Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (11): 293-302.doi: 10.13560/j.cnki.biotech.bull.1985.2020-1494
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GUAN Jun-jiao1(), YANG Xiao-hong1, ZHANG Peng1, HUANG Qing-mei1, ZHANG Jian-hua2()
Received:
2020-12-09
Online:
2021-11-26
Published:
2021-12-03
Contact:
ZHANG Jian-hua
E-mail:guanjunjiao@163.com;zhjhua748@163.com
GUAN Jun-jiao, YANG Xiao-hong, ZHANG Peng, HUANG Qing-mei, ZHANG Jian-hua. Establishment of a Method Identifying the Varieties of Hulless Barley Based on Fluorescence Detection Technology[J]. Biotechnology Bulletin, 2021, 37(11): 293-302.
序号 No. | 材料名称 Material name | 来源 Origin | 棱型 Type |
---|---|---|---|
1 | 长芒六棱露仁 | 江苏省农业科学院 | 多棱 |
2 | 藏83062 | 江苏省农业科学院 | 多棱 |
3 | 昆仑1号 | 青海省农林科学院 | 多棱 |
4 | 昆仑14号 | 青海省农林科学院 | 多棱 |
5 | 北青1号 | 青海省农林科学院 | 多棱 |
6 | 中甸紫青稞 | 云南地方品种 | 多棱 |
7 | 镇康软壳青稞 | 云南地方品种 | 多棱 |
8 | 禄劝裸黑大麦 | 云南地方品种 | 二棱 |
9 | 康青3号 | 四川省甘孜农科所 | 多棱 |
10 | 米大麦 | 云南地方品种 | 多棱 |
11 | 勾芒米大麦 | 云南地方品种 | 多棱 |
12 | 北青88 | 西北农林科技大学 | 多棱 |
13 | 阿青5号 | 青海省农林科学院 | 多棱 |
14 | 迪青3号 | 迪庆州农业科学院 | 多棱 |
15 | 黄长钩裸大麦 | 云南地方品种 | 多棱 |
16 | 紫光芒裸大麦 | 云南地方品种 | 二棱 |
17 | 开宝麦 | 云南地方品种 | 多棱 |
18 | 光芒鲜大麦 | 云南地方品种 | 二棱 |
19 | 早熟白青稞 | 云南地方品种 | 多棱 |
20 | 洋大麦 | 云南地方品种 | 多棱 |
21 | 早熟大麦 | 云南地方品种 | 多棱 |
22 | 云青3号 | 云南省农业科学院 | 多棱 |
23 | Atahualpa | 国际干旱农业研究中心 | 二棱 |
24 | MADRE SELVA | 国际干旱农业研究中心 | 二棱 |
Table 1 Informations of 24 hulless barley cultivars used in this study
序号 No. | 材料名称 Material name | 来源 Origin | 棱型 Type |
---|---|---|---|
1 | 长芒六棱露仁 | 江苏省农业科学院 | 多棱 |
2 | 藏83062 | 江苏省农业科学院 | 多棱 |
3 | 昆仑1号 | 青海省农林科学院 | 多棱 |
4 | 昆仑14号 | 青海省农林科学院 | 多棱 |
5 | 北青1号 | 青海省农林科学院 | 多棱 |
6 | 中甸紫青稞 | 云南地方品种 | 多棱 |
7 | 镇康软壳青稞 | 云南地方品种 | 多棱 |
8 | 禄劝裸黑大麦 | 云南地方品种 | 二棱 |
9 | 康青3号 | 四川省甘孜农科所 | 多棱 |
10 | 米大麦 | 云南地方品种 | 多棱 |
11 | 勾芒米大麦 | 云南地方品种 | 多棱 |
12 | 北青88 | 西北农林科技大学 | 多棱 |
13 | 阿青5号 | 青海省农林科学院 | 多棱 |
14 | 迪青3号 | 迪庆州农业科学院 | 多棱 |
15 | 黄长钩裸大麦 | 云南地方品种 | 多棱 |
16 | 紫光芒裸大麦 | 云南地方品种 | 二棱 |
17 | 开宝麦 | 云南地方品种 | 多棱 |
18 | 光芒鲜大麦 | 云南地方品种 | 二棱 |
19 | 早熟白青稞 | 云南地方品种 | 多棱 |
20 | 洋大麦 | 云南地方品种 | 多棱 |
21 | 早熟大麦 | 云南地方品种 | 多棱 |
22 | 云青3号 | 云南省农业科学院 | 多棱 |
23 | Atahualpa | 国际干旱农业研究中心 | 二棱 |
24 | MADRE SELVA | 国际干旱农业研究中心 | 二棱 |
序号 No. | 位点 Locus | 引物序列 Primer sequence(5'-3') | 退火温度 Annealing temperature/℃ | 序号 No. | 位点 Locus | 引物序列 Primer sequences(5'-3') | 退火温度 Annealing temperature/℃ | |
