PHASEOMICS TRANSFORMATION WORKGROUP

Introduction

On the 2nd Phaseomics meeting in Geneva (May 2002) it was suggested that different working groups could be formed to concentrate on certain aspect of the Phaseomics goals. One of this is a workgroup on transformation, as this is still a problematic area. The idea is to keep in touch through e-mail on recent developments and to help each other with suggestions and advice, and to avoid duplications of unsuccessful attempts. Nancy Terryn of IPBO, Ghent University, Belgium will co-ordinate these efforts. This web site will provide a literature overview on the topic and links to related sites.
 

 

Why this web site

To present the status of Phaseolus transformation to the Phaseomics group in particular, and other interested parties in general, based on discussions on the Phaseomics meeting in Geneva (may 2002) and the meeting on " Legume genetics and genomics" in Minneapolis (June2002) attended by several Phaseomics members..

Members of the workgroup in Phaseomics that discussed in Geneva: Nancy Terryn, Alvaro Mejia, Farida Shah, Gina Hernandez, Miguel Lara, Jos Van Der Leyden, Tsai Siu Mui, Alexandre Bartsev. Anybody of Phaseomics (or outside) that wants to contribute is off course welcome to join the working group. Just mail to Nancy to have you added to the list.

Status

First we should stress that our working group focuses on Phaseolus transformation through Agrobacterium mediated protocols, however an update on some other approaches will be given:

1. Agrobacterium

 We should distinguish between Phaseolus vulgaris and Phaseolusacutifolius.

For P.acutifolius it is now clear that two labs can routinely transform this, that is the group in Gent (Nancy Terryn took over from Geert Angenon) (see refs below) and the group of CIAT (Alvaro)(paper in preparation). Most work is done with NI576, a wild P.acutifolius line, whereas the Gent group is now also able to use a more cultivated variety. Both groups can be contacted if you wish to introduce your gene in Phaseolus acutifolius.
P. acutifolius can be crossed to P.vulgaris throughembryo rescue.

Work is in progress in CIAT on hybrid lines between P.acutifolius and P.vulgaris to test there transformability. This could make it easier to cross transgenics obtained in these lines with P.vulgaris varieties of interest. 

For P.vulgaris different memebers of the working group will continue their efforts. The group in Ghent will continue, based on their callus based protocol, to try to obtain transformation of P.vulgaris. The group of Cuernavaca, Mexico (Gina and Miguel) have developed a regeneration protocol via organogenesis for five varieties of P. vulgaris widely grown in Mexico, using cotyledonary nodes (cot-nodes) as explants. Based on this, they will work on Agrobacterium-based transformation, following the cot-nod transformation system established for soybean by the group of Dave Somers (U. Minnesota).
Farida and her colleague Ruslan Abdullah might work with embryonic cultures as she is successful in for oil palm.
 

Figure :  P acutifolius callus with a shoot emerging (photo W.Dillen and M. Zambre)

2. Particle bombardment

With particle bombardment EMBRAPA in Brazil has obtained stable lines that are now introgressed in breeding varieties (see refs below). The frequency of transformation is however very low, which reinforces the importance to continue working on different approaches to obtain an efficient, preferentially agrobacterium based transformation system.
 

3. Floral Transformation

In the meeting in Minneapolis Andrew Bent gave a talk on Soybean transformation, which basically came down to the fact that after 5 years he is still unsuccessful. So if anybody wants to try floral dip they should look at his work first.

4. References

Particle bombardment:

Aragao FJL, Vianna GR, Albino MMC, Rech EL Transgenic dry bean tolerant to the herbicide glufosinate ammonium CROP SCIENCE 42: (4) 1298-1302 JUL-AUG 2002

Aragao FJL, Barros LMG, de Sousa MV, et al. Expression of a methionine-rich storage albumin from the Brazil nut (Bertholletia excelsa HBK, Lecythidaceae) in transgenic bean plants (Phaseolus vulgaris L., Fabaceae) GENET MOL BIOL 22: (3) 445-449 SEP 1999

Aragao FJL, Ribeiro SG, Barros LMG, et al. Transgenic beans (Phaseolus vulgaris L.) engineered to express viral antisense RNAs show delayed and attenuated symptoms to bean golden mosaic geminivirus MOL BREEDING 4: (6) 491-499 1998

Aragao FJL, Barros LMG, Brasileiro ACM, et al. Inheritance of foreign genes in transgenic bean (Phaseolus vulgaris L) co-transformed via particle bombardment THEOR APPL GENET 93: (1-2) 142-150 JUL 1996

Kim JW, Minamikawa T (1996) Transformation and regeneration of French bean plant by the particle bombardment process. Plant Sci 117:131-138

