Enhancement of microspore culture efficiency of recalcitrant barley genotypes

被引：39

作者：

Li H. ^{[1
]}

Devaux P. ^{[1
]}

机构：

[1] Florimond Desprez, Biotechnology Laboratory, 59242 Cappelle en Pevele

来源：

Plant Cell Reports | 2001年 / 20卷 / 06期

关键词：

Barley; Embryogenesis; Isolated microspore; Ovary co-culture; Pretreatment;

D O I：

10.1007/s002990100368

中图分类号：

学科分类号：

摘要：

The culture response of isolated microspores of seven recalcitrant cultivars of barley has been largely improved by identifying an appropriate pretreatment and utilizing ovary co-cultivation. After comparison of three pretreatment media, medium B was shown to be most efficient for inducing microspore embryogenesis, while 0.3 M mannitol frequently used for the responsive cv. Igri was found to be ineffective for recalcitrant genotypes. A further significant improvement of embryogenesis was achieved by using ovary co-culture, which resulted in an overall 2.1-fold increase in embryo formation and 2.4-fold increase in green plant regeneration from all cultivars compared with the control. Optimal co-culture conditions were identified as 5 ovaries/ml medium kept over 20 days in induction culture. Microspore plating densities in cultures with and without co-culture were found to be optimal at 4×104/ml and 8-12×104/ml, respectively. The most effective and reproducible method for culturing microspores of recalcitrant genotypes appeared to be the combination of medium B pretreatment with ovary co-culture. By using this procedure, the genotypic difference in microspore embryogenesis could be reduced. It was found that medium B mainly enhanced percent live embryogenic microspores, and ovary co-culture subsequently improved cell division and embryogenic development. The method described here is important for the application of the microspore culture technique to barley breeding and biotechnology.

引用

页码：475 / 481

页数：6

共 29 条

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