Agrobacterium tumefaciens-mediated transformation and regeneration of Saaz hop - Osvald's clones 31 and 72

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F14864347%3A_____%2F21%3AN0000019" target="_blank" >RIV/14864347:_____/21:N0000019 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.actahort.org/books/1328/1328_11.htm" target="_blank" >https://www.actahort.org/books/1328/1328_11.htm</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.17660/ActaHortic.2021.1328.11" target="_blank" >10.17660/ActaHortic.2021.1328.11</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Agrobacterium tumefaciens-mediated transformation and regeneration of Saaz hop - Osvald's clones 31 and 72

Popis výsledku v původním jazyce

Humulus lupulus is a well-known economically important crop widely used in the brewing industry and in biomedical applications. Despite the importance of hop in industry, agriculture and biology, genetic transformation protocols are still not efficient enough because the regeneration of transformed plantlets is dependent on explant type, cultivars, and mainly concentration of growth regulators. Therefore, in this study suitable conditions for in vitro regeneration and Agrobacterium tumefaciens-mediated transformation for ‘Saaz’ hop explants were tested. Different explants, genotypes, and growing media with varying concentrations of indole-3-acetic acid (IAA), thidiazuron (TDZ) and zeatin were compared. Whereas shoot internodes regenerated successfully on MS-B5 medium supplemented with 1 mg L-1 zeatin and 0.25 mg L-1 IAA, callus formation and shoot regeneration frequency was lower with 2.11 mg L-1 TDZ and 0.33 mg L-1 IAA. To test the efficiency of transformation and subsequent regeneration, transformation protocols of ‘Saaz’ hop - Osvald’s clones 31 and 72 were performed using Agrobacterium tumefaciens strain GV3101 and the vector pCAMBIA 1301. As next step, data obtained from transient and stable expression were compared for pre-cultured and fresh hop explants (shoot internodes and leaves sections) of both cultivars. The highest transient transformation frequency was observed in the freshly cut or pre-cultured explants of Osvald’s clone 72, while the highest stable transformation frequency was achieved using fresh explants of Osvald’s clone 31. GUS (β-glucuronidase) positive hop plants were obtained after cultivation on MS-B5 selection medium with 1 mg L-1 zeatin, 0.25 mg L-1 IAA, 250 mg L-1 timentin and 1.5 mg L-1 hygromycin. Thus, our results show that the most suitable explants for transformation and regeneration are shoot internodes. In addition, fresh and pre-cultured explants of Osvald’s clones 31 and 72 differ in transient and stable frequency. This shows the importance of protocol optimization for studied clones and other cultivars in hop agriculture.

Název v anglickém jazyce

Agrobacterium tumefaciens-mediated transformation and regeneration of Saaz hop - Osvald's clones 31 and 72

Popis výsledku anglicky

Humulus lupulus is a well-known economically important crop widely used in the brewing industry and in biomedical applications. Despite the importance of hop in industry, agriculture and biology, genetic transformation protocols are still not efficient enough because the regeneration of transformed plantlets is dependent on explant type, cultivars, and mainly concentration of growth regulators. Therefore, in this study suitable conditions for in vitro regeneration and Agrobacterium tumefaciens-mediated transformation for ‘Saaz’ hop explants were tested. Different explants, genotypes, and growing media with varying concentrations of indole-3-acetic acid (IAA), thidiazuron (TDZ) and zeatin were compared. Whereas shoot internodes regenerated successfully on MS-B5 medium supplemented with 1 mg L-1 zeatin and 0.25 mg L-1 IAA, callus formation and shoot regeneration frequency was lower with 2.11 mg L-1 TDZ and 0.33 mg L-1 IAA. To test the efficiency of transformation and subsequent regeneration, transformation protocols of ‘Saaz’ hop - Osvald’s clones 31 and 72 were performed using Agrobacterium tumefaciens strain GV3101 and the vector pCAMBIA 1301. As next step, data obtained from transient and stable expression were compared for pre-cultured and fresh hop explants (shoot internodes and leaves sections) of both cultivars. The highest transient transformation frequency was observed in the freshly cut or pre-cultured explants of Osvald’s clone 72, while the highest stable transformation frequency was achieved using fresh explants of Osvald’s clone 31. GUS (β-glucuronidase) positive hop plants were obtained after cultivation on MS-B5 selection medium with 1 mg L-1 zeatin, 0.25 mg L-1 IAA, 250 mg L-1 timentin and 1.5 mg L-1 hygromycin. Thus, our results show that the most suitable explants for transformation and regeneration are shoot internodes. In addition, fresh and pre-cultured explants of Osvald’s clones 31 and 72 differ in transient and stable frequency. This shows the importance of protocol optimization for studied clones and other cultivars in hop agriculture.

Druh

D - Stať ve sborníku

CEP obor

—

OECD FORD obor

40402 - GM technology (crops and livestock), livestock cloning, marker assisted selection, diagnostics (DNA chips and biosensing devices for the early/accurate detection of diseases) biomass feedstock production technologies, biopharming

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název statě ve sborníku

Acta Horticulturae 1328

ISBN

978-94-62613-25-6

ISSN

0567-7572

e-ISSN

2406-6168

Počet stran výsledku

8

Strana od-do

81-88

Název nakladatele

International Society for Horticultural Science

Místo vydání

Leuven

Místo konání akce

Stuttgart

Datum konání akce

8. 3. 2021

Typ akce podle státní příslušnosti

WRD - Celosvětová akce

Kód UT WoS článku

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