---|---|---|---|---|---|---|---|---|
1 | Bmag0211 | F:ATTCATCGATCTTGTATTAGTCC R:ACATCATGTCGATCAAAGC | 58 | 15 | EBmac0701 | F:ATGATGAGAACTCTTCACCC R:TGGCACTAAAGCAAAAGAC | 55 | |
2 | Bmag0345 | F:ATGAGGAATAACTCAACCAAA R:AATATATTTTCGATGTCGAGC | 58 | 16 | Bmac0310 | F:CTACCTCTGAGATATCATGCC R:ATCTAGTGTGTGTTGCTTCCT | 55 | |
3 | Bmag0579 | F:CCTAGATAAGGAACATAGCCA R:CAAAGACCCTAACTCATGTTC | 55 | 17 | Bmag387 | F:CGATGACCATTGTATTGAAG R:CTCATGTTGATGTGTGGTTAG | 58 | |
4 | EBmac0501 | F:ACTTAAGTGCCATGCAAAG R:AGGGACAAAAATGGCTAAG | 58 | 18 | Bmac0096 | F:GCTATGGCGTACTATGTATGGTTG R:TCACGATGAGGTATGATCAAAGA | 58 | |
5 | Scssr07759 | F:GCAACTCCTCATCATCTCAGG R:CAACAGCCAGAAGGTCTACG | 62 | 19 | Scssr03907 | F:CTCCCATCACACCATCTGTC R:GACATGGTTCCCTTCTTCTTC | 60 | |
6 | Bmag0378 | F:CTTTTGTTTCCGTAGCATCTA R:ATCCAACTATAGTAGCAAAGCC | 58 | 20 | Scssr10148 | F:AAGCAGCAAAGCAAAGTACC R:TCATCAGCATCTGATCATCC | 60 | |
7 | Bmag0711 | F:GGAGAGTCACATATCAAGGAC R:CCACTCCTTCTCATACCTTTA | 55 | 21 | Bmac0018 | F:GTCCTTTACGCATGAACCGT R:ACATACGCCAGACTCGTGTG | 60 | |
8 | Bmag125 | F:AATTAGCGAGAACAAAATCAC R:AGATAACGATGCACCACC | 55 | 22 | Bmag0009 | F:AAGTGAAGCAAGCAAACAAACA R:ATCCTTCCATATTTTGATTAGGCA | 58 | |
9 | Bmag0013 | F:AAGGGGAATCAAAATGGGAG R:TCGAATAGGTCTCCGAAGAAA | 58 | 23 | Bmag0867 | F:CCCCACACTGACCTACAG R:TTACATCTGCTAGATCGAAGC | 55 | |
10 | EBmac705 | F:GTGGAAAACTGAGTGAAACTC R:TTGAGGAGAAGTAATGACGAT | 55 | 24 | Bmag0870 | F:AACCATAGGATTTGTACTAGTTTC R:TCATGACATCTCAAGAACG | 55 | |
11 | HVM60 | F:CAATGATGCGGTGAACTTTG R:CCTCGGATCTATGGGTCCTT | 58 | 25 | Bmac0064 | F:CTGCAGGTTTCAGGAAGG R:AGATGCCCGCAAAGAGTT | 58 | |
12 | HVM62 | F:TCGCGACCAGACGAGAAG R:AGCTAGCCGACGACGCAC | 60 | 26 | HVCMA | F:GCCTCGGTTTGGACATATAAAG R:GTAAAGCAAATGTTGAGCAACG | 62 | |
13 | EBmac0635 | F:TGCTGCGATGATGAGAACT R:TAGGGTAGATCCGTCCCTATG | 58 | 27 | EBmac0827 | F:CATGGTATTCAAACATACACG R:AAGGTCTTAAGGGGTGATG | 55 | |
14 | EBmac0679 | F:ATTGGAGCGGATTAGGAT R:CCCTATGTCATGTAGGAGATG | 55 | 28 | Scssr15864 | F:GCATAAACGGGTGTAAGAGC R:CATCCAGTTCAGAGGATAGAGC | 55 |
Table 2 Sequence of primer and annealing temperature
序号 No. | 位点 Locus | 引物序列 Primer sequence(5'-3') | 退火温度 Annealing temperature/℃ | 序号 No. | 位点 Locus | 引物序列 Primer sequences(5'-3') | 退火温度 Annealing temperature/℃ | |
---|---|---|---|---|---|---|---|---|
