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利用InDel 标记检测玉米互交种混杂的原理及应用

Molecular Plant Breeding

ISSN1672-416X

CN46-1068/S

2003

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Molecular Plant Breeding

ISSN1672-416X

CN46-1068/S

Started in2003

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Hainan Association for Science and Technology

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Hainan Institutes of Biotechnology

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Zhang Q.F.Ph.D.

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Fang X.J.Ph.D.

Wu W.R.Ph.D.

Zhu Y.X.Ph.D.

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Molecular Plant Breeding

Editor-in-Chief

Dr.Zhang Q.F.,National Key Laboratory of Crop Genetic Improvement,Hua-zhong Agricultural University

Executive Editor-in-Chief

Dr.Fang X.J.,Hainan Institute of Tropical Agricultural Resources.

Dr.Wu W.R.,College of Agriculture&Biotechnology,Zhejiang University

Dr.Zhu Y.X.,College of Life Sciences,Peking University

Associate Editor-in-chief

Dr.Li Z.K.,Institute of Crop Sciences,CAAS

Prof.Shi J.S.,Nanjing Forestry University

Dr.Zhang G.Q.,College of Agriculture,South China Agricultural University Field of Competence

Plant genomics&molecular breeding

Field of Competence

Applied genomics&molecular marker genetics Molecular quantitative genetics&MAS

Plant functional genomics&gene regulation Field of Competence

Molecular evolution&molecular breeding Tree molecular genetics&breeding

Plant molecular marker assited breeding

Members of Editorial Board

Field of competence Applied genomics&molecular improvement Dr.Chen L.J.,Rice Research Institute,Yunnan Agricultural

University.chenlijuan@https://www.sodocs.net/doc/ef5543516.html,

Dr.Chen Q.S.,College of Agriculture,Northeast Agricultural University.qshchen@https://www.sodocs.net/doc/ef5543516.html,

Dr.Li H.Y.,Engineering Research Center of Bioreactor and Pharmaceutical Development,Ministry of Education Jilin Agri-cultural University.hyli99@https://www.sodocs.net/doc/ef5543516.html,

Dr.Li R.B.,Rice Research Institute,Guangxi University.lirong-Dr.Liu S.K.,Alkali Soil Natural Environmental Science Center, Northeast Forestry University.shenkuiliu@https://www.sodocs.net/doc/ef5543516.html,

Dr.Mu P.,College of Life Science,Qingdao Agricultural Univer-sity.mupingcau@https://www.sodocs.net/doc/ef5543516.html,

Dr.Wang F.,Biotechnology Research Institute,Fujian Academy of Agricultural Sciences.wf@https://www.sodocs.net/doc/ef5543516.html,

Molecular Plant Breeding

Dr.Bao J.S.,College of Agriculture and Biotechnology,Zhejiang University.jsbao@https://www.sodocs.net/doc/ef5543516.html,

Dr.Cao Q.F.,Biotechnology Research Center,Shanxi Academy of Agricultural Sciences.caoqiufeng@https://www.sodocs.net/doc/ef5543516.html,

Dr.Chen X.H.,School of Horticulture and Plant Protection, Yangzhou University.Xhchen@https://www.sodocs.net/doc/ef5543516.html,

Dr.Li Z.G.,School of Life Sciences,Chongqing University. zhengguoli@https://www.sodocs.net/doc/ef5543516.html, Dr.Song R.T.,School of Life Sciences,Shanghai University. rentaosong@https://www.sodocs.net/doc/ef5543516.html,

Dr.Xie Z.M.,Center for Molecular Agrobiotechnology and breeding,Xinjiang Academy of Agricultural and Reclamation Science.xiezmchy@https://www.sodocs.net/doc/ef5543516.html,

Dr.Xu Z.F.,Xishuangbanna Tropical Botanical Garden,Chinese Academy of Sciences.zfxu@https://www.sodocs.net/doc/ef5543516.html,

Dr Zhao D.G.,College of Life Science,Guizhou University. dgzhao@https://www.sodocs.net/doc/ef5543516.html,

