摘要酸浆属(Physalis)是茄科(Solanaceae)中的一个属,小酸浆( Physalis minima L。 ) 又名灯笼草、天泡子等,为酸浆同属。小酸浆具有较高的药用价值, 其种质资源的遗传多样性和遗传结构研究直接关系到植物基因资源的鉴定、保育,现已成为世界各国科学家普遍关注和研究的焦点。但遗憾的是,国内外对酸浆属种质资源评价及遗传多样性、遗传分化的研究极少。因此,为了小酸浆的充分利用和资源保护,开展小酸浆种质遗传多样性评价及保护研究是十分急迫和必要的。74993
本研究利用SCoT分子标记技术对六种不同来源地的82份野生小酸浆样品进行遗传多样性检测与评价,旨在为野生小酸浆的品种鉴定、遗传多样性研究及遗传资源道地性保护提供可靠的参考数据,为实现最大限度的保存野生小酸浆遗传多样性的策略提供科学依据。主要研究方法与结果如下:
1、野生小酸浆基因组DNA提取:以鲜叶片为原料,采用Ezup柱式植物基因组DNA抽提试剂盒提取小酸浆的基因组DNA,并通过紫外可见分光光度计(BioSpec-nano)和1%的琼脂糖凝胶电泳检测其浓度与纯度,结果表明获得的DNA可满足后续分子生物学实验的需要。
2、试验建立SCoT反应体系:小酸浆 SCoT-PCR反应体系(10uL)含模板DNA 1uL,5X PCR mix 5。0 uL,引物1uL,ddH2O 3uL。SCoT-PCR扩增程序:94 ℃预变性5 min;35个循环(94 ℃变性1 min,引物Tm值复性1 min,72 ℃延伸1。5min);72 ℃延伸10 min,4 ℃保存。
3、基于SCoT标记的野生小酸浆遗传多样性分析:SCoT标记分析选出的23条引物共扩增出186个SCoT位点,不同引物的多态性百分比为66。67%,平均每个引物产生7。13条多态性带。各材料的遗传相似系数在0。559 ~0。989之间。
Abstract Physalis is one genus of Solanaceae, P。 minima L。has a high medicinal value。The genetic persity and genetic structure of its germplasm resources are directly related to the identification and conservation of plant genetic resources, which has become the focus of scientists in the world。 The research about the genetic persity of Physalis germplasm resources is few。 Therefore, in order to Physalis full utilization and protection of resources, to carry out P。 minima L。germplasm genetic persity evaluation and protection research is very urgent and necessary。
In this study, the SCoT molecular markers were used to study the genetic persity of six different sources of 82 samples of wild P。 minima L。 It will provide reliable reference data for wild P。 minima L。of cultivar identification, genetic persity research and genetic resources genuineness protection, provide scientific basis for the realization of maximum preservation of wild P。 minima L。 genetic persity of strategy。 The main contents and results of this experiment are as follows:
1。 Wild P。 minima L。 genomic DNA extraction: The genomic DNA was isolated using the Ezup pillar Plant Genomic DNA Extraction kit with Fresh, young P。 minima L。 leaves。 Integrity and quality of DNA was evaluated by electrophoresis on 1% agarose gels, and concentration of genomic DNA samples was determined by UV spectrometer。 The results obtained show that the DNA can meet the needs of the subsequent molecular biology experiments。
2。 The optimized SCoT Reaction System: The P。 minima L。 SCoT-PCR reaction system (10 μL) was established, including 1uL template DNA, 5。0 uL5X PCR mix, 1uL primer, 3uL ddH2O。The amplification was performed with a PCR program: 94ºC for 5 min, followed by 35 cycles of 94ºC for 1 min, 50ºC (Primer Tm depends) for 1 min, 72ºC for 1。5 min, and a final extension at 72ºC for 10 min。
3。 Genetic persity and cluster analysis by SCoT: The twenty-three SCoT primer in total produced 186 reliable loci, The percentage of polymorphism of different primers was 66。67%, and the average of 7。13 polymorphic bands were generated by each primer。 The genetic similarity coefficient of each material was between 0。559 ~0。989。