122份栽培桃品种（系）黄白肉性状的分子标记辅助鉴定

doi:10.3864/j.issn.0578-1752.2020.14.016

Abstract

Abstract:

【Objective】It shows that peach flesh color (yellow/white) is controlled by the gene CCD 4 (carotenoid cleavage dioxygenase 4). Based on three types of CCD4 allele variations, molecular markers of indel, SSR-CE, and Sanger sequence for SNP were used to analyze the genotypes of 122 peach cultivars (lines), with the aim to determine the correlation between flesh color and genetic variation. This result could provide information for parental matching and the selection of corresponding molecular markers for phenotyping in their offspring. 【Method】Three types of variations were detected via PCR. LTR transposable element insertion was detected by 1% agarose gel electrophoresis, and SSR repeat numbers were detected using CE-SSR in ABI3730XL. Nucleotide substitution was detected using the Sanger sequence and analyzed with the ContigExpress software. In total, the genotypes of 122 cultivars (lines) were analyzed, and the correlation between phenotype and genetic variation was determined.【Result】After genotyping of the 122 cultivars, it was found that 31 accessions were LTR transposable element insertions with 729-bp amplified fragments, accounting for 25.4% of the total accessions, of which eight accessions (25.8%) were homozygous. Sixty-eight cultivars (lines) were SSR repeat number variations, accounting for 55.7% of the total accessions and including 25 (36.8%) homozygous types with 2-bp insertions. Of the 122 cultivars (lines), only one cultivar (Fertilia Morettini) was caused by nucleotide substitution and SSR repeat number variation, accounting for 0.82%, which was not widely used in the breeding program. LTR transposable element insertion and SSR repeat number variation were the key types affecting flesh color. Of the 122 cultivars (lines), seven yellow-flesh cultivars (lines) were caused by SSR repeat number variation and LTR transposable element insertion, accounting for 5.7%. One homozygous or two heterozygous sequence variations were both responsible for yellow flesh. The results showed that genotypes were identical with phenotypes of the 122 accessions, with 100% accuracy. 【Conclusion】The genotypes of 122 peach cultivars (lines) (white and yellow flesh color) were identified using molecular markers, which could be applied for parental selection, offspring identification in breeding programs, and flesh color selection (white or yellow) using molecular marker-assisted selection.

Key words: peach, CCD4, MAS, flesh color

LU ZhenHua,SHEN ZhiJun,NIU Liang,PAN Lei,CUI GuoChao,ZENG WenFang,WANG ZhiQiang. Molecular Marker-Assisted Identification of Yellow/White Flesh Trait for 122 Peach Cultivars (Lines)[J].Scientia Agricultura Sinica, 2020, 53(14): 2929-2940.

Figures/Tables 7

Table 1

Allelic variants of CCD4 as related to phenotype in yellow/white flesh peach"