Members of Editorial Board

Field of competence Plant genetic engineering&transgenic breeding

Members of Editorial Board

Field of competence Plant molecular marker assited breeding

Dr.Li R.Z.,Institute of Molecular Agriculture&Bioenergy, Shanxi Agricultural University.rli2001@https://www.sodocs.net/doc/ef5543516.html,

Dr.Liu Z.S.,Oilseed Crops Institute,Hunan Agricultural Univer-sity.zsliu48@https://www.sodocs.net/doc/ef5543516.html,

Dr.Piao Z.Y.,College of Horticulture,Shenyang Agricultural University.zypiao@https://www.sodocs.net/doc/ef5543516.html,

Dr.Shen X.H.,Rice Research Institute,Jiangxi Academy of Agricultural Sciences.shen_xh2000@https://www.sodocs.net/doc/ef5543516.html,

Dr.Tian J.C.,College of Agriculture,Shandong Agricultural University.jctian@https://www.sodocs.net/doc/ef5543516.html, Dr.Xi Z.Y.,Agronomy College of Henan Agricultural Universi-ty.xizhangying@https://www.sodocs.net/doc/ef5543516.html,

Prof.Yang Y.,Institute of Vegetable,Zhejiang Academy of Agri-cultural Sciences.hzyyj@https://www.sodocs.net/doc/ef5543516.html,

Dr.Zhang H.Y.,Sesame Research Center,Henan Academy of Agricultural Sciences.zhy@https://www.sodocs.net/doc/ef5543516.html,

Dr.Zhao J.R.,Maize Research Center,Beijing Academy of Agri-culture and Forestry Sciences.maizezhao@https://www.sodocs.net/doc/ef5543516.html,

Prof.Zhao Z.G.,Academy of Agricultural and Forestry,Qinghai University.138********@https://www.sodocs.net/doc/ef5543516.html,

Molecular Plant Breeding 62-7153-61

Research Articles

Genetic Analysis and Molecular Mapping of a Thermo-sensitive Chlorosis Mutant in Rice

(Dong Y.J.),(Lin D.Z.),(Mei J.),(Su Q.Q.)(Zhang J.H.),(Ye S.H.),(Zhang X.M.)

SUSIRI RNA

Molecular and Phenotypic Analysis on Effect of Over-expression and RNA Interference of a Transcriptional Factor Gene SUSIRI on Transgenic Rice

(Chen J.),(Lian X.H.),(Yang Z.M.),(Chen B.),(Zheng S.P.),(Zhu B.Y.)

(Puccinellia tenuifolra )(PutAMT)N C NH 4+

Effect of Ammonium Transporter (PutAMT)and N-C-terminally Truncated Forms on NH 4+Uptake Capacity in Puccinellia tenuifolra

(Bu Y.Y.),(Sun B.),(Luo Y.Y.),(Zhou A.M.),(Liu S.K.)

Research Reports

123SSR

Fingerprinting Analysis of 123Rice Varieties Based on the New SSR Core Markers

(Tian D.G.),(Lin Y.),(Liu H.Q.),(Su J.),(Chen Z.J.),(Yan J.W.),(Xu Y.),(Wang F.)

SSR

Survey of SSRs in Foxtail Millet Genome and Development of SSR Markers

(Zhang H.),(Wang X.M.),(Wang D.J.),(Sung J.M.),(Yang Y.D.),(Duan L.L.),(Li H.),(Song G.A.),(Wang X.Y.),(Li R.Y.)

Insertion/Deletion (InDel)

The Principle of Distinguishing Maize Hybrids of Direct and Reciprocal Crosses Using Insertion/Deletion (InDel)Markers and Its Application

(Ge M.),(Jiang L.),(Zhang X.L.),(Zhao H.),(Zhang T.F.)

DREB2A

Transformation of Salt-tolerance Gene DREB2A into Maize Mediated by Agrobacterium Tumefaciens

(Wang Y.),(Ren X.),(Yu Z.J.),(Cai Q.A.),(Lin X.F.),(Ma R.)

