苦瓜全雌性KASP标记的开发与应用

doi:10.13560/j.cnki.biotech.bull.1985.2026-0205

摘要/Abstract

摘要：

目的苦瓜全雌系在高效杂交育种中起着重要作用，为提高全雌系材料的选择效率，通过定位苦瓜全雌性状调控基因，开发与其紧密连锁的分子标记，为开展苦瓜全雌系分子育种提供理论依据与技术支撑。方法以苦瓜全雌自交系MC52及雌雄同株自交系K8为试材，构建F₂分离群体，调查其F₁、F₂等世代的花型分化情况，并进行遗传规律分析；根据F₂分离群体构建极端性状混池，通过BSA-seq重测序和精细定位，对调控苦瓜全雌性状基因进行定位；基于定位区间内两亲本间的变异位点，开发与全雌性状紧密连锁的分子标记并验证该标记的准确性和通用性。结果 MC52与K8进行杂交，其F₂分离符合3∶1孟德尔遗传规律；成功将调控全雌性状基因gy1-1定位在分子标记G-8和G-9之间，区域大小约176 kb，区间共包含21个编码基因；开发了GY-KASP分子标记并对F₂群体进行基因型鉴定，与苦瓜花性型表型鉴定结果相比较，准确率达到100%；进一步利用GY-KASP对54份苦瓜自交系材料进行基因型检测，其中全雌材料表现为C：C型，雌雄同株材料表现为G：G或G：C型。结论苦瓜全雌性状由隐性单基因控制，命名为gy1-1，定位在1号染色体上约176 kb的物理区间；开发的GY-KASP标记能准确高效鉴定苦瓜全雌系和雌雄同株系，提高良种选择效率，降低育种成本，加快育种进程。

关键词: 苦瓜, 全雌性, 基因定位, KASP标记, 功能标记, 辅助选择育种, 遗传规律

Abstract:

Objective Gynoecious lines play a key role in efficient hybrid breeding. To enhance the selection efficiency of such lines, this study aims to map the genes regulating the gynoecious trait in bitter gourd and develop closely linked molecular markers, thereby providing a theoretical basis and technical support for molecular breeding of gynoecious lines. Method The gynoecious inbred line MC52 was crossed with the monoecious inbred line K8 to generate an F₂ segregating population. Floral type segregation in the F₁, F₂ and other generations was investigated, and genetic segregation analysis was performed. Based on the F₂ population, extreme-phenotype bulked pools were constructed for fine mapping of the regulatory genes for the gynoecious trait using BSA-seq and subsequent fine mapping. Based on variant sites between the two parental lines within the mapped interval, molecular markers closely linked to the gynoecious trait were developed and validated for accuracy and universality. Result The segregation of the F₂ population derived from MC52 × K8 followed a Mendelian ratio of 3∶1. The gynoecious gene gy1-1 was successfully mapped between markers G-8 and G-9, covering approximately 176 kb, which contained 21 coding genes. The GY-KASP marker was developed and used to genotype the F₂ population, showing 100% concordance with the floral sex phenotype data. Furthermore, GY-KASP was applied to genotype 54 bitter gourd inbred lines, in which gynoecious materials displayed the C:C genotype, whereas monoecious materials displayed the G:G or G:C genotypes. Conclusion Gynoecy in bitter gourd is controlled by a single recessive gene, designated gy1-1, which was delimited to a physical interval of approximately 176 kb on chromosome 1. The developed GY-KASP marker can accurately and efficiently distinguish gynoecious from monoecious lines of bitter gourd, thereby improving selection efficiency for elite varieties, reducing breeding costs, and accelerating the breeding process.

Key words: Momordica charantia L., gynoecy, gene mapping, KASP marker, functional marker, marker-assisted selection (MAS) breeding, inheritance pattern

黄玉辉, 黄如葵, 冯诚诚, 琚茜茜, 黄熊娟, 梁家作, 刘杏连, 陈小凤. 苦瓜全雌性KASP标记的开发与应用[J]. 生物技术通报, doi: 10.13560/j.cnki.biotech.bull.1985.2026-0205.

HUANG Yu-hui, HUANG Ru-kui, FENG Cheng-cheng, JU Xi-xi, HUANG Xiong-juan, LIANG Jia-zuo, LIU Xing-lian, CHEN Xiao-feng. Development and Application of KASP Markers for Gynoecy in Bitter Gourd[J]. Biotechnology Bulletin, doi: 10.13560/j.cnki.biotech.bull.1985.2026-0205.

