【摘要】 游离小孢子培养技术是快速创制连州菜心双单倍体纯系的一条有效途径,为建立高效的连州菜心小孢子培养体系,应用于优势杂交育种实践,以引进的3个广东省连州市地方菜心品种迟熟连州菜心(17AY007)、连州尺寸心(17AY008)和连州尖叶迟菜心(17AY009)为试材进行小孢子培养,研究不同基因型和植物生长调节剂(芸薹素内酯,BR及噻苯隆,TDZ)对小孢子胚胎发生及植株再生的影响,并对获得的连州菜心小孢子再生植株进行倍性鉴定。结果表明:3个不同基因型连州菜心的小孢子胚胎发生率无显著差异;在NLN-13培养基中添加适宜浓度的BR或TDZ可显著提高连州菜心小孢子胚胎发生率和直接成苗率,其中,当BR浓度为0.04mg·L-1时,17AY007和17AY009的胚诱导率和直接成苗率均达到最高(2.38胚·蕾-1,72.82%;2.83胚·蕾-1,54.62%);当TDZ浓度为0.1mg·L-1和0.2mg·L-1时,17AY007的胚诱导率和直接成苗率分别达到最高(2.67胚·蕾-1,57.76%);TDZ浓度为0.3mg·L-1时,17AY009的胚诱导率和直接成苗率均达到最高(2.42胚·蕾-1,57.36%);3种连州菜心再生植株的双单倍体率均在56%以上,其中,17AY007双单倍体率最高,为64.29%。综上,创制了双单倍体植株42株,为连州菜心优势杂交育种奠定了材料基础。更多还原

【Abstract】 Microspore culture is a method of creating doubled haploid lines of Lianzhou flowering Chinese cabbage. This study aims were to establish a highly efficient microspore culture protocol and applied it to the practice of heterosis breeding program.Three local varieties of flowering Chinese cabbage in Lianzhou City, Guangdong Province, were used as donor plants which were late-maturing Lianzhou flowering Chinese cabbage(17AY007), Lianzhou short flowering Chinese cabbage(17AY008) and Lianzhou late flowering Chinese cabbage with sharp leaves(17AY009). The effects of genotypes and plant growth regulators brassinolide(BR) and thidiazuron(TDZ) on microspore embryogenesis and plant regeneration were studied, and the ploidy of microspore regenerated plants was identified. The results showed that there was no significant difference in the frequency of microspore embryogenesis among the three genotypes of Lianzhou flowering Chinese cabbage. Adding appropriate concentration of BR or TDZ to NLN-13 medium could significantly increase the frequencies of microspore embryogenesis and direct conversion to plants.When the concentration of BR was 0.04 mg·L-1, both the frequencies of microspore embryogenesis and direct conversion to plants of 17AY007 and 17AY009 were the highest(2.38 embryos per bud,72.82%; 2.83 embryos per bud,54.62%). When the TDZ concentrations were 0.1 mg·L-1and 0.2 mg·L-1, the frequencies of microspore embryogenesis and direct conversion to plants of17AY007 reached the highest(2.67 embryos per bud,57.76%), respectively. When the TDZ concentration was 0.3 mg·L-1, the frequencies of microspore embryogenesis and direct conversion to plants of 17AY009 reached the highest(2.42 embryos per bud,57.36%). The doubled haploid rates of three varieties of regenerated plants were above 56%, and the doubled haploid rate of17AY007 was the highest(64.29%). 42 doubled haploid lines were obtained in this study, which were materials for heterosis breeding program for Lianzhou flowering Chinese cabbage.更多还原