通过梯牧草（Phleum pratense L.）离体小孢子培养实现高效雄核胚胎发生和植株再生的方法。

An efficient androgenic embryogenesis and plant regeneration method through isolated microspore culture in timothy (Phleum pratense L.).

作者信息

Guo Y-D, Pulli S

机构信息

Laboratory of Plant Physiology and Molecular Biology, Department of Biology, University of Turku, 20014, Turku, Finland Fax: +358-2-333,5549 e-mail:

出版信息

Plant Cell Rep. 2000 Jul;19(8):761-767. doi: 10.1007/s002990000193.

DOI:10.1007/s002990000193

PMID:30754866

Abstract

A method employing isolated microspore culture was established for the androgenic embryogenesis of timothy (Phleum pratense L). Embryos/calli were obtained and green plants regenerated. The induction medium was PG-96 (1.0 mg l 2,4-D, 0.1 mg l Kinetin) supplemented with 6% maltose monohydrate. Timothy microspore culture was genotype-dependent, among 12 genotypes, 6 produced embryos/calli and 4 produced green plants. Macerating the spikes with a blender and purifying the microspores at a mannitol/maltose monohydrate interface gave a relatively high percentage of cell vitality. The optimum microspore developmental stage was from the very late uninucleate stage to the binucleate stage. Heat shock promoted the initiation of microspore culture. Over 150 regenerated green plants were obtained; in a random sample of 32 of these 65.6% were doubled haploids (6n=42). Albinism was a problem in plant regeneration (9.3-22%). This paper is the first to describe timothy androgenic embryogenesis by isolated microspore culture.

摘要

建立了一种采用离体小孢子培养的方法用于梯牧草（Phleum pratense L）的雄核胚胎发生。获得了胚/愈伤组织并再生出绿色植株。诱导培养基为添加了6%一水麦芽糖的PG - 96（1.0 mg/l 2,4 - D，0.1 mg/l激动素）。梯牧草小孢子培养依赖于基因型，在12个基因型中，6个产生了胚/愈伤组织，4个产生了绿色植株。用搅拌器捣碎穗并在甘露醇/一水麦芽糖界面纯化小孢子可获得相对较高比例的细胞活力。小孢子发育的最佳阶段是从单核晚期到双核期。热激促进了小孢子培养的起始。获得了150多株再生绿色植株；在随机抽取的32株样本中，65.6%为双单倍体（6n = 42）。白化现象是植株再生中的一个问题（9.3 - 22%）。本文首次描述了通过离体小孢子培养进行梯牧草雄核胚胎发生的情况。