Improving the efficiency of isolated microspore culture in six-row spring barley: I-optimization of key physical factors

Improving the efficiency of isolated microspore culture in six-row spring barley: I-optimization... Doubled haploid (DH) plants are completely homozygous individuals that can be generated in just a few months via androgenesis in vitro. DHs are useful tools in genetic research and in plant breeding. Isolated microspore culture (IMC) is the most efficient way to produce DHs, but a strong genotype dependency imposes limitations to its wide application. Six-row, spring barley genotypes are considered as particularly recalcitrant due to a low frequency of embryogenesis and a high rate of albinism. Seeking to develop an efficient IMC protocol for this type of barley, we explored four important factors: (1) the harvest stage of immature spikes, (2) the type of pretreatment applied, (3) the osmotic potential in the induction medium, and (4) the plating density of microspores. This work was first performed using four barley genotypes: two typical six-row spring cultivars (ACCA and Léger), a two-row spring (Gobernadora) and a two-row winter (Igri) cultivar. First, by optimizing the harvest stage for each genotype we obtained a twofold to fourfold increase in the yield of embryogenic microspores. Second, two pretreatments (0.3 M mannitol for 2 days, or a combination of cold and heat over 15 days) both performed significantly better than the commonly used cold pretreatment (28 days at 4 °C). Third, an induction medium-containing mannitol (32 g/l) doubled green plant regeneration. Fourth, a plating density of 10 6 microspores/ml yielded the highest number of green regenerated plants. Our most important findings were then confirmed using sets of F1s from a six-row, spring-type breeding program. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Cell Reports Springer Journals

Improving the efficiency of isolated microspore culture in six-row spring barley: I-optimization of key physical factors

Plant Cell Reports, Volume 33 (6) – Jun 1, 2014

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Publisher
Springer Journals
Copyright
Copyright © 2014 by Springer-Verlag Berlin Heidelberg
Subject
Life Sciences; Cell Biology; Plant Sciences; Biotechnology; Plant Biochemistry
ISSN
0721-7714
eISSN
1432-203X
D.O.I.
10.1007/s00299-014-1583-x
Publisher site
See Article on Publisher Site

Abstract

Doubled haploid (DH) plants are completely homozygous individuals that can be generated in just a few months via androgenesis in vitro. DHs are useful tools in genetic research and in plant breeding. Isolated microspore culture (IMC) is the most efficient way to produce DHs, but a strong genotype dependency imposes limitations to its wide application. Six-row, spring barley genotypes are considered as particularly recalcitrant due to a low frequency of embryogenesis and a high rate of albinism. Seeking to develop an efficient IMC protocol for this type of barley, we explored four important factors: (1) the harvest stage of immature spikes, (2) the type of pretreatment applied, (3) the osmotic potential in the induction medium, and (4) the plating density of microspores. This work was first performed using four barley genotypes: two typical six-row spring cultivars (ACCA and Léger), a two-row spring (Gobernadora) and a two-row winter (Igri) cultivar. First, by optimizing the harvest stage for each genotype we obtained a twofold to fourfold increase in the yield of embryogenic microspores. Second, two pretreatments (0.3 M mannitol for 2 days, or a combination of cold and heat over 15 days) both performed significantly better than the commonly used cold pretreatment (28 days at 4 °C). Third, an induction medium-containing mannitol (32 g/l) doubled green plant regeneration. Fourth, a plating density of 10 6 microspores/ml yielded the highest number of green regenerated plants. Our most important findings were then confirmed using sets of F1s from a six-row, spring-type breeding program.

Journal

Plant Cell ReportsSpringer Journals

Published: Jun 1, 2014

References

  • Developmental staging of maize microspores reveals a transition in developing microspore proteins
    Bedinger, PA; Edgerton, MD
  • Differential development of plastids during microspore embryogenesis in barley
    Caredda, S; Devaux, P; Sangwan, RS; Clément, C
  • Comparison of anther and isolated microspore cultures in barley. Effects of culture density and regeneration medium
    Castillo, AM; Valdés, MP; Cistué, L
  • Influence of anther pretreatment and culture medium composition on the production of barley doubled haploids from model and low responding cultivars
    Cistué, L; Ramos, A; Castillo, AM

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