1 | Bmag0211 | F:ATTCATCGATCTTGTATTAGTCC R:ACATCATGTCGATCAAAGC | 58 | 15 | EBmac0701 | F:ATGATGAGAACTCTTCACCC R:TGGCACTAAAGCAAAAGAC | 55 | |
2 | Bmag0345 | F:ATGAGGAATAACTCAACCAAA R:AATATATTTTCGATGTCGAGC | 58 | 16 | Bmac0310 | F:CTACCTCTGAGATATCATGCC R:ATCTAGTGTGTGTTGCTTCCT | 55 | |
3 | Bmag0579 | F:CCTAGATAAGGAACATAGCCA R:CAAAGACCCTAACTCATGTTC | 55 | 17 | Bmag387 | F:CGATGACCATTGTATTGAAG R:CTCATGTTGATGTGTGGTTAG | 58 | |
4 | EBmac0501 | F:ACTTAAGTGCCATGCAAAG R:AGGGACAAAAATGGCTAAG | 58 | 18 | Bmac0096 | F:GCTATGGCGTACTATGTATGGTTG R:TCACGATGAGGTATGATCAAAGA | 58 | |
5 | Scssr07759 | F:GCAACTCCTCATCATCTCAGG R:CAACAGCCAGAAGGTCTACG | 62 | 19 | Scssr03907 | F:CTCCCATCACACCATCTGTC R:GACATGGTTCCCTTCTTCTTC | 60 | |
6 | Bmag0378 | F:CTTTTGTTTCCGTAGCATCTA R:ATCCAACTATAGTAGCAAAGCC | 58 | 20 | Scssr10148 | F:AAGCAGCAAAGCAAAGTACC R:TCATCAGCATCTGATCATCC | 60 | |
7 | Bmag0711 | F:GGAGAGTCACATATCAAGGAC R:CCACTCCTTCTCATACCTTTA | 55 | 21 | Bmac0018 | F:GTCCTTTACGCATGAACCGT R:ACATACGCCAGACTCGTGTG | 60 | |
8 | Bmag125 | F:AATTAGCGAGAACAAAATCAC R:AGATAACGATGCACCACC | 55 | 22 | Bmag0009 | F:AAGTGAAGCAAGCAAACAAACA R:ATCCTTCCATATTTTGATTAGGCA | 58 | |
9 | Bmag0013 | F:AAGGGGAATCAAAATGGGAG R:TCGAATAGGTCTCCGAAGAAA | 58 | 23 | Bmag0867 | F:CCCCACACTGACCTACAG R:TTACATCTGCTAGATCGAAGC | 55 | |
10 | EBmac705 | F:GTGGAAAACTGAGTGAAACTC R:TTGAGGAGAAGTAATGACGAT | 55 | 24 | Bmag0870 | F:AACCATAGGATTTGTACTAGTTTC R:TCATGACATCTCAAGAACG | 55 | |
11 | HVM60 | F:CAATGATGCGGTGAACTTTG R:CCTCGGATCTATGGGTCCTT | 58 | 25 | Bmac0064 | F:CTGCAGGTTTCAGGAAGG R:AGATGCCCGCAAAGAGTT | 58 | |
12 | HVM62 | F:TCGCGACCAGACGAGAAG R:AGCTAGCCGACGACGCAC | 60 | 26 | HVCMA | F:GCCTCGGTTTGGACATATAAAG R:GTAAAGCAAATGTTGAGCAACG | 62 | |
13 | EBmac0635 | F:TGCTGCGATGATGAGAACT R:TAGGGTAGATCCGTCCCTATG | 58 | 27 | EBmac0827 | F:CATGGTATTCAAACATACACG R:AAGGTCTTAAGGGGTGATG | 55 | |
14 | EBmac0679 | F:ATTGGAGCGGATTAGGAT R:CCCTATGTCATGTAGGAGATG | 55 | 28 | Scssr15864 | F:GCATAAACGGGTGTAAGAGC R:CATCCAGTTCAGAGGATAGAGC | 55 |
位点 Locus | 染色体 Chromosome | 等位基因数 Allele number | 基因多样性指数 Gene diversity | 杂合性 Heterozygosity | 多态信息量 PIC | 特异等位基因 Unique gene |
---|---|---|---|---|---|---|
Bmag0211 | 1H | 8 | 0.68 | 0.01 | 0.62 | 1 |
Bmag0345 | 1H | 5 | 0.75 | 0.00 | 0.71 | 0 |
Bmag0579 | 1H | 9 | 0.69 | 0.06 | 0.63 | 0 |
EBmac0501 | 1H | 10 | 0.45 | 0.02 | 0.44 | 2 |
Scssr07759 | 2H | 6 | 0.70 | 0.13 | 0.64 | 1 |
