岩风的遗传多样性与谱系地理学研究
发布时间：2018-08-13 21:19
【摘要】：岩风(Libanotis buchtormensis(Fisch.)DC.)为伞形科(Umbelliferae)岩风属(Libanotis Haller ex Zinn)多年生亚灌木状草本,为秦岭地区“太白七药”之一,俗称“长春七”。该物种广泛分布于欧亚草原和中亚山地,东亚的温带森林地区也有零星分布,其自然分布区呈现西北荒漠区两侧的间断分布,而且居群间存在明显的形态变异。目前,岩风的遗传多样性和遗传结构研究仍处于空白状态,其地理分布格局的成因也并不明朗。本研究在广泛采样基础上,利用多种分子遗传标记(ISSR、cpDNA和rpb2)对岩风自然居群进行了遗传多样性、遗传结构、冰期避难所和居群历史动态分析,探讨了其间断地理分布式样的形成原因。此外,从资源利用的角度,结合高效液相技术和多元统计分析,建立了岩风不同地理分布区居群的化学多样性评价体系。将遗传多样性、化学多样性数据和生态位模型结果相结合,系统地阐释了不同分布区岩风资源的异同,为岩风资源可持续开发与利用提供了有效建议。其主要研究结果如下:⑴利用ISSR分子标记对采自涵盖岩风自然分布区的19个居群进行了遗传多样性和遗传结构分析。10条ISSR引物共扩增出125条多态性条带,占总扩增带的90.58%。岩风在物种水平上表现出较高的遗传多样性(h=0.298),但是居群水平上的遗传多样性较低(h=0.110)。基于遗传距离的NJ聚类、贝叶斯算法的STRUCTURE聚类以及主坐标分析,都支持将岩风的19个自然居群分为两组,来自新疆的居群聚为NW组,来自秦岭(陕西、甘肃)和四川的居群聚为SE组,秦岭和四川地区居群可细分为两个亚支,聚类结果和地理分布格局相对应。AMOVA分析表明岩风居群间存在显著的遗传分化(68.5%)。Mantel test检验表明遗传距离和地理距离显著相关(r=0.76,P0.001),遗传分化呈现地理分布格局。⑵基于cpDNA和低拷贝核基因rpb2序列对间断分布的岩风居群进行了谱系地理分析。21个自然居群和6个标本材料共发现23个叶绿体单倍型和24个核基因单倍型。岩风表现出明显的谱系地理结构,主要分为NW和SE两个谱系分支,支持ISSR的分析结果。受西北荒漠区扩张的影响,NW和SE谱系大约在中更新世分化(c.0.51 Ma BP),然后适应不同的分布区从而独立进化和发展。西北荒漠区产生的地理隔离可能阻碍了NW和SE谱系的基因流,形成现在荒漠区两侧间断分布的地理格局,而且由于长时间的局域适应已经形成明显的形态变异。NW谱系居群受到第四纪冰期和间冰期反复作用的影响,并在冰期存在至少两个独立的避难所,间冰期从不同避难所沿着山脉向外扩张,居群可能发生了多次交汇融合,在末次盛冰期(LGM)前已经形成和现代相似的分布。SE谱系受LGM影响较大,在LGM时居群退缩至南方位于川西高原附近的避难所,间冰期条件适宜时向北回迁。居群可能经历了严重的瓶颈效应和奠基者效应,导致SE谱系遗传多样性较低。⑶利用高效液相色谱技术建立了12个不同产地岩风的化学指纹图谱,发现不同产地的岩风样品化学成分含量差异显著,特别是蛇床子素和异欧前胡素存在明显的组间差异,可以作为不同产地的指示性化学成分。多元统计分析方法都支持将不同产地的12批岩风样品划分为2组,根据蛇床子素和异欧前胡素组分比的高低,命名为两个化学型。⑷基于PWG距离法、系统树方法和相似性搜索法系统评价了五对DNA候选条形码(ITS、mat K、rbcL、trnL-trnF和ycf1b)及其主要组合片段序列对18种岩风属植物的鉴定能力。结果发现核心条形码matK+rbcL的鉴定能力小于ITS和任一叶绿体片段的组合,ITS+matK、ITS+matK+rbcL和ITS+matK+rbcL+trnL-trnF+ycf1b能成功鉴别所有的岩风属植物,出于时效和成本考虑,ITS+matK可以作为岩风属物种鉴别的适用条形码。联合片段(ITS+matK+rbcL+trnL-trnF+ycf1b)构建的系统发育树证明了岩风的种级地位。西北荒漠区两侧区域差异的岩风植株为不同的生态型。⑸最大熵生态位模型(MAXENT)的结果表明,岩风绝大部分的适生区位于新疆地区,而且该地区岩风最适生态位参数和秦岭及四川地区存在异同。刀切法显示,降水量年变化(25.8%)、一月份降雨量(25.5%)和海拔(24.2%)是影响岩风生长的关键环境因子,土壤因子对岩风生长分布影响较小。新疆地区岩风适宜分布区的降水量季节性变化系数在50左右,秦岭和四川地区的变异系数为70~80。这可能和两区域所处不同的气候带有关系,也正是如此秦岭地区的岩风成为特色的道地药材。结合遗传多样性、化学多样性数据以及生态位参数,四川地区可作为岩风药材资源的扩展分布区。在未来全球逐渐变暖的场景预设下(2050和2070年),岩风的适宜分布区逐渐缩小,影响最大的为准噶尔盆地荒漠区边缘以及秦岭地区的分布区,并表现出适宜分布区逐渐向高海拔的川西高原迁移的趋势,该地区的石质干旱河谷可能在未来成为岩风植物新的集中分布区之一,可作为今后岩风GAP种植的选址地区。秦岭地区可采取就地保护并结合近地保护的措施对岩风资源进行保护。
[Abstract]:Libanotis buchtormensis (Fisch.) DC. is a shrubby perennial herb of Umbelliferae (Libanotis Haller ex Zinn), which is one of the "Taibai Qiyao" in Qinling Mountains, commonly known as "Changchun Qi". The species is widely distributed in Eurasian grasslands and mountainous areas of Central Asia, and also sporadically distributed in temperate forest areas of East Asia. At present, the genetic diversity and genetic structure of rock breeze are still in a blank state, and the genesis of its geographical distribution pattern is not clear. In this study, a variety of molecular genetic markers (ISSR, cpDNA and rpb) were used on the basis of extensive sampling. 2) The genetic diversity, genetic structure, glacial shelter and population history of natural rock breeze populations were analyzed, and the reasons for the formation of discontinuous geographic distribution patterns were discussed. Value system. Combining genetic diversity, chemical diversity data and niche model results, the similarities and differences of rock breeze resources in different districts are systematically explained, which provides effective suggestions for sustainable development and utilization of rock breeze resources. The main results are as follows: (1) Using ISSR molecular markers, 19 natural distribution areas of rock breeze are sampled. Genetic diversity and genetic structure of the population were analyzed. 