(BnBADH -1)

The Cloning of Betaine Aldehyde Dehydrogenase Gene from Brassica napus (BnBADH -1)and the Construction of Eukaryotic Expression Vector

(Chai L.),(Li H.J.),(Pu X.B.),(Zhang J.F.),(Jiang J.),(Hu H.B.),(Zheng B.C.),(Jiang L.C.)

GmHsFA1

The Expression Response in Transgenic GmHsFA1Soybean to the Signals of Heat and Drought Stress

(Wu G.X.),(Wei L.),(Tang X.F.),(Wang W.W.),(Wang P.F.)(Wang X.Y.),1-78-1314-1920

-2930-3648-5237-47

Molecular Plant Breeding

139-14472-7677-8485-8990-9899-106KCS

Cloning of Soybean KCS Gene and Predicting of Its Structure

(Zhao Y.),(Zhai Y.),(Qiu Z.C.),(Xu H.H.),(Guo C.H.)

Establishment of Particle Bombardment-mediated Genetic Transformation System of Guzmania Bromeliad

(Shen X.L.),(Wang W.Y.),(Mao B.Z.),(Yu S.H.),(Tian D.Q.),(Yu X.Y.)ISSR

Identification of Vriesea Hybrids by ISSR

(Ge Y.Y.),(Zhang F.),(Wang W.Y.),(Shen X.L.),(Tian D.Q.),(Yu X.Y.),(Yu Y.M.)

WRKY in silico

In silico Identification and Expression Analysis of WRKY Transcription Factors in Tomato

(Wan H.J.),(Yu K.),(Yuan W.),(Liu Y.F.),(Ye Q.J.),(Wang R.Q.),(Ruan M.Y.),(Yao Z.P.),(Yang Y.J.)

SF-HY -0201

Genetic Characteristics of a New Red-flesh Kiwi Fruit Line SF-HY -0201

(Xie Y.),(Pan M.L.),(Zhuang Q.G.),(Wang L.H.),(Zheng X.Q.),(Liao M.A.),(Li M.Z.)

Fingerprints of Hybrids and Their Parents of the Qinyan Series of Chinese Cabbage and Purity Identification of Hybridss

(Yang X.Y.),(Tiana S.M.),(Zhang Q.X.),(Zhang S.X.),(Si C.G.),(Wang Y.),(Wang D.S.)

The Factors Affecting on Regeneration of Root Tips of Chinese Chive

(Zhang X.Z.),(Zhang Y.L.),(Li M.L),(Gao X.Y.),(Hou W.J.),(Ma J.P.),(Li C.L.),(Hou L.P.)

SRAP Genetic Diversity of Phalaenopsis Cultivars in Small and Medium Sizes Based on with the Markers of Morphology and SRAP

(Zhong H.Q.),(Zhong H.F.),(Lin B.),(Huang M.L.),(Wu J.S.),(Fan R.H.)

`'CHS DFR Effect of Light on the Anthocyanin Biosythesis and Expression of CHS and DFR in Rosa chinensis `Spectra '

(Luo J.J.),(Li H.),(Bai B.B.),(Yu H.Q.),(You J.)

-4(Daucus carrot )Transferred Interleukin -4Gene in Carrot (Daucus carota )by Cell Transformation

(Gao J.Q.),(Yu L.J.),(Tao L.)

