花椰菜花球发育相关基因BoCAL的KASP标记开发和应用

doi:10.3969/j.issn.1004-1524.2022.06.09

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (6): 1183-1192.DOI: 10.3969/j.issn.1004-1524.2022.06.09

花椰菜花球发育相关基因BoCAL的KASP标记开发和应用

盛小光(), 沈钰森, 虞慧芳, 王建升, 赵振卿, 顾宏辉()

浙江省农业科学院蔬菜研究所,浙江杭州 310021

收稿日期:2022-02-15 出版日期:2022-06-25 发布日期:2022-06-30
通讯作者: 顾宏辉
作者简介:*顾宏辉,E-mail: guhh@zaas.ac.cn
盛小光（1982—）,女,安徽芜湖人,博士,副研究员,从事甘蓝类蔬菜遗传育种工作。E-mail: xguang@zaas.ac.cn
基金资助:
政府间国际科技创新合作重点专项(2017YFE0114500);浙江省科技计划(2022C02051);浙江省农业（蔬菜）新品种选育重大专项——花椰菜(2021C02065-4-3)

Development and application of KASP marker of BoCAL gene related to curd development in cauliflower

SHENG Xiaoguang(), SHEN Yusen, YU Huifang, WANG Jiansheng, ZHAO Zhenqing, GU Honghui()

Institute of Vegetable, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2022-02-15 Online:2022-06-25 Published:2022-06-30
Contact: GU Honghui

摘要/Abstract

摘要：

BoCAL基因是控制花椰菜花球发生的关键基因之一。为了提高对花椰菜BoCAL基因型鉴定效率,根据该基因第5个外显子中第16个碱基（Ex5+16）G-T的功能性单核苷酸多态性（SNP）突变,开发了竞争性等位基因特异性PCR（KASP）标记(BoCAL-KASP1)。利用该标记对甘蓝类7个变种的蔬菜作物与野生种共计86份材料进行了BoCAL等位基因的分型,发现共有36份材料的基因型是T:T,包括28份花椰菜、6份青花菜与2份引进的地方种材料;43份材料的基因型是G:G,包括2份花椰菜、9份青花菜、6份结球甘蓝、5份芥蓝、3份苤蓝、7份羽衣甘蓝、2份抱子甘蓝、4份引进的地方种与5份甘蓝野生种材料;7份材料显示杂合基因型T:G,包括2份青花菜、2份羽衣甘蓝和3份引进的地方种材料。这86份材料的BoCAL基因型KASP鉴定结果和酶切扩增多态性标记(BoCAL-CAPS1)的鉴定结果完全一致。以上结果说明BoCAL-KASP1标记可以高效准确地鉴定BoCAL基因目标位点的基因型,从而为花椰菜花球发育研究和分子育种提供可靠的功能性标记。

关键词: 花椰菜, 花球发育, BoCAL基因, KASP标记

Abstract:

BoCAL is one of the key genes controlling the curd formation in cauliflower. In order to improve the efficiency of BoCAL genotype identification of cauliflower, a competitive allele specific PCR (KASP) marker (BoCAL-KASP1) was developed according to the functional single nucleotide polymorphism (SNP) mutation of the 16th base (Pe5 + 16) G-T in the fifth exon of BoCAL. Using this KASP marker, BoCAL alleles were genotyped in 86 core accessions from seven variants of Brassica. oleracea and related wild species. It turned out that a total of thirty-six accessions showed the genotype of T:T, including twenty-eight cauliflower, six broccoli and two imported landraces; forty-three accessions had the genotype of G:G, including two cauliflower, nine broccoli, six cabbage, five Chinese kale, three kohlrabi, seven kale, two Brussels sprouts, four imported landraces and five related wild species; Seven accessions showed heterozygous genotype T:G, including two broccoli, two kale and three imported landraces. The results of KASP identification of BoCAL genotype of these 86 accessions were completely consistent with those of cleaved amplified polymorphic sequences (BoCAL-CAPS1) identification. Thus, the KASP marker developed in this study could efficiently and accurately identify the genotype of BoCAL target locus, so as to provide reliable functional markers for cauliflower curd development research and molecular breeding.

Key words: cauliflower, curd development, BoCAL gene, KASP marker

中图分类号:

S635.3

盛小光, 沈钰森, 虞慧芳, 王建升, 赵振卿, 顾宏辉. 花椰菜花球发育相关基因BoCAL的KASP标记开发和应用[J]. 浙江农业学报, 2022, 34(6): 1183-1192.