125 polymorphic bands were amplified by 10 ISSR primers, accounting for 90.58% of the total amplified bands. Rock breeze showed high genetic diversity at species level (h = 0.298), but low genetic diversity at population level (h = 0.110). Based on NJ clustering of genetic distance, STR of Bayesian algorithm was used. UCTURE clustering and principal coordinate analysis support that 19 natural populations of Yanfeng can be divided into two groups. The populations from Xinjiang are clustered into NW group, the populations from Qinling (Shaanxi, Gansu) and Sichuan are clustered into SE group, and the populations from Qinling and Sichuan can be subdivided into two sub-branches. The results of cluster analysis correspond to the geographic distribution pattern. Mantel test showed that there was a significant correlation between genetic distance and geographical distance (r = 0.76, P 0.001), and the genetic differentiation showed a geographical distribution pattern. _Based on cpDNA and low copy nuclear gene rpb2 sequence, the phylogenetic analysis of discontinuous distribution of rock breeze population was carried out. There are 23 chloroplast haplotypes and 24 nuclear gene haplotypes. Rock breeze shows distinct pedigree geographic structure, mainly divided into two pedigree branches, NW and SE, which support ISSR analysis results. Geographical isolation in the northwest desert area may hinder the gene flow of the NW and SE lineages, forming a geographic pattern of discontinuous distribution on both sides of the desert area, and the population of the NW lineage is influenced by the repetition of the Quaternary glacial and interglacial periods, and exists during the glacial period. At least two independent shelters, extending outward from different shelters along the mountains during the interglacial period, may have multiple confluences and merges, forming a modern-day distribution similar to that before the Last Glacial Maximum (LGM). The SE pedigree was strongly influenced by LGM, and the population retreated to the southern shelter near the western Sichuan Plateau at LGM, where the interglacial conditions were appropriate. The population may have experienced severe bottleneck effect and founder effect, resulting in low genetic diversity of SE lineage. _Chemical fingerprints of rock breeze from 12 different habitats were established by high performance liquid chromatography, and significant differences in chemical composition were found among rock breeze samples from different habitats, especially Osthol and isoeurope. There are obvious inter-group differences in praeruptorin, which can be used as indicative chemical constituents of different habitats. Multivariate statistical analysis supports dividing 12 batches of rock breeze samples from different habitats into two groups, named two chemical types according to the ratio of Osthol and isoimperatorin. _Based on PWG distance method, system tree method and similarity. Five pairs of DNA candidate barcodes (ITS, mat K, rbcL, trnL-trnF and ycf1b) and their major combinations were systematically evaluated to identify 18 species of genus Lyophyllum. The results showed that the identification ability of core barcodes matK + rbcL was less than that of ITS and any combination of chloroplast segments, ITS + matK, ITS + matK + rbcL and ITS + K + rbcL + matrnL-trnF + f1b. For the sake of time and cost, ITS+matK can be used as a suitable bar code for the identification of the genus Lyophylla. The phylogenetic tree constructed by the combined fragment (ITS+matK+rbcL+trnL-trnF+ycf1b) proved the species status of Lyophylla. The results of the maximum entropy niche model (MAXENT) show that most of the suitable areas for rock breeze are located in Xinjiang, and the optimum niche parameters of rock breeze in this area are different from those in Qinling and Sichuan areas. The seasonal variation coefficient of precipitation in the suitable distribution area of rock breeze in Xinjiang is about 50, and the variation