摘要
该研究以6~8月上午10点左右摘取的新鲜黄瓜花朵为材料,采用渗透压冲击的方法分离黄瓜生殖细胞,并应用竞争型定量PCR技术测定其线粒体DNA数量,分析生殖细胞在发育过程中线粒体DNA的变化,以明确高丰度线粒体DNA的来源,为进一步研究被子植物调控线粒体DNA扩增的分子机制奠定基础。结果显示：（1）DAPI染色观察发现,黄瓜生殖细胞的细胞核周围存在大量的细胞器DNA荧光点,表明黄瓜生殖细胞的细胞质中存在大量的线粒体DNA。（2）成熟黄瓜生殖细胞平均包含（1 037±126）个线粒体DNA拷贝。（3）成熟生殖细胞内线粒体DNA含量为早期生殖细胞的14.5倍,表明成熟生殖细胞中的线粒体DNA主要来自于生殖细胞形成后其内活跃的线粒体DNA扩增。研究认为,黄瓜生殖细胞内活跃的线粒体DNA是黄瓜线粒体父系遗传的基础。
In this study,we collected fresh flowers fromCucumissativusaround 10：00amfrom June to August,isolated the generative cells（GC）of C.sativus with the method of osmotic shock,quantified its mitochondrial DNA（mtDNA）contents through employing the competitive quantitative PCR technology,and analyzed the change of mtDNA contents in GC during its development,in order to investigate the origin of the abundant mtDNA,and lay the foundation for further study on the molecular mechanism of regulation of mitochondrial DNA amplification in angiosperm.The results showed that：（1）DAPI staining represents abundant fluorescence points of organellar DNA around the nucleus of GC of C.sativus,indicating the presence of a large number of mtDNA in cytoplasm of cucumber GC.（2）The single mature GC of C.sativus averagely owns（1 037±126）mtDNA copies.（3）Compared with early GC,the mature GC up-regulates the mtDNA by 14.5times,suggesting that mtDNA in mature GC is mainly from its active amplification after GC formation.The study suggests that the active increase of mtDNA in GC is the basis of the paternal mitochondrial inheritance of C.sativus.
出处
《西北植物学报》
CAS
CSCD
北大核心
2016年第3期429-434,共6页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家自然科学基金(31200234)
陕西省教育厅科研计划(14JS089)
关键词
线粒体DNA
拷贝数
父系遗传
黄瓜
生殖细胞
mtDNA
copies
paternal inheritance
Cucumis sativus
generative cell（GC）