编号 Code	品种（系） Cultivar (line)	来源 Origin	果肉类型 Flesh color	微卫星序列 SSR	SNP位点 SNP	CCD4/LTR CCD4/LTR
1	雨花2号 Yuhua 2	NFGRN	白 White	177/177	AA	1/1
2	霞脆 Xiacui	NFGRN	白 White	177/177	AA	1/1
3	雨花1号 Yuhua 1	NFGRN	白 White	177/177	AA	1/0
4	瑞光18号 Ruiguang 18	NFGRN	黄Yellow	179/179	AA	1/0
5	瑞光19号 Ruiguang 19	NFGRN	白 White	177/179	AA	1/0
6	早美Zaomei	NFGRN	白 White	177/177	AA	1/0
7	红粉佳人 Pinklady	NFGRN	白 White	177/177	AA	1/0
8	瑞光美玉 Ruiguangmeiyu	NFGRN	白 White	177/179	AA	1/0
9	早花露 Zaohualu	NFGRN	白 White	177/177	AA	1/0
10	大久保 Okubo	NFGRN	白 White	177/177	AA	1/0
11	金陵黄露 Jinlinghuanglu	NFGRN	黄 Yellow	179/179	AA	1/0
12	沪油002号 Huyou 002	NFGRN	白 White	177/179	AA	1/0
13	锦绣 Jinxiu	NFGRN	黄 Yellow	177/177	AA	0/1
14	金童7号 Babygold 7	NFGRN	黄 Yellow	177/179	AA	1/1
15	沪油003号 Huyou 003	NFGRN	黄 Yellow	177/179	AA	1/1
16	玉霞蟠桃 Yuxiapantao	NFGRN	白 White	177/179	AA	1/0
17	五月火 Mayfire	NFGRN	黄 Yellow	179/179	AA	1/0
18	早凤玉 Zaofengyu	NFGRN	白 White	177/179	AA	1/0
19	沪021号 Hu 021	NFGRN	白 White	177/177	AA	1/1
20	日川白凤 Richuanbaifeng	NFGRN	白 White	177/177	AA	1/1
21	瑞光7号 Ruiguang 7	NFGRN	白 White	177/179	AA	1/0
22	瑞光22号 Ruiguang 22	NFGRN	黄 Yellow	179/179	AA	1/0
23	阿姆肯 Armking	NFGRN	黄 Yellow	179/179	AA	1/0
24	霞晖2号 Xiahui 2	NFGRN	白 White	177/177	AA	1/0
25	瑞蟠3号 Ruipan 3	NFGRN	白 White	177/177	AA	1/0
26	早金露 Zaojinlu	NFGRN	黄 Yellow	177/179	AA	1/1
27	瑞蟠5号 Ruipan 5	NFGRN	白 White	177/177	AA	1/1
28	晚硕蜜 Wanshuomi	NFGRN	白 White	177/177	AA	1/0
29	早露蟠桃 Zaolupantao	NFGRN	白 White	177/177	AA	1/0
30	霞晖8号 Xiahui 8	NFGRN	白 White	177/179	AA	1/0
31	晚白花 Wanbaihua	NFGRN	白 White	177/177	AA	1/1
32	连黄 Lianhuang	NFGRN	黄 Yellow	177/179	AA	0/1
33	春花 Chunhua	NFGRN	白 White	177/177	AA	1/1
34	黄露蟠桃 Huanglupantao	NFGRN	白 White	177/177	AA	1/1
35	金童5号 Babygold 5	NFGRN	黄 Yellow	177/179	AA	1/1
36	砂子早生 Sunago Wase	NFGRN	白 White	177/177	AA	1/1
37	早上海水蜜 Zaoshanghaishuimi	NFGRN	白 White	177/177	AA	1/0
38	金童9号 Babygold 9	NFGRN	黄 Yellow	177/177	AA	0/1
39	紫金红2号 Zijinghong 2	NFGRN	黄 Yellow	179/179	AA	1/0
40	银花露 Yinhualu	NFGRN	白 White	177/177	AA	1/1
编号 Code	品种（系） Cultivar （line）	来源 Origin	果肉类型 Flesh color	微卫星序列 SSR	SNP位点 SNP	CCD4/LTR CCD4/LTR
41	弗雷德里克 Frederica	NFGRN	黄 Yellow	179/179	AA	1/0
42	新白凤 Early Hakuho	NFGRN	白 White	177/177	AA	1/0
43	奉化玉露早 Fenghuayuluzao	NFGRN	白 White	177/177	AA	1/0
44	霞晖4号 Xiahui 4	NFGRN	白 White	177/179	AA	1/0
45	早硕蜜 Zaoshuomi	NFGRN	白 White	177/177	AA	1/0
46	早魁蜜 Zaokuimi	NFGRN	白 White	177/177	AA	1/0
47	霞光 Xiaguang	NFGRN	黄 Yellow	177/179	AA	1/1
48	瑞蟠4号 Ruipan 4	NFGRN	白 White	177/177	AA	1/0
49	京玉 Jingyu	NFGRN	白 White	177/177	AA	1/1
50	金霞蟠桃 Jinxiapantao	NFGRN	黄 Yellow	179/179	AA	1/1
51	京红 Jinghong	NFGRN	白 White	177/177	AA	1/0
52	银河 Galaxy	NFGRN	白 White	177/179	AA	1/0
53	瑞光28号 Ruiguang 28	NFGRN	黄 Yellow	179/179	AA	1/0
54	瑞蟠2号 Ruipan 2	NFGRN	白 White	177/177	AA	1/0
55	金晖 Jinhui	NFGRN	黄 Yellow	177/177	AA	0/1
56	雨花露 Yuhualu	NFGRN	白 White	177/177	AA	1/0
57	白花水蜜 Baihuashuilu	NFGRN	白 White	177/177	AA	1/1
58	丰黄 Fenghuang	NFGRN	黄 Yellow	177/177	AA	0/1
59	晖雨露 Huiyulu	NFGRN	白 White	177/177	AA	1/0
60	奉化玉露 Fenghuayulu	NFGRN	白 White	177/177	AA	1/0