coefficient in Qinling and Sichuan is 70-80. This may be related to the different climatic zones in the two regions, and it is also the rock breeze in Qinling that has become a characteristic genuine medicinal material. The genetic diversity, chemical diversity data and niche parameters can be used as the extended distribution areas of rock breeze medicinal materials in Sichuan. Under the scenario of global warming in the future (2050 and 2070), the suitable distribution areas of rock breeze will gradually shrink, and the most influential areas are the desert margin of Junggar Basin and the distribution areas of Qinling Mountains. The arid valleys in this area may become one of the new concentrated distribution areas of rock breeze plants in the future, and can be used as the site selection area for future rock breeze GAP planting. The Qinling Mountains can take measures of local protection and near-ground protection to protect rock breeze resources. Protect.
【学位授予单位】：西北农林科技大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S567.239
，
本文编号：2182220
[Abstract]:Libanotis buchtormensis (Fisch.) DC. is a shrubby perennial herb of Umbelliferae (Libanotis Haller ex Zinn), which is one of the "Taibai Qiyao" in Qinling Mountains, commonly known as "Changchun Qi". The species is widely distributed in Eurasian grasslands and mountainous areas of Central Asia, and also sporadically distributed in temperate forest areas of East Asia. At present, the genetic diversity and genetic structure of rock breeze are still in a blank state, and the genesis of its geographical distribution pattern is not clear. In this study, a variety of molecular genetic markers (ISSR, cpDNA and rpb) were used on the basis of extensive sampling. 2) The genetic diversity, genetic structure, glacial shelter and population history of natural rock breeze populations were analyzed, and the reasons for the formation of discontinuous geographic distribution patterns were discussed. Value system. Combining genetic diversity, chemical diversity data and niche model results, the similarities and differences of rock breeze resources in different districts are systematically explained, which provides effective suggestions for sustainable development and utilization of rock breeze resources. The main results are as follows: (1) Using ISSR molecular markers, 19 natural distribution areas of rock breeze are sampled. Genetic diversity and genetic structure of the population were analyzed. 125 polymorphic bands were amplified by 10 ISSR primers, accounting for 90.58% of the total amplified bands. Rock breeze showed high genetic diversity at species level (h = 0.298), but low genetic diversity at population level (h = 0.110). Based on NJ clustering of genetic distance, STR of Bayesian algorithm was used. UCTURE clustering and principal coordinate analysis support that 19 natural populations of Yanfeng can be divided into two groups. The populations from Xinjiang are clustered into NW group, the populations from Qinling (Shaanxi, Gansu) and Sichuan are clustered into SE group, and the populations from Qinling and Sichuan can be subdivided into two sub-branches. The results of cluster analysis correspond to the geographic distribution pattern. Mantel test showed that there was a significant correlation between genetic distance and geographical distance (r = 0.76, P 0.001), and the genetic differentiation showed a geographical distribution pattern. _Based on cpDNA and low copy nuclear gene rpb2 sequence, the phylogenetic analysis of discontinuous distribution of rock breeze population was carried out. There are 23 chloroplast haplotypes and 24 nuclear gene haplotypes. Rock breeze shows distinct pedigree geographic structure, mainly divided into two pedigree branches, NW and SE, which support