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1.
Synergistic Function of rolB, rolC, ORF13 and ORF14 of TL-DNA of Agrobacterium rhizogenes in Hairy Root Induction in Nicotiana tabacum 总被引:1,自引:0,他引:1
Comparison of the frequency of rooting in the tobacco leaf segmentsinoculated with Agrobacterium tumefaciens harboring variouscombinations of rolB, rolC, ORF13 and ORF14 of TL-DNA of Riplasmid (pRiHRI) revealed that the genes differ in their functionto stimulate adventitious root induction. A single gene rolBinduced roots, while rolC, ORF13 and ORF14 independently promotedthe root induction by the rolB gene. The effects of these geneson the rolB-mediated rooting were in the order of ORF13>rolCORF14.
Present address: Laboratory of Phylogenetic Botany, Departmentof Biology, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba,263-8522 Japan.
2 Present address: Department of Chemical and Biological Sciences,Faculty of Science, Japan Women's University, 2-8-1 Mejirodai,Bunkyo-ku, Tokyo, 112-8681 Japan. 相似文献
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The protein encoded by the rolB plant oncogene hydrolyses indole glucosides. 总被引:8,自引:0,他引:8
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The rolB gene of Agrobacterium rhizogenes, whose expression stimulates the formation of roots by transformed plant tissues and other growth alterations in transgenic plants, codes for a beta-glucosidase able to hydrolyse indole-beta-glucosides. Indeed, we show that extracts of bacteria and/or plant tissue expressing the rolB protein hydrolyse indoxyl-beta-glucoside (plant indican). Because of the structural similarity between indoxyl-beta-glucoside and indole-3-acetyl-beta-glucoside (IAA-beta-glucoside), we propose that the physiological and developmental alterations in transgenic plants expressing the rolB gene could be the result of an increased intracellular auxin activity caused by the release of active auxins from inactive beta-glucosides. Thus two of the oncogenes carried by the T-DNA of the plant pathogen Agrobacterium rhizogenes (rolB and rolC) perturb plant growth and development by coding for beta-glucosidases with distinct specificities. Whereas the rolC beta-glucosidase releases cytokinins from their glucoside conjugates, the rolB encoded protein hydrolyses indole-beta-glucosides. The combined action of these two genes therefore is expected to modulate the intracellular concentration of two of the main growth factors active in plants. 相似文献
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Summary Growth characteristics of tobacco protoplasts containing rolA linked to its own promoter, or the rolB, or rolC genes of Agrobacterium rhizogenes linked to the Cauliflower Mosaic Virus 35S RNA promoter were compared with those from untransformed plants. RolA protoplasts require auxin and cytokinin for callus formation. Protoplasts overexpressing rolB and C form callus in the absence of exogenously applied auxin and cytokinin, respectively. Long term callus growth requires auxin, but the requirement for cytokinin is not critical. Optimal transient expression of an auxin responsive promoter element occurred at lower external levels of auxin in rolB and rolC protoplasts compared with untransformed protoplasts. Addition of putrescine was required for auxin responsive transient gene expression in rolA protoplasts suggesting that polyamines, or their products affect gene expression in rolA plants.Abbreviations T-DNA
transferred DNA
- TL-DNA
left transferred DNA
- NAA
naphthalene acetic acid
- PEG
polyethylene glycol
- GUS
glucuronidase
- CaMV
cauliflower mosaic virus 相似文献
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Different promoter regions control level and tissue specificity of expression of Agrobacterium rhizogenes rolB gene in plants 总被引:6,自引:0,他引:6
I. Capone M. Cardarelli D. Mariotti M. Pomponi A. De Paolis P. Costantino 《Plant molecular biology》1991,16(3):427-436
Expression of the rolB gene of A. rhizogenes T-DNA triggers root differentiation in transformed plant cells. In order to study the regulation of this morphogenetic gene, the GUS reporter gene was placed under the control of several deleted fragments of the rolB 5 non-coding region: carrot disc transformations and the analysis of transgenic tobacco plants containing these constructions identified the presence of distinct regulatory domains in the rolB promoter. Two regions (located from positions –623 to –471 and from –471 to –341, from the translation start codon) control the level but not the tissue specificity of rolB expression: progressive deletions of the rolB promoter starting from position –1185 to –341, although at different levels, maintained the same pattern of GUS expression — maximal in root meristems and less pronounced in the vascular tissue of aerial organs. Further deletion of 35 bp, from –341 to –306, drastically affected tissue specificity: GUS activity was still clearly detectable in the vascular tissue of the aerial organs while expression in the root meristem was totally suppressed. Analysis of transgenic embryos and seedlings confirmed that distinct promoter domains are responsible for meristematic (root) and differentiated (vascular) expression of rolB. Finally, we present data concerning the effects of plant hormones on the expression of rolB-GUS constructions. 相似文献