ISSR-PCR Optimization of ISSR-PCR Reaction System and Primer Selection for Hyacinth

(Hu F.R.),(Wang F.),(Wang Z.Q.),(Ren C.),(Bao R.L.)107-112119-125113-118126-131132-138

201311137-47

Molecular Plant Breeding,2013,Vol.11,No.1,37-47

Research Report

Insertion/Deletion(InDel)

1*1*211**

1,,,210014;2,,210095

**,zhangtifu_82@https://www.sodocs.net/doc/ef5543516.html,

Insertion/Deletion(InDel)PCR

B73Mo178InDel5 PCR(B73Mo17Mo17B73)

PCR3737

3DNA1

6.50E-08 1.000

4InDel

110

/,,,,,

The Principle of Distinguishing Maize Hybrids of Direct and Reciprocal Crosses Using Insertion/Deletion(InDel)Markers and Its Application

Ge Min1*Jiang Lu1*Zhang Xiaolin2Zhao Han1Zhang Tifu1**

1Institute of Biotechnology,Jiangsu Academy of Agricultural Sciences,Provincial Key Laboratory of Agrobiolgy,Nanjing,210014;2Nanking Agri-cultural University,Nanjing,210095

*These authors contributed equally to this work

**Corresponding author,zhangtifu_82@https://www.sodocs.net/doc/ef5543516.html,

DOI:10.3969/mpb.011.000037

Abstract The molecular markers have been extensively employing in the purity identification of maize hybrid. However,the application in distinguishing the hybrids of direct and reciprocal crosses was limited because of the same genomic content contained in the two hybrids.In consideration of endosperm carrying two copies of the maternal alleles and one copy of the paternal allele in its genome,we intended to differentiate the maize hybrids of direct and reciprocal crosses using the statistical probability of chi-square test between the observed value of relative quantification of allelic amplified products(RQAAP)and the expected value indicated by Insertion/Deletion (InDel)in the exponential amplification phase of PCR.Five out of eight co-dominant polymorphic InDel markers between parents B73and Mo17were screened for identification of hybrids of B73Mo17and Mo17B73.The amplification results of InDel markers with different PCR cycles showed that37cycles was the best time point for analysis during the exponential amplification stage with three technical replications.The statistical probability of

2012-09-042012-09-232012-11-07

URL:https://www.sodocs.net/doc/ef5543516.html,/abstract-1005-mpbopa

Molecular Plant Breeding

2011,,3:103-104)

(,2003;,2011)

AA aa,

AAa Aaa,

A:a =2:1a:A =2:1)

DNA PCR

PCR

RQAAP in the leaf genome of hybrid from 6.50E -08to 1.000exhibited that the amplification rates of different markers between alleles could be inconsistent.Statistical analyses of RQAAP in the hybrid endosperm genome of direct and reciprocal crosses revealed that four out of five markers could distinguish the direction of crosses accurately.The probability measured from the remaining one marker was capable for detecting the direct cross,but has its ineffectiveness for reciprocal cross.The results show that the proposed methodology is a promising strategy to identify maize hybrids of direct and reciprocal crosses using molecular markers.

Keywords InDel,Molecular marker,Direct cross,Reciprocal cross,Purity identification,Maize

11.1Insertion/Deletion (InDel)B73Mo1711400100bp InDel (>3)20bp InDel 20bp InDel 143InDel 118PCR B73Mo1782.5%B73InDel 20~80bp PCR 141~361bp InDel InDel 10(1)8B73Mo17InDel (2)BM MB (3)ImageJ (1)(2)318546117~301bp InDel 38

1.2PCR

BM DNA

PCR

PCR PCR

2931333537PCR

27~37

PCR (4)

ImageJ 37PCR 1.3637

Insertion/Deletion (InDel)

Maize Hybrids of Direct and Reciprocal Crosses Using Insertion/Deletion (InDel)Markers and Its Application 2InDel

:M:100bp DNA ladder marker;

Mo17:B73

Figure 2The polymorphism amplified by InDel markers between

Mo17and B73

Note:M:100bp DNA ladder marker;Odd-numbered lane repre-

1InDel

:InDel ,InDel (Mb)

Figure 1Physical distribution of InDels in 10chromosomes of Maize

Note:The number in the right and left sides of chromosome indicates the InDel marker and its physical distance

(Mb)

Molecular Plant Breeding

PCR 5BM 5

3BM MB

:M:100bp DNA ladder marker;

;

