Effect of urea-type cytokinins on the adventitious shoots regeneration from cotyledonary node explant in the common ice plant, Mesembryanthemum crystallinum

Haruki Sunagawa, Sakae Agarie, Makiko Umemoto, Yuko Makishi, Akihiro Nose

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Mesembryanthemum crystallinum (common ice plant) was used as a model plant to study the regulatory properties of Crassulacean acid metabolism (CAM) and tolerance to abiotic stresses. Although transformation is a useful genetic approach, it has not been established in this species due to recalcitrancy for regeneration. To establish an efficient procedure for regeneration of M. crystallinum, we examined the effects of urea-type cytokinins, thidiazuron (TDZ) and forchlorofenuron (CPPU) on the adventitious shoot induction. Adventitious shoots were generated only from explants obtained from the cotyledonary node, not from explants obtained from the cotyledon, hypocotyl and roots. Urea-type cytokinins, TDZ and CPPU were more effective for the induction and the morphogenesis of adventitious shoots than adenine-type cytokinin, 6-benzyladenopurine (BA). We have found that the 2.5 mg L-1 TDZ induced the largest number of multiple shoots and the highest frequency of adventitious shoot induction from single explant. In addition, fewer hyperhydric shoots were produced on the medium containing TDZ than in that containing BA and CPPU in the presence of 1.0 mg L-1 NAA. The regenerated shoots rooted on the MS medium within one month, and the rooting was promoted by replacing the agar medium with vermiculite or Florialite. The fertile plant with normal morphological properties was harvested for four months after sowing. Using the improved regeneration procedure with TDZ, we successfully introduced a kanamycin-resistant gene (nptII-HPH) into the cotyledonary node mediated by Agrobacterium tumefaciens. These results indicated that this regeneration procedure using cotyledonary node explants and TDZ could be useful for the genetic engineering of M. crystallinum.

Original languageEnglish
Pages (from-to)47-56
Number of pages10
JournalPlant Production Science
Volume10
Issue number1
DOIs
Publication statusPublished - Feb 1 2007
Externally publishedYes

Fingerprint

Mesembryanthemum crystallinum
adventitious shoots
cytokinins
explants
urea
shoots
Crassulacean acid metabolism
vermiculite
kanamycin
adenine
Agrobacterium radiobacter
genetic engineering
naphthaleneacetic acid
hypocotyls
abiotic stress
morphogenesis
thidiazuron
cotyledons
rooting
sowing

All Science Journal Classification (ASJC) codes

  • Agronomy and Crop Science

Cite this

Effect of urea-type cytokinins on the adventitious shoots regeneration from cotyledonary node explant in the common ice plant, Mesembryanthemum crystallinum. / Sunagawa, Haruki; Agarie, Sakae; Umemoto, Makiko; Makishi, Yuko; Nose, Akihiro.

In: Plant Production Science, Vol. 10, No. 1, 01.02.2007, p. 47-56.

Research output: Contribution to journalArticle

@article{9b06b795d6594b0fa3d217a463b4dd1c,
title = "Effect of urea-type cytokinins on the adventitious shoots regeneration from cotyledonary node explant in the common ice plant, Mesembryanthemum crystallinum",
abstract = "Mesembryanthemum crystallinum (common ice plant) was used as a model plant to study the regulatory properties of Crassulacean acid metabolism (CAM) and tolerance to abiotic stresses. Although transformation is a useful genetic approach, it has not been established in this species due to recalcitrancy for regeneration. To establish an efficient procedure for regeneration of M. crystallinum, we examined the effects of urea-type cytokinins, thidiazuron (TDZ) and forchlorofenuron (CPPU) on the adventitious shoot induction. Adventitious shoots were generated only from explants obtained from the cotyledonary node, not from explants obtained from the cotyledon, hypocotyl and roots. Urea-type cytokinins, TDZ and CPPU were more effective for the induction and the morphogenesis of adventitious shoots than adenine-type cytokinin, 6-benzyladenopurine (BA). We have found that the 2.5 mg L-1 TDZ induced the largest number of multiple shoots and the highest frequency of adventitious shoot induction from single explant. In addition, fewer hyperhydric shoots were produced on the medium containing TDZ than in that containing BA and CPPU in the presence of 1.0 mg L-1 NAA. The regenerated shoots rooted on the MS medium within one month, and the rooting was promoted by replacing the agar medium with vermiculite or Florialite. The fertile plant with normal morphological properties was harvested for four months after sowing. Using the improved regeneration procedure with TDZ, we successfully introduced a kanamycin-resistant gene (nptII-HPH) into the cotyledonary node mediated by Agrobacterium tumefaciens. These results indicated that this regeneration procedure using cotyledonary node explants and TDZ could be useful for the genetic engineering of M. crystallinum.",
author = "Haruki Sunagawa and Sakae Agarie and Makiko Umemoto and Yuko Makishi and Akihiro Nose",
year = "2007",
month = "2",
day = "1",
doi = "10.1626/pps.10.47",
language = "English",
volume = "10",
pages = "47--56",
journal = "Plant Production Science",
issn = "1343-943X",
publisher = "Crop Science Society of Japan",
number = "1",

