基于粪便DNA的黄河三角洲丹顶鹤越冬种群个体识别及遗传多样性分析
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1.辽宁大学生命科学院 沈阳 110036;2.吉林省民航机场集团有限公司 长春 130000;3.辽宁大学司法鉴定中心 沈阳 110036

作者简介:

李楠,女,硕士研究生;研究方向:鸟类学;E-mail:ln991012@163.com。

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基金项目:

国家自然科学基金项目(No. 31672316,31911540468);


Individual identification and genetic diversity analysis of the wintering population of Red-crowned Cranes (Grus japonensis) based on fecal DNA in the Yellow River Delta, China
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Affiliation:

1.School of Life Sciences, Liaoning University, Shenyang 110036; 2.Jilin Province Civil Aviation Airport Group Co., Ltd., Changchun 130000; 3.Liaoning University Judicial Authentication Center, Shenyang 110036, China

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    摘要:

    丹顶鹤(Grus japonensis)为我国一级重点保护野生动物,被世界自然保护联盟(IUCN)列为易危级(VU)物种。受自然栖息地不断恶化的影响,我国野生丹顶鹤种群数量持续处于低位。黄河三角洲作为我国丹顶鹤的重要迁徙停歇地和第二大越冬地,近些年越冬种群数量持续增加,了解该种群的遗传背景及多样性水平对于开展其种群遗传保护具有重要意义。本研究采用非损伤性取样方法,于2021年12月至2022年2月在山东黄河三角洲国家级自然保护区采集了264份丹顶鹤粪样,并成功提取127份样品的DNA。选用7个微卫星位点和线粒体Cyt b基因进行个体识别和遗传多样性分析。经微卫星基因分型,通过个体识别确认127份样品来自于115只个体。检测到60个微卫星等位基因,有效等位基因数Ne在1.640 ~ 6.414之间,平均观测杂合度HO为0.617,平均多态信息含量PIC为0.578,平均期望杂合度HE为0.628,其中,期望杂合度显著低于扎龙保护区种群(P < 0.05)。有6个位点偏离哈迪-温伯格平衡,近交系数为0.128。基于Cyt b基因进行线粒体遗传多样性分析,共成功扩增53份序列,检测到13个变异位点和12个单倍型,单倍型多样性Hd为0.756,核苷酸多样性Pi为0.001 92,表现为较低水平。微卫星群体遗传结构分析显示,该种群可分为4个亚群,但遗传分化程度较低,群体间遗传分化指数FST为0.020 ~ 0.056。Cyt b单倍型网络图未见明显种内遗传分化。综上,黄河三角洲丹顶鹤越冬种群的遗传多样性水平较低,可能反映出该种群的遗传结构相对单一。本文利用粪便DNA进行野生丹顶鹤个体识别和遗传多样性研究,此方法将为进一步开展野生丹顶鹤种群的遗传多样性评估及其他濒危动物的遗传学研究提供新思路。

    Abstract:

    [Objectives] The Red-crowned Crane (Grus japonensis) is listed as a National Class I Key Protected Species in China and evaluated as Vulnerable (VU) by the International Union for Conservation of Nature (IUCN). It has maintained a small wild population in China due to the deterioration of its natural habitats. As an important migratory stopover site and the second-largest wintering ground for this species in China, the Yellow River Delta has witnessed a continuous increase in its wintering population size in recent years. Understanding the genetic characteristics and diversity of this population is crucial for implementing effective conservation strategies. [Methods] We employed a non-invasive sampling method to collect 264 fecal samples of Red-crowned Cranes from the Yellow River Delta National Nature Reserve from December 2021 to February 2022 (Fig. 1), and extracted DNA from 127 samples. Individual identification and genetic diversity assessment were conducted based on seven microsatellite loci and the mitochondrial Cyt b gene. CERVUS 3.0 was utilized for individual identification and calculation of genetic diversity parameters, including the effective number of alleles (Ne), observed heterozygosity (HO), expected heterozygosity (HE), polymorphism information content (PIC), and Hardy-Weinberg equilibrium (HWE). GenAlEx V6.51b2 was used to quantify private alleles, inbreeding coefficient (FIS), probability of identity (PID), and probability of identity among siblings (PIDsib). The population genetic structure was analyzed via Structure 2.3.4, and the genetic differentiation index (FST) and gene flow (Nm) between populations were calculated. Mitochondrial genetic diversity analysis was performed with DNASP 5.0 to calculate haplotype diversity (Hd) and nucleotide diversity (Pi). A median-joining haplotype network was constructed by POPART 1.7. [Results] Individual identification confirmed that 127 samples originated from 115 distinct individuals (Fig. 2). A total of 60 microsatellite alleles were detected, with Ne ranging from 1.640 to 6.414. The population exhibited low genetic diversity, with HO, PIC, and HE values of 0.617, 0.578, and 0.628, respectively. Its expected heterozygosity was significantly lower than that of the Zhalong population (P = 0.002), but not significantly different from those of the Yancheng and Hokkaido wintering populations (P > 0.05; Tables 2 and 4). The population showed a high inbreeding coefficient (FIS = 0.128). Cyt b gene sequences were successfully amplified from 53 samples, revealing 13 variable sites and 12 distinct haplotypes. Haplotype diversity (Hd = 0.756) and nucleotide diversity (Pi = 0.001 92) indicated low genetic diversity (Pi < 0.005). Microsatellite-based population structure analysis divided the population into four subpopulations, while the genetic differentiation was minimal (FST 0.020 ~ 0.056, Nm 4.198 ~ 12.329; Figs. 3 and 4, Table 3). The haplotype network constructed from Cyt b sequences showed no significant intraspecific genetic differentiation (Fig. 5). [Conclusion] These findings suggest that the wintering population in the Yellow River Delta exhibits low genetic diversity and a relatively homogeneous genetic structure. This study pioneers the use of fecal DNA for individual identification and genetic analysis in wild Red-crowned Cranes, providing a methodological application for genetic diversity assessments for wild Red-crowned Crane populations and other endangered species.

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李楠,孙娇,李东来. 2026.基于粪便DNA的黄河三角洲丹顶鹤越冬种群个体识别及遗传多样性分析. 动物学杂志, 61(4): 564-575.

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  • 收稿日期:2025-05-07
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  • 在线发布日期: 2026-08-25
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