61	花玉露 Huayulu	NFGRN	白 White	177/177	AA	1/0
62	瑞光23 号 Ruiguang 23	NFGRN	白 White	177/179	AA	1/0
63	瑞蟠1号 Ruipan 1	NFGRN	白 White	177/177	AA	1/0
64	霞晖3号 Xiahui 3	NFGRN	白 White	177/177	AA	1/0
65	金童8号 Babygold 8	NFGRN	白 White	177/177	AA	0/1
66	霞晖1号 Xiahui 1	NFGRN	白 White	177/177	AA	1/0
67	雨花3号 Yuhua 3	NFGRN	白 White	177/177	AA	1/1
68	金陵锦桃 Jinlingjintao	NFGRN	白 White	177/177	AA	1/0
69	源东白桃 Yuandongbaitao	NFGRN	白 White	177/177	AA	1/1
70	新白花 Xinbaihua	NFGRN	白 White	177/177	AA	1/1
71	金童6号 Babygold 6	NFGRN	黄 Yellow	177/179	AA	1/1
72	紫金红3号 Zijinhong 3	NFGRN	黄 Yellow	179/179	AA	1/0
73	金山早红 Jinshanzaohong	NFGRN	黄 Yellow	179/179	AA	1/0
74	早红港 Early Redhaven	NFGRN	黄 Yellow	177/177	AA	0/1
75	霞晖5号 Xiahui 5	NFGRN	白 White	177/177	AA	0/0
76	南山甜桃 Nanshantiantao	NFGRN	白 White	177/177	AA	1/0
77	锦香 Jinxiang	NFGRN	黄 Yellow	177/177	AA	0/1
78	弗尔蒂尼莫蒂尼 Fertilia Morettini	NFGRN	黄 Yellow	177/179	AT	1/0
79	京春 Jingchun	NFGRN	白 White	177/177	AA	1/0
80	中蟠1号 Zhongpan 1	ZFRI	白 White	177/179	AA	1/0
81	99-54-36	ZFRI	白 White	177/179	AA	1/0
编号 Code	品种（系） Cultivar （line）	来源 Origin	果肉类型 Flesh color	微卫星序列 SSR	SNP位点 SNP	CCD4/LTR CCD4/LTR
82	36-3	ZFRI	白 White	177/179	AA	1/0
83	黄金蜜1号 Huangjinmi 1	ZFRI	黄 Yellow	179/179	AA	1/0
84	09南9-22 09nan9-22	ZFRI	白 White	177/177	AA	1/0
85	09南10-5 09nan10-5	ZFRI	白 White	177/179	AA	1/0
86	4-3-11	ZFRI	黄 Yellow	179/179	AA	1/0
87	5-3-7	ZFRI	黄 Yellow	179/179	AA	1/0
88	中油桃13号 Zhongyoutao 13	ZFRI	白 White	177/179	AA	1/0
89	春美 Chunmei	ZFRI	白 White	177/179	AA	1/0
90	中桃9号 Zhongtao 9	ZFRI	白 White	177/179	AA	1/0
91	中桃10号 Zhogntao 10	ZFRI	黄 Yellow	179/179	AA	1/0
92	08北-12-1 08bei-12-1	ZFRI	白 White	177/177	AA	1/0
93	双喜红 Shuangxihong	ZFRI	黄 Yellow	179/179	AA	1/0
94	中油20号 Zhognyou 20	ZFRI	白 White	177/179	AA	1/0
95	中桃红玉 Zhongtaohongyu	ZFRI	白 White	177/179	AA	1/0
96	06-3-113	ZFRI	黄 Yellow	179/179	AA	1/0
97	05-3-102	ZFRI	黄 Yellow	179/179	AA	1/0
98	晚油桃 Wanyoutao	ZFRI	黄 Yellow	179/179	AA	1/0
99	中油18号 Zhongyou 18	ZFRI	白 White	177/179	AA	1/0
100	1区井蟠 Yiqujingpan	ZFRI	白 White	177/179	AA	1/0
101	中桃22号 Zhongtao 22	ZFRI	白 White	177/179	AA	1/0
102	春瑞 Chunrui	ZFRI	白 White	177/179	AA	1/0
103	9-6-180	ZFRI	白 White	177/179	AA	1/0
104	04-7-13	ZFRI	白 White	177/179	AA	1/0
105	中桃白玉 Zhongtaobaiyu	ZFRI	白 White	177/177	AA	1/0
106	中油19号 Zhongyou 19	ZFRI	黄 Yellow	179/179	AA	1/0
107	中油27号 Zhongyou 27	ZFRI	白 White	177/179	AA	1/0
108	中油12号 Zhongyou 12	ZFRI	白 White	177/179	AA	1/0
109	4-1-6	ZFRI	黄 Yellow	177/179	AA	1/1
110	枣油桃 Zaoyoutao	ZFRI	白 White	177/179	AA	1/0
111	红不软 Hongburuan	ZFRI	白 White	177/177	AA	1/0
112	6-7-6	ZFRI	白 White	177/179	AA	1/0
113	13-33	ZFRI	白 White	177/179	AA	1/0
114	4-9-16	ZFRI	白 White	177/179	AA	1/0
115	小花红芒果 Xiaohuahongmangguo	ZFRI	黄 Yellow	179/179	AA	1/0
116	中桃8号 Zhongtao 8	ZFRI	白 White	177/179	AA	1/0
117	中油15号 Zhongyou 15	ZFRI	白 White	177/179	AA	1/0
118	红芒果 Hongmangguo	ZFRI	黄 Yellow	179/179	AA	1/0
119	中蟠2号 Zhongpan 2	ZFRI	黄 Yellow	179/179	AA	1/0
120	春蜜 Chunmi	ZFRI	白 White	177/179	AA	1/0
121	中农金辉 Zhongnongjinhui	ZFRI	黄 Yellow	179/179	AA	1/0
122	中桃5号 Zhongtao 5	ZFRI	白 White	177/179	AA	1/0

