an important role for induction of a-amylase in germinating rice seeds. Northern blot analysis showed that HvGA3ox2 gene was expressed only in germinating seeds, not in other organs. Typical feedback regulation was not observed in seedling treated with either biologically active GA (GA 3 ) or uniconazole, an inhibitor of GA biosynthesis. HvGA3ox2 mRNA was detected from 12 h after imbibition just before expression of a-amylase gene, Amy1. In situ hybridization of germinating seeds revealed that HvGA3ox2 mRNA was localized in the epithelium up to at least 3 days after imbibition. Our results suggest that GA biosynthesis in epithelium is important for expression of a-amylase in germinating barley seeds, and that HvGA3ox2 encodes the key enzyme of this event.Key words: a-amylase, epithelium, gibberellin, Hordeum vulgare. Original Paper Copyright © 2005 The Japanese Society for Plant Cell and Molecular BiologyAbbreviations: DIG, dioxigenin; PBS, phosphate-buffered saline; RACE, rapid amplification of cDNA end. The nucleotide sequence reported in this paper has been submitted to DDBJ/GenBank/EMBL nucleotide sequence database under the accession number AB189152. a TEPresent address: Research Institute for Bioresources, Okayama University, Kurashiki, Okayama 710-0046, Japan 1998; Yamaguchi et al. 1998;Yamaguchi et al. 2001;Itoh et al. 2001). Itoh et al. (2001) cloned and analyzed two GA3ox genes (OsGA3ox1 and OsGA3ox2) from rice. Their study indicated that OsGA3ox1 was preferentially expressed in unopened flower and OsGA3ox2 in elongating leaves. Kaneko et al. (2002) reported that OsGA3ox2 production in epithelium cells was important for a-amylase induction in early stages of germinating rice seeds. On the other hand, little is known about the molecular mechanism of GA biosynthesis in germinating barley seeds. Recently, Spielmeyer et al. (2004) reported isolation and chromosome mapping of two genes encoding GA 3b-hydroxylase, HvGA3ox1 and HvGA3ox2, from barley. However, there is no information about the expression pattern of these genes, and their role in a-amylase expression during germination remains to be clarified. The research described here predicts that HvGA3ox2 plays an important role in GA biosynthesis for the induction of hydrolytic enzymes, including a-amylase, in germinating barley seeds. Materials and methods Plant materialsBarley seeds (Hordeum vulgare cv. Harrington) were surface-sterilized in 1% NaClO for 10 min and rinsed in distilled water. The seeds were imbibed in distilled water for 8 h, plated on petri dishes with wetted filter paper, and grown in a culture chamber at 20°C in the dark. Seedling s...

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Isolation of gibberellin metabolic pathway genes from barley and comparative mapping in barley, wheat and rice

Cited by 82 publications

References 20 publications

Regulation of Flowering in the Long-Day Grass Lolium temulentum by Gibberellins and the FLOWERING LOCUS T Gene

Regulation of Flowering in the Long-Day Grass Lolium temulentum by Gibberellins and the FLOWERING LOCUS T Gene

EFFECTS OF BLUE LIGHT AND PACLOBUTRAZOL ON SEED GERMINATION, VEGETATIVE GROWTH AND YIELD OF BLACK RICE (Oryza sativa L. 'Cempo Ireng')

Specific expression of the gibberellin 3.BETA.-hydroxylase gene, HvGA3ox2, in the epithelium is important for Amy1 expression in germinating barley seeds

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