Bmag0378 | 2H | 6 | 0.55 | 0.02 | 0.45 | 2 |
Bmag0711 | 2H | 11 | 0.77 | 0.14 | 0.74 | 0 |
Bmag125 | 2H | 11 | 0.66 | 0.12 | 0.62 | 0 |
Bmag0013 | 3H | 16 | 0.86 | 0.09 | 0.84 | 3 |
EBmac705 | 3H | 11 | 0.72 | 0.19 | 0.68 | 3 |
HVM60 | 3H | 10 | 0.72 | 0.09 | 0.69 | 3 |
HVM62 | 3H | 11 | 0.78 | 0.18 | 0.75 | 3 |
EBmac0635 | 4H | 8 | 0.64 | 0.07 | 0.62 | 0 |
EBmac0679 | 4H | 11 | 0.72 | 0.15 | 0.70 | 1 |
EBmac0701 | 4H | 9 | 0.66 | 0.10 | 0.64 | 1 |
Bmac0310 | 4H | 12 | 0.83 | 0.14 | 0.81 | 0 |
Bmag0387 | 5H | 10 | 0.79 | 0.08 | 0.76 | 3 |
Bmac0096 | 5H | 10 | 0.71 | 0.07 | 0.68 | 2 |
Scssr03907 | 5H | 13 | 0.78 | 0.11 | 0.75 | 3 |
Scssr10148 | 5H | 9 | 0.80 | 0.13 | 0.77 | 1 |
Bmac0018 | 6H | 5 | 0.53 | 0.12 | 0.50 | 0 |
Bmag0009 | 6H | 5 | 0.56 | 0.10 | 0.51 | 0 |
Bmag0867 | 6H | 8 | 0.50 | 0.12 | 0.45 | 2 |
Bmag0870 | 6H | 7 | 0.82 | 0.15 | 0.80 | 0 |
Bmac0064 | 7H | 9 | 0.54 | 0.08 | 0.50 | 1 |
HVCMA | 7H | 6 | 0.63 | 0.16 | 0.58 | 1 |
EBmac0827 | 7H | 10 | 0.75 | 0.13 | 0.73 | 1 |
Scssr15864 | 7H | 9 | 0.48 | 0.15 | 0.46 | 3 |
平均(Average) | - | 9 | 0.68 | 0.10 | 0.65 | 1.32 |
Table 3 Information of 28 core SSR primers of hulless barley
位点 Locus | 染色体 Chromosome | 等位基因数 Allele number | 基因多样性指数 Gene diversity | 杂合性 Heterozygosity | 多态信息量 PIC | 特异等位基因 Unique gene |
---|---|---|---|---|---|---|
Bmag0211 | 1H | 8 | 0.68 | 0.01 | 0.62 | 1 |
Bmag0345 | 1H | 5 | 0.75 | 0.00 | 0.71 | 0 |
Bmag0579 | 1H | 9 | 0.69 | 0.06 | 0.63 | 0 |
EBmac0501 | 1H | 10 | 0.45 | 0.02 | 0.44 | 2 |
Scssr07759 | 2H | 6 | 0.70 | 0.13 | 0.64 | 1 |
Bmag0378 | 2H | 6 | 0.55 | 0.02 | 0.45 | 2 |
Bmag0711 | 2H | 11 | 0.77 | 0.14 | 0.74 | 0 |
Bmag125 | 2H | 11 | 0.66 | 0.12 | 0.62 | 0 |
Bmag0013 | 3H | 16 | 0.86 | 0.09 | 0.84 | 3 |
EBmac705 | 3H | 11 | 0.72 | 0.19 | 0.68 | 3 |
HVM60 | 3H | 10 | 0.72 | 0.09 | 0.69 | 3 |
HVM62 | 3H | 11 | 0.78 | 0.18 | 0.75 | 3 |
EBmac0635 | 4H | 8 | 0.64 | 0.07 | 0.62 | 0 |
EBmac0679 | 4H | 11 | 0.72 | 0.15 | 0.70 | 1 |
EBmac0701 | 4H | 9 | 0.66 | 0.10 | 0.64 | 1 |
Bmac0310 | 4H | 12 | 0.83 | 0.14 | 0.81 | 0 |
Bmag0387 | 5H | 10 | 0.79 | 0.08 | 0.76 | 3 |
Bmac0096 | 5H | 10 | 0.71 | 0.07 | 0.68 | 2 |
Scssr03907 | 5H | 13 | 0.78 | 0.11 | 0.75 | 3 |
Scssr10148 | 5H | 9 | 0.80 | 0.13 | 0.77 | 1 |
Bmac0018 | 6H | 5 | 0.53 | 0.12 | 0.50 | 0 |
Bmag0009 | 6H | 5 | 0.56 | 0.10 | 0.51 | 0 |
