Transgenic barley by particle bombardment. Inheritance of the transferred gene and characteristics of transgenic barley plants

Anneli Ritala, Reino Aikasalo, Kristian Aspegren, Marjatta Salmenkallio-Marttila, Satu Åkerman, Leena Mannonen, Ulrika Kurten, Riitta Puupponen-Pimiä, Teemu Teeri, Veli Kauppinen

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Transgenic barley plants (Hordeum vulgare L. cv. Kymppi) were obtained by particle bombardment of various tissues. Immature embryos and microspore-derived cultures were bombarded with gold particles coated with plasmid DNA carrying the gene coding for neomycin phosphotransferase II (NPTII), together with plasmid DNA containing the gene for β-glucuronidase (GUS).

Bombarded immature embryos were grown to plants without selection and NPTII activity was screened in small plantlets. One plant proved to be transgenic (T0). This chimeric plant passed the transferred nptII gene to its T1 progeny. The presence of the nptII gene was demonstrated by the PCR technique and enzyme activity was analyzed by an NPTII gel assay. Four T0 spikes and 15 T1 offspring were transgenic. The integration and inheritance was confirmed by Southern blot hybridization. Transgenic T2 and T3 plants were produced by isolating embryos from green grains of transgenic T1 and T2 plants, respectively and growing them to plants. After selfing, the ratio of transgenic to non-transgenic T2 offspring was shown to follow the rule of Mendelian inheritance. The general performance of transgenic plants was normal and no reduction in fertility was observed.

Microspore-derived cultures were bombarded one and four weeks after microspore isolation. After bombardment, cultures were grown either with or without antibiotic selection (geneticin R or kanamycin). When cultures were grown without selection and regenerated plants were transferred to kanamycin selection in rooting phase, one out of a total of about 1500 plants survived. This plant both carried and expressed the transferred nptII gene. The integration was confirmed by Southern blot hybridization. This plant was not fertile.
Original languageEnglish
Pages (from-to)81-88
JournalEuphytica
Volume85
Issue number1-3
DOIs
Publication statusPublished - 1995
MoE publication typeA1 Journal article-refereed

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Genetically Modified Plants
Hordeum
inheritance (genetics)
barley
genetically modified organisms
Genes
Kanamycin Kinase
genes
kanamycin kinase
microspores
embryo (plant)
Kanamycin
Embryonic Structures
Southern Blotting
kanamycin
Southern blotting
Plasmids
plasmids
hybridization
immatures

Cite this

Ritala, A., Aikasalo, R., Aspegren, K., Salmenkallio-Marttila, M., Åkerman, S., Mannonen, L., ... Kauppinen, V. (1995). Transgenic barley by particle bombardment. Inheritance of the transferred gene and characteristics of transgenic barley plants. Euphytica, 85(1-3), 81-88. https://doi.org/10.1007/BF00023933
Ritala, Anneli ; Aikasalo, Reino ; Aspegren, Kristian ; Salmenkallio-Marttila, Marjatta ; Åkerman, Satu ; Mannonen, Leena ; Kurten, Ulrika ; Puupponen-Pimiä, Riitta ; Teeri, Teemu ; Kauppinen, Veli. / Transgenic barley by particle bombardment. Inheritance of the transferred gene and characteristics of transgenic barley plants. In: Euphytica. 1995 ; Vol. 85, No. 1-3. pp. 81-88.
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title = "Transgenic barley by particle bombardment. Inheritance of the transferred gene and characteristics of transgenic barley plants",
abstract = "Transgenic barley plants (Hordeum vulgare L. cv. Kymppi) were obtained by particle bombardment of various tissues. Immature embryos and microspore-derived cultures were bombarded with gold particles coated with plasmid DNA carrying the gene coding for neomycin phosphotransferase II (NPTII), together with plasmid DNA containing the gene for β-glucuronidase (GUS).Bombarded immature embryos were grown to plants without selection and NPTII activity was screened in small plantlets. One plant proved to be transgenic (T0). This chimeric plant passed the transferred nptII gene to its T1 progeny. The presence of the nptII gene was demonstrated by the PCR technique and enzyme activity was analyzed by an NPTII gel assay. Four T0 spikes and 15 T1 offspring were transgenic. The integration and inheritance was confirmed by Southern blot hybridization. Transgenic T2 and T3 plants were produced by isolating embryos from green grains of transgenic T1 and T2 plants, respectively and growing them to plants. After selfing, the ratio of transgenic to non-transgenic T2 offspring was shown to follow the rule of Mendelian inheritance. The general performance of transgenic plants was normal and no reduction in fertility was observed.Microspore-derived cultures were bombarded one and four weeks after microspore isolation. After bombardment, cultures were grown either with or without antibiotic selection (geneticin R or kanamycin). When cultures were grown without selection and regenerated plants were transferred to kanamycin selection in rooting phase, one out of a total of about 1500 plants survived. This plant both carried and expressed the transferred nptII gene. The integration was confirmed by Southern blot hybridization. This plant was not fertile.",
author = "Anneli Ritala and Reino Aikasalo and Kristian Aspegren and Marjatta Salmenkallio-Marttila and Satu {\AA}kerman and Leena Mannonen and Ulrika Kurten and Riitta Puupponen-Pimi{\"a} and Teemu Teeri and Veli Kauppinen",
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Ritala, A, Aikasalo, R, Aspegren, K, Salmenkallio-Marttila, M, Åkerman, S, Mannonen, L, Kurten, U, Puupponen-Pimiä, R, Teeri, T & Kauppinen, V 1995, 'Transgenic barley by particle bombardment. Inheritance of the transferred gene and characteristics of transgenic barley plants', Euphytica, vol. 85, no. 1-3, pp. 81-88. https://doi.org/10.1007/BF00023933