Figure 3Amplification of co-dominant markers in the maize leaf

DNA of direct and reciprocal crosses

Note:M:100bp DNA ladder marker;The number in the upper

right corner of waveform plot corresponds to the number of agarose

gel electrophoresis lane and waveform plot demonstrates amplifi-

cation quality of markers between alleles;The No.below the

waveform is the InDel marker name

DNA DNA PCR PCR ImageJ (5)DNA 12(DNA 1)(2)110DNA 1.0032P 1.000181DNA 3.6032P 6.50E -0811108(83DNA ImageJ ,)DNA 134043811

Table 1The information of co-dominant markers

Markers

110Primer sequence F:TCATGGATCGATTTGTGCTGAC R:TCATTTCCCAACTGACGCTACC F:GCTAACCGTATCACTCCCATTG R:GGGCCGCCTGTTACTTCC F:CCCATGTGATTGGTCCACTT R:ATCCACAATCCCTGTGGAAA F:TGCCCAAAAATGTTTCCTGT R:GTCAGTGTGCACCGCTTTC F:TGTGTCTCCCTTTCCGCTTA R:TCCATCTGTCCGACACAAAA F:TGCCTCAGTTCAGTGTGCTC

R:ATGTATGGGCTTTGGTTTCG B73233150214220229186Mo17301107141142372117Length difference

6843737814369Amplified product length 40

581

:M:100bp DNA ladder marker;1,281B73,Mo17;;B73,Mo17ImageJ

Figure 5Identification of samples from direct and reciprocal crosses with marker 81

Note:M:100bp DNA ladder marker;Lane 1,2shows the amplification band of parent B73,Mo17with marker 81;The No.in the waveform plot corresponds to the No.of lane;The left and right crest in the waveform plot displays the image of amplification band from B73and Mo17identified by

ImageJ

4PCR

:M:100bp DNA ladder marker;PCR

Figure 4The agarose gel electrophoresis amplification products from different PCR cycles

Note:M:100bp DNA ladder marker;The Number of lane represents number of PCR cycles

(2)

110

22.1Insertion/Deletion (InDel)

Maize Hybrids of Direct and Reciprocal Crosses Using Insertion/Deletion (InDel)Markers and Its Application 41

Molecular Plant Breeding

P a 0.566

0.102P b

0.079

0.777

0.255

0.307

0.519

0.025

0.039

0.022

0.028

0.040

0.626

0.529

0.697

0.679

0.674

0.015

0.016

P c

1.63E-19

2.56E-03

2.19E-14

4.86E-14

0.040

0.820

0.906

0.787

0.837

0.964

1.64E-03

4.55E-03

7.15E-04

8.60E-04

6.05E-04

0.948

0.962

2InDel

Table2The results of direct and reciprocal crosses with five co-dominant InDel markers

Markers 13

40Meterial

BM leaf

MB leaf

BM1

BM endosperm1

BM2

BM endosperm2

BM3

BM endosperm3

BM4

BM endosperm4

BM5

BM endosperm5

MB1

MB endosperm1

MB2

MB endosperm2

MB3

MB endosperm3

MB4

MB endosperm4

MB5

MB endosperm5

BM leaf

MB leaf

BM1

BM endosperm1

BM2

BM endosperm2

BM3

BM endosperm3

BM4

BM endosperm4

BM5

BM endosperm5

MB1

MB endosperm1

MB2

MB endosperm2

Replicate1

1.644

1.392

6.280

2.795

4.250

4.616

2.270

0.451

0.550

0.400

0.765

0.704

2.291

2.545

3.109

3.435

3.303

3.395

3.029

0.911

0.861

Replicate2

2.389

1.312

5.435

2.858

4.780

5.620

2.127

0.529

0.628

0.560

0.418

0.799

2.231

2.100

3.541

2.846

3.636

3.614

3.360

0.978

0.982

Replicate3

1.917

1.669

5.772

2.453

6.081

4.858

2.249

0.608

0.997

0.528

0.541

0.676

1.984

2.194

3.204

3.007

3.335

3.168

3.897

0.865

1.032

Mean SD

1.7210.392

5.8300.426

2.7020.218

5.0370.942

5.0310.524

2.2150.077

0.5290.078

0.7250.239

0.4960.085

0.5740.176

0.7260.064

2.2240.191

3.2840.227

3.0960.304

3.4250.184

3.3930.223

3.4290.438

0.9180.057

0.9580.088

Chi-square test

RQAAP(B73/Mo17)

