scholarly journals Tumor Formation and β-Glucuronidase Expression in Phaseolus vulgaris Inoculated with Agrobacterium tumefaciens

1994 ◽  
Vol 119 (2) ◽  
pp. 361-366 ◽  
Author(s):  
Mark E. Lewis ◽  
Fred A. Bliss
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Phaseolus Vulgaris ◽  
Tumor Formation ◽  
Gus Activity ◽  
Shoot Tips ◽  
High Rate ◽  
Naphthaleneacetic Acid ◽  
Genotypic Differences ◽  
Germinating Seeds ◽  
Gus Assays

Ten common bean (Phaseolus vulgaris L.) lines—including cultivars, breeding lines, and one wild line—were evaluated for susceptibility to Agrobacterium tumefaciens strain C58 by stab-inoculating intact shoot tips of germinating seeds. Significant differences for tumor frequency and size were found on the resulting 3-week-old seedlings. UW 325, a wild bean, had the highest rate of tumorigenesis; `Olathe', a dry bean cultivar, had the lowest. Uninoculated excised shoot tips cultured in media with BA or BA plus NAA exhibited differences in phytohormone sensitivity, as evidenced by callusing and root initiation. The cultivar Montcalm seemed to be highly sensitive, while `Olathe' was relatively insensitive. Fluorometric GUS assays of shoot tips from germinating seeds inoculated with the disarmed GUS-containing A. tumefaciens strain C58C1(pGV3850/pKIWI105) showed that UW 325 had the highest level of GUS activity. `Montcalm' had a high rate of tumorigenesis but a low level of GUS activity; this anomaly was attributed to its high phytohormone sensitivity. The use of the virulence-inducing compound acetosyringone in the inoculum culture medium did not alter genotypic differences (ranks) in susceptibility. Histochemical GUS assays of inoculated UW 325 shoot tips showed that 60% of the apexes exhibited one or more transformation events. Chemical names used: β-glucuronidase (GUS); α-naphthaleneacetic acid (NAA); N-(phenylmethyl)-1H-purin-6-amine (BA).

scholarly journals Induction of In Vitro Cultured Masses of Shoot Primordia of Hybrid Statice and Its Cryopreservation by Vitrification

HortScience ◽  
1997 ◽  
Vol 32 (2) ◽  
pp. 309-311 ◽  
Author(s):  
Toshikazu Matsumoto ◽  
Yoji Nako ◽  
Chiaki Takahashi ◽  
Akira Sakai
Keyword(s):  
Large Scale ◽  
Shoot Tips ◽  
High Rate ◽  
Naphthaleneacetic Acid ◽  
Scale Production ◽  
Ms Medium ◽  
Shoot Primordia ◽  
Large Scale Production

Bulbous structures consisting of meristematic clumps (designated “shoot primordia”) were induced from a meristematic culture of a hybrid statice (Limonium altaica Mill. × L. caspium Mill., cv. Blue Symphonet). The shoot tips were cultured in 25 mL of liquid 1/2 Murashige & Skoog (MS) medium supplemented with 0.44 μm BA and 0.054 μm NAA and 3% sucrose at pH 5.8 by vertically shaking at 2 rpm on rotating stages (1 m in diameter) at 25 °C. One month after inoculation of shoot tips, numerous small globular structures were formed and propagated vegetatively at a high rate following subculture. Segments of shoot primordia had developed into plantlets 2 weeks after transfer to solidified 1/2 MS medium supplemented with 0.44 μm BA and 1% sucrose. Plantlets successfully acclimated and grew into normal plants in a greenhouse. Cold-hardened, precultured small segments of shoot primordia were successfully cryopreserved in liquid N by vitrification. Vitrified and warmed segments plated on solidified 1/2 MS medium produced shoots about 21 d after plating. Cultured masses of shoot primordia appear promising for large-scale production and cryopreservation of annual and biennial statice. Chemical names used: 6-benzyladenine (BA); 1-naphthaleneacetic acid (NAA).

scholarly journals Plant regeneration and transformation of Trachyspermum ammi using Agrobacterium tumefaciens and zygotic embryos

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Masoumeh Nomani ◽  
Masoud Tohidfar
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Plant Regeneration ◽  
Genetic Transformation ◽  
Zygotic Embryo ◽  
Plant Biotechnology ◽  
Hplc Method ◽  
Zygotic Embryos ◽  
Naphthaleneacetic Acid ◽  
Transformed Plants ◽  
Trachyspermum Ammi