}

TY - JOUR

T1 - Effect of urea-type cytokinins on the adventitious shoots regeneration from cotyledonary node explant in the common ice plant, Mesembryanthemum crystallinum

AU - Sunagawa, Haruki

AU - Agarie, Sakae

AU - Umemoto, Makiko

AU - Makishi, Yuko

AU - Nose, Akihiro

PY - 2007/2/1

Y1 - 2007/2/1

N2 - Mesembryanthemum crystallinum (common ice plant) was used as a model plant to study the regulatory properties of Crassulacean acid metabolism (CAM) and tolerance to abiotic stresses. Although transformation is a useful genetic approach, it has not been established in this species due to recalcitrancy for regeneration. To establish an efficient procedure for regeneration of M. crystallinum, we examined the effects of urea-type cytokinins, thidiazuron (TDZ) and forchlorofenuron (CPPU) on the adventitious shoot induction. Adventitious shoots were generated only from explants obtained from the cotyledonary node, not from explants obtained from the cotyledon, hypocotyl and roots. Urea-type cytokinins, TDZ and CPPU were more effective for the induction and the morphogenesis of adventitious shoots than adenine-type cytokinin, 6-benzyladenopurine (BA). We have found that the 2.5 mg L-1 TDZ induced the largest number of multiple shoots and the highest frequency of adventitious shoot induction from single explant. In addition, fewer hyperhydric shoots were produced on the medium containing TDZ than in that containing BA and CPPU in the presence of 1.0 mg L-1 NAA. The regenerated shoots rooted on the MS medium within one month, and the rooting was promoted by replacing the agar medium with vermiculite or Florialite. The fertile plant with normal morphological properties was harvested for four months after sowing. Using the improved regeneration procedure with TDZ, we successfully introduced a kanamycin-resistant gene (nptII-HPH) into the cotyledonary node mediated by Agrobacterium tumefaciens. These results indicated that this regeneration procedure using cotyledonary node explants and TDZ could be useful for the genetic engineering of M. crystallinum.

AB - Mesembryanthemum crystallinum (common ice plant) was used as a model plant to study the regulatory properties of Crassulacean acid metabolism (CAM) and tolerance to abiotic stresses. Although transformation is a useful genetic approach, it has not been established in this species due to recalcitrancy for regeneration. To establish an efficient procedure for regeneration of M. crystallinum, we examined the effects of urea-type cytokinins, thidiazuron (TDZ) and forchlorofenuron (CPPU) on the adventitious shoot induction. Adventitious shoots were generated only from explants obtained from the cotyledonary node, not from explants obtained from the cotyledon, hypocotyl and roots. Urea-type cytokinins, TDZ and CPPU were more effective for the induction and the morphogenesis of adventitious shoots than adenine-type cytokinin, 6-benzyladenopurine (BA). We have found that the 2.5 mg L-1 TDZ induced the largest number of multiple shoots and the highest frequency of adventitious shoot induction from single explant. In addition, fewer hyperhydric shoots were produced on the medium containing TDZ than in that containing BA and CPPU in the presence of 1.0 mg L-1 NAA. The regenerated shoots rooted on the MS medium within one month, and the rooting was promoted by replacing the agar medium with vermiculite or Florialite. The fertile plant with normal morphological properties was harvested for four months after sowing. Using the improved regeneration procedure with TDZ, we successfully introduced a kanamycin-resistant gene (nptII-HPH) into the cotyledonary node mediated by Agrobacterium tumefaciens. These results indicated that this regeneration procedure using cotyledonary node explants and TDZ could be useful for the genetic engineering of M. crystallinum.

UR - http://www.scopus.com/inward/record.url?scp=33846540651&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33846540651&partnerID=8YFLogxK

U2 - 10.1626/pps.10.47

DO - 10.1626/pps.10.47

M3 - Article

AN - SCOPUS:33846540651

VL - 10

SP - 47

EP - 56

JO - Plant Production Science

JF - Plant Production Science

SN - 1343-943X

IS - 1

ER -