木薯原产于巴西，随着现在经济的快速发展，木薯在世界各地的热带及亚热带地区都被广泛种植。目前，在中国的广西，广东，福建，云南，贵州及海南等省均广泛种植。木薯的块根富含淀粉，在我国工业淀粉产业中，木薯淀粉占到市场份额的10%左右，同时是重要的燃料乙醇和绿色化学工业材料。木薯是热带地区低收入农民的主要粮食作物，全球总产量的65％都被用来制作人类食品，木薯粉以及木薯叶则是高能量的饲料原料。木薯淀粉以及木薯干片也可作为制造酒精，柠檬酸，赖氨酸，谷氨酸，木薯蛋白，果糖，葡萄糖等的重要原料，生产的木薯制品在食品，饮料，药品，纺织品，造纸等各个方面都有极大的用途。木薯的工业利用和食用市场广大，我国年综合产值在200亿元人民币，并且保持数倍于产量的进口额度，保持和发展木薯产业意义重大。在木薯的科学研究和品种推广利用中，涉及万余份种质资源的保存、管理和性状鉴定评价，是木薯产业利用的物质基础，随着木薯育种和新品种选育工作的迅速发展，培育出了许多优良品种，构建基于SNP标记技术的指纹图谱有助于木薯遗传资源的进一步挖掘和利用，也有利于品种的登记保护,品种更新和品种资源的充分利用，与此同时，对于木薯品种特异性和真实性鉴别、种子纯度鉴定具有重要意义。

本研究将采用自有专利技术的AFSM简化重测序方法，获取544份木薯种质材料的SNP标记，经过一系列算法筛选以及群体结构分析，最终挑选出100条核心SNP位点来构建木薯品种指纹图谱, 期望应用于木薯品种特异性和真实性鉴别、种子纯度鉴定等，为木薯遗传资源的进一步挖掘和利用，品种的登记保护提供新的技术依据。

其主要研究结果如下：

1. 针对544份木薯样本，依照AFSM简化重测序建库要求，分别构建两个EcoRI-MspI和EcoRI-HpaII文库，经过最后的单克隆检测合格后，将对应的EcoRI-MspI和EcoRI-HpaII文库混合成2个测序文库。经过对两个文库的双端测序，总共获得了650G的数据。经过统计我们选择了1641026个变异位点，相对均匀的分布在木薯的18条染色体上。
2. 从原始的1641026个SNP位点中，通过两步过滤，去除缺失率过高以及单一变异率过高的SNP，最终保留了6886条SNP位点作为指纹图谱备选位点。
3. 经过遗传算法对6886条备选SNP计算，最终得到了在染色体上分布均匀的100条SNP组成的指纹图谱库去区分所有的544个样本。
4. 经检测，100个SNP变异位点能有效区分544份样品，并且通过计算遗传距离进行聚类分析，可以发现，相似来源的样本可以根据这100条SNP的信息聚类到一起，说明根据遗传算法计算得到的指纹图谱真实有效。
5. 根据本研究的AFSM测序及遗传算法计算指纹图谱的方法，我们对284份马铃薯成功构建了由100条SNP构成的马铃薯指纹图谱，验证了该指纹图谱技术的实用性。

Cassava originated from Brazil. With the rapid economic development, cassava is widely cultivated in tropical and subtropical regions around the world. At present, it is widely planted in Guangxi, Guangdong, Fujian, Yunnan, Guizhou and Hainan provinces in China. Cassava storage roots are rich in starch. In China's industrial starch market, cassava starch shared about 10%and also is an important fuel ethanol and green chemical industrial material. Cassava is the main food crop for low-income farmers in tropical areas. 65% of the world's total output is used to make human food, while cassava flour and cassava leaves are high-energy feed materials. Cassava starch and dried cassava slices can also be used as important raw materials for producing alcohol, citric acid, lysine, glutamic acid, tapioca protein, fructose, glucose, etc. The produced cassava products have great applications in various aspects such as food, beverage, medicine, textiles, paper making, etc. Cassava has a vast industrial use and food market. China's annual comprehensive output value is 20 billion yuan, and the import quota is several times the output. It is of great significance to maintain and develop the cassava industry. In the scientific research and variety promotion and utilization of cassava, it involves the preservation, management and character identification and evaluation of more than ten thousand germplasm resources, which is the material basis of cassava industrial utilization. With the rapid development of cassava breeding and new variety breeding, many excellent varieties have been cultivated. The construction of fingerprint based on SNP marker technology is helpful for further excavation and utilization of cassava genetic resources, as well as for the registration and protection of varieties, variety renewal and full utilization of variety resources. At the same time, it is of great significance for cassava variety specificity and authenticity identification and seed purity identification.

In this study, AFSM simplified resequencing method with proprietary technology will be adopted to obtain SNP markers of 544 cassava germplasm materials. After a series of algorithm screening and population structure analysis, 100 core SNP and INDEL loci were finally selected to construct cassava variety fingerprint. It is expected to be applied to cassava variety specificity and authenticity identification, seed purity identification, etc., that will provide new technical basis for further mining and utilization of cassava genetic resources and variety registration and protection. The main research results are as follows:

1. For 544 cassava samples, two EcoRI-MspI and EcoRI-HpaII libraries were respectively constructed according to AFSM simplified resequencing database requirements. After passing the final monoclonal detection, the corresponding EcoRI-MspI and EcoRI-HpaII libraries were mixed into two sequencing libraries. After double-ended sequencing of the two libraries, we obtained 650G of data in total. According to statistics, we selected  a total of 1,641,026 SNP  sites, which are relatively evenly distributed on 8 chromosomes of cassava genome.

2. From the original 1641026 SNP and INDEL sites, SNPs with too high deletion rate and too high single mutation rate were removed through two-step filtration, and 6,886 SNPs and INDELs sites were finally retained to be fingerprint candidate sites.

3. After calculating 6886 candidate SNPs by genetic algorithm, a fingerprint database composed of 100 SNPs and INDELs evenly distributed on chromosomes was finally obtained to distinguish all 544 samples.

4. Through detection, 100 SNPs and INDELs mutation sites can effectively distinguish 544 samples, and through clustering analysis by calculating genetic distance, it can be found that samples from similar sources can be clustered together according to the information of 100 SNPs and INDELs, which shows that the fingerprint calculated according to genetic algorithm is true and effective.

5. According to the same AFSM genotyping method and  the genetic algorithm obtained in this study, by calculating 284 potato SNP sites, we successfully constructed a potato fingerprint consisting of 100 SNPs, which verified the practicability of this fingerprint technique.

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