This research aimed to study the androgenesis and spontaneous chromosome doubling of five barley genotypes using an isolated in vitro microspore culture technique, involving a completely randomized design (CRD) with three replications. Statistical analysis of embryogenesis and cytogenetic results showed that genotype had a significant effect on haploid embryogenesis (P<0.01) and on spontaneous chromosome doubling (P<0.05). The genotype Igri was found to have the highest potential to produce haploid embryos (1577 embryos from 100 anthers), followed by the genotypes Boyer/Rojo, Afzal/Turkman/Kavir, Ashar/Hebo and Agrigashar/Matico with 369, 304, 278 and 150 embryos from 100 anthers, respectively. The highest percentage of spontaneous chromosome doubling (76%) was observed for the genotype which had the lowest embryogenesis (Agrigashar/Matico) and the lowest (65%) for the genotype with the highest androgenic capacity (Igri). Microspore embryogenesis also showed comparatively higher genotypic (109.2) and phenotypic (109.5) coefficients of variation, heritability (99.62) and genetic advance (1206.77), indicating the pre-dominance of additive gene action in the control of this character in the material studied. Estimates of genetic parameters (PCV, GCV and heritability) for microspore embryogenesis were higher than for spontaneous doubled haploids. These results indicated that selection for androgenic capacity would be more effective than for spontaneous doubled haploids. The findings showed a negative relationship (r= −0.68) between embryogenesis and spontaneous chromosome doubling in the barley genotypes studied. All the large embryos used had high regenerability and good plantlet formation.
Barnabás, B., Pfahler, P. L., Kovács, G. (1991): Direct effect of colchicine on the microspore embryogenesis to produce dihaploid plants in wheat ( Triticum aestivum L.). Theor. Appl. Genet. , 81 , 675–678.
Kovács G. , 'Direct effect of colchicine on the microspore embryogenesis to produce dihaploid plants in wheat (Triticum aestivum L.) ' (1991 ) 81 Theor. Appl. Genet. : 675 -678 .
Burton, G. W., de Vane, R. W. (1953): Estimating heritability in tall fescue ( Festuca arundinaceia ) from replicated clonal material. Agron. J. , 45 , 478–481
Vane R. W. , 'Estimating heritability in tall fescue (Festuca arundinaceia) from replicated clonal material ' (1953 ) 45 Agron. J. : 478 -481 .
Carlson, A. R. (1998): Visual selection of transgenic barley (Hordeum vulgare) structures and their regeneration into green plants . Thesis, University of Guelph.
Castillo, A. M., Valles, M. P., Cistue, L. (2000): Comparison of anther and isolated microspore cultures in barley. Effects of culture density and regeneration medium. Euphytica , 113 , 1–8.
Cistue L. , 'Comparison of anther and isolated microspore cultures in barley. Effects of culture density and regeneration medium ' (2000 ) 113 Euphytica : 1 -8 .
Hagio, T., Hirabayashi, T., Machii, H., Tomotsune, H. (1995): Production of fertile transgenic barley ( Hordeum vulgare L.) plants using the hygomycin-resistance marker. Plant Cell Rep. , 14 , 329–334.
Tomotsune H. , 'Production of fertile transgenic barley (Hordeum vulgare L.) plants using the hygomycin-resistance marker ' (1995 ) 14 Plant Cell Rep. : 329 -334 .
Hansen, N. J. P., Anderson, S. B. (1998a): In vitro chromosome doubling with colchicine during microspore culture in wheat ( Triticum aestivum L.). Euphytica , 102 , 101–108.
Anderson S. B. , 'In vitro chromosome doubling with colchicine during microspore culture in wheat (Triticum aestivum L.) ' (1998 ) 102 Euphytica : 101 -108 .
Hansen, N. J. P., Anderson, S. B. (1998b): In vitro chromosome doubling potential of colchicine, oryzaline, trifuralin and APM in Brassica napus microspore culture. Euphytica , 88 , 159–164.
Anderson S. B. , 'In vitro chromosome doubling potential of colchicine, oryzaline, trifuralin and APM in Brassica napus microspore culture ' (1998 ) 88 Euphytica : 159 -164 .
Hoekstra, S., Van Zijderwold, M. H., Heidekamap, F. L., Roue, C. (1993): Microspore culture of Hordeum vulgare L.: the influence of density and osmolality. Plant Cell Rep. , 12 , 661–665.
Roue C. , 'Microspore culture of Hordeum vulgare L.: the influence of density and osmolality ' (1993 ) 12 Plant Cell Rep. : 661 -665 .
