The Brolga (Antigone rubicunda) is a large Australian crane species with a broad distribution spanning from the tropical north to the south-eastern regions of the continent. Brolga populations throughout New South Wales, Victoria and South Australia have been in decline since the early twentieth century, with the species being listed as vulnerable in each state. To aid future conservation of the species, its taxonomic status needs to be validated, and patterns of gene flow and population connectivity across the species distribution need to be understood. To assist future genetic studies, we developed a suite of polymorphic microsatellite markers and the complete mitochondrial genome sequence by next-generation sequencing. A total of 18 polymorphic loci were characterised using DNA extractions from 47 individuals, comprising 30 and 17 individuals from Victoria and northern Australia, respectively. We observed moderate genetic variation across loci with only a single locus deviating significantly from Hardy–Weinberg equilibrium. De novo and reference-based genome assemblies were used to assemble the A. rubicunda mitochondrial genome sequence, which consists of 16,700 base pairs, and a typical metazoan mitochondrial gene content and arrangement. We test these new markers by conducting a preliminary analysis of genetic structure between south-eastern and northern Australian Brolga populations. Mitochondrial analyses provided evidence of shared haplotypes across the species range supporting the conspecific status of extant populations, while microsatellite markers indicated weak but significant genetic differentiation suggesting gene flow is limited. We discuss the implications of these findings and the benefits that these genetic markers will provide for future population genetic research on this iconic Australian bird species.

中文翻译：

---

通过下一代测序和从头基因组组装表征的微卫星位点和完整的线粒体 DNA 序列，以及对澳大利亚鹤安提戈涅 rubicunda 种群遗传结构的初步评估

Brolga (Antigone rubicunda) 是一种大型的澳大利亚鹤类，分布广泛，从热带北部到非洲大陆的东南部地区。自 20 世纪初以来，新南威尔士州、维多利亚州和南澳大利亚州的 Brolga 种群数量一直在下降，该物种在每个州都被列为易危物种。为了帮助未来保护该物种，需要验证其分类地位，并需要了解整个物种分布的基因流动和种群连通性模式。为了协助未来的遗传研究，我们通过下一代测序开发了一套多态性微卫星标记和完整的线粒体基因组序列。使用来自 47 个个体的 DNA 提取物表征了总共 18 个多态性位点，分别由来自维多利亚州和澳大利亚北部的 30 人和 17 人组成。我们观察到跨位点的中等遗传变异，只有一个位点显着偏离 Hardy-Weinberg 平衡。使用从头和基于参考的基因组组装来组装 A. rubicunda 线粒体基因组序列，该序列由 16,700 个碱基对组成，以及典型的后生动物线粒体基因含量和排列。我们通过对澳大利亚东南部和北部 Brolga 种群之间的遗传结构进行初步分析来测试这些新标记。线粒体分析提供了支持现存种群同种状态的物种范围内共享单倍型的证据，而微卫星标记表明弱但显着的遗传分化表明基因流有限。