Regeneration and Agrobacterium-mediated transformation of multiple lily cultivars

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Abstract

To pursue genetic improvement of lily, efficiency of both regeneration and transformation from callus cultures induced from different explants were evaluated in multiple cultivars. Thirty-five callus lines induced from filaments or styles and one control callus line derived from bulb scales of in total twenty lily cultivars representing Lilium longiflorum, Oriental × Trumpet and Longiflorum × Asiatic hybrids were maintained on a medium with 8.3 µM picloram (PIC). In this study, they were tested for their regeneration potential by transferring them onto a regeneration medium supplemented with 0.4 µM PIC and 0.044 µM 6-benzyladenine. Regeneration was obtained in all cultivars examined and the percentage varied from zero to 89 % in the 36 callus lines. Regeneration frequency was significantly influenced by the genotype (cultivar). Subculturing the calli every 4 weeks by refreshing the regeneration medium contributed positively to bulblet formation, when compared to an eight week subculture frequency. It was found that the regeneration ability generally decreased with an increasing age of the callus cultures for all cultivars. The origin of the callus (style or filament) did not lead to significant differences in regeneration frequency, but there was an interaction between callus origin and genotype. Calli of eight randomly chosen cultivars were co-cultivated with Agrobacterium tumefaciens strain AGL0 carrying binary vectors with the gus gene as reporter and putative transgenic plants were produced. GUS histochemical assays demonstrated transient and stable expression of the gus gene in both calli and regenerated lily plants. Transient expression frequencies ranged from 0.3 to 20.6 % while stable transformation was much lower, only 1.4 % as the maximum
Original languageEnglish
Pages (from-to)113-122
JournalPlant Cell, Tissue and Organ Culture: an international journal on in vitro culture of higher plants
Volume111
Issue number1
DOIs
Publication statusPublished - 2012

Fingerprint

Lilium
Agrobacterium
callus
cultivars
picloram
callus culture
Lilium longiflorum
genotype
Agrobacterium radiobacter
benzyladenine
bulbs
genetic improvement
transgenic plants
explants
gene expression
assays

Keywords

  • triticum-aestivum l
  • lilium-longiflorum
  • plant-regeneration
  • beta-glucuronidase
  • transgenic wheat
  • inhibitor(s)
  • floriculture
  • expression
  • cultures
  • gene

Cite this

@article{d019551011194fd1a753d4a23367a27a,
title = "Regeneration and Agrobacterium-mediated transformation of multiple lily cultivars",
abstract = "To pursue genetic improvement of lily, efficiency of both regeneration and transformation from callus cultures induced from different explants were evaluated in multiple cultivars. Thirty-five callus lines induced from filaments or styles and one control callus line derived from bulb scales of in total twenty lily cultivars representing Lilium longiflorum, Oriental × Trumpet and Longiflorum × Asiatic hybrids were maintained on a medium with 8.3 µM picloram (PIC). In this study, they were tested for their regeneration potential by transferring them onto a regeneration medium supplemented with 0.4 µM PIC and 0.044 µM 6-benzyladenine. Regeneration was obtained in all cultivars examined and the percentage varied from zero to 89 {\%} in the 36 callus lines. Regeneration frequency was significantly influenced by the genotype (cultivar). Subculturing the calli every 4 weeks by refreshing the regeneration medium contributed positively to bulblet formation, when compared to an eight week subculture frequency. It was found that the regeneration ability generally decreased with an increasing age of the callus cultures for all cultivars. The origin of the callus (style or filament) did not lead to significant differences in regeneration frequency, but there was an interaction between callus origin and genotype. Calli of eight randomly chosen cultivars were co-cultivated with Agrobacterium tumefaciens strain AGL0 carrying binary vectors with the gus gene as reporter and putative transgenic plants were produced. GUS histochemical assays demonstrated transient and stable expression of the gus gene in both calli and regenerated lily plants. Transient expression frequencies ranged from 0.3 to 20.6 {\%} while stable transformation was much lower, only 1.4 {\%} as the maximum",
keywords = "triticum-aestivum l, lilium-longiflorum, plant-regeneration, beta-glucuronidase, transgenic wheat, inhibitor(s), floriculture, expression, cultures, gene",
author = "Yue Wang and {van Kronenburg-van de Ven}, B.C.E. and T.R. Menzel and C.A. Maliepaard and X. Shen and F.A. Krens",
year = "2012",
doi = "10.1007/s11240-012-0172-3",
language = "English",
volume = "111",
pages = "113--122",
journal = "Plant Cell, Tissue and Organ Culture: an international journal on in vitro culture of higher plants",
issn = "0167-6857",
publisher = "Springer Verlag",
number = "1",

}

TY - JOUR

T1 - Regeneration and Agrobacterium-mediated transformation of multiple lily cultivars

AU - Wang, Yue

AU - van Kronenburg-van de Ven, B.C.E.

