联合基因组重测序和10× Genomics scRNA-Seq解析乌骨鸡胸肌黑色素转运的分子机制

我国部分乌骨鸡品种缺乏科学、系统的选育，导致在“乌色”这一重要性状上存在群体整齐度低的共性不足，制约种质资源的保护利用效率和养殖经济效益的提高。胸肌是乌骨鸡黑色素沉积的重要部位，证据表明其黑色素转运调控可能与皮肤组织不同，但相关机制仍不清楚，阻碍遗传选育进展、种质创新和保护。本项目拟以雪峰乌骨鸡为研究对象，联用基因组重测序、10× Genomics scRNA-Seq技术、基因过表达、shRNA沉默和Crispr/Cas9等技术，挖掘黑色素转运关联突变和候选基因，构建雪峰乌骨鸡胸肌各细胞亚群/子亚群的基因调控网络，研究关键突变和基因在胸肌黑色素转运中的作用，阐明遗传变异和基因表达调控胸肌黑色素转运的分子机制。研究结果对于认识乌骨鸡个体间胸肌黑色素含量差异的遗传基础和发生机制具有重要的理论价值，而且也为我国乌骨鸡乌色度的选育提高和营养调控提供新的分子标记和分子靶标，具有重要的应用价值。

A poor uniformity of melanin pigmentation, probably result from a deficiency of scientific and systematic breeding, has been observed widely in some native black-boned chicken breeds. It can partly account for the decrease in preservation and utilization efficiency of genetic resources and the economic benefit from raising industry of black-bone chickens. Pectoral muscle is one of the major tissues for melanin deposition in black-bone chickens. It has been reported that the genes involving in the melanin transfer of pectoral muscle is distinct from that involving in cutaneous tissue. However, the detailed molecular mechanism remains yet to be elucidated. Investigation of the key genetic variations and genes affecting melanin transferring in pectoral muscle has been proposed as the optimal periodic strategy, and should be served as the prerequisite for the subsequent genetic breeding program, innovation and preservation of genetic resources. In this study, Xuefeng black-boned chickens will be used to screen the key genes and allelic genetic variations, which make up the gene regulatory network and manipulate the capacity of melanin transferring in pectoral muscles. We plan to construct the gene regulatory network in level of cell types/subtypes by integrating the genome re-sequencing and 10× genomic RNA-Seq. Furthermore, the subsequent functional validation of screened genes will also be performed mediated by overexpression profiles, shRNA knocking-down and Crispr/Cas9. The genetic basis and molecular mechanism will be elucidated, by which make the capacity of melanin transferring different in pectoral muscles. The results will not only shed light on understanding the genetic basis and the causal mechanisms resulting in difference of melanin content in pectoral muscles, but also provide practically valuable molecular markers and potential targeted genes for improvements of blackness by means of genetics breeding and/or nutrition regulation.