Hoekstra, S., van Zijderwold, M. H., Louwerse, J. D., Heidekamp, F., Vandermark, F. (1992): Anther and microspore culture of Hordeum vulgare L. cv. Igri. Plant Sci. , 86 , 89–96.
Vandermark F. , 'Anther and microspore culture of Hordeum vulgare L. cv. Igri ' (1992 ) 86 Plant Sci. : 89 -96 .
Hu, T. C., Kasha, K. J. (1999): A cytological study of pretreatment effects on isolated microspore culture of wheat Triticum aestivum cv. Chris. Genome , 42 , 432–441.
Kasha K. J. , 'A cytological study of pretreatment effects on isolated microspore culture of wheat Triticum aestivum cv. Chris ' (1999 ) 42 Genome : 432 -441 .
Inagaki, M. (1985): Chromosome doubling of wheat haploids obtained from crosses with Hordeum bulbosum L. Jpn. J. Breed. , 35 , 193–195.
Inagaki M. , 'Chromosome doubling of wheat haploids obtained from crosses with Hordeum bulbosum L ' (1985 ) 35 Jpn. J. Breed. : 193 -195 .
Jähne, A., Lörz, H. (1995): Cereal microspore culture (Review). Plant Sci. , 109 , 1–12.
Lörz H. , 'Cereal microspore culture (Review) ' (1995 ) 109 Plant Sci. : 1 -12 .
Johnson, H. W., Robinson, H. F., Comstock, R. W. (1955): Estimates of genetic and environmental variability in soybeans. Agron. J. , 47 , 314–318.
Comstock R. W. , 'Estimates of genetic and environmental variability in soybeans ' (1955 ) 47 Agron. J. : 314 -318 .
Kahrizi, D. (1998): Effect of donor plant genotype and media composition on androgenesis of hexaploid wheat (Triticum aestivum L. ). (MSc. Thesis) Razi University, Kermanshah, Iran.
Kahrizi D. , '', in Effect of donor plant genotype and media composition on androgenesis of hexaploid wheat (Triticum aestivum L.) , (1998 ) -.
Kahrizi, D. (2001): Study on gametophytic embryogenesis in barley ( Hordeum vulgare L.) genotypes via androgenesis. The Second National Biotechnology Congress Islamic Republic of Iran . Oct. 9–11, 2001, Karaj, Iran.
Kahrizi, D., Arminian, A., Masumi Asl, A. (2007a): In vitro Plant Breeding . Razi University Press, Razi, Iran (in Persian).
Masumi Asl A. , '', in In vitro Plant Breeding , (2007 ) -.
Kahrizi, D., Moieni, A., Bozorgipour, R. (2000): Effect of genotype and carbohydrate source on androgenesis of hexaploid wheat ( Triticum aestivum L.). Seed Plant , 16 , 41–51 (in Persian).
Bozorgipour R. , 'Effect of genotype and carbohydrate source on androgenesis of hexaploid wheat (Triticum aestivum L.) ' (2000 ) 16 Seed Plant : 41 -51 .
Kahrizi, D., Salmanian, A. H., Afshari, A., Moieni, A., Mousavi, A. (2007b): Simultaneous substitution of Gly96 to Ala and Ala183 to Thr in 5-enolpyruvylshikimate-3-phosphate synthase gene of E. coli (k12) and transformation of rapeseed ( Brassica napus L.) in order to make tolerance to glyphosate. Plant Cell Rep. , 26 , 95–104.
Mousavi A. , 'Simultaneous substitution of Gly96 to Ala and Ala183 to Thr in 5-enolpyruvylshikimate-3-phosphate synthase gene of E. coli (k12) and transformation of rapeseed (Brassica napus L.) in order to make tolerance to glyphosate ' (2007 ) 26 Plant Cell Rep. : 95 -104 .
Kasha, K. J., Hu, T. C., Oro, R., Simion, E., Shim, Y. S. (2001): Nuclear fusion leads to chromosome doubling during mannitol pretreatment of barley ( Hordeum vulgare L.) microspores. J. Exp. Bot. , 52 , 1227–1238.
Shim Y. S. , 'Nuclear fusion leads to chromosome doubling during mannitol pretreatment of barley (Hordeum vulgare L.) microspores ' (2001 ) 52 J. Exp. Bot. : 1227 -1238 .
Kasha, K. J., Ziauddin, A., Cho, U., Simion, E., Petroski, R., Cistué, L. (1997): Anther and microspore cultures of barley and wheat. J. Appl. Genet. , 38 , 373–380.
Cistué L. , 'Anther and microspore cultures of barley and wheat ' (1997 ) 38 J. Appl. Genet. : 373 -380 .
Kasha, K. J., Ziauddin, A., Simion, E. (1991): Plant regeneration from single cell (microspore) cultures of wheat. ASA Abstracts (p. 196). American Society of Agronomy, Madison, WI.
