Rhododendron is famous for its high ornamental value.However,the genus is taxonomically difficult and the relationships within Rhododendron remain unresolved.In addition,the origin of key morphological characters with...Rhododendron is famous for its high ornamental value.However,the genus is taxonomically difficult and the relationships within Rhododendron remain unresolved.In addition,the origin of key morphological characters with high horticulture value need to be explored.Both problems largely hinder utilization of germplasm resources.Most studies attempted to disentangle the phylogeny of Rhododendron,but only used a few genomic markers and lacked large-scale sampling,resulting in low clade support and contradictory phylogenetic signals.Here,we used restriction-site associated DNA sequencing(RAD-seq)data and morphological traits for 144 species of Rhododendron,representing all subgenera and most sections and subsections of this species-rich genus,to decipher its intricate evolutionary history and reconstruct ancestral state.Our results revealed high resolutions at subgenera and section levels of Rhododendron based on RAD-seq data.Both optimal phylogenetic tree and split tree recovered five lineages among Rhododendron.Subg.Therorhodion(cladeⅠ)formed the basal lineage.Subg.Tsutsusi and Azaleastrum formed cladeⅡand had sister relationships.CladeⅢincluded all scaly rhododendron species.Subg.Pentanthera(cladeⅣ)formed a sister group to Subg.Hymenanthes(cladeⅤ).The results of ancestral state reconstruction showed that Rhododendron ancestor was a deciduous woody plant with terminal inflorescence,ten stamens,leaf blade without scales and broadly funnelform corolla with pink or purple color.This study shows significant distinguishability to resolve the evolutionary history of Rhododendron based on high clade support of phylogenetic tree constructed by RAD-seq data.It also provides an example to resolve discordant signals in phylogenetic trees and demonstrates the application feasibility of RAD-seq with large amounts of missing data in deciphering intricate evolutionary relationships.Additionally,the reconstructed ancestral state of six important characters provides insights into the innovation of key characters in Rhododendron.展开更多
Prunus is an economically important genus widely distributed in the temperate Northern Hemisphere.Previous studies on the genus using a variety of loci yielded conflicting phylogenetic hypotheses.Here,we generated nuc...Prunus is an economically important genus widely distributed in the temperate Northern Hemisphere.Previous studies on the genus using a variety of loci yielded conflicting phylogenetic hypotheses.Here,we generated nuclear reduced representation sequencing data and plastid genomes for 36 Prunus individuals and two outgroups.Both nuclear and plastome data recovered a well-resolved phylogeny.The species were divided into three main clades corresponding to their inflorescence types,-the racemose group,the solitary-flower group and the corymbose group-with the latter two sister to one another.Prunus was inferred to have diversified initially in the Late Cretaceous around 67.32 million years ago.The diversification of the three major clades began between the Paleocene and Miocene,suggesting that paleoclimatic events were an important driving force for Prunus diversification.Ancestral state reconstructions revealed that the most recent common ancestor of Prunus had racemose inflorescences,and the solitary-flower and corymb inflorescence types were derived by reduction of flower number and suppression of the rachis,respectively.We also tested the hybrid origin hypothesis of the racemose group proposed in previous studies.Prunus has undergone extensive hybridization events,although it is difficult to identify conclusively specific instances of hybridization when using SNP data,especially deep in the phylogeny.Our study provides well-resolved nuclear and plastid phylogenies of Prunus,reveals substantial cytonuclear discord at shallow scales,and sheds new light on inflorescence evolution in this economically important lineage.展开更多