Bmag0867 | 6H | 8 | 0.50 | 0.12 | 0.45 | 2 |
Bmag0870 | 6H | 7 | 0.82 | 0.15 | 0.80 | 0 |
Bmac0064 | 7H | 9 | 0.54 | 0.08 | 0.50 | 1 |
HVCMA | 7H | 6 | 0.63 | 0.16 | 0.58 | 1 |
EBmac0827 | 7H | 10 | 0.75 | 0.13 | 0.73 | 1 |
Scssr15864 | 7H | 9 | 0.48 | 0.15 | 0.46 | 3 |
平均(Average) | - | 9 | 0.68 | 0.10 | 0.65 | 1.32 |
位点 Locus | 染色体 Chromosome | 参照品种(等位变异大小,bp) Reference variety(size of allele,bp) |
---|---|---|
Bmag0211 | 1H | 温泉青稞(174),S-16(178),小哨洋大麦(180),ICB01-1145(182),米麦114(184),97-14(186),八十天紫青稞(188),羊毛青稞(190) |
Scssr07759 | 2H | 米麦114(179),15YD-3(185),S-63(209),S-22(213),羊毛青稞(221),ICB02-0490(224) |
HVM62 | 3H | 小哨洋大麦(225),米麦114(231),S-22(247),Giza130(251),15YD-3(255),K35(259),羊毛青稞(261),八十天紫青稞(263),ICB02-0611(265),洋大麦(267) |
EBmac0679 | 4H | S-63(116),迪青3号(128),羊毛青稞(130),白青稞(132),川12259(136),米麦114(142),97-14(144),ICB01-0555(146),K35(148),黄长光大麦(150),ICB02-0546(154) |
Scssr03907 | 5H | S-63(108),(110),米麦114(112),ICB01-0555(117),K35(122),S-22(124),羊毛青稞(126),S-16(130),Giza130(132),15YD-3(138),ICB02-0611(142),宽颖大麦(149) |
Bmag0870 | 6H | S-63(116),S-10(120),S-16(122),羊毛青稞(124),米麦114(126),洋大麦(128),K35(130) |
EBmac0827 | 7H | 宽颖大麦(102),羊毛青稞(104),八十天紫青稞(106),小哨洋大麦(111),黄长光大麦(113),青稞1(115),米麦114(117),K35(119),S-63(121) |
Table 4 Details of 7 SSR molecular markers for molecular fingerprinting in hulless barley varieties
位点 Locus | 染色体 Chromosome | 参照品种(等位变异大小,bp) Reference variety(size of allele,bp) |
---|---|---|
Bmag0211 | 1H | 温泉青稞(174),S-16(178),小哨洋大麦(180),ICB01-1145(182),米麦114(184),97-14(186),八十天紫青稞(188),羊毛青稞(190) |
Scssr07759 | 2H | 米麦114(179),15YD-3(185),S-63(209),S-22(213),羊毛青稞(221),ICB02-0490(224) |
HVM62 | 3H | 小哨洋大麦(225),米麦114(231),S-22(247),Giza130(251),15YD-3(255),K35(259),羊毛青稞(261),八十天紫青稞(263),ICB02-0611(265),洋大麦(267) |
EBmac0679 | 4H | S-63(116),迪青3号(128),羊毛青稞(130),白青稞(132),川12259(136),米麦114(142),97-14(144),ICB01-0555(146),K35(148),黄长光大麦(150),ICB02-0546(154) |
Scssr03907 | 5H | S-63(108),(110),米麦114(112),ICB01-0555(117),K35(122),S-22(124),羊毛青稞(126),S-16(130),Giza130(132),15YD-3(138),ICB02-0611(142),宽颖大麦(149) |
Bmag0870 | 6H | S-63(116),S-10(120),S-16(122),羊毛青稞(124),米麦114(126),洋大麦(128),K35(130) |
EBmac0827 | 7H | 宽颖大麦(102),羊毛青稞(104),八十天紫青稞(106),小哨洋大麦(111),黄长光大麦(113),青稞1(115),米麦114(117),K35(119),S-63(121) |
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