Abstract Background Trachyspermum ammi is one of the key medicinal plant species with many beneficial properties. Thymol is the most important substance in the essential oil of this plant. Thymol is a natural monoterpene phenol with high anti-microbial, anti-bacterial, and anti-oxidant properties. Thymol in the latest research has a significant impact on slowing the progression of cancer cells in human. In this research, embryos were employed as convenient explants for the fast and effectual regeneration and transformation of T. ammi. To regenerate this plant, Murashige and Skoog (MS) and Gamborg's B5 (B5) media were supplemented with diverse concentrations of plant growth regulators, such as 6-benzyladenine (BA), 1-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), and kinetin (kin). Transgenic Trachyspermum ammi plants were also obtained using Agrobacterium-mediated transformation and zygotic embryos explants. Moreover, two Agrobacterium tumefaciens strains (EHA101 and LBA4404) harboring pBI121-TPS2 were utilized for genetic transformation to Trachyspermum ammi. Results According to the obtained results, the highest plant-regeneration frequency was obtained with B5 medium supplemented with 0.5 mg/l BA and 1 mg/l NAA. The integrated gene was also approved using the PCR reaction and the Southern blot method. Results also showed that the EHA101 strain outperformed another strain in inoculation time (30 s) and co-cultivation period (1 day) (transformation efficiency 19.29%). Furthermore, HPLC method demonstrated that the transformed plants contained a higher thymol level than non-transformed plants. Conclusions In this research, a fast protocol was introduced for the regeneration and transformation of Trachyspermum ammi, using zygotic embryo explants in 25–35 days. Our findings confirmed the increase in the thymol in the aerial part of Trachyspermum ammi. We further presented an efficacious technique for enhancing thymol content in Trachyspermum ammi using Agrobacterium-mediated plant transformation system that can be beneficial in genetic transformation and other plant biotechnology techniques.

scholarly journals The Presence and Characterization of a virF Gene on Agrobacterium vitis Ti Plasmids

1998 ◽  
Vol 11 (5) ◽  
pp. 429-433 ◽  
Author(s):  
B. Schrammeijer ◽  
J. Hemelaar ◽  
P. J. J. Hooykaas
Keyword(s):  
Amino Acids ◽  
Agrobacterium Tumefaciens ◽  
Molecular Mass ◽  
Promoter Region ◽  
Consensus Sequence ◽  
Tumor Formation ◽  
Nicotiana Glauca ◽  
Agrobacterium Vitis ◽  
Ti Plasmids

Octopine and nopaline strains of Agrobacterium tumefaciens differ in their ability to induce tumors on Nicotiana glauca. The presence of a virF locus on the octopine Ti plasmid makes N. glauca a host plant for these strains, indicating that the VirF protein is a host-range determinant. Here we show the presence of a virF locus not only on the Agrobacterium vitis octopine/cucumopine plasmids pTiAg57 and pTiTm4, but also on the nopaline Ti plas-mids pTiAT1, pTiAT66a, and pTiAT66b. On the octopine Ti plasmids from A. tumefaciens the virF gene is located between the virE locus and the left border of the T-region. In contrast, the virF gene on Ti plasmids of A. vitis is located at the very left end of the vir-region near the virA locus. The virF gene of pTiAg57 has been sequenced and codes for a protein of 202 amino acids with a molecular mass of 22,280 Da. Comparison showed that the virF gene from A. vitis strain Ag57 is almost identical to that from A. tumefaciens octopine strains. The transcription of the pTiAg57 virF is inducible by the plant phenolic compound acetosyringone through the presence of a vir-box consensus sequence in its promoter region. The VirF protein from pTiAg57 can complement octopine A. tumefaciens strains deleted for virF as shown by tumor formation on N. glauca.

scholarly journals Of Mice, Dogs, Pigs, and Men: Choosing the Appropriate Model for Immuno-Oncology Research

ILAR Journal ◽  
2018 ◽  
Vol 59 (3) ◽  
pp. 247-262 ◽  
Author(s):  
Nana H Overgaard ◽  
Timothy M Fan ◽  
Kyle M Schachtschneider ◽  
Daniel R Principe ◽  
Lawrence B Schook ◽  
...  
Keyword(s):  
Immune System ◽  
Immune Cell ◽  
Tumor Formation ◽  
Genetically Engineered ◽  
Model Systems ◽  
High Rate ◽  
Host Immune System ◽  
Cancer Immunoediting ◽  
Oncology Research ◽  
Experimental Cancer

