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LacDWARF1 基因突变导致丝瓜(Luffa acutangula)出现 GA 缺乏型矮化表型。

A mutation in LacDWARF1 results in a GA-deficient dwarf phenotype in sponge gourd (Luffa acutangula).

机构信息

Guangdong Key Laboratory for New Technology Research of Vegetables, Vegetable Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, 510640, Guangdong, China.

College of Agriculture & Biology, Zhongkai University of Agriculture and Engineering, Guangzhou, 510225, Guangdong, China.

出版信息

Theor Appl Genet. 2021 Oct;134(10):3443-3457. doi: 10.1007/s00122-021-03938-4. Epub 2021 Aug 14.

DOI:10.1007/s00122-021-03938-4
PMID:34390352
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8440308/
Abstract

A dwarfism gene LacDWARF1 was mapped by combined BSA-Seq and comparative genomics analyses to a 65.4 kb physical genomic region on chromosome 05. Dwarf architecture is one of the most important traits utilized in Cucurbitaceae breeding because it saves labor and increases the harvest index. To our knowledge, there has been no prior research about dwarfism in the sponge gourd. This study reports the first dwarf mutant WJ209 with a decrease in cell size and internodes. A genetic analysis revealed that the mutant phenotype was controlled by a single recessive gene, which is designated Lacdwarf1 (Lacd1). Combined with bulked segregate analysis and next-generation sequencing, we quickly mapped a 65.4 kb region on chromosome 5 using F segregation population with InDel and SNP polymorphism markers. Gene annotation revealed that Lac05g019500 encodes a gibberellin 3β-hydroxylase (GA3ox) that functions as the most likely candidate gene for Lacd1. DNA sequence analysis showed that there is an approximately 4 kb insertion in the first intron of Lac05g019500 in WJ209. Lac05g019500 is transcribed incorrectly in the dwarf mutant owing to the presence of the insertion. Moreover, the bioactive GAs decreased significantly in WJ209, and the dwarf phenotype could be restored by exogenous GA treatment, indicating that WJ209 is a GA-deficient mutant. All these results support the conclusion that Lac05g019500 is the Lacd1 gene. In addition, RNA-Seq revealed that many genes, including those related to plant hormones, cellular process, cell wall, membrane and response to stress, were significantly altered in WJ209 compared with the wild type. This study will aid in the use of molecular marker-assisted breeding in the dwarf sponge gourd.

摘要

矮化基因 LacDWARF1 通过联合 BSA-Seq 和比较基因组学分析,被定位到 05 号染色体上一个 65.4kb 的物理基因组区域。矮化结构是葫芦科作物育种中最重要的性状之一,因为它可以节省劳动力并提高收获指数。据我们所知,之前没有关于丝瓜矮化的研究。本研究报道了第一个具有细胞变小和节间缩短的矮化突变体 WJ209。遗传分析表明,突变体表型由一个隐性单基因控制,该基因被命名为 Lacdwarf1(Lacd1)。结合分离群体分组分析和下一代测序,我们使用 F2 分离群体中的 InDel 和 SNP 多态性标记,快速定位到 5 号染色体上的一个 65.4kb 区域。基因注释表明,Lac05g019500 编码赤霉素 3β-羟化酶(GA3ox),它是 Lacd1 最可能的候选基因。DNA 序列分析表明,WJ209 中 Lac05g019500 的第一个内含子中存在一个约 4kb 的插入。由于插入的存在,Lac05g019500 在矮化突变体中转录异常。此外,WJ209 中的生物活性 GAs 显著减少,外源 GA 处理可恢复矮化表型,表明 WJ209 是一个 GA 缺陷突变体。所有这些结果都支持 Lac05g019500 是 Lacd1 基因的结论。此外,RNA-Seq 显示,与野生型相比,WJ209 中有许多基因,包括与植物激素、细胞过程、细胞壁、膜和应激反应相关的基因,发生了显著改变。本研究将有助于利用分子标记辅助育种培育矮化丝瓜。

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