番茄尖果脐突变体的生理特性及其候选基因分析

doi:10.16420/j.issn.0513-353x.2023-0060

摘要/Abstract

摘要：

在栽培番茄Money Maker（MM）背景下获得的尖果脐（sharp blossom-end，shb）突变体果实的果脐突出，果实较小，呈心形，果实纵横比增加，说明SHB调控番茄果实大小和形状。徒手切片和石蜡切片观察发现，shb突变体果皮较MM果皮细胞层数显著减少，果皮变薄。在花柱发育早期，MM花柱基部的细胞小而紧密，但shb突变体花柱细胞持续膨大，花柱基部不断扩张，并在脱落后形成尖果脐。以MM和shb为亲本杂交构建了F₂代分离群体，遗传分析表明尖果脐表型是由隐性单基因控制的质量性状。将尖果脐和平果脐极端表型单株进行DNA混池，并利用BSA-seq（Bulked Segregant Analysis Coupled with Whole-Genome Sequencing）的方法初步筛选到4个候选基因。

关键词: 番茄, 果实脐部, 果实形状, 细胞分裂

Abstract:

The sharp blossom-end mutant was obtained from cultivar tomato Money Maker（MM），named as shb. Fruits of shb mutants had a pointed tip and a predominantly increased fruit shape index，shown heart-shaped with smaller size，suggesting that SHB regulates tomato fruit size and shape. The freehand section and paraffin section observations showed that the pericarp thickness and cell layers of shb mutants were significantly decreased compared with MM. During early style development，different from the small and dense cells at the base of the MM style，the shb mutant style base and the cells at this region continued to expand，eventually forming a pointed end after style dropping. An F₂population which was derived from a cross between MM（flat blossom end）and shb（sharp blossom end）was generated. Genetic analysis indicated the sharp blossom end was a qualitative trait controlled by a single recessive gene. The individual plants with extreme fruit phenotypes of the pointed end and the flat one were divided into two groups for DNA pooling，and four candidate genes were preliminarily screened by using Bulked Segregant Analysis Coupled with Whole-Genome Sequencing（BSA-Seq）.

Key words: tomato, blossom-end, fruit shape, cell division

刘泽营, 孙帅, 刘志强, 崔霞, 李仁. 番茄尖果脐突变体的生理特性及其候选基因分析[J]. 园艺学报, 2024, 51(5): 982-992.

LIU Zeying, SUN Shuai, LIU Zhiqiang, CUI Xia, LI Ren. Mapping of the Sharp Blossom-end Gene and Screening of Candidate Gene in Tomato[J]. Acta Horticulturae Sinica, 2024, 51(5): 982-992.

图/表 7

图1 栽培番茄MM及其shb突变体果实表型分析数据采用Student’s t-test检验。*，**表示显著差异（* P < 0.05；** P < 0.01）。 H：最大高度；W：最大宽度。

Fig. 1 Phenotype analysis of the cultivated tomato MM and its mutant shb fruits Data analysis was performed by student’s t-test. *，** means statistically significant differences（* P < 0.05；** P < 0.01）. H：Maximum height；W：Maximum width.

图2 栽培番茄MM和突变体shb果皮细胞形态学观察黑色直线a部分用于统计果皮细胞层数；红色矩形部分用来统计细胞数量；橙色直线b、c分别用来统计细胞长度和宽度。数据采用Student’s t-test检验。** 表示差异极显著（P < 0.01）。

Fig. 2 The morphological observation of pericarp cells in the cultivated tomato MM and the mutant shb fruits The black line a indicates the cell layer number，the red rectangle indicates the total cell number，and the orange lines b and c indicate the cell length and width，respectively. Data analysis was performed by student’s t-test. ** means statistically extremely significant differences（P < 0.01）.

图3 栽培番茄MM和shb突变体果实在花后0 d和6 d的脐部形态红箭头代表花柱将来脱落的位置。

Fig. 3 The morphology of the blossom end in the cultivated tomato MM and the mutant shb at 0 and 6 days post-anthesis The red arrow indicates the location where the style would drop.

图4 栽培番茄MM和shb突变体花后0、3和6 d的果实花柱基部组织切片红箭头指的是花柱将来脱落的位置。

Fig. 4 The cell morphological observation of the style base in the cultivated tomato MM and the shb mutant fruits at 0，3，6 days post-anthesis The red arrow indicates the location where the style would drop.

图5 栽培番茄MM、shb突变体以及其F1代果实果脐表型

Fig. 5 The phenotype of the blossom end of the cultivated tomato MM，the shb mutant line，and MM × shb F1 generation fruits

图6 利用BSA-seq绘制的影响番茄尖果脐性状的Δ(SNP-index)在全基因组上的分布图中散点代表Δ(SNP-index)，橙色直线代表|Δ(SNP-index)| > 0.8。红箭头指示筛选到的位于基因附近的5个SNP/InDel，其中2个Δ(SNP-index)值相等。

Fig. 6 Δ(SNP-index) from BSA-seq for mapping the genomic regions controlling tomato sharp blossom end The scattered points in the figure represent the calculated values of Δ(SNP-index). The orange straight line represents the |Δ(SNP-index)| > 0.8. The red arrows indicate the five screened SNP/InDels located near genes，two of which have equal delta SNP-index values.

表1 与番茄尖脐相关SNP位点及候选基因列表

Table 1 SNP and candidate genes list associated with the sharp bloosom end in tomato

单核苷酸多态性 SNP	突变在基因上位置 Location of mutation	候选基因 Candidate gene	基因注释 Annotation
SL4.0ch06_ 38607063	启动子 Promoter	Solyc06g065570	N-α-乙酰转移酶50 N-α-Acetyltransferase 50
SL4.0ch06_ 38607063	3′端非编码区 3′UTR	Solyc06g065580	尿苷-胞苷激酶 Uridine kinase
SL4.0ch08_ 55974557	启动子 Promoter	Solyc08g068740	未知蛋白 Unknown protein
SL4.0ch08_ 55974560	启动子 Promoter	Solyc08g068740	未知蛋白 Unknown protein
SL4.0ch09_ 1411527	启动子 Promoter	Solyc09g007860	钙依赖的磷脂结合蛋白Calcium-dependent lipid-binding

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