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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science, Ltd
    Physiologia plantarum 114 (2002), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: 1L-myo-inositol-1-phosphate [Ins(1)P1] synthase (EC 5.5.1.4) catalyses the formation of Ins(1)P1 from glucose-6-phosphate, the first step in the biosynthesis of myo-inositol. Ins(1)P1 is a precursor of phytin (inositol hexakisphosphate), a storage form of phosphate and cations in seeds. Since sucrose and abscisic acid (ABA) are known to affect synthesis of storage compounds in seeds, we investigated the effects of ABA and sucrose on Ins(1)P1 synthase gene (RINO1) expression in cultured cells derived from the scutellum of mature rice seeds. Higher levels of RINO1 transcript accumulation were evident after treatment with either sucrose (10–100 mM) or ABA (10−8M to 10−4M). Glucose was also effective in the upregulation, whereas mannitol was not, suggesting that sucrose and glucose acted as metabolizable sugars and not as osmotica. Treatment with ABA and sucrose together resulted in much higher levels of transcript accumulation, suggesting a synergistic induction of the Ins(1)P1 synthase gene.
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  • 2
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] To the editor: As concerned plant scientists at major plant science research institutions in Japan, we would like to express our collective concern over the impact of Japanese public resistance to plant genetic engineering on the actions of local and national government. We are concerned that ...
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  • 3
    ISSN: 1432-2048
    Subject(s): Key wordsO-Acetyl-l-serine ; Arabidopsis (transgenic) ; β-Conglycinin (β-subunit) ; Glycine (storage protein) ; Storage protein ; Sulfur deficiency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. The composition of seed storage proteins is regulated by sulfur and nitrogen supplies. Under conditions of a low sulfur-to-nitrogen ratio, accumulation of the β-subunit of β-conglycinin, a sulfur-poor seed storage protein of soybean (Glycine max [L.] Merr.), is elevated, whereas that of glycinin, a sulfur-rich storage protein, is reduced. Using transgenic Arabidopsis thaliana [L.] Heynh., it was found that the promoter from the gene encoding the β-subunit of β-conglycinin up-regulates gene expression under sulfur deficiency and down-regulates gene expression under nitrogen deficiency. To obtain an insight into the metabolic control of this regulation, the concentrations of metabolites related to the sulfur assimilation pathway were determined. Among the metabolites, O-acetyl-l-serine (OAS), one of the precursors of cysteine biosynthesis, accumulated to higher levels under low-sulfur and high-nitrogen conditions in siliques of transgenic A. thaliana. The pattern of OAS accumulation in response to various levels of sulfur and nitrogen was similar to that of gene expression driven by the β-subunit promoter. Elevated levels of OAS accumulation were also observed in soybean cotyledons cultured under sulfur deficiency. Moreover, OAS applied to in-vitro cultures of immature soybean cotyledons under normal sulfate conditions resulted in a high accumulation of the β-subunit mRNA and protein, whereas the accumulation of glycinin was reduced. These changes were very similar to the responses observed under conditions of sulfur deficiency. Our results suggest that the level of free OAS mediates sulfur- and nitrogen-regulation of soybean seed storage-protein composition.