Russell DR, Wallece KM, Bathe JH, Martinell BJ, McCabe DE (1993) Stable transformation of Phaseolus vulgaris L. via electric-discharge mediated particle acceleration. Plant Cell Rep 12: 165-169

P. acutifolius transformation:

Dillen W, DeClercq J, Goossens A, Van Montagu M and Angenon Gl. Agrobacterium-mediated transformation of Phaseolus acutifolius A Gray THEOR APPL GENET 94: (2) 151-158 FEB 1997

Goossens A, Dillen W, De Clercq J, Van Montagu M and Angenon G .The arcelin-5 gene of Phaseolus vulgaris directs high seed-specific expression in transgenic Phaseolus acutifolius and Arabidopsis plants PLANT PHYSIOL 120: (4) 1095-1103 AUG 1999

Goossens A, Quintero C, Dillen W, Van Montagu M and Angenon G. Analysis of bruchid resistance in the wild common bean accession G02771: no evidence for insecticidal activity of arcelin 5 J EXP BOT 51: (348) 1229-1236 JUL 2000

De Jaeger G., Scheffer S., Jacobs A., Zambre M., Zobell O., Goossens A, Depicker A and Angenon G. Boosting heterologous protein production in transgenic dicotyledenous seeds using Phaseolsus vulgaris regulatory sequences (2002) NATURE BIOTECHNOLOGY Vol20, p1265

De Clerq J., Zambre M., Van Montagu M., Dillen W.  and Angenon G. An optimised Agrobacterium-mediated transformation procedure for Phaseolsus acutifolius A Gray (tepary bean) Plant Cell reports 2002  21(4):333-340

Zambre M., Terryn N., De Clercq J., De Buck S., Dillen W., Van Montagu M., Van Der Straeten D. And Angenon G (2002) Light strongly promotes gene transfer from Agrobacterium tumefaciens to plant cells. Planta (2003) 216: 580-586

Zambre M., Goossens M., Van Montagu M. and Angenon G. An efficient and reproducible Agrobacterium-mediated trasnformation sytems for cultivated Phaseolus acutifolius (tepary bean) submitted

De Clercq J., Hamelryck T., Zambre M., Goossens A., Van Montagu M., Terryn N.and Angenon G. (2003). Increasing the methionine content of Phaseolus beans
through modification of the P. vulgaris seed protein Arcelin 5a. In preparation.
 
 

P.vulgaris regeneration:

Zambre MA, De Clercq J, Vranova E, et al. Plant regeneration from embryo derived callus in Phaseolus vulgaris L. (common bean) and P-acutifolius A Gray (tepary bean) PLANT CELL REP 17: (8) 626-630 MAY 1998

de Carvalho MHC, Le BV, Zuily-Fodil Y, et al. Efficient whole plant regeneration of common bean (Phaseolus vulgaris L.) using thin-cell-layer culture and silver nitrate PLANT SCI 159: (2) 223-232 NOV 6 2000

Mejia-Jimenez, A., H. J. Jacobsen and W. M. Roca (1998). Development of an in vitro regeneration system in common bean suitable for genetic transformation. EUCARPIA International symposium on breeding of protein and oil crops, Pontevedra - Spain 1-4 April.

Franklin CI, Trieu TN, Gonzales RA and Dixon RA (1991) Plant regeneration from seedling explants of green bean (Phaseolus vulgaris L.) via organogenesis. Plant Cell Tiss Org Cult 24: 199-206

Santalla, Marta, J. Brian Power, Michael R. Davey. 1998. Efficient in vitro shoot regeneration responses of Phaseolus vulgaris and Phaseolus coccineus. Enphytica 102: 195-202

P.vulgaris transformation through Agrobacterium:

Factors affecting Agrobacterium-mediated transformation of common bean. Zhang ZY, Coyne DP, Mitra A JOURNAL OF THE AMERICAN SOCIETY FOR HORTICULTURAL SCIENCE 122: (3) 300-305 MAY 1997

McClean P, Chee P, Held B, Simental J, Drong RF, Slightom J (1991) Susceptibility of dry bean (Phaseolus vulgaris L.) to Agrobacterium infection: Transformation of cotyledonary hypocotyl tissues. Plant Cell Tiss Org Cult 24: 131-138

P.vulgaris-P.acutifolius hybridization:

Mejía-Jiménez, A., Muñoz, C. Jacobsen, H.J., Roca, W.M. and Singh, S.P. 1994. Interspecific hybridization between common bean and tepary bean: Increased hybrid embryo growth, fertility, and efficiency of hybridization through recurrent and congruity backcrossing. Theor. Appl. Genet. 88: 324-331.
 
 

A list with transformation papers from other legumes will be added later.
Any suggestions or additions please mail to nater@gengenp.rug.ac.be