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Indole-3-acetic acid homeostasis in transgenic tobacco plants expressing the Agrobacterium rhizogenes rolB gene 总被引:5,自引:1,他引:4
Ove Nilsson Alan Crozier Thomas Schmülling Göran Sandberg Olof Olsson 《The Plant journal : for cell and molecular biology》1993,3(5):681-689
The rolB gene of the plant pathogen Agrobacterium rhizogenes has an important role in the establishment of hairy root disease in infected plant tissues. When expressed as a single gene in transgenic plants the RolB protein gives rise to effects indicative of increased auxin activity. It has been reported that the RolB product is a β-glucosidase and proposed that the physiological and developmental alterations in transgenic plants expressing the rolB gene are the result of this enzyme hydrolysing bound auxins, in particular (indole-3-acetyl)-β-D-glucoside (IAGluc), and thereby bringing about an increase in the intracellular concentration of indole-3-acetic acid (IAA). Using tobacco plants as a test system, this proposal has been investigated in detail. Comparisons have been made between the RolB phenotype and that of IaaM/iaaH transformed plants overproducing IAA. In addition, the levels of IAA and IAA amide and IAA ester conjugates were determined in wild-type and transgenic 35S-rolB tobacco plants and metabolic studies were carried out with [13C6]IAA [2′-14C]IAA, [14C]IAGluc, [5-3H]-2-o-(indole-3-acetyl)-myo-inositol and [14C]indole-3-acetylaspartic acid. The data obtained demonstrate that expression of the rolB encoded protein in transgenic tobacco does not produce a phenotype that resembles that of IAA over producing plants, does not alter the size of the free IAA pool, has no significant effect on the rate of IAA metabolism, and, by implication, appears not to influence the overall rate of IAA biosynthesis. Furthermore, the in vivo hydrolysis of IAGluc, and that of the other IAA conjugates that were tested, is not affected. On the basis of these findings, it is concluded that the RolB phenotype is not the consequence of an increase in the size of the free IAA pool mediated by an enhanced rate of hydrolysis of IAA conjugates. 相似文献
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Lise Jouanin David Bouchez Roger F. Drong David Tepfer Jerry L. Slightom 《Plant molecular biology》1989,12(1):75-85
A Charon 4A phage library, containing insert DNA isolated from a morning glory (Convolvulus arvensis) plant genetically transformed by Ri T-DNA from Agrobacterium rhizogenes strain A4, was used to isolate a lambda clone that contains part of the Ri TL-DNA and the complete TR-DNA. The two Ri T-DNAs were recovered adjacent to each other in a tail-to-tail configuration (i.e. with the TR-DNA inverted with respect to the TL-DNA). Comparison of nucleotide sequences from this lambda clone with the corresponding sequences from the Ri plasmid allowed us to determine the location of the T-DNA/plant junction for the right end of the TL-DNA and the left and right ends of the TR-DNA. We located, near each of these borders, a 24 bp sequence that is similar to the 24 bp consensus sequence found near the pTi T-DNA extremities. In addition, sequences similar to the core overdrive sequence from pTi are located near each right border. Hybridization and nucleotide sequence analysis of the DNA adjacent to the TL/TR junction shows that no plant DNA is located between the TL and TR-DNAs and suggests that the plant DNA adjacent to the end of the TR-DNA may have been rearranged during the integration into the plant genome. 相似文献
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Filippini Francesco; Schiavo Fiorella Lo; Terzi Mario; Costantino Paolo; Trovato Maurizio 《Plant & cell physiology》1994,35(5):767-771
