分子标记辅助构建甜瓜矮化基因Cmerecta近等基因系

doi:10.16420/j.issn.0513-353x.2023-0989

摘要/Abstract

摘要：

前期通过图位克隆的方法精细定位到一个控制节间长度的CmERECTA基因，并开发了一个和甜瓜矮化共分离的标记Cmerecta-dCAPS2。为了进一步研究其功能，以甜瓜自交系TopMark为轮回亲本，矮化自交系M406为供体亲本，利用Cmerecta-dCAPS2为前景选择标记，对筛选出的102对在亲本间多态性好、染色体上分布相对均匀的SSR标记为背景选择标记，在TopMark遗传背景下构建Cmerecta近等基因系，并统计分析其表型。结果显示，BC₁F₁和BC₂F₁世代最高背景回复率单株的背景回复率分别为80.61%、98.57%。背景回复率最高的3个BC₂F₂代Cmerecta纯系植株的表型显示其株高均为矮化，叶片变圆。该近等基因系可作为后续研究甜瓜矮化基因CmERECTA的调控机理以及应用的基础材料。

关键词: 甜瓜, 分子标记, 近等基因系, 矮化基因, CmERECTA, 辅助育种

Abstract:

In the early stage，a CmERECTA gene that controls internode length was finely mapped by map cloning，and a marker Cmerecta-dCAPS2 co-separated from melon dwarf was developed. In order to further study its function，the 102 pairs of SSR markers with good polymorphism and relatively uniform distribution on chromosomes were detected by using the melon inbred line TopMark as the reincarnation parent，the dwarf inbred line M406 as the donor parent，and the Cmerecta-dCAPS2 as the foreground selection marker，the background recovery rate of the SSR markers with good polymorphism and relatively uniform distribution on the chromosomes was detected，and the Cmerecta near-isogenic line was constructed under the TopMark genetic background，and its phenotype was statistically analyzed. The results showed that the highest background response rates per plant of BC₁F₁ and BC₂F₁ generations were 80.61% and 98.57%，respectively. The phenotypes of the three BC₂F₂ generation Cmerecta pure lines with the highest background recovery rate showed that the plant height was dwarf and the leaves were rounded. The construction of this near-isogenic line can be used as a basic material for subsequent research on the regulatory mechanism and application of CmERECTA，a dwarf gene in melon.

Key words: melon, molecular marker, near-isogenic line, dwarfing gene, CmERECTA, assisted breeding

刘港运, 段世享, 许娜娜, 郭姚淼, 豆峻岭, 杨森, 牛欢欢, 刘东明, 杨路明, 胡建斌, 朱华玉. 分子标记辅助构建甜瓜矮化基因Cmerecta近等基因系[J]. 园艺学报, 2024, 51(9): 2048-2062.

LIU Gangyun, DUAN Shixiang, XU Nana, GUO Yaomiao, DOU Junling, YANG Sen, NIU Huanhuan, LIU Dongming, YANG Luming, HU Jianbin, ZHU Huayu. Molecular Markers Assisted Construction of Cmerecta Near-Isogenic Line of Melon Dwarfing Gene[J]. Acta Horticulturae Sinica, 2024, 51(9): 2048-2062.

图/表 9

图1 Cmerecta近等基因系（NIL）构建过程

Fig. 1 Cmerecta near-isogenic line（NIL）construction process

图2 dCAPS2前景选择标记和102对多态性SSR标记在甜瓜染色体组上的分布左侧数字代表背景选择标记在染色体上的实际物理位置（Mb）。

Fig. 2 dCAPS2 distribution of foreground selection markers and 102 pairs of polymorphic SSR markers on the melon chromosome group The number on the left side represents the actual physical position of the background selection marker on the chromosome（Mb）.

表1 甜瓜近等基因系背景选择标记在染色体上的分布

Table 1 Distribution of background selection markers in melon near isogenic lines on the chromosome

染色体 Chromosome	多态性标记数量 Number of polymorphism markers	平均距离/Mb Average distance	标记间最大距离/Mb The maximum distance between markers
Chr1	7	5.5	17.6
Chr2	8	3.4	12.1
Chr3	9	3.0	10.0
Chr4	9	2.7	14.5
Chr5	11	2.5	8.7
Chr6	8	3.8	11.7
Chr7	9	3.0	12.3
Chr8	7	3.7	14.6
Chr9	9	2.9	13.5
Chr10	8	2.0	3.7
Chr11	7	4.2	10.5
Chr12	10	2.7	8.5
总计 Total	102	3.1	11.5

图3 BC2F1群体前景选择和部分背景选择结果 A：dCAPS2在前景选择中的酶切结果。B：背景选择标记SSR07253的扩增结果。

Fig. 3 BC2F1 population foreground selection and partial background selection results A：Digestion results of dCAPS2 in prospect selection. B：Amplification of background selection marker SSR07253.

图4 BC2F1群体候选单株基因型图示红色和蓝色区域分别代表轮回亲本遗传背景和供体亲本导入的染色体片段，灰色代表杂合片段，绿色代表背景回复率最高的单株。

Fig. 4 Graphrepresentation of candidate genotypes of individual strains of the BC2F1 population The red and blue regions of the genotype represent the genetic background of the reincarnation parent and the chromosome fragments introduced by the donor parent. Gray represents heterozygous fragment，and green represents the individual plant with the highest background recovery rate.

图5 BC2F1背景回复率最高单株背景选择标记在染色体上的分布红色和蓝色区域分别代表轮回亲本遗传背景和供体亲本导入的染色体片段，灰色代表杂合片段。

Fig. 5 Distribution of individual plant background selection markers across chromosomes with the highest BC2F1 background response rate The red and blue regions of the genotype represent the genetic background of the reincarnation parent and the chromosome fragments introduced by the donor parent. Gray represents heterozygous fragment.

图6 BC2F2群体候选单株基因型图示红色和蓝色区域分别代表轮回亲本遗传背景和供体亲本导入的染色体片段，灰色代表杂合片段，绿色代表背景回复率最高的单株。

Fig. 6 Graphrepresentation of candidate genotypes of individual strains of the BC2F2 population The red and blue regions of the genotype represent the genetic background of the reincarnation parent and the chromosome fragments introduced by the donor parent. Gray represents heterozygous fragment，and green represents the individual plant with the highest background recovery rate.

图7 BC2F2背景回复率最高单株在染色体上的基因型分布红色和蓝色区域分别代表轮回亲本遗传背景和供体亲本导入的染色体片段，灰色代表杂合片段。

Fig. 7 Genotype distribution on the chromosome with the highest BC2F2 background response rate The red and blue regions of the genotype represent the genetic background of the reincarnation parent and the chromosome fragments introduced by the donor parent. Gray represents heterozygous fragment.

图8 定植30 d Cmerecta-NIL表型观察（A、B）与性状统计（C）柱状图统计方法为方差分析，不同字母表示差异显著。P < 0.05。

Fig. 8 Cmerecta-NIL phenotype observation（A，B）and trait statistics（C）at 30 days The statistical method of the histogram is ANOVA. The difference lowercase letters indicate significant differences at 0.05 level.

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