SHENG Xiaoguang, SHEN Yusen, YU Huifang, WANG Jiansheng, ZHAO Zhenqing, GU Honghui. Development and application of KASP marker of BoCAL gene related to curd development in cauliflower[J]. Acta Agriculturae Zhejiangensis, 2022, 34(6): 1183-1192.

图/表 6

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编号 No.	种质名称 Name	类型 Type	SNP	编号 No.	种质名称 Name	类型 Type	SNP	编号 No.	种质名称 Name	类型 Type	SNP
1/1	松花55-4 SH55-4	花椰菜 Cauliflower	T:T	30/8	极晚180 JW180	花椰菜 Cauliflower	G:G	59/35	苤蓝1号 PL1	苤蓝 Kohlrabi	G:G
2/2	农乐55 NL55	花椰菜 Cauliflower	T:T	31/9	青花60-2 QH60-2	青花菜 Broccoli	G:G	60/49	苤蓝2号 PL2	苤蓝 Kohlrabi	G:G
3/3	松花58-1 SH58-1	花椰菜 Cauliflower	T:T	32/10	阳光60-1 YG60-1	青花菜 Broccoli	G:G	61/39	苤蓝3号 PL3	苤蓝 Kohlrabi	G:G
4/4	松花60-2 SH60-2	花椰菜 Cauliflower	T:T	33/12	报春60 BC60	青花菜 Broccoli	G:G	62/41	羽甘3号 YG3	羽衣甘蓝 Kale	G:G
5/5	优松60-8 YS60-8	花椰菜 Cauliflower	T:T	34/13	秀美65 XM65	青花菜 Broccoli	G:G	63/38	羽甘6号YG6	羽衣甘蓝 Kale	G:T
6/6	松花65-1 SH65-1	花椰菜 Cauliflower	T:T	35/72	浙青65-4 ZQ65-4	青花菜 Broccoli	T:T	64/48	粉欧1号 FOU1	羽衣甘蓝 Kale	G:G
7/11	松花65-4 SH65-4	花椰菜 Cauliflower	T:T	36/73	青美70 QM70	青花菜 Broccoli	T:T	65/42	白鸥1号 BOU1	羽衣甘蓝 Kale	G:G
8/23	庆农65-3 QN65-3	花椰菜 Cauliflower	T:T	37/14	绿贵75 LG75	青花菜 Broccoli	G:G	66/80	紫鸽1号 ZGE1	羽衣甘蓝 Kale	G:G
9/24	佳丽佳65 JLJ65	花椰菜 Cauliflower	T:T	38/74	浙青75-9 ZQ75-9	青花菜 Broccoli	T:T	67/50	白珊瑚 White coral	羽衣甘蓝 Kale	G:T
10/28	松花68 SH68	花椰菜 Cauliflower	T:T	39/15	浙青80 ZQ80	青花菜 Broccoli	G:G	68/47	红珊瑚 Red coral	羽衣甘蓝 Kale	G:G
11/29	松花70-2 SH70-2	花椰菜 Cauliflower	T:T	40/16	喜鹊80 XQ80	青花菜 Broccoli	G:G	69/44	Kale-2	羽衣甘蓝 Kale	G:G
12/30	松花70-3 SH70-3	花椰菜 Cauliflower	T:T	41/17	报春85 BC85	青花菜 Broccoli	G:G	70/46	Kale-8	羽衣甘蓝 Kale	G:G
13/37	农胜70-1 NS70-1	花椰菜 Cauliflower	T:T	42/75	野崎90-6 YQ90-6	青花菜 Broccoli	T:T	71/45	王子 Prince	抱子甘蓝 Brussels sprout	G:G
