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Puddephat, I. J.; Robinson, Helen T.; Fenning, T. M.; Barbara, D. J.; Morton, A.; Pink, David

doi:10.1023/a:1011338322000

Cited by 45 publications

(7 citation statements)

References 30 publications

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“…Our observations were consistent with literature data (David and Tempé 1988; Puddephat et al 2001) concerning the morphology of plants. The transgenic plants exhibited an altered phenotype, although it varied between individuals from the same transformation event.…”

Section: Discussionsupporting

confidence: 94%

“…The transgenic plants exhibited an altered phenotype, although it varied between individuals from the same transformation event. It must be pointed out, however, that Puddephat et al (2001) noticed such alteration only in transgenic cauliflower, we observed a certain degree of dwarfism, leaf wrinkling or lancet-like leaves and hairy root-like rooting also in wild-type plants rooted on a high concentration of NAA. It seems that the phenotype of newly regenerated transgenic plants after the R. rhizogenes mediated transformation changed in a way similar to that caused by a high dose of auxin (or maybe just NAA).…”

Section: Discussioncontrasting

confidence: 45%

“…A similar protocol utilizing R. rhizogenes (A4T) was reported by Puddephat et al (2001). It was successfully applied to produce transgenic broccoli and cauliflower.…”

Section: Discussionmentioning

confidence: 83%

“…Our parameters yielded the transformation efficiency of 72% (the number of seedlings with at least 1 hairy root) and of 55% (measured as the number of inoculated hypocotyls producing at least 1 transgenic hairy-root determined by the GUS assay). In comparison, Puddephat et al (2001) obtained the maximum transformation efficiency of 8.7% for histochemically confirmed hairy-roots. Establishing the aforementioned parameters we relied on earlier studies reporting that e.g.…”

Section: Discussionmentioning

confidence: 87%

“…To obtain callus tissue for transgenic plant regeneration, selected hairy roots were transferred to the MS medium with 5 mg/L BAP and NAA, 3% sucrose, solidified with 0.8% agar, partly based on Puddephat et al (2001). Petri dishes with the roots were incubated in the dark at 24 °C and the proliferating tissue was transferred to a new medium every 3 weeks.…”

Section: Methodsmentioning

confidence: 99%

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High efficiency transformation of Brassica oleracea var. botrytis plants by Rhizobium rhizogenes

et al. 2018

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Brassica oleracea var. botrytis, a very popular crop grown for its edible inflorescence, is bred only as a mutated annual cultivar and does not naturally occur in environment. Since cauliflower is still described as the most troublesome of all the B. oleracea vegetables regarding transformation processes, it is fully justified to focus on the improvement of tools for its genetic modifications. Here, we present a successful protocol for genetic transformation of cauliflower employing the process of agroinfection. The primary analysis of in vitro response of five cultivars allowed us to have chosen Pionier as the most promising cultivar; in consequence the Pionier was transformed via Rhizobium-mediated techniques in order to evaluate both, R. radiobacter (EHA 105, LBA 4404) and R. rhizogenes (ATCC 18534, A4) species. However, the latter system turned out to be more effective and, the A4 strain, in particular (72% transformation efficiency, 55% confirmed by GUS assay). That shows a promising technical advance especially when compared to the results of previous literature reports (e.g. 8.7% reported efficiency using R. rhizogenes). The transgenic cauliflower was obtained from hairy roots via organogenic callus induction. The potential transformants were analysed at the genomic and proteomic levels and their transgenic character was fully confirmed.Electronic supplementary materialThe online version of this article (10.1186/s13568-018-0656-6) contains supplementary material, which is available to authorized users.

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Section: Discussionsupporting

confidence: 94%

Section: Discussioncontrasting

confidence: 45%

“…A similar protocol utilizing R. rhizogenes (A4T) was reported by Puddephat et al (2001). It was successfully applied to produce transgenic broccoli and cauliflower.…”

Section: Discussionmentioning

confidence: 83%

Section: Discussionmentioning

confidence: 87%

Section: Methodsmentioning

confidence: 99%

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High efficiency transformation of Brassica oleracea var. botrytis plants by Rhizobium rhizogenes

et al. 2018

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Vegetable Brassicas

Christey

Braun

2008

Compendium of Transgenic Crop Plants

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Vegetable Brassicas are economically important and grown worldwide for consumption as both fresh and frozen produce. In addition, they are highly nutritious and are gaining in popularity as they contain compounds with anticancer properties. Transformation systems have been developed for several vegetable Brassica oleracea and Brassica rapa types and a range of genes introduced. Agrobacterium tumefaciens ‐mediated transformation is the method of choice but remains difficult with rates low and genotype dependent. In this chapter, we review factors to enhance transformation rates and discuss the range of genes of agronomic importance that have been introduced including pest and disease resistance. Research is also being conducted to produce plants with altered adaptation to water and salt stress and with altered health benefits. While no transgenic vegetable Brassicas have yet been released commercially, field testing and nontarget impacts research of transgenic vegetable Brassicas is in progress. In the future, commercial release could occur if environmental and public concerns are adequately addressed.

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MALDI‐TOF characterization of hGH1 produced by hairy root cultures of Brassica oleracea var. italica grown in an airlift with mesh bioreactor

López

Ramírez

Guzmán

et al. 2013

Biotechnology Progress

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Expression systems based on plant cells, tissue, and organ cultures have been investigated as an alternative for production of human therapeutic proteins in bioreactors. In this work, hairy root cultures of Brassica oleracea var. italica (broccoli) were established in an airlift with mesh bioreactor to produce isoform 1 of the human growth hormone (hGH1) as a model therapeutic protein. The hGH1 cDNA was cloned into the pCAMBIA1105.1 binary vector to induce hairy roots in hypocotyls of broccoli plantlets via Agrobacterium rhizogenes. Most of the infected plantlets (90%) developed hairy roots when inoculated before the appearance of true leaves, and keeping the emerging roots attached to hypocotyl explants during transfer to solid Schenk and Hildebrandt medium. The incorporation of the cDNA into the hairy root genome was confirmed by PCR amplification from genomic DNA. The expression and structure of the transgenic hGH1 was assessed by ELISA, western blot, and MALDITOF-MS analysis of the purified protein extracted from the biomass of hairy roots cultivated in bioreactor for 24 days. Production of hGH1 was 5.1 ± 0.42 µg/g dry weight (DW) for flask cultures, and 7.8 ± 0.3 µg/g DW for bioreactor, with productivity of 0.68 ± 0.05 and 1.5 ± 0.06 µg/g DW*days, respectively, indicating that the production of hGH1 was not affected by the growth rate, but might be affected by the culture system. These results demonstrate that hairy root cultures of broccoli have potential as an alternative expression system for production of hGH1, and might also be useful for production of other therapeutic proteins.

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Untitled

Cited by 45 publications

References 30 publications

High efficiency transformation of Brassica oleracea var. botrytis plants by Rhizobium rhizogenes

High efficiency transformation of Brassica oleracea var. botrytis plants by Rhizobium rhizogenes

Vegetable Brassicas

MALDI‐TOF characterization of hGH1 produced by hairy root cultures of Brassica oleracea var. italica grown in an airlift with mesh bioreactor

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