State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Horticulture, Nanjing Agricultural University, Weigang Street No. 1, Nanjing, 210095, China.
Theor Appl Genet. 2020 Feb;133(2):371-382. doi: 10.1007/s00122-019-03468-0. Epub 2019 Nov 16.
Via bulked segregant analysis sequencing combined with linkage mapping, the ist gene responsible for the irregularly striped rind mutation was delimited to a 144-kb region in cucumber. Sequencing and expression analysis identified Csa1G005490 as the candidate gene. The rind appearance of cucumber is one of the most important commercial quality traits. Usually, an immature cucumber fruit has a uniform rind that varies from green to yellow to white among different cultivated varieties. In the present paper, we isolated a novel fruit appearance cucumber mutant, ist, that has an irregularly striped rind pattern. The mutant displayed green irregular stripes on a yellow-green background at the immature fruit stage. Genetic analysis revealed that a single recessive gene, ist, is responsible for this mutation. A BSA (bulked segregant analysis) sequencing approach combined with genetic mapping delimited the ist locus to an interval with a length of 144 kb, and 21 predicted genes were annotated in the region. Based on mutation site screening and expression analysis, two single-nucleotide polymorphisms within the candidate gene, Csa1G005490, were identified as constituting the mutation. Csa1G005490 encodes a polygalacturonase-1 noncatalytic subunit beta protein (PG1β) known to be involved in fruit softening. The expression of Csa1G005490 was significantly lower in the ist mutant than in the wild type. Transcriptome analysis identified 1796 differentially expressed genes (DEGs) between the ist mutant and wild type. Gene ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that these DEGs were enriched mostly in photosynthesis and chlorophyll metabolism pathways. Decreased expression patterns of several chlorophyll synthesis genes in the mutant suggest that ist plays a key role in chlorophyll biosynthesis. These results will provide new insight into the molecular mechanism underlying rind appearance polymorphisms in cucumber.
通过 bulked segregant analysis sequencing 结合连锁图谱定位,负责果皮不规则条纹突变的 ist 基因被限定在黄瓜的 144kb 区域内。测序和表达分析鉴定 Csa1G005490 为候选基因。黄瓜的果皮外观是最重要的商业品质性状之一。通常,未成熟的黄瓜果实具有均匀的果皮,不同栽培品种之间从绿色到黄色到白色变化。在本研究中,我们分离出一种新型果实外观黄瓜突变体 ist,其果皮具有不规则的条纹图案。在未成熟果实阶段,突变体显示出绿色的不规则条纹,背景为黄绿色。遗传分析表明,单个隐性基因 ist 负责该突变。BSA(bulked segregant analysis)测序方法与遗传图谱相结合,将 ist 基因座定位到 144kb 的区间内,该区域注释了 21 个预测基因。基于突变位点筛选和表达分析,在候选基因 Csa1G005490 内鉴定出两个单核苷酸多态性构成突变。Csa1G005490 编码多聚半乳糖醛酸酶-1 非催化亚基 β 蛋白(PG1β),已知其参与果实软化。在 ist 突变体中,Csa1G005490 的表达明显低于野生型。转录组分析鉴定出 ist 突变体和野生型之间有 1796 个差异表达基因(DEGs)。GO 注释和 KEGG 富集分析表明,这些 DEGs 主要富集在光合作用和叶绿素代谢途径中。突变体中几个叶绿素合成基因的表达下调表明,ist 在叶绿素生物合成中起关键作用。这些结果将为黄瓜果皮外观多态性的分子机制提供新的见解。