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  • 4
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic calli of Pharbitis nil which grow in the presence of kanamycin were obtained by introduction of plasmid pBI121, which carries kanamycin resistance and the gene for beta-glucuronidase. The calli were shown to have fragments of vector DNA in their genome and high levels of beta-glucuronidase activity. This is the first report of the introduction of T-DNA into P. nil and the T-DNA has been shown to be integrated without apparent rearrangement in its genome. The range of copy numbers was between 3 and 5. The beta-glucuronidase activities measured were about 10 times higher than those of transgenic tobacco by introduction of the same plasmid as previously described by Jefferson et al. (1987). Thus, the widely used CaMV 35S promoter also appears to be very active in P. nil.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 9 (1991), S. 602-606 
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The soybean 7S seed storage protein, β-conglycinin, is comprised of three major subunits, α′, α, and β. Chimeric genes having β-conglycinin α′ and β subunit promoters and the β-glucuronidase gene coding sequence were constructed and electroporated into protoplasts prepared from three cultured cell lines and from tobacco mesophyll cells. The β-conglycinin promoters were active in all protoplasts examined, and their activities were 10–60% of that of the cauliflower mosaic virus 35S promoter. In electroporated protoplasts isolated from tobacco suspension cultures the time course of expression and the pattern of cell cycle dependency of the β-conglycinin promoters were similar to those of the 35S promoter. The responses to exogenously added L-methionine and abscisic acid, which are known to have differential effects on the expression of β-conglycinin promoters in cultured soybean cotyledons, were essentially the same among the promoters used. The results indicate that β-conglycinin promoters are expressed in electroporated protoplasts, but their regulation is relaxed.
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  • 6
    ISSN: 1432-2048
    Subject(s): β-Conglycinin ; Glycine (β-conglycinin) ; Nicotiana (transgenic) ; Petunia (transgenic) ; Seed development ; Storage protein (seed) ; Transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Soybean (Glycine max (L.) Merr.) seeds contain the storage protein β-conglycinin, encoded by a multigene family. β-Conglycinin consists of three subunits; α′, α, and β. A genomic clone for a β-subunit of β-conglycinin has been characterized by restriction-enzyme mapping and hybrid selected in-vitro translation followed by immunoprecipitation. In order to determine the developmental regulation of this β-subunit gene, its expression was studied in seeds of transgenic petunia (Petunia hybrida) and tobacco (Nicotiana tabacum L.) plants. The β-subunit expressed in seeds of petunia and tobacco was recognized by anti-β-conglycinin serum at a relative molecular mass of 53 000, equivalent to that of the native protein. Separation of the petunia-seed proteins by isoelectric focusing followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis showed that multiple isoelectric forms of the β-subunit were produced. There was approximately a twofold variation in the accumulation of the β-subunit protein in the mature seeds of transgenic petunia plants, each containing a single β-subunit gene. However, the level of protein accumulation in mature seeds and the amount of β-subunit mRNA in developing seeds was not correlated. Accumulation of the β-subunit protein in transgenic seeds was less than the α′-subunit protein that accumulated in transgenic petunia seeds containing a single α′-subunit gene and less than the amount of the β-subunit in mature soybean seeds which contain 8–13 β-subunit genes. In transgenic tobacco plants, the accumulation of the β-subunit protein in seeds was generally well correlated with the number of genes that were incorporated in the different transformants.
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  • 7
    ISSN: 1572-994X
    Subject(s): tobamoviruses ; crucifer strain ; amino acid sequence comparison ; 130 kD protein ; Arabidopsis thaliana ; plant-virus interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Tobacco mosaic virus (TMV)-Cg is a crucifer-infecting tobamovirus that was isolated from field-grown garlic. We determined the complete nucleotide sequence of the genomic RNA of TMV-Cg. The genomic RNA of TMV-Cg consists of 6303 nucleotides and encodes four large open reading frames, organized basically in the same way as that of other tobamoviruses. The nucleotide and deduced amino acid sequences are very similar to those of the other crucifer-infecting tobamoviruses that have been sequenced so far.
