Protocol for wheat (Triticum aestivum L.) anther culture
Protocol for wheat (Triticum aestivum L.) anther culture
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小麦（Triticum aestivum L.）花药培养方案
DOI：
10.1007/978-94-017-1293-4_10
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发表时间：
2003
期刊：
影响因子：
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通讯作者：
M. Puolimatka
中科院分区：
文献类型：
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作者：
J. Pauk;R. Mihály;M. Puolimatka
经过多年的不同实验在世界各地的单倍体生产不同的方法已成为可供育种。在塞格德，花药培养是产生纯合系的最有效的系统之一。利用双单倍体（DH）系进行小麦育种，可以节省时间，提高选择效率，在小麦新品种选育中具有重要的应用价值。在将双单倍体生产系统发展为常规技术的过程中，许多问题已经得到解决（诱导培养基、基因型依赖性、碳水化合物来源、育种应用等）。诱导培养基是诱导雄核发育以及胚状体和植株发育的重要因素之一。Ouyanget al.（1983）发表的马铃薯（P-4）培养基经过我们的微小修改后，似乎是最有效的单倍体诱导培养基（P-4 mf，表2.9-1）。已显示氨基酸谷氨酰胺和天冬酰胺增强雄核发育，并且它们可以部分地替代马铃薯培养基中的马铃薯提取物。在烟草花药培养中，丝氨酸对花药壁有促进作用。Marsolaiset等（1986）等描述了一种通过大麦（Hordeum vulcanum L.）花药培养生产绿色植物的有效方法。和小麦（Triticum aestivumL.）在媒体中使用Ficoll。Ficoll可防止花药沉入培养基中，并避免琼脂中存在抑制剂，从而改善雄激素反应。它的应用为小麦花药培养开辟了新的篇章。在花药培养体系中，培养温度和小孢子发育阶段是影响花药培养的重要因素。冷预处理打破了正常的小孢子发育（配子体途径），增加了对称的有丝分裂的频率，这导致了自发的双单倍体形成的外观进一步增加。在培养开始时进行短时间的热休克处理也是有效的。用其他糖，特别是麦芽糖代替蔗糖，提高了花药培养的成功率。对谷物的实验通常表明，通过使用麦芽糖代替蔗糖，胚胎诱导增加或胚胎发育和绿色植物再生得到改善（Orshinskyet等人，1990年）。
After many years of different experiments around the world on haploid production different methods have become available to the breeders. In Szeged,in vitroandrogenesisviaanther culture is one of the most efficient systems of homozygous line production. It can be widely applied in wheat and could make a significant contribution to the breeding of new varieties by saving time and increasing selection efficiency via doubled haploid (DH) lines. During the development of the doubled haploid production system into a routine technique, many problems have been solved (induction medium, genotype dependence, carbohydrate source, breeding applications, etc.). Induction medium is one of the most important factors for the induction of androgenesis and the subsequent development of embryoids and plantlets. The potato (P-4) medium published by Ouyanget al. (1983), seems to be the most effective haploid induction medium, after our minor modifications (P-4mf, Table 2.9-1). The amino acids glutamine and asparagine have been shown to enhance androgenesis and they can partially replace the potato extract in potato medium. Serine has reputedly a positive effect on the anther wall inNicotianaanther culture. Marsolaiset al. (1986) and others, have described an efficient method of green plant production by anther culture of barley (Hordeum vulgareL.) and wheat (Triticum aestivumL.) using Ficoll in the media. Ficoll prevents anthers from sinking into the medium and avoids the presence of inhibitors from agar leading to improved androgenic response. Its application has opened up a new chapter in wheat anther culture. Among the conditions and variables used in anther culture system, incubation temperature and stage of microspore development are important factors. Cold pretreatment breaks down normal microspore development (gametophytic pathway) and increases the frequency of symmetrical mitoses, which causes a further increase in the appearance of spontaneous doubled haploid formation. Short heat shock treatments at the initiation of culture are also effective. The replacement of sucrose by other sugars, notably by maltose, enhances the success of anther culture. Experiments on cereals have generally shown an increase in embryo induction or an improvement in embryo development and green plant regeneration by the use of maltose instead of sucrose (Orshinskyet al., 1990).