Optimizing somatic embryogenesis and particle bombardment of barley (Hordeum vulgare L.) immature embryos

1994 ◽  
Vol 30 (2) ◽  
pp. 117-123 ◽  
Author(s):  
Steven P. King ◽  
Ken J. Kasha
Keyword(s):  
Somatic Embryogenesis ◽  
Hordeum Vulgare ◽  
Particle Bombardment ◽  
Immature Embryos ◽  
Hordeum Vulgare L

Transformation of isolated barley (Hordeum vulgare L.) microspores: I. The influence of pretreatments and osmotic treatment on the time of DNA synthesis

Genome ◽  
2009 ◽  
Vol 52 (2) ◽  
pp. 166-174 ◽  
Author(s):  
Youn-Seb Shim ◽  
K. Peter Pauls ◽  
Ken J. Kasha
Keyword(s):  
Cell Cycle ◽  
Hordeum Vulgare ◽  
Transgenic Plants ◽  
Dna Synthesis ◽  
Particle Bombardment ◽  
Osmotic Adjustment ◽  
Hordeum Vulgare L ◽  
Cell Cycle Stage ◽  
Dna Density ◽  
Homozygous Transgenic Plants

The objective of this study was to determine when DNA synthesis occurred during pretreatments of cultured barley ( Hordeum vulgare L.) microspores and during their preparation for particle bombardment. Based on this information, an investigation of the influence of cell cycle stage on the ability to obtain homozygous transgenic plants by particle bombardment will be presented in paper II of this series. It was hypothesized that the introduction of foreign genes at the G1 cell cycle stage in cultured uninucleate microspores would produce homozygous transgenic plants. Experiments were conducted with two different commonly used pretreatments to induce microspore embryogenesis: cold (4 °C) for 21days and cold plus 0.3 mol/L mannitol for 4 days. After pretreatment, the microspores were placed in a higher osmotic medium for 4 h prior to and for 18 h following bombardment. It was confirmed that during the cold plus mannitol pretreatment, there was no apparent change in the cell cycle stage, with the majority of the microspores remaining at the G1 stage. While in the cold for 21 days, the microspores progressed slowly through to G2, with a few progressing further into the mitosis and binucleate stages. Hourly DNA density measurements that were taken during the 4 h osmotic adjustment period following the cold plus mannitol pretreatment indicated that DNA synthesis began during this period at 25 °C, while at 4 °C, there was no apparent change in cell cycle stage or in DNA density. Thus, one might expect to find a higher frequency of homozygous doubled haploids by maintaining the temperature low during the 4 h osmotic adjustment period following the cold plus mannitol pretreatment than following the 21 day cold pretreatment. However, it is also not known what effect the temperatures during the whole high-osmotic treatments will have on the rate and time of incorporation of the transgene.

scholarly journals Somatic Embryogenesis and Plant Regeneration in Barley (Hordeum vulgare L.)

2006 ◽  
Vol 33 (4) ◽  
pp. 237-242 ◽  
Author(s):  
Tae-Il Park ◽  
Mi-Ok Um ◽  
Young-Jin Kim ◽  
Jong-Jin Hwang ◽  
Kyung-Gu Choi ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Hordeum Vulgare ◽  
Plant Regeneration ◽  
Hordeum Vulgare L
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