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    基于多基因rDNA标记的重庆地区椭圆四极虫遗传多样性研究

    GENETIC DIVERSITY ANALYSIS OF CHLOROMYXUM ELLIPTICUM FROM CHONGQING BASED ON MULTI-GENE rDNA MARKERS

    • 摘要: 为补充椭圆四极虫(Chloromyxum ellipticum)的分子数据并解析其种内遗传变异特征, 本研究对采自重庆市合川、大足、江津、长寿、万州5个区县81尾养殖草鱼(Ctenopharyngodon idellus)胆囊中的阳性样本进行了形态学观察与多基因rDNA分子系统学分析。形态学观察记录了壳面条纹(10—16条)、极丝圈数(4—5圈)及孢子量度, 孢子长7.58±0.64 (6.26—9.83) μm, 孢子宽6.02±1.08 (5.36—7.99) μm, 极囊长2.29±0.25 (1.38—2.79) μm, 极囊宽 1.35±0.26 (0.93—1.55) μm。基于18S rDNA、5.8S rDNA、ITS2 rDNA、5.8S-ITS2 rDNA及18S-5.8S-ITS2 rDNA联合序列, 开展了系统发育、单倍型、主成分及遗传距离分析。结果表明, 重庆地区椭圆四极虫存在明显的种内遗传变异, 且5个区县的单倍型分布与地理来源无显著关联, 推测与养殖草鱼苗种的跨地区流通及混合养殖模式有关。在所用分子标记中, 18S-5.8S-ITS2 rDNA联合序列的分辨率最高, 共检出10个变异位点, 定义16个单倍型(Hd = 0.8929), 种内遗传距离为0.001—0.003。此外, 本研究首次报道了椭圆四极虫ITS2 rDNA基因序列, 并发现重庆种群与湖北种群在18S rDNA上存在4个固定碱基差异, 提示两地种群可能存在更深层次的遗传分化。本研究为椭圆四极虫的准确鉴定及种群遗传学研究提供了新的分子标记与基础数据。

       

      Abstract: To supplement molecular data of Chloromyxum ellipticum and elucidate its intraspecific genetic variation, this study conducted morphological observations and multi-gene rDNA molecular phylogenetic analyses on positive samples isolated from the gallbladders of 81 cultured grass carp (Ctenopharyngodon idellus) from five districts (Hechuan, Dazu, Jiangjin, Changshou, and Wanzhou). Morphological observation recorded 10—16surface striations, 4—5 polar filament coils, a myxospore length of 7.58±0.64 (6.26—9.83) μm, a myxospore width of 6.02±1.08 (5.36—7.99) μm, a polar capsule length of 2.29±0.25(1.38—2.79) μm, and a polar capsule width of 1.35±0.26(0.93—1.55) μm. Phylogenetic, haplotype, principal component, and genetic distance analyses were performed based on 18S rDNA, 5.8S rDNA, ITS2 rDNA, 5.8S-ITS2 rDNA, as well as the combined 18S-5.8S-ITS2 rDNA sequence. The results indicated that C. ellipticum in Chongqing exhibits significant intraspecific genetic variation, and haplotype distribution from the five districts exhibited no significant correlation with geographical origins, presumably owing to cross-regional circulation and mixed farming practices of grass carp fingerlings. Among all molecular markers used, the concatenated 18S-5.8S-ITS2 rDNA sequence showed the highest resolution, detecting 10 variable sites and defined 16haplotypes (Hd =0.8929), with intraspecific genetic distances ranging from 0.001 to 0.003. Furthermore, the ITS2 rDNA sequence of C. ellipticum was reported for the first time in the present study, and four fixed nucleotide substitutions were detected in the 18S rDNA sequences between Chongqing and Hubei population, indicating potential deep genetic divergence between them. This study provides new molecular markers and fundamental data for the accurate identification and future population genetic studies of C. ellipticum.

       

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