Shoot meristem

An ideal explant for Zea mays L. transformation

Sairam V. Rudrabhatla, M. Parani, G. Franklin, Z. Lifeng, B. Smith, J. MacDougall, C. Wilber, H. Sheikhi, N. Kashikar, K. Meeker, D. Al-Abed, K. Berry, R. Vierling, S. L. Goldman

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

We report on a rapid high-frequency somatic embryogenesis and plant regeneration protocol for Zea mays. Maize plants were regenerated from complete shoot meristem (3-4 mm) explants via organogenesis and somatic embryogenesis. In organogenesis, the shoot meristems were directly cultured on a high-cytokinin medium comprising 5-10 mg·L-1 6-benzylaminopurine (BAP). The number of multiple shoots produced per meristem varied from six to eight. Plantlet regeneration through organogenesis resulted in just four weeks. Callus was induced in five days of incubation on an auxin-modified Murashige and Skoog (MS) medium. Prolific callus, with numerous somatic embryos, developed within 3-4 weeks when cultured on an auxin medium containing 5 mg 2,4-dichlorophenoxyacetic acid·L-1. The number of multiple shoots varied from three to six per callus. Using R23 (Pioneer, Hi-Bred, Johnston, Iowa), the frequency of callus induction was consistently in excess of 80% and plant regeneration ranged between 47 and 64%. All regenerated plantlets survived in the greenhouse and produced normal plants. Each transgenic plant produced leaves, glumes, and anthers that uniformly expressed green fluorescent protein (GFP). The GFP gene segregated in the pollen. Based on this data it is concluded that the transgenics arose from single-cell somatic embryos. The rate of transfer DNA (T-DNA) transfer to complete shoot meristems of Zea mays was high on the auxin medium and was independent of using super-virulent strains of Agrobacterium.

Original languageEnglish (US)
Pages (from-to)323-329
Number of pages7
JournalGenome
Volume46
Issue number2
DOIs
StatePublished - Apr 2003

Fingerprint

Meristem
Bony Callus
Zea mays
Indoleacetic Acids
Organogenesis
Regeneration
N-benzyladenine
Green Fluorescent Proteins
Plant Somatic Embryogenesis Techniques
Embryonic Structures
Cytokinins
Agrobacterium
Plant Leaves
Genetically Modified Plants
Pollen
Embryonic Development
DNA
Genes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Molecular Biology
  • Genetics

Cite this

Rudrabhatla, S. V., Parani, M., Franklin, G., Lifeng, Z., Smith, B., MacDougall, J., ... Goldman, S. L. (2003). Shoot meristem: An ideal explant for Zea mays L. transformation. Genome, 46(2), 323-329. https://doi.org/10.1139/g02-120
Rudrabhatla, Sairam V. ; Parani, M. ; Franklin, G. ; Lifeng, Z. ; Smith, B. ; MacDougall, J. ; Wilber, C. ; Sheikhi, H. ; Kashikar, N. ; Meeker, K. ; Al-Abed, D. ; Berry, K. ; Vierling, R. ; Goldman, S. L. / Shoot meristem : An ideal explant for Zea mays L. transformation. In: Genome. 2003 ; Vol. 46, No. 2. pp. 323-329.
@article{7dd36676b11547bca97945b64ac63bb8,
title = "Shoot meristem: An ideal explant for Zea mays L. transformation",
abstract = "We report on a rapid high-frequency somatic embryogenesis and plant regeneration protocol for Zea mays. Maize plants were regenerated from complete shoot meristem (3-4 mm) explants via organogenesis and somatic embryogenesis. In organogenesis, the shoot meristems were directly cultured on a high-cytokinin medium comprising 5-10 mg·L-1 6-benzylaminopurine (BAP). The number of multiple shoots produced per meristem varied from six to eight. Plantlet regeneration through organogenesis resulted in just four weeks. Callus was induced in five days of incubation on an auxin-modified Murashige and Skoog (MS) medium. Prolific callus, with numerous somatic embryos, developed within 3-4 weeks when cultured on an auxin medium containing 5 mg 2,4-dichlorophenoxyacetic acid·L-1. The number of multiple shoots varied from three to six per callus. Using R23 (Pioneer, Hi-Bred, Johnston, Iowa), the frequency of callus induction was consistently in excess of 80{\%} and plant regeneration ranged between 47 and 64{\%}. All regenerated plantlets survived in the greenhouse and produced normal plants. Each transgenic plant produced leaves, glumes, and anthers that uniformly expressed green fluorescent protein (GFP). The GFP gene segregated in the pollen. Based on this data it is concluded that the transgenics arose from single-cell somatic embryos. The rate of transfer DNA (T-DNA) transfer to complete shoot meristems of Zea mays was high on the auxin medium and was independent of using super-virulent strains of Agrobacterium.",
author = "Rudrabhatla, {Sairam V.} and M. Parani and G. Franklin and Z. Lifeng and B. Smith and J. MacDougall and C. Wilber and H. Sheikhi and N. Kashikar and K. Meeker and D. Al-Abed and K. Berry and R. Vierling and Goldman, {S. L.}",
year = "2003",
month = "4",
doi = "10.1139/g02-120",
language = "English (US)",
volume = "46",
pages = "323--329",
journal = "Genome",
issn = "0831-2796",
publisher = "National Research Council of Canada",
number = "2",

