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通过两个果皮颜色存在差异的基因型的 RNA-Seq 数据,探索砂梨果皮红锈色的候选基因。

Exploring candidate genes for pericarp russet pigmentation of sand pear (Pyrus pyrifolia) via RNA-Seq data in two genotypes contrasting for pericarp color.

机构信息

Institute of Horticulture, Zhejiang Academy of Agricultural Sciences, Hangzhou, Zhejiang Province, China.

出版信息

PLoS One. 2014 Jan 6;9(1):e83675. doi: 10.1371/journal.pone.0083675. eCollection 2014.

Abstract

Sand pear (Pyrus pyrifolia) russet pericarp is an important trait affecting both the quality and stress tolerance of fruits. This trait is controlled by a relative complex genetic process, with some fundamental biological questions such as how many and which genes are involved in the process remaining elusive. In this study, we explored differentially expressed genes between the russet- and green-pericarp offspring from the sand pear (Pyrus pyrifolia) cv. 'Qingxiang' × 'Cuiguan' F1 group by RNA-seq-based bulked segregant analysis (BSA). A total of 29,100 unigenes were identified and 206 of which showed significant differences in expression level (log2fold values>1) between the two types of pericarp pools. Gene Ontology (GO) analyses detected 123 unigenes in GO terms related to 'cellular_component' and 'biological_process', suggesting developmental and growth differentiations between the two types. GO categories associated with various aspects of 'lipid metabolic processes', 'transport', 'response to stress', 'oxidation-reduction process' and more were enriched with genes with divergent expressions between the two libraries. Detailed examination of a selected set of these categories revealed repressed expressions of candidate genes for suberin, cutin and wax biosynthesis in the russet pericarps.Genes encoding putative cinnamoyl-CoA reductase (CCR), cinnamyl alcohol dehydrogenase (CAD) and peroxidase (POD) that are involved in the lignin biosynthesis were suggested to be candidates for pigmentation of sand pear russet pericarps. Nine differentially expressed genes were analyzed for their expressions using qRT-PCR and the results were consistent with those obtained from Illumina RNA-sequencing. This study provides a comprehensive molecular biology insight into the sand pear pericarp pigmentation and appearance quality formation.

摘要

砂梨(Pyrus pyrifolia)锈斑果皮是影响果实品质和抗逆性的重要性状。该性状受相对复杂的遗传过程控制,一些基本的生物学问题,如涉及多少个基因以及哪些基因,仍不清楚。在这项研究中,我们通过 RNA-seq 基于 bulked segregant analysis (BSA) 方法,探索了砂梨(Pyrus pyrifolia)cv. 'Qingxiang' × 'Cuiguan' F1 群体中锈斑果和绿皮果之间差异表达的基因。共鉴定了 29100 个 unigenes,其中 206 个在两种果皮池之间的表达水平差异显著(log2fold 值>1)。GO 分析检测到与“细胞组分”和“生物过程”相关的 GO 术语中的 123 个 unigenes,表明两种果皮之间存在发育和生长分化。与“脂质代谢过程”、“运输”、“应激反应”、“氧化还原过程”等方面相关的 GO 类别与两种文库之间差异表达的基因富集在一起。对这些类别中的一个选定集合进行详细检查,发现锈斑果皮中参与角质素、蜡质生物合成的候选基因表达受到抑制。推测编码肉桂酰辅酶 A 还原酶(CCR)、肉桂醇脱氢酶(CAD)和过氧化物酶(POD)的基因是砂梨锈斑果皮色素沉着的候选基因,这些基因参与木质素生物合成。使用 qRT-PCR 分析了 9 个差异表达基因的表达情况,结果与 Illumina RNA-seq 一致。本研究为砂梨果皮色素沉着和外观品质形成提供了全面的分子生物学见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd35/3882208/18ae7534f1e4/pone.0083675.g001.jpg

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