限制性酶切位点关联DNA测序技术(restriction-site-associated DNA sequencing,RAD-seq)是在二代测序技术基础上发展起来的一种简化基因组技术(reduced-representation genome sequencing,RRGS)。RAD-seq利用限性核酸内切酶使基因组片段...限制性酶切位点关联DNA测序技术(restriction-site-associated DNA sequencing,RAD-seq)是在二代测序技术基础上发展起来的一种简化基因组技术(reduced-representation genome sequencing,RRGS)。RAD-seq利用限性核酸内切酶使基因组片段化,经过修饰后连接含标记的接头构建文库并进行测序。因其具有操作简单、实验成本低、通量高等优点,在分子生态学、进化基因组学、保护遗传学等领域得到应用。目前发展起来的RAD-seq技术主要包括利用稀有酶和物理打断方式进行基因组片段化的mbRAD、利用稀有酶和常见酶组合酶切的方式进行基因组片段化的ddRAD、利用IIB型限制性内切酶得到短片段文库的2bRAD、利用同裂酶和Illumina试剂盒建库的ezRAD和完全利用Nextera试剂盒建库的nextRAD。本文对每种RAD-seq技术的特点及其在昆虫种群遗传学、群落生态学、物种界定、系统发育和遗传连锁图谱构建等研究领域的应用进行了综述。展开更多
[目的]筛选一种能够得到高质量桑树基因组DNA的方法。[方法]采用天根试剂盒法(Plant Genomic DNA Kit离心柱型),并稍加改进,从11个桑种的新鲜嫩叶中提取DNA。[结果]该方法简单快速,不需要复杂仪器,且能有效地除去桑叶中的多糖、多酚、...[目的]筛选一种能够得到高质量桑树基因组DNA的方法。[方法]采用天根试剂盒法(Plant Genomic DNA Kit离心柱型),并稍加改进,从11个桑种的新鲜嫩叶中提取DNA。[结果]该方法简单快速,不需要复杂仪器,且能有效地除去桑叶中的多糖、多酚、蛋白质等杂质,得到的DNA能够满足RAD-Seq高通量的测序要求。[结论]建立了一种用于桑RAD-Seq高通量测序的基因组DNA提取方法。展开更多
Restriction-site associated DNA sequencing(RAD-seq)技术是在二代测序基础上发展起来的一项基于全基因组酶切位点的简化基因组测序技术。该方法技术流程简单,不受有无参考基因组的限制,可大大简化基因组的复杂性,减少实验费用,通过...Restriction-site associated DNA sequencing(RAD-seq)技术是在二代测序基础上发展起来的一项基于全基因组酶切位点的简化基因组测序技术。该方法技术流程简单,不受有无参考基因组的限制,可大大简化基因组的复杂性,减少实验费用,通过一次测序就可以获得数以万计的多态性标记。目前,RAD-seq技术已成功应用于超高密度遗传图谱的构建、重要性状的精细定位、辅助基因组序列组装、群体基因组学以及系统发生学等基因组研究热点领域。文章主要介绍了RAD-seq的技术原理、技术发展及其在基因组研究中的广泛应用。鉴于RAD-seq方法的独特性,该技术必将在复杂基因组研究领域具有广泛的应用前景。展开更多
旨在分析静原鸡白羽、麻羽、黑羽保种群之间的遗传变异,挖掘调控特征性状的关键候选基因。本研究利用RAD-seq技术对180日龄特征明显、发育正常且健康的白羽、麻羽、黑羽静原鸡(每个羽色选取60个个体,40只母鸡,20只公鸡)进行测序,基于SN...旨在分析静原鸡白羽、麻羽、黑羽保种群之间的遗传变异,挖掘调控特征性状的关键候选基因。本研究利用RAD-seq技术对180日龄特征明显、发育正常且健康的白羽、麻羽、黑羽静原鸡(每个羽色选取60个个体,40只母鸡,20只公鸡)进行测序,基于SNP标记计算观察杂合度(Ho)、群体内核酸多态性(Pi)、群体的平均近交系数(Fis)等指标,分析静原鸡3个类群的遗传多样性和群体结构,并通过选择性清除分析和全基因组关联分析(GWAS)筛选出候选基因,利用KOBAS对候选基因进行KEGG(Kyoto Encyclopedia of Genes and Genomes)通路富集,最终筛选出调控静原鸡羽色的候选基因。测序结果表明,静原鸡180个样品共产生198.83 Gb Clean Data,Q30达到93%以上。遗传多样性分析表明,白羽、麻羽、黑羽静原鸡分别鉴定出238533、233562和240820个SNPs标记,Ho、Pi和Fis分别在0.2732~0.2782、0.3049~0.3096和0.0961~0.1098之间。群体结构分析表明,静原鸡根据不同羽色分为不同类群。通过选择性清除分析和全基因组关联分析共筛选出11个(FZD4、WNT16、EDNRB、TYR、KRAS、CTNNB1、DDC、MC1R、CAMK2A、PRKCB、PRKCA)与静原鸡羽色相关的候选基因,富集结果显示这些基因主要与黑色素生成、酪氨酸代谢和Wnt信号传导等通路相关。综上所述,本研究利用SNPs标记信息可以全面地评价静原鸡的保种现状,为静原鸡的遗传资源保护奠定理论基础。同时,筛选出了与静原鸡羽色性状相关的候选基因,为静原鸡不同羽色的品系化培育提供新的遗传标记和基因靶点。展开更多
限制性酶切位点相关DNA测序(restriction-site associated DNA sequencing,RAD-seq)是在二代测序技术上发展起来的简化基因组测序技术,该技术操作简单、实验成本低、并且简化了基因组的复杂度.鱼类是脊椎动物中物种数最多的类群,由于生...限制性酶切位点相关DNA测序(restriction-site associated DNA sequencing,RAD-seq)是在二代测序技术上发展起来的简化基因组测序技术,该技术操作简单、实验成本低、并且简化了基因组的复杂度.鱼类是脊椎动物中物种数最多的类群,由于生活环境的不同,造就了丰富的物种多样性,常作为基因组学的研究对象.目前,在鱼类基因组学研究中,RAD-seq技术广泛应用于鱼类的系统发育、物种分化、遗传图谱、群体遗传和适应性进化等方面的研究.研究主要基于RAD-seq技术在鱼类基因组学中的研究进展进行综述,以期为鱼类资源的保护和合理利用提供参考依据.展开更多
基金supported by Ten Thousand Talent Program of Yunnan Province(Grant No.YNWR-QNBJ-2018-174)the Key Basic Research Program of Yunnan Province,China(Grant No.202101BC070003)+3 种基金National Natural Science Foundation of China(Grant No.31901237)Conservation Program for Plant Species with Extremely Small Populations in Yunnan Province(Grant No.2022SJ07X-03)Key Technologies Research for the Germplasmof Important Woody Flowers in Yunnan Province(Grant No.202302AE090018)Natural Science Foundation of Guizhou Province(Grant No.Qiankehejichu-ZK2021yiban 089&Qiankehejichu-ZK2023yiban 035)。