Transgenic barley by particle bombardment. Inheritance of the transferred gene and characteristics of transgenic barley plants. / Ritala, Anneli; Aikasalo, Reino; Aspegren, Kristian; Salmenkallio-Marttila, Marjatta; Åkerman, Satu; Mannonen, Leena; Kurten, Ulrika; Puupponen-Pimiä, Riitta; Teeri, Teemu; Kauppinen, Veli.

In: Euphytica, Vol. 85, No. 1-3, 1995, p. 81-88.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Transgenic barley by particle bombardment. Inheritance of the transferred gene and characteristics of transgenic barley plants

AU - Ritala, Anneli

AU - Aikasalo, Reino

AU - Aspegren, Kristian

AU - Salmenkallio-Marttila, Marjatta

AU - Åkerman, Satu

AU - Mannonen, Leena

AU - Kurten, Ulrika

AU - Puupponen-Pimiä, Riitta

AU - Teeri, Teemu

AU - Kauppinen, Veli

N1 - Project code: BEL3030

PY - 1995

Y1 - 1995

N2 - Transgenic barley plants (Hordeum vulgare L. cv. Kymppi) were obtained by particle bombardment of various tissues. Immature embryos and microspore-derived cultures were bombarded with gold particles coated with plasmid DNA carrying the gene coding for neomycin phosphotransferase II (NPTII), together with plasmid DNA containing the gene for β-glucuronidase (GUS).Bombarded immature embryos were grown to plants without selection and NPTII activity was screened in small plantlets. One plant proved to be transgenic (T0). This chimeric plant passed the transferred nptII gene to its T1 progeny. The presence of the nptII gene was demonstrated by the PCR technique and enzyme activity was analyzed by an NPTII gel assay. Four T0 spikes and 15 T1 offspring were transgenic. The integration and inheritance was confirmed by Southern blot hybridization. Transgenic T2 and T3 plants were produced by isolating embryos from green grains of transgenic T1 and T2 plants, respectively and growing them to plants. After selfing, the ratio of transgenic to non-transgenic T2 offspring was shown to follow the rule of Mendelian inheritance. The general performance of transgenic plants was normal and no reduction in fertility was observed.Microspore-derived cultures were bombarded one and four weeks after microspore isolation. After bombardment, cultures were grown either with or without antibiotic selection (geneticin R or kanamycin). When cultures were grown without selection and regenerated plants were transferred to kanamycin selection in rooting phase, one out of a total of about 1500 plants survived. This plant both carried and expressed the transferred nptII gene. The integration was confirmed by Southern blot hybridization. This plant was not fertile.

AB - Transgenic barley plants (Hordeum vulgare L. cv. Kymppi) were obtained by particle bombardment of various tissues. Immature embryos and microspore-derived cultures were bombarded with gold particles coated with plasmid DNA carrying the gene coding for neomycin phosphotransferase II (NPTII), together with plasmid DNA containing the gene for β-glucuronidase (GUS).Bombarded immature embryos were grown to plants without selection and NPTII activity was screened in small plantlets. One plant proved to be transgenic (T0). This chimeric plant passed the transferred nptII gene to its T1 progeny. The presence of the nptII gene was demonstrated by the PCR technique and enzyme activity was analyzed by an NPTII gel assay. Four T0 spikes and 15 T1 offspring were transgenic. The integration and inheritance was confirmed by Southern blot hybridization. Transgenic T2 and T3 plants were produced by isolating embryos from green grains of transgenic T1 and T2 plants, respectively and growing them to plants. After selfing, the ratio of transgenic to non-transgenic T2 offspring was shown to follow the rule of Mendelian inheritance. The general performance of transgenic plants was normal and no reduction in fertility was observed.Microspore-derived cultures were bombarded one and four weeks after microspore isolation. After bombardment, cultures were grown either with or without antibiotic selection (geneticin R or kanamycin). When cultures were grown without selection and regenerated plants were transferred to kanamycin selection in rooting phase, one out of a total of about 1500 plants survived. This plant both carried and expressed the transferred nptII gene. The integration was confirmed by Southern blot hybridization. This plant was not fertile.

U2 - 10.1007/BF00023933

DO - 10.1007/BF00023933

M3 - Article

VL - 85

SP - 81

EP - 88

JO - Euphytica

JF - Euphytica

SN - 0014-2336

IS - 1-3

ER -