Wang Baotian, Li Weijia, Xu Kexin, Lei Yingying, Zhao Di, Li Xue, Zhang Junxiang, Zhang Zhihong
Liaoning Key Laboratory of Strawberry Breeding and Cultivation, College of Horticulture, Shenyang Agricultural University, Shenyang, 110866, China.
Laboratory of Protected Horticulture (Shenyang Agricultural University), Ministry of Education, Shenyang, People's Republic of China.
Hortic Res. 2022 Nov 10;10(1):uhac249. doi: 10.1093/hr/uhac249. eCollection 2023.
Leaves and flowers are crucial for the growth and development of higher plants. In this study we identified a mutant with narrow leaflets and early flowering () in an ethyl methanesulfonate-mutagenized population of woodland strawberry () and aimed to identify the candidate gene. Genetic analysis revealed that a single recessive gene, , controlled the mutant phenotype. We found that FvH4_1g25470, which encodes a putative DNA polymerase α with a polymerase and histidinol phosphatase domain (PHP), might be the candidate gene, using bulked segregant analysis with whole-genome sequencing, molecular markers, and cloning analyses. A splice donor site mutation (C to T) at the 5' end of the second intron led to an erroneous splice event that reduced the expression level of the full-length transcript of in mutant plants. FvePHP was localized in the nucleus and was highly expressed in leaves. Silencing of using the virus-induced gene silencing method resulted in partial developmental defects in strawberry leaves. Overexpression of the gene can largely restore the mutant phenotype. The expression levels of , , , , and were higher in the mutants than those in 'Yellow Wonder' plants, probably contributing to the early flowering phenotype in mutant plants. Our results indicate that mutation in is associated with multiple developmental pathways. These results aid in understanding the role of DNA polymerase in strawberry development.
叶片和花朵对于高等植物的生长和发育至关重要。在本研究中,我们在林地草莓(Fragaria vesca)经甲磺酸乙酯诱变的群体中鉴定出一个小叶狭窄且开花早的突变体(),旨在鉴定候选基因。遗传分析表明,一个单隐性基因,控制了突变体表型。通过全基因组测序的混合分组分析法、分子标记和克隆分析,我们发现FvH4_1g25470可能是候选基因,该基因编码一种具有聚合酶和组氨醇磷酸酶结构域(PHP)的推定DNA聚合酶α。第二个内含子5'端的剪接供体位点突变(C到T)导致错误的剪接事件,降低了突变体植株中全长转录本的表达水平。FvePHP定位于细胞核,在叶片中高度表达。使用病毒诱导基因沉默方法沉默导致草莓叶片出现部分发育缺陷。基因的过表达可在很大程度上恢复突变体表型。突变体中、、、和的表达水平高于‘Yellow Wonder’植株,这可能是突变体植株开花早表型的原因。我们的结果表明,中的突变与多个发育途径相关。这些结果有助于理解DNA聚合酶在草莓发育中的作用。