Markers

43 81

P a

1.000

6.50E-08

P b

0.011

0.021

0.015

0.640

0.846

0.927

0.949

0.712

8.01E-03

2.20E-03

2.49E-03

2.93E-03

4.37E-03

0.854

0.527

0.865

0.974

0.552

P c

0.872

0.997

0.953

3.28E-05

5.14E-06

1.75E-06

6.35E-09

5.19E-05

0.943

0.537

0.582

0.639

0.776

2.39E-08

1.07E-05

4.64E-13

8.79E-12

8.54E-06

Continuing table2 Meterial

MB3

MB endosperm3

MB4

MB endosperm4

MB5

MB endosperm5

BM leaf

MB leaf

BM1

BM endosperm1

BM2

BM endosperm2

BM3

BM endosperm3

BM4

BM endosperm4

BM5

BM endosperm5

MB1

MB endosperm1

MB2

MB endosperm2

MB3

MB endosperm3

MB4

MB endosperm4

MB5

MB endosperm5

BM leaf

MB leaf

BM1

BM endosperm1

BM2

BM endosperm2

BM3

BM endosperm3

BM4

BM endosperm4

BM5

BM endosperm5

Replicate1

0.860

1.074

1.015

2.668

2.335

3.693

4.257

4.721

5.427

4.316

1.148

0.569

0.572

0.591

0.768

3.250

2.851

6.489

5.116

7.255

7.305

5.894

Replicate2

0.833

1.071

0.910

2.637

2.726

3.554

4.169

4.440

5.684

3.542

0.984

0.452

0.684

0.717

0.774

3.755

3.733

6.671

5.360

6.911

7.743

5.393

Replicate3

0.709

1.062

0.873

2.346

2.697

5.229

5.041

4.788

4.740

4.608

1.081

0.657

0.561

0.695

0.928

4.210

3.821

5.991

5.996

8.619

6.914

5.324

Mean SD

0.8010.081

1.0690.006

0.9330.074

2.5680.179

4.1590.930

4.4890.480

4.6490.185

5.2840.488

4.1550.551

1.0710.083

0.5600.103

0.6060.068

0.6690.067

0.8240.091

3.6030.479

6.3840.353

5.4910.455

7.5950.904

7.3210.415

5.5370.311

RQAAP(B73/Mo17)Chi-square test

Insertion/Deletion(InDel)

Maize Hybrids of Direct and Reciprocal Crosses Using Insertion/Deletion(InDel)Markers and Its Application

Molecular Plant Breeding

Mean SD 0.5510.0880.4120.0210.4990.1620.3540.0250.4270.0691.0030.0651.0260.7312.8460.5031.9701.0613.2200.1613.4180.3962.5000.0920.5570.0630.3910.0370.4140.0290.4110.107Replicate 30.6210.3880.6720.3430.3520.9171.0952.0701.5414.2703.2722.8440.5090.4570.4230.4440.695

Replicate 2

0.452

0.429

0.352

0.337

0.488

0.976

3.522

2.523

3.243

3.391

2.067

0.663

0.332

0.374

0.391

0.505Replicate 10.5800.4200.4710.3820.4431.0211.0052.9461.8462.1493.5902.5860.5000.3850.4450.3980.684Meterial

MB 1MB endosperm 1MB 2MB endosperm 2MB 3MB endosperm 3MB 4MB endosperm 4MB 5MB endosperm 5BM BM leaf MB MB leaf BM 1BM endosperm 1BM 2BM endosperm 2BM 3BM endosperm 3BM 4BM endosperm 4BM 5BM endosperm 5MB 1MB endosperm 1MB 2MB endosperm 2MB 3MB endosperm 3MB 4MB endosperm 4MB 5