AU - Menzel, T.R.

AU - Maliepaard, C.A.

AU - Shen, X.

AU - Krens, F.A.

PY - 2012

Y1 - 2012

N2 - To pursue genetic improvement of lily, efficiency of both regeneration and transformation from callus cultures induced from different explants were evaluated in multiple cultivars. Thirty-five callus lines induced from filaments or styles and one control callus line derived from bulb scales of in total twenty lily cultivars representing Lilium longiflorum, Oriental × Trumpet and Longiflorum × Asiatic hybrids were maintained on a medium with 8.3 µM picloram (PIC). In this study, they were tested for their regeneration potential by transferring them onto a regeneration medium supplemented with 0.4 µM PIC and 0.044 µM 6-benzyladenine. Regeneration was obtained in all cultivars examined and the percentage varied from zero to 89 % in the 36 callus lines. Regeneration frequency was significantly influenced by the genotype (cultivar). Subculturing the calli every 4 weeks by refreshing the regeneration medium contributed positively to bulblet formation, when compared to an eight week subculture frequency. It was found that the regeneration ability generally decreased with an increasing age of the callus cultures for all cultivars. The origin of the callus (style or filament) did not lead to significant differences in regeneration frequency, but there was an interaction between callus origin and genotype. Calli of eight randomly chosen cultivars were co-cultivated with Agrobacterium tumefaciens strain AGL0 carrying binary vectors with the gus gene as reporter and putative transgenic plants were produced. GUS histochemical assays demonstrated transient and stable expression of the gus gene in both calli and regenerated lily plants. Transient expression frequencies ranged from 0.3 to 20.6 % while stable transformation was much lower, only 1.4 % as the maximum

AB - To pursue genetic improvement of lily, efficiency of both regeneration and transformation from callus cultures induced from different explants were evaluated in multiple cultivars. Thirty-five callus lines induced from filaments or styles and one control callus line derived from bulb scales of in total twenty lily cultivars representing Lilium longiflorum, Oriental × Trumpet and Longiflorum × Asiatic hybrids were maintained on a medium with 8.3 µM picloram (PIC). In this study, they were tested for their regeneration potential by transferring them onto a regeneration medium supplemented with 0.4 µM PIC and 0.044 µM 6-benzyladenine. Regeneration was obtained in all cultivars examined and the percentage varied from zero to 89 % in the 36 callus lines. Regeneration frequency was significantly influenced by the genotype (cultivar). Subculturing the calli every 4 weeks by refreshing the regeneration medium contributed positively to bulblet formation, when compared to an eight week subculture frequency. It was found that the regeneration ability generally decreased with an increasing age of the callus cultures for all cultivars. The origin of the callus (style or filament) did not lead to significant differences in regeneration frequency, but there was an interaction between callus origin and genotype. Calli of eight randomly chosen cultivars were co-cultivated with Agrobacterium tumefaciens strain AGL0 carrying binary vectors with the gus gene as reporter and putative transgenic plants were produced. GUS histochemical assays demonstrated transient and stable expression of the gus gene in both calli and regenerated lily plants. Transient expression frequencies ranged from 0.3 to 20.6 % while stable transformation was much lower, only 1.4 % as the maximum

KW - triticum-aestivum l

KW - lilium-longiflorum

KW - plant-regeneration

KW - beta-glucuronidase

KW - transgenic wheat

KW - inhibitor(s)

KW - floriculture

KW - expression

KW - cultures

KW - gene

U2 - 10.1007/s11240-012-0172-3

DO - 10.1007/s11240-012-0172-3

M3 - Article

VL - 111

SP - 113

EP - 122

JO - Plant Cell, Tissue and Organ Culture: an international journal on in vitro culture of higher plants

JF - Plant Cell, Tissue and Organ Culture: an international journal on in vitro culture of higher plants

SN - 0167-6857

IS - 1

ER -