五章分子标记辅助育种课件.ppt

1、第五章分子标记辅助育种Single base mutation Insertion/deletion RearrangementBased on variation in the DNA sequenceMolecular MarkerCAPACITY1985199019952000DNA ChipsISSRAFLPsSSRsRAPDsRFLPs?Advanced in Molecular Markers 1 RAPDsAnonymous markers-but can be convertedto SCARsDominant markers-homozygotes cannot be dis

2、tinguished from heterozygotesFast,easy and cheap-commercial primer sets availablePCR-based markers微卫星分子标记对微卫星分子标记对18份山羊草基因型的扩增结果份山羊草基因型的扩增结果 设计探针，筛选重组克隆设计探针，筛选重组克隆根据根据SSRSSR两侧序列设计并合成引物两侧序列设计并合成引物 PCRPCR扩增反应扩增反应 建立基因组建立基因组DNADNA的质粒文库的质粒文库对阳性克隆对阳性克隆DNADNA插入序列测序插入序列测序凝胶电泳检测多态性凝胶电泳检测多态性http:/www.agron.m

3、issouri.edu双酶切连接节头预扩增 选择性扩增The specific band in Xianyou 63 amplified withAFLP primer combination E-AAC/M-CAT CAP 标记Indica (1)CGGCATATGCTATTTTCTTTGCATCTTTTAACTCTTTATGGTTCTCTCATJaponica(1)CGGCATATGCTATTTTCTTTGCATCTTTTAACTCTTTATGGTTCTCTCATIndica (51)TACTTGTTTTGTAGGCTATTTGAAGTTTGGGATTAATACAGTTGATGGTGJap

4、onica(51)TACTAGTTTTGTAGGCTATTTGAAGTTTGGGATTAATACAGTTGATGGTGIndica (101)CCACAATATATCGTGAATGGGCGCCTGCTGCACAGTAAGTTCTAATGTAGJaponica(101)CCACAATATATCGTGAATGGGCGCCTGCTGCACAGTAAGTTCTAATGTAGIndica (151)TCATCCAGCTATTCGCTAATGTTTATGTGTTGTAGGAAATATGGATCACTJaponica(151)TCATCCAGCTATTCGCTAATGTTTATGTGTTGTAGGAAATA

5、TGGATCACTSBE1片段片段Left primerRight primerSNP标记Indica (1)CGGCATATGCTATTTTCTTTGCATCTTTTAACTCTTTATGGTTCTCTCATJaponica(1)CGGCATATGCTATTTTCTTTGCATCTTTTAACTCTTTATGGTTCTCTCATIndica (51)TCCATGTTTTGTAGGCTATTTGAAGTTTGGGATTAATACAGTTGATGGTGJaponica(51)TCCATc TTTTGTAGGCTATTTGAAGTTTGGGATTAATACAGTTGATGGTGIndica (10

6、1)CCACAATATATCGTGAATGGGCGCCTGCTGCACAGTAAGTTCTAATGTAGJaponica(101)CCACAATATATCGTGAATGGGCGCCTGCTGCACAGTAAGTTCTAATGTAGIndica (151)TCATCCAGCTATTCGCTAATGTTTATGTGTTGTAGGAAATATGGATCACTJaponica(151)TCATCCAGCTATTCGCTAATGTTTATGTGTTGTAGGAAATATGGATCACTSBE1片段片段Left primerRight primerGCATPCR变性，退火变性，退火GCATCEL I变性变

7、性走胶，检测走胶，检测自花授粉作物作图群体的构建方法自花授粉作物作图群体的构建方法P PFFFFFB1:BCF PF PF PB2:BCDHL异花授粉作物作图群体的构建方法异花授粉作物作图群体的构建方法ABCDEfGABCDEfGAbcdEfgAbcdEfgAbCDEfGAbCDEfGaBCdefgaBCdefg杂合杂合F对对B、D、G位点来说，相当于位点来说，相当于F对对A、C、E位点来说，相当于测交位点来说，相当于测交F位点不分离位点不分离The specific band in Xianyou 63 amplified withAFLP primer combination E-AAC/M-CAT 1 2 3Complementary pattern amplified with AFLP primer pair EcoRI-AAG/MseI-CAA.1.Female parent Zhenxian 97A;2.Shanyou 63(Hybrid);3.Male parent Minghui63.SSR检测杂交种郑单958纯度P1:父本；P2：母本；1-22：杂交种M P1P21 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16171819202122 提莫菲维小麦提莫菲维小麦G祖染色体特异性祖染色体特异性RAPD 标记和标记和SCAR标记标记