图/表 5

表1 苦瓜全雌性状遗传规律

Table 1 Inheritance of the gynoecious trait in bitter gourd

世代 Generation	雌雄同株植株 Monoecious plants	全雌植株 Gynoecious plants	分离比 Segregation ratio	χ²	χ²（0.05）
P₁（MC52）	0	20
P₂（K8）	20	0
F₁（P₁×P₂）	30	0
F₂（F_{1（P1×P2）}）	371	134	3∶1	0.59	3.84

图1 基于BSA-Seq预测对苦瓜全雌基因gy1-1的定位A：Δ（SNP-index）值的分布图；B：ED值的全基因组分布图；散点图为原始值，黑色曲线为拟合值，蓝线和红线分别为95%和99%置信水平阈值线。横坐标表示染色体的位置

Fig. 1 Localization of gynoecious gene gy1-1 based on BSA-Seq predictionA: Δ(SNP-index) distribution map. B: Whole-genome distribution map of ED values. The scatter plot represents the original values, the black curve represents the corresponding fitted values, the blue and red lines represent the threshold lines for 95% and 99% confidence level respectively. The horizontal axis represents the position of chromosomes

图2 苦瓜全雌基因gy1-1的物理图谱A：gy1-1初定位于1号染色体18.7‒21.1 cM之间；B：利用505个单株精细定位gy1-1于G-8和G-9标记约176 kb范围内，标记下面的数字代表交换单株数

Fig. 2 Physical map of gynoecious gene gy1-1A: The gy1-1 gene is located on chromosome 1 between markers 18.7 cM and 21.1 cM. B: The gy1-1 gene was mapped to a 176 kb region between markers G-8 and G-9, number of recombinant individuals are shown below the markers

图3 GY-KASP标记在部分F2代群体（A）和54份苦瓜材料（B）中的基因分型结果蓝色信号：全雌型，基因型为C︰C；红色信号：雌雄同株型，基因型为G︰G；绿色：雌雄同株型，基因型为G︰C

Fig. 3 Genotyping results of the GY-KASP marker in some F2 populations (A) and 54 bitter gourd materials (B)Blue signal: Gynoecious type with genotype C:C. Red signal: Monoecious type with genotype G:G. Green: Monoecious type with genotype G:C

表2 供试苦瓜材料表型和基因型鉴定

Table 2 Phenotypic and genotypic identification of bitter gourd materials

编号 No.	材料名称 Material name	花性型 Sexual phenotype	基因型 Genotype	编号 No.	材料名称 Material name	花性型 Sexual phenotype	基因型 Genotype
1	MC52	全雌型	C：C	29	GY-27	全雌型	C：C
2	K8	雌雄同株型	G：G	30	GY-28	全雌型	C：C
3	GY-1	全雌型	C：C	31	GY-29	全雌型	C：C
4	GY-2	全雌型	C：C	32	GY-30	全雌型	C：C
5	GY-3	全雌型	C：C	33	GY-31	全雌型	C：C
6	GY-4	全雌型	C：C	34	GY-32	全雌型	C：C
7	GY-5	全雌型	C：C	35	GY-33	全雌型	C：C
8	GY-6	全雌型	C：C	36	GY-34	全雌型	C：C
9	GY-7	全雌型	C：C	37	GY-35	全雌型	C：C
10	GY-8	全雌型	C：C	38	GY-36	全雌型	C：C
11	GY-9	全雌型	C：C	39	GY-37	全雌型	C：C
12	GY-10	全雌型	C：C	40	K1	雌雄同株型	G：G
13	GY-11	全雌型	C：C	41	K2	雌雄同株型	G：G
14	GY-12	全雌型	C：C	42	K3	雌雄同株型	G：G
15	GY-13	全雌型	C：C	43	K4	雌雄同株型	G：G
16	GY-14	全雌型	C：C	44	K5	雌雄同株型	G：G
17	GY-15	全雌型	C：C	45	K6	雌雄同株型	G：G
18	GY-16	全雌型	C：C	46	K7	雌雄同株型	G：C
19	GY-17	全雌型	C：C	47	K8	雌雄同株型	G：G
20	GY-18	全雌型	C：C	48	K9	雌雄同株型	G：G
21	GY-19	全雌型	C：C	49	K10	雌雄同株型	G：G
22	GY-20	全雌型	C：C	50	K11	雌雄同株型	G：G
23	GY-21	全雌型	C：C	51	K12	雌雄同株型	G：C
24	GY-22	全雌型	C：C	52	K13	雌雄同株型	G：G
25	GY-23	全雌型	C：C	53	K14	雌雄同株型	G：G
26	GY-24	全雌型	C：C	54	K15	雌雄同株型	G：G
27	GY-25	全雌型	C：C	55	K16	雌雄同株型	G：C
28	GY-26	全雌型	C：C	56	K17	雌雄同株型	G：G

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