MB endosperm 5

Markers 110P a 1.000P b 9.90E-056.72E-058.53E-055.69E-057.01E-050.4520.8810.1890.2220.7710.2070.1420.1500.1490.240P c 0.2710.2000.2400.1750.2072.49E -089.56E -042.64E -118.44E -124.77E -060.9740.9570.9750.9740.932

Continuing table 2

:P a DNA 1;P b DNA 2X A /X a :P c DNA 0.5X A /X a ;-:

Note:P a indicates the statistical probability of chi-square test measured the alignment between the observed value of relative quantifica-tion of allelic amplified products (RQAAP)in the leaf genome of hybrid and its expected value 1;P b indicates the statistical probability of chi-square test measured the alignment between the observed value of RQAAP in the endosperm genome of hybrid and its expected value 2X A /X a ;P c indicates the statistical probability of chi-square test measured the alignment between the observed value of RQAAP RQAAP (B73/Mo17)Chi-square test 44

(Phillips and Evans2011;Nodine and Bartel,2012)

DNA 2.2

(1)

SSR(single sequence repeat)(Sundaram et al.,2008;Wu et al., 2010)

SSR(DB11/T507-2007) SSR

DNA(

1:1)

(2)PCR

PCR

100%PCR X=X0 (1+EX)n

PCR

DNA

ImageJ

PCR

(3)PCR

PCR

(DNA2:1,DNA

1:1)

(4)InDel

InDel

2.3InDel

InDel

(Bhattramakki et al.,2002)

SSR InDel

(, 2012)InDel( ,2005;,2011)

InDel

309bp InDel(Vroh et al.,2006)

6.04Kb SSR

(Linda et al.,2000)InDel

InDel

3'UTR InDel(Fu et al.,2006;Lai et al.,2010)

InDel

InDel InDel

PCR

InDel

117bp301bp InDel

43~78bp

Insertion/Deletion(InDel)

Maize Hybrids of Direct and Reciprocal Crosses Using Insertion/Deletion(InDel)Markers and Its Application

Molecular Plant Breeding InDel

2.4

8InDel3

110

InDel InDel

InDel

3.1

B73Mo17

B73Mo17(BM)Mo17B73(MB)

3.2DNA

B73Mo17BM MB DNA5BM5 MB DNA Karroten DNA

DNA1%

3.3PCR

PCR25L2mmol/L Mg2+ 100mol/L dNTP0.2mol/L1U Taq100g DNA PCR953min9540s5840s, 7240s37725min2%

(0.5g/ml)80V

45min

3.4

ImageJ 3.5

InDel

https://www.sodocs.net/doc/ef5543516.html, Mo17

JGI-DOE emboss eprimer3InDel B73 Mo17

3.6

PCR X=X0(1+Ex)n n

X n X

Ex Ex=1

PCR

A a PCR

A=X A0(1+E A)n X a=X a0(1+E a)n

X A X a A a n

A a

E A E a A a PCR

A a X A0:X a0=1n

(1+E

)n/(1+E a)n=X A/X a

AAa

aaA A:a=2 (X A0:X a0=2)

A:a=0.5(

X A0:X a0=0.5)

DNA DNA PCR

'/X a'=2X A/X a(X A/X a

/X a

);X

'/X a'= 0.5/X A/X a

3.7

(2)

P>0.05

P0.05

(BK2012375)

[CX(12)5020]

Bhattramakki D.,Dolan M.,Hanafey M.,Wineland R.,Vaske D., Register J.C.,Tingey S.V.,and Rafalski A.,2002,Inser-tion-deletion polymorphisms in3'regions of maize genes occur frequently and can be used as highly informative genetic markers,Plant.Mol.Biol.,48:539-547

Feng F.J.,Luo L.J.,Li Y.,Zhou L.G.,Xu X.Y.,Wu J.H.,Chen H.