14/55	优松70-1 YS70-1	花椰菜 Cauliflower	T:T	43/22	浙青95-4 ZQ95-4	青花菜 Broccoli	G:T	72/53	绿宝石 Emerald	抱子甘蓝 Brussels sprout	G:G
15/56	松花75-1 SH75-1	花椰菜 Cauliflower	T:T	44/18	野崎100 YQ100	青花菜 Broccoli	G:G	73/51	SG1913	地方种 Landrace	G:T
16/57	宝岛75 BD75	花椰菜 Cauliflower	T:T	45/40	浙青100-6 ZQ100-6	青花菜 Broccoli	G:T	74/64	SG1915	地方种 Landrace	G:G
17/58	松花78 SH78	花椰菜 Cauliflower	T:T	46/76	台绿3号 TL3	青花菜 Broccoli	T:T	75/54	SG1917	地方种 Landrace	G:T
18/59	橙松80-3 CS80-3	花椰菜 Cauliflower	T:T	47/82	台绿5号 TL5	青花菜 Broccoli	T:T	76/77	SG1401	地方种 Landrace	G:T
19/60	越优80-9 YY80-9	花椰菜 Cauliflower	T:T	48/19	中甘15号 ZG15	甘蓝 Cabbage	G:G	77/52	SG1501	地方种 Landrace	G:G
20/61	浙松80-5 ZS80-5	花椰菜 Cauliflower	T:T	49/20	中甘21号 ZG21	甘蓝 Cabbage	G:G	78/68	SG1521	地方种 Landrace	G:G
21/62	越优85-2 YY85-2	花椰菜 Cauliflower	T:T	50/21	浙甘85 ZG85	甘蓝 Cabbage	G:G	79/85	SG1528	地方种 Landrace	T:T
22/63	浙松88 ZS88	花椰菜 Cauliflower	T:T	51/25	苏甘8号 SG8	甘蓝 Cabbage	G:G	80/78	SG1539	地方种 Landrace	G:G
23/65	庆农90-4 QN90-4	花椰菜 Cauliflower	T:T	52/26	紫甘1号 ZG1	甘蓝 Cabbage	G:G	81/86	CR06	地方种 Landrace	T:T
24/66	松花90-7 SH90-7	花椰菜 Cauliflower	T:T	53/27	紫甘2号 ZG2	甘蓝 Cabbage	G:G	82/43	SP01	野生种 Wild species	G:G
25/67	浙松100 ZS100	花椰菜 Cauliflower	T:T	54/31	顺宝芥蓝 SB Kale	芥蓝 Chinese kale	G:G	83/81	SP9	野生种 Wild species	G:G
26/69	浙塔120-1 ZT120-2	花椰菜 Cauliflower	T:T	55/36	绿芥蓝 Green Kale	芥蓝 Chinese kale	G:G	84/83	SP12	野生种 Wild species	G:G
27/70	副将140-3 FJ140-3	花椰菜 Cauliflower	T:T	56/34	秋绿芥蓝 QL Kale	芥蓝 Chinese kale	G:G	85/84	SP16	野生种 Wild species	G:G
28/71	大圣140-3 DS140-3	花椰菜 Cauliflower	T:T	57/32	J1401	芥蓝 Chinese kale	G:G	86/79	SP20	野生种 Wild species	G:G
29/7	CR01	花椰菜 Cauliflower	G:G	58/33	J1402	芥蓝 Chinese kale	G:G