文摘Rhododendron is famous for its high ornamental value.However,the genus is taxonomically difficult and the relationships within Rhododendron remain unresolved.In addition,the origin of key morphological characters with high horticulture value need to be explored.Both problems largely hinder utilization of germplasm resources.Most studies attempted to disentangle the phylogeny of Rhododendron,but only used a few genomic markers and lacked large-scale sampling,resulting in low clade support and contradictory phylogenetic signals.Here,we used restriction-site associated DNA sequencing(RAD-seq)data and morphological traits for 144 species of Rhododendron,representing all subgenera and most sections and subsections of this species-rich genus,to decipher its intricate evolutionary history and reconstruct ancestral state.Our results revealed high resolutions at subgenera and section levels of Rhododendron based on RAD-seq data.Both optimal phylogenetic tree and split tree recovered five lineages among Rhododendron.Subg.Therorhodion(cladeⅠ)formed the basal lineage.Subg.Tsutsusi and Azaleastrum formed cladeⅡand had sister relationships.CladeⅢincluded all scaly rhododendron species.Subg.Pentanthera(cladeⅣ)formed a sister group to Subg.Hymenanthes(cladeⅤ).The results of ancestral state reconstruction showed that Rhododendron ancestor was a deciduous woody plant with terminal inflorescence,ten stamens,leaf blade without scales and broadly funnelform corolla with pink or purple color.This study shows significant distinguishability to resolve the evolutionary history of Rhododendron based on high clade support of phylogenetic tree constructed by RAD-seq data.It also provides an example to resolve discordant signals in phylogenetic trees and demonstrates the application feasibility of RAD-seq with large amounts of missing data in deciphering intricate evolutionary relationships.Additionally,the reconstructed ancestral state of six important characters provides insights into the innovation of key characters in Rhododendron.
基金This work was supported by grants from National Natural Science Foundation of China(No.32170381 and 31770200).
文摘Prunus is an economically important genus widely distributed in the temperate Northern Hemisphere.Previous studies on the genus using a variety of loci yielded conflicting phylogenetic hypotheses.Here,we generated nuclear reduced representation sequencing data and plastid genomes for 36 Prunus individuals and two outgroups.Both nuclear and plastome data recovered a well-resolved phylogeny.The species were divided into three main clades corresponding to their inflorescence types,-the racemose group,the solitary-flower group and the corymbose group-with the latter two sister to one another.Prunus was inferred to have diversified initially in the Late Cretaceous around 67.32 million years ago.The diversification of the three major clades began between the Paleocene and Miocene,suggesting that paleoclimatic events were an important driving force for Prunus diversification.Ancestral state reconstructions revealed that the most recent common ancestor of Prunus had racemose inflorescences,and the solitary-flower and corymb inflorescence types were derived by reduction of flower number and suppression of the rachis,respectively.We also tested the hybrid origin hypothesis of the racemose group proposed in previous studies.Prunus has undergone extensive hybridization events,although it is difficult to identify conclusively specific instances of hybridization when using SNP data,especially deep in the phylogeny.Our study provides well-resolved nuclear and plastid phylogenies of Prunus,reveals substantial cytonuclear discord at shallow scales,and sheds new light on inflorescence evolution in this economically important lineage.
