枸杞花药离体培养获得单倍体植株

Wolfberry Anther Culture to Obtain Haploid Plants

利用枸杞花药离体培养获得单倍体植株,枸杞单倍体植株是新品种选育和基因组测序、制作遗传图谱、研究遗传规律的良好材料。以"宁杞1号"花药为试验材料,采用MS+5%蔗糖+0.7%琼脂+0.8%活性炭为基本培养基,添加不同浓度的6-BA和NAA,进行花药离体培养,获得花粉植株。利用流式细胞仪检测花培植株的倍性,再经根尖细胞染色体数目鉴定花粉植株的倍性。结果表明,在基本培养基中添加1.0 mg/L 6-BA和0.1 mg/L NAA时,花药离体培养花粉胚状体诱导率最高。花粉植株经过流式细胞仪倍性鉴定,其DNA在荧光通道值100处有一条很明显的尖峰,说明获得的花粉植株为单倍体。又经根尖细胞染色体数目鉴定,染色体数为12条,验证了来自花药中花粉发育成的花粉植株为单倍体植株。

Anther culture technique was used in this paper to obtain haploid plants which could be used in new varieties breeding, genome sequencing, genetic map drawing, and inheritance study. This paper took ＂Ning Qi No.1＂ anther as subject, adopted MS ＋ 5% sucrose plus 0.7% Agarose plus 0.8% activated carbon as basic medium, mixed with different concentrations of 6-BA and NAA to get pollen plants. Then identify plant pollen tablet ploidy through flow cytometry method and chromosome number. Results showed that anther culture pollen embryoid induction rate with 1.0 mg/L 6-BA and 0.1mg/L NAA in the basic medium was the highest. Though flow cytometry test, the DNA of ploidy showed a very significant spike in fluorescence channel 100, indicating that pollen haploid plant was obtained. The number of chromosomes, which was 12, verified that the pollen haploid plant was developed from the pollen of anthers.