Table 1

Table 2

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Table 3

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名称 Code	引物序列 Primer sequence	退火温度 Annealing temperature (℃)	片段长度 Size (bp)
CCD4-F	5'-ACCACCTGTTTGACGGAGAC-3'	55	594
CCD4-R	5'-TGCTCATGAAGAGCTTGCCA-3'
LTR-F（CCD4）	5'-TACCTGAGAGCTTCTCGTGC-3'	55	729
CCD4-SSR-F	5'- ROX -CCCATTTTGCAGTGAAGGGC-3'	55	177
CCD4-SSR-R	5'-GCTGTGGTGCTTTTGTGGAG-3'
CCD4-SNP-F	5′-GGGTGATCCAATGCCTAAGA-3′	55	510
CCD4-SNP-R	5′-GGCTCTCTAGCCACGAAAAA-3′

Molecular Marker-Assisted Identification of Yellow/White Flesh Trait for 122 Peach Cultivars (Lines)

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基因型 Genotype	品种（系） Cultivar (line)
CT插入 CT insertion	瑞光18号 Ruiguang 18
	金陵黄露 Jinlinghuanglu
	五月火 Mayfire
	瑞光22号 Ruiguang 22
	阿姆肯 Armking
	连黄 Lianhuang
	紫金红2号 Zijinghong 2
	弗雷德里克 Frederica
	金霞蟠桃Jinxiapantao
	瑞光28号 Ruiguang 28
	紫金红3号 Zijinhong 3
	金山早红 Jinshanzaohong
	黄金蜜1号 Huangjinmi 1
	4-3-11
	5-3-7
	中桃10号 Zhongtao 10
	双喜红 Shuangxihong
	06-3-113
	05-3-102
	晚油桃 Wanyoutao
	中油19号 Zhongyou 19
	小花红芒果 Xiaohuahongmangguo
	红芒果 Hongmangguo
	中蟠2号 Zhongpan 2
	中农金辉 Zhongnongjinhui
LTR插入 LTR insertion	锦绣 Jinxiu
	连黄 Lianhuang
	金童9号 Babygold 9
	金晖 Jinhui
	丰黄 Fenghuang
	早红港 Early Redhaven
	锦香 Jinxiang 金童8号 Babygold 8
CT重复+LTR插入 CT insertion + LTR insertion	金童7号 Babygold 7
	沪油003号 Huyou 003
	早金露 Zaojinlu
	金童5号 Babygold 5
	霞光 Xiaguang
	金童6号 Babygold 6
	04-7-13
CT插入+SNP替换 CT insertion +SNP subsitution	弗尔蒂尼莫蒂尼 Fertilia Morettini

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