文摘限制性酶切位点关联DNA测序技术(restriction-site-associated DNA sequencing,RAD-seq)是在二代测序技术基础上发展起来的一种简化基因组技术(reduced-representation genome sequencing,RRGS)。RAD-seq利用限性核酸内切酶使基因组片段化,经过修饰后连接含标记的接头构建文库并进行测序。因其具有操作简单、实验成本低、通量高等优点,在分子生态学、进化基因组学、保护遗传学等领域得到应用。目前发展起来的RAD-seq技术主要包括利用稀有酶和物理打断方式进行基因组片段化的mbRAD、利用稀有酶和常见酶组合酶切的方式进行基因组片段化的ddRAD、利用IIB型限制性内切酶得到短片段文库的2bRAD、利用同裂酶和Illumina试剂盒建库的ezRAD和完全利用Nextera试剂盒建库的nextRAD。本文对每种RAD-seq技术的特点及其在昆虫种群遗传学、群落生态学、物种界定、系统发育和遗传连锁图谱构建等研究领域的应用进行了综述。
文摘[目的]筛选一种能够得到高质量桑树基因组DNA的方法。[方法]采用天根试剂盒法(Plant Genomic DNA Kit离心柱型),并稍加改进,从11个桑种的新鲜嫩叶中提取DNA。[结果]该方法简单快速,不需要复杂仪器,且能有效地除去桑叶中的多糖、多酚、蛋白质等杂质,得到的DNA能够满足RAD-Seq高通量的测序要求。[结论]建立了一种用于桑RAD-Seq高通量测序的基因组DNA提取方法。
文摘Restriction-site associated DNA sequencing(RAD-seq)技术是在二代测序基础上发展起来的一项基于全基因组酶切位点的简化基因组测序技术。该方法技术流程简单,不受有无参考基因组的限制,可大大简化基因组的复杂性,减少实验费用,通过一次测序就可以获得数以万计的多态性标记。目前,RAD-seq技术已成功应用于超高密度遗传图谱的构建、重要性状的精细定位、辅助基因组序列组装、群体基因组学以及系统发生学等基因组研究热点领域。文章主要介绍了RAD-seq的技术原理、技术发展及其在基因组研究中的广泛应用。鉴于RAD-seq方法的独特性,该技术必将在复杂基因组研究领域具有广泛的应用前景。
文摘旨在分析静原鸡白羽、麻羽、黑羽保种群之间的遗传变异,挖掘调控特征性状的关键候选基因。本研究利用RAD-seq技术对180日龄特征明显、发育正常且健康的白羽、麻羽、黑羽静原鸡(每个羽色选取60个个体,40只母鸡,20只公鸡)进行测序,基于SNP标记计算观察杂合度(Ho)、群体内核酸多态性(Pi)、群体的平均近交系数(Fis)等指标,分析静原鸡3个类群的遗传多样性和群体结构,并通过选择性清除分析和全基因组关联分析(GWAS)筛选出候选基因,利用KOBAS对候选基因进行KEGG(Kyoto Encyclopedia of Genes and Genomes)通路富集,最终筛选出调控静原鸡羽色的候选基因。测序结果表明,静原鸡180个样品共产生198.83 Gb Clean Data,Q30达到93%以上。遗传多样性分析表明,白羽、麻羽、黑羽静原鸡分别鉴定出238533、233562和240820个SNPs标记,Ho、Pi和Fis分别在0.2732~0.2782、0.3049~0.3096和0.0961~0.1098之间。群体结构分析表明,静原鸡根据不同羽色分为不同类群。通过选择性清除分析和全基因组关联分析共筛选出11个(FZD4、WNT16、EDNRB、TYR、KRAS、CTNNB1、DDC、MC1R、CAMK2A、PRKCB、PRKCA)与静原鸡羽色相关的候选基因,富集结果显示这些基因主要与黑色素生成、酪氨酸代谢和Wnt信号传导等通路相关。综上所述,本研究利用SNPs标记信息可以全面地评价静原鸡的保种现状,为静原鸡的遗传资源保护奠定理论基础。同时,筛选出了与静原鸡羽色性状相关的候选基因,为静原鸡不同羽色的品系化培育提供新的遗传标记和基因靶点。
文摘限制性酶切位点相关DNA测序(restriction-site associated DNA sequencing,RAD-seq)是在二代测序技术上发展起来的简化基因组测序技术,该技术操作简单、实验成本低、并且简化了基因组的复杂度.鱼类是脊椎动物中物种数最多的类群,由于生活环境的不同,造就了丰富的物种多样性,常作为基因组学的研究对象.目前,在鱼类基因组学研究中,RAD-seq技术广泛应用于鱼类的系统发育、物种分化、遗传图谱、群体遗传和适应性进化等方面的研究.研究主要基于RAD-seq技术在鱼类基因组学中的研究进展进行综述,以期为鱼类资源的保护和合理利用提供参考依据.