}

Rudrabhatla, SV, Parani, M, Franklin, G, Lifeng, Z, Smith, B, MacDougall, J, Wilber, C, Sheikhi, H, Kashikar, N, Meeker, K, Al-Abed, D, Berry, K, Vierling, R & Goldman, SL 2003, 'Shoot meristem: An ideal explant for Zea mays L. transformation', Genome, vol. 46, no. 2, pp. 323-329. https://doi.org/10.1139/g02-120

Shoot meristem : An ideal explant for Zea mays L. transformation. / Rudrabhatla, Sairam V.; Parani, M.; Franklin, G.; Lifeng, Z.; Smith, B.; MacDougall, J.; Wilber, C.; Sheikhi, H.; Kashikar, N.; Meeker, K.; Al-Abed, D.; Berry, K.; Vierling, R.; Goldman, S. L.

In: Genome, Vol. 46, No. 2, 04.2003, p. 323-329.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Shoot meristem

T2 - An ideal explant for Zea mays L. transformation

AU - Rudrabhatla, Sairam V.

AU - Parani, M.

AU - Franklin, G.

AU - Lifeng, Z.

AU - Smith, B.

AU - MacDougall, J.

AU - Wilber, C.

AU - Sheikhi, H.

AU - Kashikar, N.

AU - Meeker, K.

AU - Al-Abed, D.

AU - Berry, K.

AU - Vierling, R.

AU - Goldman, S. L.

PY - 2003/4

Y1 - 2003/4

N2 - We report on a rapid high-frequency somatic embryogenesis and plant regeneration protocol for Zea mays. Maize plants were regenerated from complete shoot meristem (3-4 mm) explants via organogenesis and somatic embryogenesis. In organogenesis, the shoot meristems were directly cultured on a high-cytokinin medium comprising 5-10 mg·L-1 6-benzylaminopurine (BAP). The number of multiple shoots produced per meristem varied from six to eight. Plantlet regeneration through organogenesis resulted in just four weeks. Callus was induced in five days of incubation on an auxin-modified Murashige and Skoog (MS) medium. Prolific callus, with numerous somatic embryos, developed within 3-4 weeks when cultured on an auxin medium containing 5 mg 2,4-dichlorophenoxyacetic acid·L-1. The number of multiple shoots varied from three to six per callus. Using R23 (Pioneer, Hi-Bred, Johnston, Iowa), the frequency of callus induction was consistently in excess of 80% and plant regeneration ranged between 47 and 64%. All regenerated plantlets survived in the greenhouse and produced normal plants. Each transgenic plant produced leaves, glumes, and anthers that uniformly expressed green fluorescent protein (GFP). The GFP gene segregated in the pollen. Based on this data it is concluded that the transgenics arose from single-cell somatic embryos. The rate of transfer DNA (T-DNA) transfer to complete shoot meristems of Zea mays was high on the auxin medium and was independent of using super-virulent strains of Agrobacterium.

AB - We report on a rapid high-frequency somatic embryogenesis and plant regeneration protocol for Zea mays. Maize plants were regenerated from complete shoot meristem (3-4 mm) explants via organogenesis and somatic embryogenesis. In organogenesis, the shoot meristems were directly cultured on a high-cytokinin medium comprising 5-10 mg·L-1 6-benzylaminopurine (BAP). The number of multiple shoots produced per meristem varied from six to eight. Plantlet regeneration through organogenesis resulted in just four weeks. Callus was induced in five days of incubation on an auxin-modified Murashige and Skoog (MS) medium. Prolific callus, with numerous somatic embryos, developed within 3-4 weeks when cultured on an auxin medium containing 5 mg 2,4-dichlorophenoxyacetic acid·L-1. The number of multiple shoots varied from three to six per callus. Using R23 (Pioneer, Hi-Bred, Johnston, Iowa), the frequency of callus induction was consistently in excess of 80% and plant regeneration ranged between 47 and 64%. All regenerated plantlets survived in the greenhouse and produced normal plants. Each transgenic plant produced leaves, glumes, and anthers that uniformly expressed green fluorescent protein (GFP). The GFP gene segregated in the pollen. Based on this data it is concluded that the transgenics arose from single-cell somatic embryos. The rate of transfer DNA (T-DNA) transfer to complete shoot meristems of Zea mays was high on the auxin medium and was independent of using super-virulent strains of Agrobacterium.

UR - http://www.scopus.com/inward/record.url?scp=12444293405&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=12444293405&partnerID=8YFLogxK

U2 - 10.1139/g02-120

DO - 10.1139/g02-120

M3 - Article

VL - 46

SP - 323

EP - 329

JO - Genome

JF - Genome

SN - 0831-2796

IS - 2

ER -

Rudrabhatla SV, Parani M, Franklin G, Lifeng Z, Smith B, MacDougall J et al. Shoot meristem: An ideal explant for Zea mays L. transformation. Genome. 2003 Apr;46(2):323-329. https://doi.org/10.1139/g02-120