We measured auxin-binding capacity of the membrane preparationsfrom tobacco cells transformed by rolB as compared to untransformedcontrols. In the transformed cells, the overall auxin-bindingactivity is severalfold enhanced through an increase in a bindingactivity removable from the membranes at 0.5 M salt, while thebinding activity still attached to the membranes after saltwashes remains unchanged. Antibodies against the 22 kDa maizeauxin binding protein (ABP) depress most of the membrane-attachedbinding activity in both normal and rolB-transformed cells,while they do not affect the salt-washable binding activity.In contrast, antibodies against the RolB protein prevent completelybinding of auxin to the latter activity in both normal and transformedcells, while substantially unaffecting the membrane-associatedbinding. These results point to the presence, in untransformedmembranes, of an auxin-binding activity associated with a proteinimmunologically related to RolB. This activity is much increasedin rolB cells. In contrast, the auxin-binding protein analogousto maize ABP present in tobacco membranes does not increasein the rolB-transformed cells. (Received October 1, 1993; Accepted April 22, 1993) 相似文献
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L. Jouanin P. Guerche N. Pamboukdjian C. Tourneur F. Casse Delbart J. Tourneur 《Molecular & general genetics : MGG》1987,206(3):387-392
Summary The structure of the T-DNA in Ri-transformed plants of Brassica napus, Nicotiana plumbaginifolia and Nicotiana tabacum was analysed. All the plants studied present a particular phenotype with wrinkled leaves. The T-DNA is composed of two parts: TL and TR. The size of the TL-DNA (19–20 kb) seems to be almost constant, except in N. tabacum where it is shorter. The TR-DNA can be absent, and its size varies from about 5–28 kb, with two predominant lengths. The smaller size does not include the region homologous to the tms genes of the pTi T-DNA. The copy number varies from one to four copies per plant genome. TL and TR-DNA are not always present in the same copy number, but in some cases are linked together. 相似文献
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Frank T. Röder Thomas Schmülling Christiane Gatz 《Molecular genetics and genomics : MGG》1994,243(1):32-38
We have investigated the use of the tetracycline-dependent gene expression system to regenerate and propagate tobacco plants transformed with a gene whose product — when highly expressed — interferes with regeneration and/or further reproduction. Plants transformed with the Agrobacterium rhizogenes rolB gene under the control of the tetracycline-dependent expression system were phenotypically indistinguishable from wild type owing to efficient repression of the promoter. Induction of the rolB gene with tetracycline led to high-level expression of the rolB mRNA, which resulted in extremely stunted plants with necrotic and wrinkled leaves that did not develop a floral meristem. Upon cessation of tetracycline treatment healthy shoots developed even from severely affected meristems. Data on the dose response of the rolB phenotype as a function of tetracycline concentration demonstrate that the tetracycline-dependent gene expression system can be used to modulate the manifestation of a particular phenotype. 相似文献
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MCINNES E.; DAVEY M. R.; MULLIGAN B. J.; DAVIES K.; SARGENT A. W.; J.MORGAN A. 《Journal of experimental botany》1989,40(10):1135-1144
Nicotiana glauca, N. tabacum, Solanian dulcamara and S. nigrumwere transformed by Agrobacteriun rhizogenes strain BN1010 (TLTR+).The TR-DNA stimulated agropine-positive root induction and wastransformation competent in the absence of the TL-DNA. An unusualpattern of root induction was seen when stem explants were inoculatedwith this strain; occasionally, agropine-positive roots wereinduced at the inoculation sites, but prolific agropine-negativeroots were formed in profusion down the stems. The utility ofBN1010 as an efficient co-integrating vector was demonstratedby the separate transfer of a fragment containing rol ABC (BN1010::pEM15) and of a chimeric nopaline synthase-kanamycin resistancegene (BN1010:: Neo) into plants. Root cultures of S. dulcamaratransformed with BN1010:: Neo had an unusual, positively geotropicphenotype. Strain BN1010:: pEM15 (rol ABC+DTR+) incitedmore roots down stem explants than strain A4T. This indicatesthat rol D may act to suppress agropine-negative root productionin N. glauca and N. tabacum. Key words: Agrobacterium rhizogenes, TL-DNA, TR-DNA, disarmed Ri vector, transformed roots, Nicotiana glauca, N. tabacun, Solatium dulcamara, S. nigrum 相似文献
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Waheed Arshad Ihsan-ul- Haq Mohammad Tahir Waheed Kirankumar S. Mysore Bushra Mirza 《PloS one》2014,9(5)
