Agrobacterium tumefaciens -mediated transformation of Pinus pinea L. cotyledons: an assessment of factors influencing the efficiency of uidA gene transfer

Humara, Jaime M.; López, Marián; Ordás, Ricardo J.

doi:10.1007/s002990050709

Cited by 63 publications

(31 citation statements)

References 39 publications

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“…These are in agreement with Humara et al [14]. Ultrasonication has been reported to mediate gene transfer in plant suspension cells and intact pieces of tissues [14]. Our results demonstrate the advantages of combined s o n i c a t i o n o f l o b l o l l y p i n e e m b r y o s a n d Agrobacterium tumefaciens to achieve high levels of further expression of the uidA gene, similar to the reports by Trick and Finer [15].…”

Section: Discussionsupporting

confidence: 94%

“…Increasing co-cultivation time improving transient GUS expression was also observed. These are in agreement with Humara et al [14]. Ultrasonication has been reported to mediate gene transfer in plant suspension cells and intact pieces of tissues [14].…”

Section: Discussionsupporting

confidence: 91%

“…Scraped mature zygotic embryos, as well as particle bombardment wounded and ultrasound wounded embryos were tested. These wounding methods were performed following the protocols published by Humara et al [14] and Trick and Finer [15] and by using seed source E-22 as target explants.…”

Section: Co-cultivation and Selectionmentioning

confidence: 99%

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Agrobacterium-mediated transformation and assessment of factors influencing transgene expression in loblolly pine (Pinus taeda L.)

Tang

2001

Cell Res

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This investigation reports a protocol for transfer and expression of foreign chimeric genes in loblolly pine (Pinus taeda L.). Transformation was achieved by co-cultivation of mature zygotic embryos with Agrobacterium tumefaciens strain LBA4404 which harbored a binary vector (pBI121) including genes for b-glucuronidase (GUS) and neomycin phosphotransferase (NPTII). Factors influencing transgene expression including seed sources of loblolly pine, concentration of bacteria, and the wounding procedures of target explants were investigated. The expression of foreign gene was confirmed by the ability of mature zygotic embryos to produce calli in the presence of kanamycin, by histochemical assays of GUS activity, by PCR analysis, and by Southern blot. The successful expression of the GUS gene in different families of loblolly pine suggests that this transformation system is probably useful for the production of the genetically modified conifers.

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Section: Discussionsupporting

confidence: 94%

Section: Discussionsupporting

confidence: 91%

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Agrobacterium-mediated transformation and assessment of factors influencing transgene expression in loblolly pine (Pinus taeda L.)

Tang

2001

Cell Res

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“…Briefly, approximately 50 explants were vacuum-infiltrated in the buffer solution for 5 min and thereafter incubated for 24 hrs at 37ºC in the dark (Humara et al 1999). Prior to counting the number of blue spots, explants were washed in methanol:acetone (3:1, v/v).…”

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confidence: 99%

Agrobacterium-mediated transient transformation of Mexican prickly poppy (Argemone mexicana L.)

Godoy-Hernandez¹,

Aviles-Berzunza²,

Carrillo-Pech³

et al. 2008

Electron. J. Biotechnol.

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“…Studies of transient gene expression also provide a fast and useful tool for studying trans-acting factors and other molecules that modulate gene expression (Goff et al 1991;Gourrierec et al 1999;Gubler et al 1999) and for studying the expression of novel proteins in various plants (Tian et al 1997;Hibberd et al 1998;Ponappa et al 1999). Investigation and evaluation of transient expression of introduced genes is also necessary for the development of genetic transformation methodologies for plants; for example, the determination of optimal conditions for gene delivery, determination of types of explants, tissues and developmental stages of the tissues suitable for the transformation, identification and location of cells that are competent for transformation, evaluation and determination of genes and constructs suitable for transformation, monitoring development and selective growth of transformed cells (Ellis et al 1993;Ritala et al 1994;Hagio et al 1995;Jain et al 1996;Cabrera-Ponce et al 1997;Cheng et al 1997;Dillen et al 1997;Chen et al 1998;Humara et al 1999;Ponappa et al 1999).…”

Section: Mots Clésmentioning

confidence: 99%

Transient expression of a reporter gene changes significantly during somatic embryogenesis in alfalfa

Tian¹,

Brown²,

Webb³

2000

Can. J. Plant Sci.

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Transient expression of a reporter gene changes significantly during somatic embryogenesis in alfalfa. Can. J. Plant Sci. 80: 765-771. Transient expression of the β-glucuronidase (GUS) gene driven by a 35S constitutive promoter was investigated during somatic embryo development in alfalfa (Medicago sativa L.) following microprojectile bombardment. The level of gene expression varied with the different developmental stages and changed dramatically within the stages. During embryogenic callus induction (stage I), expression was low in explants freshly transferred to culture, but increased when cells started to divide and show competence for somatic embryogenesis. Gene expression, however, decreased when extensive callus formation began and reached the lowest observed level when callus was fully developed. Reporter gene expression was consistently low during embryogenic cell proliferation (stage II). In contrast, high levels of reporter gene expression were detected in non-embryogenic suspension cultures during cell proliferation. Extremely low reporter gene expression was detected in embryonal tissues and young embryos at the embryo development and maturation stage (stage III). Expression increased as the somatic embryos developed and reached the highest level when the embryos reached maturity. Transient expression levels were similar in excised petiole tissue across 10 alfalfa genotypes, which included both diploid and tetraploid genotypes. In general, reporter gene expression was found to be higher in more organised mature tissues and organs than in less-organised or young tissues. The developmental status of the cells and tissues appears to be an important factor in the degree of transient expression of the introduced gene. Changes of gene expression during embryogenesis and factors affecting gene expression are discussed. Nous avons examiné l'expression transitoire du gène codant pour la β-glucuronidase (GUS) entraîné par un gène promoteur constituant 35S après bombardement par micro-projectiles durant le développement somato-embryonnaire de la luzerne (Medicago sativa L.). En plus de varier selon le stade de développe-ment, le niveau d'expression du gène changeait de façon spectaculaire à l'intérieur d'un même stade. Pendant l'induction du cal embryogénique (stade I), il était faible dans les explantats fraîchement transférés dans le milieu de culture, mais il augmentait lorsque les cellules commençaient à se diviser et à manifester une compétence d'embryogenèse somatique. En revanche, il diminuait au début de la phase de callogénèse, extensive pour enfin atteindre son plus bas niveau quand le cal arrivait à son plein développement. L'expression du gène rapporteur demeurait faible durant la phase de prolifération des cellules embryogéniques (stade II), alors qu'elle atteignait un degré élevé durant la phase de prolifération des cellules non-embryogéniques. Un très faible degré d'expression du gène rapporteur était observé dans les tissus embryonnaires et dans les jeunes embryons aux stades de développement et...

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Agrobacterium tumefaciens -mediated transformation of Pinus pinea L. cotyledons: an assessment of factors influencing the efficiency of uidA gene transfer

Cited by 63 publications

References 39 publications

Agrobacterium-mediated transformation and assessment of factors influencing transgene expression in loblolly pine (Pinus taeda L.)

Agrobacterium-mediated transformation and assessment of factors influencing transgene expression in loblolly pine (Pinus taeda L.)

Agrobacterium-mediated transient transformation of Mexican prickly poppy (Argemone mexicana L.)

Transient expression of a reporter gene changes significantly during somatic embryogenesis in alfalfa

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