Abstract
A rapid regeneration protocol for proembryos of Phaseolus angustissimus as young as 1 day after pollination (DAP) involving pod culture for 1 week followed by embryo culture for 2 weeks and embryo germination for 1 or 2 weeks is provided. Optimization of the media was conducted with pods collected 3 DAP. The best pod culture medium was composed of basal medium [(Phillips and Collins 1979) salts with (Geerts et al. 2001) vitamins], 1000 mg l−1 glutamine, 1000 mg l−1 casein hydrolysate, 3% sucrose and 0.5% agar. Embryo culture medium consisted of basal medium with 500 mg l−1 glutamine, 250 mg l−1 casein hydrolysate, 1.9 μM ABA, 3% sucrose and 0.5% bacto-agar. Embryos developed into plantlets on germination medium containing basal medium with 0.25 μM BA, 3% sucrose and 0.7% bacto-agar. Fertile, normal plants were recovered from direct embryogenesis and from micrografted embryo-derived shoots. Embryos obtained from pods collected 3 DAP regenerated plantlets at a rate of 29.3%, while embryos from pods collected 2 DAP and 1 DAP regenerated at rates of 20.2 and 4%, respectively. A second accession of P. angustissimusregenerated at a rate of 26.2%. Using this 5-week protocol for P. vulgaris resulted in a plantlet regeneration rate of 12.5%.
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Abbreviations
- ABA:
-
abscisic acid
- BA:
-
6-benzylaminopurine
- BM:
-
basal medium
- DAP:
-
days after pollination
- ECM:
-
embryo culture medium
- GM:
-
germination medium
- HAP:
-
hours after pollination
- PCM:
-
pod culture medium
References
P Balasubramanian (2002) Selection for chilling and freezing resistance in common bean University of Saskatchewan Saskatoon Canada
P Balasubramanian A Vandenberg P Hucl L Gusta (2002) ArticleTitlePhysiology of freezing resistance in the genus Phaseolus Annu. Rep. Bean Improv. Coop. 45 6–7
P Balasubramanian A Vandenberg P Hucl L Gusta (2004) ArticleTitleResistance of Phaseolus species to ice crystallization at subzero temperature Physiol. Plant 120 451–457 Occurrence Handle10.1111/j.0031-9317.2004.00257.x Occurrence Handle15032842
T Belivanis C Doré (1986) ArticleTitleInterspecific hybridization of Phaseolus vulgaris L. and P. angustissimus A Gray using in vitro embryo culture. Plant Cell Rep. 5 329–331 Occurrence Handle10.1007/BF00268593
MP Bridgen (1994) ArticleTitleA review of plant embryo culture HortScience 29 1243–1246
P Geerts JP Baudoin G Mergeai (1999) ArticleTitleRescue of early heart-shaped embryos and plant regeneration of Phaseolus polyanthus Greenm and Phaseolus vulgaris L Biotechnol. Agron. Soc. Environ. 3 141–148
P Geerts K Sassi G Mergeai JP Baudoin (2000) ArticleTitleDevelopment of an in vitro pod culture technique for young pods of Phaseolus vulgaris In Vitro Cell. Dev. Biol., Plant 36 481–487
P Geerts A Toussaint G Mergeai JP Baudoin (2001) ArticleTitleCulture of very young Phaseolus vulgaris L. pods and plantlet regeneration Acta Hort. 560 411–415
A Gulati P Schryer A McHughen (2001) ArticleTitleRegeneration and micrografting of lentil shoots In Vitro Cell. Dev. Biol., Plant 37 798–802
GB PhillipsGCand Collins (1979) ArticleTitleIn vitro tissue culture of selected legumes and plant regeneration from callus cultures of red clover Crop Sci. 19 59–64
V Raghavan (1994) In vitro methods for the control of fertilization and embryo development IK Vasil TA Thorpe (Eds) Plant Cell and Tissue Culture. Kluwer Academic Publishers Dordrecht Netherlands 173–194
V Raghavan JG Torrey (1963) ArticleTitleGrowth and morphogenesis of globular and older embryos of Capsella in culture Am. J. Bot. 50 540–551
J Rietsema S Satina AF Blakeslee (1953) ArticleTitleThe effect of sucrose on the growth of Datura stamonium embryos in vitro Am. J. Bot. 40 538–545
AM Sabja DWS Mok MC Mok (1990) ArticleTitleSeed and embryo growth in pod cultures of Phaseolus vulgaris and P.␣vulgaris × P. acutifolius HortScience 25 1288–1291
EC Yeung IM Sussex (1979) ArticleTitleEmbryogeny of Phaseolus coccineus: the suspensor and the growth of the embryo-proper in vitro Z. Pflanzenphysiol. Bd. 91 S 423–433
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Schryer, P.A., Lu, Q., Vandenberg, A. et al. Rapid regeneration of Phaseolus angustissimus and P. vulgaris from very young zygotic embryos. Plant Cell Tiss Organ Cult 83, 67–74 (2005). https://doi.org/10.1007/s11240-005-2586-7
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DOI: https://doi.org/10.1007/s11240-005-2586-7