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  • 8
    ISSN: 1573-5028
    Subject(s): alternative oxidase ; Arabidopsis thaliana/ ; cyanide-insensitive respiration ; differential expression ; gene family
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We investigated the copy number of the gene for alternative oxidase (AOX) of Arabidopsis thaliana by amplification by PCR and Southern hybridization. These studies indicated that there are at least four copies of the AOX gene in Arabidopsis. We isolated genomic clones containing individual copies (designated as AOX1a, AOX1b, AOX1c and AOX2) of the AOX genes. Interestingly, two of the AOX genes (AOX1a and AOX1b) were located in tandem in a ca. 5 kb region on one of the chromosomes of Arabidopsis. Comparison between genomic and cDNA sequences of the four AOX genes showed that all AOX genes are divided by three introns and the positions of the introns in AOX1a, AOX1b, AOX1c and AOX2 are the same. We examined whether expression of Arabidopsis AOX genes, like the tobacco AOX1a gene, is enhanced by treatment with antimycin A, an inhibitor of complex III in the mitochondrial respiratory chain. We found that, in young plants, the amount of Arabidopsis AOX1a mRNA was dramatically increased by addition of antimycin A, while the transcription of the other three genes (AOX1b, AOX1c and AOX2) did not respond to antimycin A. Amplification by RT-PCR showed that AOX1a and AOX1c were expressed in all organs examined (flowers and buds, stems, rosette, and roots of 8-week old plants). In contrast, transcripts of AOX1b were detected only in the flowers and buds, and transcripts of AOX2 were detected mainly in stems, rosette and roots. These results suggested that transcriptions of the four genes for alternative oxidase of Arabidopsis are differentially regulated.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 11 (1988), S. 109-123 
    ISSN: 1573-5028
    Subject(s): embryo specific genes ; enhanced expression ; seed storage protein ; soybean ; transgenic petunia ; temporal regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract β-conglycinin or 7S protein, one of the two major storage proteins of soybean (Glycine max), is comprised of three subunits, α′, α and β. Genomic clones encoding the α′ and β subunit genes were ligated together in intermediate plasmids, either in direct orientation (← ←) or in divergent orientation (← →). In the latter instance the transcriptional promoters of the two genes were approximately 2.0 kb apart. The DNAs were introduced into leaf disks of Petunia hybrida on a disarmed Ti plasmid in Agrobacterium tumefaciens and transgenic plants were regenerated. Expression of the introduced genes, limited to specific stages in developing seeds, was analyzed by quantitating the accumulated mRNA and protein, and compared with expression in transgenic plants that contained either gene alone. Development time course studies showed that the α′ subunit mRNA and protein were first detected at 10 days after pollination while accumulation of the β subunit protein ensued at day 14 regardless of the orientation of the two genes. The temporally regulated expression of these two genes in transgenic plants mimics their temporal regulation during soybean embryo development. In transgenic plants that contain the α′ and β genes in divergent orientation, expression of the β gene was enhanced several-fold, while the expression of the α′ gene was slightly decreased. These results suggest that (i) sequences of the α′ gene enhance the level of expression of a nearby storage protein gene and that (ii) the signal(s) operating in the temporal regulation of seed storage protein genes is (are) common to soybean and petunia, and are different from those that govern the final amounts of mRNA and protein.
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  • 10
    ISSN: 1573-5028
    Subject(s): proteinase inhibitor ; RY sequence ; transgenic plants ; transcription regulation ; winged bean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A winged bean Kunitz-type chymotrypsin inhibitor (WCI) is expressed in seeds and tuberous roots. In seeds, the expression of WCI is restricted to the period between the mid- and late-maturation stage. To understand the mechanisms that regulate the expression of WCI genes, we analyzed the promoter activity of the upstream region of the WCI-3b gene, which encodes a major WCI protein, in transgenic tobacco plants. By using a series of constructs with 5′ deletions in the upstream sequences, the region between -882 and -623, relative to the transcription start site, was shown to contain multiple sequences which are responsible for high level expression in mid-maturation stage seeds. However, when this region was fused to the cauliflower mosaic virus 35S core promoter in both orientations, the chimeric promoters showed only a weak transcription activity in transgenic tobacco plants. Further analyses using internal deletion constructs revealed that the region between -882 and -174 is required for the transcription activation. Disruption of the RY sequence at -517, which is conserved in many seed protein genes, resulted in a drastic reduction of the transcription activity in seeds. These results suggest that sequences necessary for high level induction of the WCI-3b gene transcription in developing seeds are dispersed in the region between -882 and -174, and that the RY sequence is one of these sequences.
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