W.,Chen L.,and Mei H.W.,2005,Comparative analysis of polymorphism of InDel and SSR markers in rice,Fenzi Zhi-wu Yuzhong(Molecular Plant Breeding),3(5):725-730( ,,,,,,, ,,2005,InDel SSR

,,3(5):725-730)

Fu Y.,Wen T.J.,Ronin Y.I.,Chen H.D.,Guo.L.,Mester D.I., Yang Y.,Lee M.,Korol A.B.,Ashlock D.A.,and Schnable P.S.,2006,Genetic dissection of intermated recombinant inbred lines using a new genetic map of maize,Genetics, 174(3):1671-1683

Lai J.,Li R.,Xu X.,Jin W.,Xu M.,Zhao H.,Xiang Z.,Song W., Ying K.,Zhang M.,Jiao Y.,Ni P.,Zhang J.,Li D.,Guo X., Ye K.,Jian M.,Wang B.,Zheng H.,Liang H.,Zhang X., Wang S.,Chen S.,Li J.,Fu Y.,Springer N.M.,Yang H., Wang J.,Dai J.,Schnable P.S.,and Wang J.,2010,Genome-

lines,Nat.Genet.,42(11):1027-1030

Li X.H.,Li X.H.,Li W.H.,Wang Z.H.,Ma F.M.,Yuan L.X.,and Zhang S.H.,2003,Application of SSR markers in hybrid seed purity test of maize,Zuowu Xuebao(Acta Agronomica Sinica),29(1):63-68(,,,, ,,,2003,SSR

,,29(1):63-68)

Linda C.,Luke R.,Dan M.,Malcolm M.,David M.,and Robbie W.,2000,Computational and experimental characterization of physically clustered simple sequence repeats in plants, Genetics,156(2):847-854

Nodine M.D.,and Bartel D.P.,2012,Maternal and paternal genomes contribute equally to the transcriptome of early plant embryos,Nature,482(7383):94-97

Phillips A.R.,and Evans M.M.,2011,Analysis of stunter1,a maize mutant with reduced gametophyte size and maternal effects on seed development,Genetics,187(4):1085-1097 Shen L.,Shen H.L.,Chai M.,Wang Y.L.,and Yang W.C.,2011, Genomic DNA polymorphisms in tomato varieties revealed by InDel and SSR markers,Zhongguo Nongye Daxue Xue-bao(Journal of China Agricultural University),16(2):34-42 (,,,,,2011,InDel

SSR DNA,

,16(2):34-42)

Sundaram R.M.,Naveenkumar B.,Biradar S.K.,Balachandran S.

M.,Mishra B.,IlyasAhmed M.,Viraktamath B.C.,Ramesha M.S.,and Sarma N.P.,2008,Identification of informative SSR markers capable of distinguishing hybrid rice parental lines and their utilization in seed purity assessment,Euphyti-ca,163(2):215-224

Vroh B.I.,McMullen M.,Sanchez-Villeda H.,Schroeder S.,Gar-diner J.,Schaeffer M.,Soderlund C.,Wing R.,Fang Z.,and Coe J.E.,2006,Single nucleotide polymorphisms and inser-tion-deletions for genetic markers and anchoring the maize fingerprint contig physical map,Crop Sci.,6:12-21

Wu M.S.,Jia X.H.,Tian L.,and Lv B.C.,2010,Rapid and reli-able purity identification of F1hybrids of maize(Zea mays L.) using SSR markers,Maize Genomics and Genetics,1(1):1-4 Zhang H.S.,Wang F.G.,Zhao J.R.,Yi H.M.,Li R.Y.,Yang G.H., and Wang L.,2011,Identification of distinctness of maize varieties using morphological characters and SSR molecular markers,Yumi Kexue(Journal of Maize Science),19(3):51-55 (,,,,,,, 2011,SSR

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(2):64-68(,,,,2012,

Insertion/Deletion(InDel)

Maize Hybrids of Direct and Reciprocal Crosses Using Insertion/Deletion(InDel)Markers and Its Application

利用InDel 标记检测玉米互交种混杂的原理及应用

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