为建立多态性高、稳定性好的洋葱RAPD-PCR反应体系,采用正交设计,研究了Taq酶、Mg2+、引物和dNTP 4种RAPD-PCR反应组分浓度变化对扩增结果的影响,在此基础上对模板DNA用量、扩增程序中退火温度和反应循环次数进行了筛选。试验结果表明,...为建立多态性高、稳定性好的洋葱RAPD-PCR反应体系,采用正交设计,研究了Taq酶、Mg2+、引物和dNTP 4种RAPD-PCR反应组分浓度变化对扩增结果的影响,在此基础上对模板DNA用量、扩增程序中退火温度和反应循环次数进行了筛选。试验结果表明,洋葱20μl RAPD-PCR优化反应体系为1×Buffer、2.0 mmo1/L Mg2+、1.0 UTaqDNA聚合酶、200μmo1/L dNTP、0.6μmo1/L引物、2%甘油和15 ng DNA模板;PCR扩增程序为94℃预变性4 m in;94℃变性30 s,35℃退火40 s,72℃延长1.5 m in,45个循环;72℃保温延伸7 m in。展开更多
Onion （Allium cepa L.） was an economic vegetable and a strictly biennial herb, which was widely distributed in the world. In the past, it was a strictly biennial plant, the studies had shown that FT （FLOWERING LOCU...Onion （Allium cepa L.） was an economic vegetable and a strictly biennial herb, which was widely distributed in the world. In the past, it was a strictly biennial plant, the studies had shown that FT （FLOWERING LOCUS T） gene was involved in the photoperiod pathway to regulate fowering in the model plant. In this study, transcriptome sequencing method was used to obtain cDNA sequence of FT homologous gene in onion, named AcFT3 （KF864665）. AcFT3 had a full-length of 540 bp, encoded 179 amino acids, with 98.31% homology to AfFT （Allium fstulosum）, and 63.0%-84.0% homology to other higher plants. Phylogenetic tree analysis indicated that AcFT3 had the closest relationship with AfFT. The results of quantitative RT-PCR showed the expression pattern of AcFT3 both in vegetative growth of onion and in different organs of bolting and fowering, the expression level of AcFT3 reached the highest in the leaves before bolting and in the fower organs after bolting.展开更多
文摘为建立多态性高、稳定性好的洋葱RAPD-PCR反应体系,采用正交设计,研究了Taq酶、Mg2+、引物和dNTP 4种RAPD-PCR反应组分浓度变化对扩增结果的影响,在此基础上对模板DNA用量、扩增程序中退火温度和反应循环次数进行了筛选。试验结果表明,洋葱20μl RAPD-PCR优化反应体系为1×Buffer、2.0 mmo1/L Mg2+、1.0 UTaqDNA聚合酶、200μmo1/L dNTP、0.6μmo1/L引物、2%甘油和15 ng DNA模板;PCR扩增程序为94℃预变性4 m in;94℃变性30 s,35℃退火40 s,72℃延长1.5 m in,45个循环;72℃保温延伸7 m in。
基金Supported by Fund Project of Heilongjiang Province(C2015017)Heilongjiang Provincial Science and Technology Commissioner Project(GC13B09)
文摘Onion （Allium cepa L.） was an economic vegetable and a strictly biennial herb, which was widely distributed in the world. In the past, it was a strictly biennial plant, the studies had shown that FT （FLOWERING LOCUS T） gene was involved in the photoperiod pathway to regulate fowering in the model plant. In this study, transcriptome sequencing method was used to obtain cDNA sequence of FT homologous gene in onion, named AcFT3 （KF864665）. AcFT3 had a full-length of 540 bp, encoded 179 amino acids, with 98.31% homology to AfFT （Allium fstulosum）, and 63.0%-84.0% homology to other higher plants. Phylogenetic tree analysis indicated that AcFT3 had the closest relationship with AfFT. The results of quantitative RT-PCR showed the expression pattern of AcFT3 both in vegetative growth of onion and in different organs of bolting and fowering, the expression level of AcFT3 reached the highest in the leaves before bolting and in the fower organs after bolting.