编号 No.	种质名称 Name	类型 Type	SNP	编号 No.	种质名称 Name	类型 Type	SNP	编号 No.	种质名称 Name	类型 Type	SNP
1/1	松花55-4 SH55-4	花椰菜 Cauliflower	T:T	30/8	极晚180 JW180	花椰菜 Cauliflower	G:G	59/35	苤蓝1号 PL1	苤蓝 Kohlrabi	G:G
2/2	农乐55 NL55	花椰菜 Cauliflower	T:T	31/9	青花60-2 QH60-2	青花菜 Broccoli	G:G	60/49	苤蓝2号 PL2	苤蓝 Kohlrabi	G:G
3/3	松花58-1 SH58-1	花椰菜 Cauliflower	T:T	32/10	阳光60-1 YG60-1	青花菜 Broccoli	G:G	61/39	苤蓝3号 PL3	苤蓝 Kohlrabi	G:G
4/4	松花60-2 SH60-2	花椰菜 Cauliflower	T:T	33/12	报春60 BC60	青花菜 Broccoli	G:G	62/41	羽甘3号 YG3	羽衣甘蓝 Kale	G:G
5/5	优松60-8 YS60-8	花椰菜 Cauliflower	T:T	34/13	秀美65 XM65	青花菜 Broccoli	G:G	63/38	羽甘6号YG6	羽衣甘蓝 Kale	G:T
6/6	松花65-1 SH65-1	花椰菜 Cauliflower	T:T	35/72	浙青65-4 ZQ65-4	青花菜 Broccoli	T:T	64/48	粉欧1号 FOU1	羽衣甘蓝 Kale	G:G
7/11	松花65-4 SH65-4	花椰菜 Cauliflower	T:T	36/73	青美70 QM70	青花菜 Broccoli	T:T	65/42	白鸥1号 BOU1	羽衣甘蓝 Kale	G:G
8/23	庆农65-3 QN65-3	花椰菜 Cauliflower	T:T	37/14	绿贵75 LG75	青花菜 Broccoli	G:G	66/80	紫鸽1号 ZGE1	羽衣甘蓝 Kale	G:G
9/24	佳丽佳65 JLJ65	花椰菜 Cauliflower	T:T	38/74	浙青75-9 ZQ75-9	青花菜 Broccoli	T:T	67/50	白珊瑚 White coral	羽衣甘蓝 Kale	G:T
10/28	松花68 SH68	花椰菜 Cauliflower	T:T	39/15	浙青80 ZQ80	青花菜 Broccoli	G:G	68/47	红珊瑚 Red coral	羽衣甘蓝 Kale	G:G
11/29	松花70-2 SH70-2	花椰菜 Cauliflower	T:T	40/16	喜鹊80 XQ80	青花菜 Broccoli	G:G	69/44	Kale-2	羽衣甘蓝 Kale	G:G
12/30	松花70-3 SH70-3	花椰菜 Cauliflower	T:T	41/17	报春85 BC85	青花菜 Broccoli	G:G	70/46	Kale-8	羽衣甘蓝 Kale	G:G
13/37	农胜70-1 NS70-1	花椰菜 Cauliflower	T:T	42/75	野崎90-6 YQ90-6	青花菜 Broccoli	T:T	71/45	王子 Prince	抱子甘蓝 Brussels sprout	G:G
14/55	优松70-1 YS70-1	花椰菜 Cauliflower	T:T	43/22	浙青95-4 ZQ95-4	青花菜 Broccoli	G:T	72/53	绿宝石 Emerald	抱子甘蓝 Brussels sprout	G:G
15/56	松花75-1 SH75-1	花椰菜 Cauliflower	T:T	44/18	野崎100 YQ100	青花菜 Broccoli	G:G	73/51	SG1913	地方种 Landrace	G:T
16/57	宝岛75 BD75	花椰菜 Cauliflower	T:T	45/40	浙青100-6 ZQ100-6	青花菜 Broccoli	G:T	74/64	SG1915	地方种 Landrace	G:G
17/58	松花78 SH78	花椰菜 Cauliflower	T:T	46/76	台绿3号 TL3	青花菜 Broccoli	T:T	75/54	SG1917	地方种 Landrace	G:T
18/59	橙松80-3 CS80-3	花椰菜 Cauliflower	T:T	47/82	台绿5号 TL5	青花菜 Broccoli	T:T	76/77	SG1401	地方种 Landrace	G:T
19/60	越优80-9 YY80-9	花椰菜 Cauliflower	T:T	48/19	中甘15号 ZG15	甘蓝 Cabbage	G:G	77/52	SG1501	地方种 Landrace	G:G
20/61	浙松80-5 ZS80-5	花椰菜 Cauliflower	T:T	49/20	中甘21号 ZG21	甘蓝 Cabbage	G:G	78/68	SG1521	地方种 Landrace	G:G
21/62	越优85-2 YY85-2	花椰菜 Cauliflower	T:T	50/21	浙甘85 ZG85	甘蓝 Cabbage	G:G	79/85	SG1528	地方种 Landrace	T:T
22/63	浙松88 ZS88	花椰菜 Cauliflower	T:T	51/25	苏甘8号 SG8	甘蓝 Cabbage	G:G	80/78	SG1539	地方种 Landrace	G:G
23/65	庆农90-4 QN90-4	花椰菜 Cauliflower	T:T	52/26	紫甘1号 ZG1	甘蓝 Cabbage	G:G	81/86	CR06	地方种 Landrace	T:T
24/66	松花90-7 SH90-7	花椰菜 Cauliflower	T:T	53/27	紫甘2号 ZG2	甘蓝 Cabbage	G:G	82/43	SP01	野生种 Wild species	G:G
25/67	浙松100 ZS100	花椰菜 Cauliflower	T:T	54/31	顺宝芥蓝 SB Kale	芥蓝 Chinese kale	G:G	83/81	SP9	野生种 Wild species	G:G
26/69	浙塔120-1 ZT120-2	花椰菜 Cauliflower	T:T	55/36	绿芥蓝 Green Kale	芥蓝 Chinese kale	G:G	84/83	SP12	野生种 Wild species	G:G
27/70	副将140-3 FJ140-3	花椰菜 Cauliflower	T:T	56/34	秋绿芥蓝 QL Kale	芥蓝 Chinese kale	G:G	85/84	SP16	野生种 Wild species	G:G
28/71	大圣140-3 DS140-3	花椰菜 Cauliflower	T:T	57/32	J1401	芥蓝 Chinese kale	G:G	86/79	SP20	野生种 Wild species	G:G
29/7	CR01	花椰菜 Cauliflower	G:G	58/33	J1402	芥蓝 Chinese kale	G:G

花椰菜花球发育相关基因BoCAL的KASP标记开发和应用

Development and application of KASP marker of BoCAL gene related to curd development in cauliflower

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