Abstract The immune system plays dual roles in response to cancer. The host immune system protects against tumor formation via immunosurveillance; however, recognition of the tumor by immune cells also induces sculpting mechanisms leading to a Darwinian selection of tumor cell variants with reduced immunogenicity. Cancer immunoediting is the concept used to describe the complex interplay between tumor cells and the immune system. This concept, commonly referred to as the three E’s, is encompassed by 3 distinct phases of elimination, equilibrium, and escape. Despite impressive results in the clinic, cancer immunotherapy still has room for improvement as many patients remain unresponsive to therapy. Moreover, many of the preclinical results obtained in the widely used mouse models of cancer are lost in translation to human patients. To improve the success rate of immuno-oncology research and preclinical testing of immune-based anticancer therapies, using alternative animal models more closely related to humans is a promising approach. Here, we describe 2 of the major alternative model systems: canine (spontaneous) and porcine (experimental) cancer models. Although dogs display a high rate of spontaneous tumor formation, an increased number of genetically modified porcine models exist. We suggest that the optimal immuno-oncology model may depend on the stage of cancer immunoediting in question. In particular, the spontaneous canine tumor models provide a unique platform for evaluating therapies aimed at the escape phase of cancer, while genetically engineered swine allow for elucidation of tumor-immune cell interactions especially during the phases of elimination and equilibrium.

Tumor formation on plants by mixtures of attenuated Agrobacterium tumefaciens T-DNA mutants

1984 ◽  
Vol 3 (6) ◽  
pp. 337-344 ◽  
Author(s):  
G. M. S. van Slogteren ◽  
P. J. J. Hooykaas ◽  
R. A. Schilperoort
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Tumor Formation

β-Glucuronidase activity in seedlings of the parasitic angiosperm Cusctua pentagona: developmental impact of the β-glucuronidase inhibitor saccharic acid 1,4-lactone

2007 ◽  
Vol 34 (9) ◽  
pp. 811 ◽  
Author(s):  
Mark A. Schoenbeck ◽  
Gabriel A. Swanson ◽  
Sydney J. Brommer
Keyword(s):  
Seedling Growth ◽  
Early Stage ◽  
Shoot Elongation ◽  
Gus Activity ◽  
Ph Optimum ◽  
Parasitic Angiosperm ◽  
Glucuronidase Activity ◽  
Germinating Seeds ◽  
Cuscuta Pentagona

Endogenous plant β-glucuronidase (β-GUS) activity was detected in germinating seeds, seedlings, stems, flowers and haustoria of the parasitic angiosperm Cuscuta pentagona Engelm. In vitro characterisation of this activity showed it to have an acidic pH optimum, similar to previously characterised plant activities, and a sensitivity to the β-GUS inhibitor saccharic acid 1,4-lactone (SAL). Application of SAL to seeds immediately after chemical scarification resulted in a significant developmental delay and, frequently, in the total arrest of seedling growth. In contrast, application of SAL subsequent to the emergence of the radicle did not produce a significant effect on the development of the seedling. Thus, the distribution of activity and the developmentally contingent potency of SAL in inhibiting growth suggest a role for β-GUS at an early stage of seed germination or seedling growth. Further, the inability of the inhibitor to prevent subsequent shoot elongation suggests that at least some plant growth processes do not require this activity, or that it is required only at minimal levels and is unaffected by the application of SAL.

scholarly journals Micropropagation of Inula germanica L. from the Seedlings Explants

2018 ◽  
Vol 46 (1) ◽  
pp. 52-57 ◽  
Author(s):  
Alina TREJGELL ◽  
Monika KAMIŃSKA ◽  
Karolina LISOWSKA ◽  
Andrzej TRETYN
Keyword(s):  
Solid Medium ◽  
Positive Impact ◽  
Shoot Tips ◽  
Fluorescent Light ◽  
First Year ◽  
Naphthaleneacetic Acid ◽  
Multiplication Rate ◽  
Indolebutyric Acid ◽  
Regenerated Plants

This is the first communication of micropropagation system for Inula germanica using seedling explants germinated in vitro. The development of this system gives the possibility of future reintroduction of I. germanica providing a way to stabilize or re-establish its population. Shoot tips and fragments of cotyledons, hypocotyls and roots were isolated from ten-day-old seedlings. Explants were put on MS medium containing 1.0 mg l-1 benzylaminopurine and 0.1 mg l-1 naphthaleneacetic acid and cultured under continuous white fluorescent light (45 μmol.m-2.s-1) at 26 ± 1 °C. The highest percentage of shoot organogenesis (83.3%) was recorded for hypocotyl, while the highest average number of shoots per explant (12.0) was recorded for shoot tips. In subsequent subcultures, multiplication rate decreased to 3.0-4.9 shoots per explant. Less than 19% shoots were able to root on the solid medium without auxins. The highest rooting efficiency (69.3%) was recorded for solid medium supplemented with indolebutyric acid, but growth of roots was inhibited. The percentage of rooted shoots (62.2%) and number of roots per shoot (2.4 per shoot) into the liquid medium were comparable to medium with 0.1 mg·l-1 indolebutyric acid. showing a positive impact on the process of acclimatization. The regenerated plants were able to flowering in the first year after acclimatization. Developed micropropagation system for I. germanica is efficient and can be a useful tool for the active protection of this species.

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