Protoplast culture and plant regeneration in Glycine canescens F.J. Herm

Newell, C. A.; Luu, H. T.

doi:10.1007/bf00042272

Cited by 38 publications

(10 citation statements)

References 17 publications

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“…Protoplasts isolated from a wild relative of soybean, G. canescens, by this method behaved similarly to those isolated from the cultivars in assays of transient gene expression. This observation is significant because regeneration of this species from protoplasts has been reported (22).…”

Section: Resultsmentioning

confidence: 85%

Stable transformation of soybean by electroporation and root formation from transformed callus

Christou¹,

Murphy²,

Swain³

1987

Proc. Natl. Acad. Sci. U.S.A.

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Soybean protoplasts from a number of commercially important cultivars have been genetically engineered by way of electroporation using chimeric genes coding for resistance to the aminoglycoside antibiotics kanamycin and G418. Effective electroporation conditions were determined by monitoring transient expression from aminoglycoside 3'-phosphotransferase II (APHII) expression plasmids. Electroporation of protoplasts with a chimeric APHII gene and subsequent selection on media supplemented with kanamycin resulted in the recovery of calli resistant to the antibiotic. Enzyme assays for APHII activity and Southern blot hybridization confirmed the expression of the foreign DNA and its stable integration into the soybean genome. Root formation was induced from transformed calli, and these roots maintained expression of the APHII gene.Soybean (Glycine max) is one of the world's most important agronomic crops. Accordingly, a great deal of effort has been directed towards its genetic improvement by both conventional breeding techniques and genetic engineering approaches. Successful application of standard genetic engineering procedures to soybean has been limited by the lack of an efficient transformation system and the inability to regenerate transformed tissues. Oncogenic transformation of soybean by virulent Agrobacterium strains and the axenic culture of excised tumors on hormone-free media have been reported (1-5), but disarmed Ti vectors have not yet been used successfully. Agrobacterium strains have been isolated that show increased virulence toward soybean and other plants that are not particularly susceptible to crown gall infection, and "disarmed" hosts for Agrobacterium vector systems have been developed from these strains (6). However, even with these strains, the transformation frequency is very low relative to that obtained in alternative plant hosts such as tobacco and petunia. Electroporation has been shown to be an effective technique for the transformation of mammalian cells (7). Fromm et al. (8) have shown that with appropriate modification the technique is also applicable to plant protoplasts, and they have achieved the stable transformation of maize tissue cultures by this method (9). These results, together with our own experience with electroporation of tobacco, prompted us to investigate this method as a means of achieving the transformation of soybean. MATERIALS AND METHODSPlasmids. pCMC1021 was constructed from a fragment of pBR322 [nucleotides 2522-4361 (10)] containing the replication origin and the 8-lactamase gene, a chimeric gene consisting of 5' and 3' regulatory regions from the nopaline synthase gene of pTiT37 [nucleotides -265 to +36 and + 1298 to +1550, respectively (11)], and the aminoglycoside 3'-phosphotransferase II (APHII; kanamycin kinase, EC 2.7.1.95) coding region from TnS [nucleotides 1541-2517 (12)]. The coding region deletes an AUG triplet present in the TnS sequence upstream and out-of-frame with respect to the initiator AUG (13). The junctions between the pBR32...

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Section: Resultsmentioning

confidence: 85%

Stable transformation of soybean by electroporation and root formation from transformed callus

Christou¹,

Murphy²,

Swain³

1987

Proc. Natl. Acad. Sci. U.S.A.

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“…While perennial Glycine species have been regenerated from complex explants [8,10,11,23] and also from protoplasts [17,18], until recently attempts to regenerate G. max had yielded only non-functional somatic embryos [4,19]. Using immature embryos as explants, however, several instances of somatic embryogenesis [5,14,15] and whole plant regeneration [3,12,20] have now been reported.…”

Section: Introductionmentioning

confidence: 99%

Soybean somatic embryogenesis: Effects of hormones and culture manipulations

Lazzeri

Hildebrand

Collins

1987

Plant Cell Tiss Organ Cult

132

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Somatic embryos were induced in cultures of immature soybean (Glycine max (L.) Merr) embryos, or isolated cotyledons on MS modified medium supplemented with NAA and 2,4-D, BAP and ABA. When NAA and 2,4-D were compared at similar concentrations (25 and 23 #M), 2,4-D produced larger number of somatic embryos, however, embryogenesis efficiency was improved in media containing NAA by using higher levels (100-150 #M) of the auxin. Somatic embryo morphology varied with auxin type: NAA-induced embryos more closely resembled zygotic embryos than did 2,4-D-induced embryos. Additions of BAP or ABA to auxin-containing media had either no effect or reduced embryo production, although ABA altered the morphology of 2,4-D-induced embryos. In media containing both NAA and 2,4-D, the latter was dominant in terms of embryo morphology. The effects of subculture frequency and of transfers between 2,4-D and NAA media were investigated: Subculture frequency influenced mainly the frequency of normal embryos, while preculture on 2,4-D increased subsequent embryogenesis efficiency on NAA medium but reduced the frequency of normal embryos.

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“…Somatic embryogenesis has also been described as a pathway for plant regeneration in embryo cotyledon callus of G. canescens (Grant 1984). Recently, Newell and Luu (1985) made the first report of shoot regeneration from protoplasts in the genus Glycine, using hypocotyl protoplasts from G. canescens. This paper describes rapid, high frequency regeneration of shoots and whole plants from seedling cotyledon and adult leaf and petiole tissues of G. clandestina, a perennial, climbing relative of the soybean.…”

Section: Introductionmentioning

confidence: 99%

Plant regeneration from seedling cotyledons, leaves and petioles of Glycine clandestina

Hammatt

Davey

Nelson

1986

Physiologia Plantarum

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Hammatt, N., Davey, M. R. and Nelson, R. S. 1986. Plant regeneration from seedling cotyledons, leaves and petioles of Glycine clandestina. -Physiol. Plant. 68: 125-128.A B5-based culture medium containing 4.4 (iM N'-benzyladenine (BA) and 0.025 \JM indole-3-butyric add {IBA) indueed <^us from seedling cotyledons, leaves and petioles of Glycine ciandestina Wendl. Only hard, green, nodular callus tissues were capable of producing shoot buds and of five accessions examined, only two (G1231 and G1145) were morphogenetically competent. Callus that did not regenerate could often be induced to produce shoot buds after subculture to fresh regeneration medium. Buds developed into shoots following transfer of callus to a medium containing 0.9 (iM BA and 0.025 fiAf IBA. Shoots were rooted in hormone-free, half-strengtb B5 medium supplemented with 0.2% activated charcoal. The application of these results is discussed in relation to somatic hybridisation between the cultivated soybean and wild Glycine species.

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Protoplast culture and plant regeneration in Glycine canescens F.J. Herm

Cited by 38 publications

References 17 publications

Stable transformation of soybean by electroporation and root formation from transformed callus

Stable transformation of soybean by electroporation and root formation from transformed callus

Soybean somatic embryogenesis: Effects of hormones and culture manipulations

Plant regeneration from seedling cotyledons, leaves and petioles of Glycine clandestina

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