Development of haploid embryos and plants of Lactuca sativa induced by distant pollination with Helianthus annuus and H. tuberosus

详细信息    查看全文

• 作者：Ł. Piosik ; E. Zenkteler ; M. Zenkteler
• 关键词：Haploid ; Lettuce ; Parthenogenesis ; Callus proliferation ; Distant pollination ; In vitro culture
• 刊名：Euphytica
• 出版年：2016
• 出版时间：April 2016
• 年：2016
• 卷：208
• 期：3
• 页码：439-451
• 全文大小：2,886 KB
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Plant Physiology
  Plant Sciences
  Ecology
  
• 出版者：Springer Netherlands
• ISSN：1573-5060

文摘

Haploidisation is a biotechnological method used to obtain plants with improved traits that are of use to humans. Lettuce (Lactuca sativa L.), a well-known and popular leafy vegetable, is consumed worldwide. Its haploid form would provide a good basis for producing a pure line of plants (doubled haploids) allowing new varieties to be regenerated. The main aim of this work was to develop an effective haploidisation method for this economically important species. In order to stimulate the development of haploid embryos of lettuce based on our previous experience, we conducted in vivo distant pollination with fresh pollen grains of Helianthus annuus L. or H. tuberosus L. Because the haploid proembryos obtained after pollination did not develop further (despite the presence of cellular endosperm), we implemented the technique of in vitro culture of an isolated embryo sacs (surrounded by endothelium) with parthenogenetic embryos on various, modified Murashige and Skoog media. During the in vitro culture, we observed the formation of callus tissue and, after subsequent cultures of calluses, 23 haploid L. sativa plants were regenerated. The haploid status of the regenerated plantlets was confirmed by estimation of the genome size by flow cytometry, chromosome counting in root tips, stomata cell size and by disturbances in pollen formation resulting from abnormal microsporogenesis. This paper contains the complete protocol for obtaining haploid plants of L. sativa.