基于COS Marker分析柑橘属及其近缘、远缘属植物的遗传与进化

doi:10.3864/j.issn.0578-1752.2017.02.011

摘要/Abstract

摘要： 【目的】研究柑橘属及其近缘、远缘属种质资源的遗传多样性及亲缘关系，为柑橘类果树的遗传和进化关系提供新的视点，也为遗传育种以及种质资源深入的收集、保护及评价利用提供新工具和参考。【方法】下载Phytozome网站公布的全部克里迈丁Conserved ortholog sequences，并在NCBI中与测序甜橙序列比对，获得具有Gap的序列，从中设计60对COS引物，筛选并利用多态性标记对柑橘属及其近缘、远缘属植物进行扩增检测，通过Structure、PowerMarker和GenAlEx等软件进行数据分析。【结果】从设计引物中筛选出谱带清晰、稳定性好、具有多态性且分布在柑橘不同连锁群的引物25对，对45份柑橘属及其近缘、远缘属种质资源进行扩增检测，共得到584条谱带，平均每对引物约为23.36条；基因多样性为0.26—0.88，平均0.49；PIC为0.26—0.87，平均0.48。群体遗传结构分析显示，在K=9时，群体结构图清晰地揭示了柑橘属及其近缘、远缘属植物间的遗传组成关系，可区分为宽皮柑橘类群、柚类群、枸橼类群、大翼橙类群、金柑类群、枳及澳沙檬澳圆檬类群、花椒类群、黄皮酒饼簕九里香类群以及印度野橘和莽山野柑杂合类群，类群关系与聚类分析结果总体比较一致。UPGMA聚类分析显示，COS Marker能够明确地区分柑橘属及其近缘、远缘属种质资源，柑橘属在较高分类层次与金柑属、枳属、澳沙檬属、澳圆檬属、花椒属以及黄皮、酒饼簕、九里香等近、远缘植物分离；在柑橘属内，枸橼和大翼橙类首先聚类，然后与宽皮柑橘、柚类等其他柑橘属植物聚类；宽皮柑橘类群里，道县野橘、岑溪酸橘等半野生宽皮柑橘与椪柑等栽培宽皮柑橘可以明确区分，而莽山野柑和印度野橘不在本类群。【结论】根据甜橙和克里迈丁基因测序信息设计的COS引物，在柑橘近、远缘属植物中均能获得有效扩增，能够有效区分柑橘属及其近缘、远缘属种质资源；莽山野柑和印度野橘的遗传组成表现杂合，聚类分析中与宽皮柑橘距离比较远，可能并非宽皮橘类群最原始类型。

关键词: 柑橘, COS Marker, 亲缘关系, 种质资源, 遗传与进化

Abstract: 【Objective】To reveal genetic diversity and phylogenetic relationships of Citrus and its close and distant relatives with newly developed conserved ortholog sequence (COS) Marker.【Method】According to the published C. sinensis and C. clementina genomic sequencing information, COS Markers were developed and screened. Polymorphic markers were used to amplify and detect Citrus and its close and distant relatives. Structure, Power Marker and GenAlEx were used to analyze the relevant data.【Result】A total of 60 COS primers were designed, and 25 primer pairs distributed in different linkage groups were selected based on clear and good DNA polymorphic bands. In 45 samples from Citrus and its relatives, 25 COS markers produced 584 bands, with an average of about 23.36 bands per marker, and genetic diversity ranged from 0.26 to 0.88, with an average of 0.49; PIC value was between 0.26 and 0.87, with an average of 0.48. When K = 9, a clear genetic structure relationship of citrus and its relatives were revealed. They were divided into C. reticulata group, C. grandis group, C. medica group, Papeda group, Fortunella group, P. trifoliate and Eremocitrus and M. australis group, Z. bungeanum group, Clausena lansium and Atalantia buxifolia and Murraya paniculata group as well as C. indica and C. mangshanensis hybrid groups. This result was globally consistent with that in the UPGMA clustering analysis. UPGMA clustering analysis revealed that COS Marker could effectively distinguish Citrus from its close and distant germplasm resources. At higher taxonomy level, Citrus was first separated from P. trifoliate, Eremocitrus, M. australis, Z. bungeanum, Clausena lansium, Atalantia buxifolia and Murraya paniculata; In Citrus, C. medica and Papeda group were first clustered, and then they were clustered with C. reticulata and C. grandis and other plants of Citrus. In C. reticulata group, Daoxian yeju and Cenxi suanju and other half wild C. reticulata were clearly distinguished from ponkan and the rest cultivated C. reticulata, but C. mangshanensis and C. indica were not in the group.【Conclusion】COS primers based on Citrus genomic information could get effective amplification and distinguish Citrus from its close and distant relatives effectively. The genetic makeup of C. mangshanensis and C. indica were heterozygous and different from C. reticulata, so they may not be the most primitive types of C. reticulata.

Key words: Citrus, COS marker, phylogenetic relationship, germplasm, genetics and evolution

王炯，龚桂芝，彭祝春，李一兵，王艳杰，洪棋斌. 基于COS Marker分析柑橘属及其近缘、远缘属植物的遗传与进化[J]. 中国农业科学, 2017, 50(2): 320-331.

WANG Jiong, GONG GuiZhi, PENG ZhuChun, LI YiBing, WANG YanJie, HONG QiBin. Genetic and Phylogenetic Relationships Among Citrus and Its Close and Distant Relatives Based on COS Marker[J]. Scientia Agricultura Sinica, 2017, 50(2): 320-331.

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