Simion E. , '', in ASA Abstracts , (1991 ) -.
Kasha, K.J., Simion, E., Oro, R., Yao, Q. A., Hu, T. C., Carlson, A. R. (2004): An improved in vitro technique for isolated microspore culture of barley. Euphytica , 120 , 379–385.
Carlson A. R. , 'An improved in vitro technique for isolated microspore culture of barley ' (2004 ) 120 Euphytica : 379 -385 .
Kester, D. E., Hansche, P. E., Beres, V., Asay, R. N. (1977): Variance components and heritability of nut and kernel traits in almond. J. Am. Soc. Hort. Sci. , 102 , 264–266.
Asay R. N. , 'Variance components and heritability of nut and kernel traits in almond ' (1977 ) 102 J. Am. Soc. Hort. Sci. : 264 -266 .
Li, H. C., Devaux, P. (2003): High frequency regeneration of barley doubled haploid plants from isolated microspore culture. Plant Sci. , 164 , 379–386.
Devaux P. , 'High frequency regeneration of barley doubled haploid plants from isolated microspore culture ' (2003 ) 164 Plant Sci. : 379 -386 .
Mozafari, J., Wolyn, D. J., Ali-Khan, S. T. (1997): Chromosome doubling via tuber disc culture in dihaploid potato as determined by confocal microscopy. Plant Cell Rep. , 16 , 329–333.
Ali-Khan S. T. , 'Chromosome doubling via tuber disc culture in dihaploid potato as determined by confocal microscopy ' (1997 ) 16 Plant Cell Rep. : 329 -333 .
Mujeeb-Kazi, A., Miranda, J. L. (1985): Enhanced resolution of somatic chromosome constrictions as aid to identifying intergeneric hybrids among some Triticeae. Cytologia , 50 , 701–710.
Miranda J. L. , 'Enhanced resolution of somatic chromosome constrictions as aid to identifying intergeneric hybrids among some Triticeae ' (1985 ) 50 Cytologia : 701 -710 .
Nägeli, M., Schmid, J. E., Stamp, P., Büter, B. (1999): Improved formation of regenerable callus in isolated microspore culture of maize: impact of carbohydrates, plating density and time of transfer. Plant Cell Rep. , 19 , 177–184.
Büter B. , 'Improved formation of regenerable callus in isolated microspore culture of maize: impact of carbohydrates, plating density and time of transfer ' (1999 ) 19 Plant Cell Rep. : 177 -184 .
Navarro-Alvarez, W., Baenziger, P. S., Eskridge, K. M., Hugo, M., Gustafson, V. D. (1994): Addition of colchicine to wheat anther culture media to increase doubled haploid plant production. Plant Breeding , 112 , 192–198.
Gustafson V. D. , 'Addition of colchicine to wheat anther culture media to increase doubled haploid plant production ' (1994 ) 112 Plant Breeding : 192 -198 .
Seguí-Simarro, J. M., Nuez, F. (2008): Pathways to doubled haploidy: chromosome doubling during androgenesis. Cytogenet. Genome Res. , 120 , 358–369.
Nuez F. , 'Pathways to doubled haploidy: chromosome doubling during androgenesis ' (2008 ) 120 Cytogenet. Genome Res. : 358 -369 .
Sunderland, N. (1974): Anther culture as a means of haploid induction. pp. 91–122. In: Kasha, K. J. (ed.), Haploids in Higher Plants: Advances and Potential . University of Guelph, Guelph, Canada.
Sunderland N. , '', in Haploids in Higher Plants: Advances and Potential , (1974 ) -.
Sunderland, N., Collins, G. B., Dunwell, J. M. (1974): The role of nuclear fusion in pollen embryogenesis of Datura innoxia Mill. Planta , 117 , 227–241.
Dunwell J. M. , 'The role of nuclear fusion in pollen embryogenesis of Datura innoxia Mill ' (1974 ) 117 Planta : 227 -241 .
Taji, A., Kumar, P., Lakshmanan, P. (2002): In vitro Plant Breeding . Food Products Press, New York.
Lakshmanan P. , '', in In vitro Plant Breeding , (2002 ) -.
Tiwari, S., Rahimbaves, I. (1992): Comparison of glucose, sucrose and maltose for Hordeum vulgare L. isolated microspore culture using different methods. Indian J. Exp. Biol. , 30 , 624–627.
Rahimbaves I. , 'Comparison of glucose, sucrose and maltose for Hordeum vulgare L. isolated microspore culture using different methods ' (1992 ) 30 Indian J. Exp. Biol. : 624 -627 .