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אסיף מאגר המחקר החקלאי
פותח על ידי קלירמאש פתרונות בע"מ -
Efficient genetic transformation of lilium longiflorum and ornithogalum dubium by particle acceleration followed by prolonged selection in liquid medium
Year:
2004
Source of publication :
Acta Horticulturae
Authors :
ארזי, צחי
;
.
ברג, רבקה
;
.
גאבה, ויקטור
;
.
גרה, עבדאללה
;
.
זלץ, יחיעם
;
.
יון, האורל
;
.
כהן, אבנר
;
.
ליפסקי, אלכסנדר
;
.
סנדלר-זיו, דורית
;
.
סתו, רן
;
.
פינטה, קורנליו
;
.
Volume :
651
Co-Authors:
Cohen, A., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Lipsky, A., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Arazi, T., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Ion, A., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Stav, R., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Sandler-Ziv, D., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Pintea, C., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Barg, R., Dept. Plant Genetics, Volcani Center, ARO, Bet Dagan 50250, Israel
Salts, Y., Dept. Plant Genetics, Volcani Center, ARO, Bet Dagan 50250, Israel
Shabtai, S., Dept. Plant Genetics, Volcani Center, ARO, Bet Dagan 50250, Israel
Gaba, V., Dept. Virology, Volcani Center, ARO, Bet Dagan 50250, Israel
Gera, A., Dept. Virology, Volcani Center, ARO, Bet Dagan 50250, Israel
Facilitators :
From page:
131
To page:
138
(
Total pages:
8
)
Abstract:
Genetic transformation mediated by bombardment with microscopic metal particles carrying target genes is the preferred method for the introduction of foreign genes into monocotyledonous plants. The fact that most flower bulbs are monocotyle-donous and that almost all commercial cultivars are propagated vegetatively makes them good candidates for molecular breeding through microprojectile bombard-ment. We report here on a development of a reliable method for an efficient genetic transformation of both Lilium longiflorum and Ornithogalum dubium using a particle inflow gun to deliver gene constructs into the target plant tissue, followed by a prolonged selection in the dark in liquid medium supplemented with kanamycin. The system was first optimized for Lilium longiflorum 'Snow Queen'. Based on the level of transient GUS expression, liquidgrown cell clumps are more competent than leaves. Large cell clusters (2-10mm) maintain their organogenic potential while smaller clusters (<2mm) cease to grow and die. The liquid-grown tissue cultures have a level of competence for transformation about 50-70 times greater than that of solid-grown callus cultures, and compact cell clusters are more competent than loose clusters. The cells were bombarded with a pCAMBIA2301 vector, carrying nptII gene conferring kanamycin resistance and GUS reporter gene. Following selection for 4-6 months in a liquid medium supplemented with 80 mg l-1 kanamycin in the dark, the cell clusters were transferred to a regeneration medium in the light where hundreds of transgenic plantlets developed. The plants retained their stable transgenic state when grown in the greenhouse for two seasons. The transformation of O. dubium was similar in principle to that of L. longiflorum with three major differences: lily liquid-grown cultures grew more rapidly and had a higher potential for somatic embryo development. Ornithogalum cultures under selection took longer to develop into semi-organized cell clumps of sufficient size to allow continued shoot regeneration, were mostly organogenic, and the regenerated plantlets had higher rate of vitrification.
Note:
Related Files :
LILY
Monocotyledonous plants
Ornithogalum
Particle bombardment
transformation
עוד תגיות
תוכן קשור
More details
DOI :
Article number:
Affiliations:
Database:
סקופוס
Publication Type:
מאמר מתוך כינוס
;
.
Language:
אנגלית
Editors' remarks:
ID:
26634
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:24
Scientific Publication
Efficient genetic transformation of lilium longiflorum and ornithogalum dubium by particle acceleration followed by prolonged selection in liquid medium
651
Cohen, A., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Lipsky, A., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Arazi, T., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Ion, A., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Stav, R., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Sandler-Ziv, D., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Pintea, C., Dept. Ornamental Horticulture, Volcani Center, ARO, Bet Dagan 50250, Israel
Barg, R., Dept. Plant Genetics, Volcani Center, ARO, Bet Dagan 50250, Israel
Salts, Y., Dept. Plant Genetics, Volcani Center, ARO, Bet Dagan 50250, Israel
Shabtai, S., Dept. Plant Genetics, Volcani Center, ARO, Bet Dagan 50250, Israel
Gaba, V., Dept. Virology, Volcani Center, ARO, Bet Dagan 50250, Israel
Gera, A., Dept. Virology, Volcani Center, ARO, Bet Dagan 50250, Israel
Efficient genetic transformation of lilium longiflorum and ornithogalum dubium by particle acceleration followed by prolonged selection in liquid medium
Genetic transformation mediated by bombardment with microscopic metal particles carrying target genes is the preferred method for the introduction of foreign genes into monocotyledonous plants. The fact that most flower bulbs are monocotyle-donous and that almost all commercial cultivars are propagated vegetatively makes them good candidates for molecular breeding through microprojectile bombard-ment. We report here on a development of a reliable method for an efficient genetic transformation of both Lilium longiflorum and Ornithogalum dubium using a particle inflow gun to deliver gene constructs into the target plant tissue, followed by a prolonged selection in the dark in liquid medium supplemented with kanamycin. The system was first optimized for Lilium longiflorum 'Snow Queen'. Based on the level of transient GUS expression, liquidgrown cell clumps are more competent than leaves. Large cell clusters (2-10mm) maintain their organogenic potential while smaller clusters (<2mm) cease to grow and die. The liquid-grown tissue cultures have a level of competence for transformation about 50-70 times greater than that of solid-grown callus cultures, and compact cell clusters are more competent than loose clusters. The cells were bombarded with a pCAMBIA2301 vector, carrying nptII gene conferring kanamycin resistance and GUS reporter gene. Following selection for 4-6 months in a liquid medium supplemented with 80 mg l-1 kanamycin in the dark, the cell clusters were transferred to a regeneration medium in the light where hundreds of transgenic plantlets developed. The plants retained their stable transgenic state when grown in the greenhouse for two seasons. The transformation of O. dubium was similar in principle to that of L. longiflorum with three major differences: lily liquid-grown cultures grew more rapidly and had a higher potential for somatic embryo development. Ornithogalum cultures under selection took longer to develop into semi-organized cell clumps of sufficient size to allow continued shoot regeneration, were mostly organogenic, and the regenerated plantlets had higher rate of vitrification.
Scientific Publication
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