Tomato (Solanum lycopersicum L.) is the second most important cultivated crop next to potato, worldwide. Tomato serves as an important source of antioxidants in human diet. Alternaria solani and Fusarium oxysporum cause early blight and vascular wilt of tomato, respectively, resulting in severe crop losses. The foremost objective of the present study was to generate transgenic tomato plants with rolB gene and evaluate its effect on plant morphology, nutritional contents, yield and resistance against fungal infection. Tomato cv. Rio Grande was transformed via Agrobacterium tumefaciens harbouring rolB gene of Agrobacterium rhizogenes. rolB. Biochemical analyses showed considerable improvement in nutritional quality of transgenic tomato fruits as indicated by 62% increase in lycopene content, 225% in ascorbic acid content, 58% in total phenolics and 26% in free radical scavenging activity. Furthermore, rolB gene significantly improved the defence response of leaves of transgenic plants against two pathogenic fungal strains A. solani and F. oxysporum. Contrarily, transformed plants exhibited altered morphology and reduced fruit yield. In conclusion, rolB gene from A. rhizogenes can be used to generate transgenic tomato with increased nutritional contents of fruits as well as improved foliar tolerance against fungal pathogens. 相似文献
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Tatiana Y. Gorpenchenko Dmitry L. Aminin Yuliya V. Vereshchagina Yuri N. Shkryl Galina N. Veremeichik Galina K. Tchernoded Victor P. Bulgakov 《Plant signaling & behavior》2012,7(9):1058-1061
The rolB oncogene was previously identified as an important player in ROS metabolism in transformed plant cells. Numerous reports indicate a crucial role for animal oncogenes in apoptotic cell death. Whether plant oncogenes such as rolB can induce programmed cell death (PCD) in transformed plant cells is of particular importance. In this investigation, we used a single-cell assay based on confocal microscopy and fluorescent dyes capable of discriminating between apoptotic and necrotic cells. Our results indicate that the expression of rolB in plant cells was sufficient to decrease the proportion of apoptotic cells in steady-state conditions and diminish the rate of apoptotic cells during induced PCD. These data suggest that plant oncogenes, like animal oncogenes, may be involved in the processes mediating PCD. 相似文献
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Brian H. Taylor Richard M. Amasino Frank F. White Eugene W. Nester Milton P. Gordon 《Molecular & general genetics : MGG》1985,201(3):554-557
Summary Plants regenerated from hairy root tumors induced on Nicotiana glauca and Nicotiana tabacum by Agrobacterium rhizogenes strain A4 were examined for the presence of T-DNA. Regenerated N. tabacum plants contained intact copies of both TL-DNA and TR-DNA. However, plants regenerated from N. glauca tumors did not contain the TR-DNA region corresponding to the tms (auxin synthesis) genes. Some of the regenerants exhibited an abnormal phenotype which is characterized by severe leaf wrinkling. This phenotype is correlated with the presence of TL-DNA, but not TR-DNA. 相似文献
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The objective was to determine whether the protein of
rolB affects shoot formation and whether this
potential relationship depends on the developmental stages of the plant
and/or on the culture conditions. Thin cell layers (TCL) and leaf explants
were excised from tobacco plants in the vegetative and flowering stages and
cultured under various hormonal conditions. In TCLs of vegetative-stage
plants, the expression of rolB enhanced the formation
of the shoot buds under hormone-free conditions and with specific
concentrations of auxin and/or cytokinin. Histological examination showed
that the induction of the shoot meristemoids was particularly enhanced by
rolB protein and that meristemoid growth was
accelerated. In leaf explants from vegetative-stage plants, the expression
of rolB increased the formation of shoot buds in the
presence of 1 M IAA plus 1 or 10 M cytokinin. With BA alone, at a 0.1 M concentration, shoot formation occurred in the
transgenic explants only, whereas with concentrations ranging from 0.5 to
10 M, it was higher in these explants than in
controls.RolB protein enhanced the formation of shoot
buds in TCLs from flowering plants under all hormonal conditions. In the
presence of 1 M IAA and kinetin, the protein also increased the
flowering response. In leaf explants from flowering plants, the expression
of rolB increased the number of shoot buds in the
presence of 1 M IAA with 10 M BA.In conclusion,
rolB protein promotes shoot formation; it seems to
have a positive interaction with cytokinin and an effect on the induction
of the meristematic condition. 相似文献