ISSR analysis results. Geographical isolation in the northwest desert area may hinder the gene flow of the NW and SE lineages, forming a geographic pattern of discontinuous distribution on both sides of the desert area, and the population of the NW lineage is influenced by the repetition of the Quaternary glacial and interglacial periods, and exists during the glacial period. At least two independent shelters, extending outward from different shelters along the mountains during the interglacial period, may have multiple confluences and merges, forming a modern-day distribution similar to that before the Last Glacial Maximum (LGM). The SE pedigree was strongly influenced by LGM, and the population retreated to the southern shelter near the western Sichuan Plateau at LGM, where the interglacial conditions were appropriate. The population may have experienced severe bottleneck effect and founder effect, resulting in low genetic diversity of SE lineage. _Chemical fingerprints of rock breeze from 12 different habitats were established by high performance liquid chromatography, and significant differences in chemical composition were found among rock breeze samples from different habitats, especially Osthol and isoeurope. There are obvious inter-group differences in praeruptorin, which can be used as indicative chemical constituents of different habitats. Multivariate statistical analysis supports dividing 12 batches of rock breeze samples from different habitats into two groups, named two chemical types according to the ratio of Osthol and isoimperatorin. _Based on PWG distance method, system tree method and similarity. Five pairs of DNA candidate barcodes (ITS, mat K, rbcL, trnL-trnF and ycf1b) and their major combinations were systematically evaluated to identify 18 species of genus Lyophyllum. The results showed that the identification ability of core barcodes matK + rbcL was less than that of ITS and any combination of chloroplast segments, ITS + matK, ITS + matK + rbcL and ITS + K + rbcL + matrnL-trnF + f1b. For the sake of time and cost, ITS+matK can be used as a suitable bar code for the identification of the genus Lyophylla. The phylogenetic tree constructed by the combined fragment (ITS+matK+rbcL+trnL-trnF+ycf1b) proved the species status of Lyophylla. The results of the maximum entropy niche model (MAXENT) show that most of the suitable areas for rock breeze are located in Xinjiang, and the optimum niche parameters of rock breeze in this area are different from those in Qinling and Sichuan areas. The seasonal variation coefficient of precipitation in the suitable distribution area of rock breeze in Xinjiang is about 50, and the variation coefficient in Qinling and Sichuan is 70-80. This may be related to the different climatic zones in the two regions, and it is also the rock breeze in Qinling that has become a characteristic genuine medicinal material. The genetic diversity, chemical diversity data and niche parameters can be used as the extended distribution areas of rock breeze medicinal materials in Sichuan. Under the scenario of global warming in the future (2050 and 2070), the suitable distribution areas of rock breeze will gradually shrink, and the most influential areas are the desert margin of Junggar Basin and the distribution areas of Qinling Mountains. The arid valleys in this area may become one of the new concentrated distribution areas of rock breeze plants in the future, and can be used as the site selection area for future rock breeze GAP planting. The Qinling Mountains can take measures of local protection and near-ground protection to protect rock breeze resources. Protect.
【学位授予单位】：西北农林科技大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S567.239
，
本文编号：2182220
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