葡萄属野生种麦黄葡萄抗白粉病候选基因序列及应用

摘要

本发明涉及葡萄常规杂交育种、分子标记辅助育种、基因定位与作图及基因克隆的方法与技术。本发明采用抗病基因同源序列法(Resistant geneanalogus，RGAs)获得葡萄属野生种麦黄葡萄(V.bashanica P.C.He)旬阳－8抗白粉病候选基因DNA片段8个，对这些片段克隆测序后，其大小介于510bp～525bp之间。以这些序列为依据，可以进行抗白粉病标记辅助育种，用作葡萄育种中检测葡萄抗白粉病基因的探针；可以进行抗白粉病基因定位与作图；也可以采用cDNA末端快速扩增技术(Rapid amplification ofcDNA ends，RACE)(Frohmanet al，Proc.Natl.Acad.Sci，1988)克隆野生葡萄麦黄葡萄抗白粉病基因，为通过基因工程改良葡萄品种的抗病性奠定基础。

主权项

1.中国葡萄属野生种麦黄葡萄抗白粉病候选基因序列，其特征在于，包括中国葡萄属野生种麦黄葡萄旬阳-8的8个抗白粉病候选基因序列；这些抗白粉病候选基因序列是：麦黄葡萄旬阳-8(1)1 GGGGTGGGTA AGACAACTCT GGCCAAAAAA GTTTACGACA ACCAGAGAGT GGTGGGACAC61 TTCGATTGCC ATGCTTGGGT CACTGTGTCT CAATCATTCA AGATGGAGGA GCTACTACGG121 AACATGACAA AGAAATTCTA TCAGGGAAGA AAGGAAGCAG TTCCTGAGGG TATCGATACA181 ATAGACGAGA TGTCACTGAT TTCCTTAATC AGGCAGTATT TACAAGATAA AAGGTATGTG241 GTTGTTTTTG ATGATGTATG GAAACTAGAT TTCTGGGGAT TTATTAAATA TGTTTTACCA301 GAAAATGGAA ACGGCAGTAG GATAATAATC ACTACACGTA ATGATGAGGT AGCCTCTTCC361 TGCAAAGAGT CGTCATTTGA TTACATCCAC AAGCTGCAAC CCCTGTCCCC AAAGAGTTCC421 TGGGAACTCT TTTGCAAGAA GACATTCCAG GGTGGATGTC CTCCAGAATT GGAGAAACTG481 TCTCTTCATA TTGTCAAAAG ATGTGGAGGA TTGCCACTAG CGTTG麦黄葡萄旬阳-8(13)1 GGGGTAGGCA AGACTACTCT CGCACGGCAG GTTTACAAGT ATGATTTGGC CAAAATTTTG61 AACTAAAAGC GTGGGTTTGT GTGACTGATG TGTTTGATGT GGAGAATATA ACAAAAGCAA121 TTCTCAATTC GGTATTAGAG TCTGATGCAA GTGGCTCTTT GGACTTCCAA CAAGTTCAAA181 AGAAATTGAC AGACACGTTA GCAGGGAAAA CTTTTTTACT CGTTATAGAC GACGTGTGGA241 ACGAAAATTG TGGTCATTGG GATCTTTTGC GGGCTCCTTT TAGTGTAGGG TCCAAAGGAA301 GCAAAGTCAT AGTTACAACA CGCAATAAAA ATGTCGCATT GATGATGGGA GCTGCCAAGA361 ATGTTCATGA ACTAAACCCT TTATCTGAAG ATGCTTGTTG GTCTGTGTTT CAGAAGCATG421 CATTTGAACA TAGAAATATC AATGATCATC CGAACCTGGT GTCCATTGGC AGGAAAATTG481 TTGGCAAATG TGGTGGATTG CCACTAGCGT TG麦黄葡萄旬阳-8(15)1 GGGGTTGGGA AGACAACGCT CGCACGGCTG GTTTACAATG ATGATTTGGC CCAAAATTTT61 GACCTAAAAG CGTGGGTTTG TGTGACTGAT GTGTTTGATC TGGAGAATAT AACAAAAGCA121 ATTCTCAATT CGGTGTTAAA TTCTGATGCA AGTGGGTCTT TGGACTTCCA ACAAGTTCAA181 AGGAAATTGA CAGACACGTT AGCAGGGAAA ACTCTTTTAC TCATTCTAGA TGACGTGTGG241 AACGAAAATT ATTGTAATTG GGATCGTTTG CGGGCTCCTT TGAGTGTAGT GGCAAAAGGA301 AGCAAAGTCA TAGTTACAAC ACGCAATAAA AATGTTGCAT TGATGATGGG AGCTCCCCAG361 AATGTTCATG AACTAAACCC TTTATCTGAA GATGATTGTT GGTCTGTGTT TGAGAAGCAT421 GCATTTGAAC ATAGAAATAT CAATGATCAT CCGAACCTGG AGTCCATTGG TAGGAAAATT481 GTTGCCAAAT GTGGTGGATT GCCACTAGCG TTG麦黄葡萄旬阳-8(2)1 GGTGTGGGTA AGACTACGCT CGCACGGCTG GTTTACAATG ATGATTTGGC CCAAAATTTT61 GACCTAAAAG CGTGGGTTTG TGTGACTGAT GTGTTTGATC TGGAGAATAT AACAAAAGCA121 ATTCTCAATT CGGTGTTAAA TTCTGATGCA AGTGGGTCTT TGGACTTCCA ACAAGTTCAA181 AGGAAATTGA CAGACACGTT AGCAGGGAAA ACTCTTTTAC TCATTCTAGA TGACGTGTGG241 AACGGAAATT ATTGTAATTG GGATCGTTTG CGGGCTCCTT TGAGTGTGGT GGCAAAAGGA301 AGCAAAGTCA TAGTTACAAC ACGCAATAAA AATGTTGCAT TGATGATGGG AGCTCCCCAG361 AATGTTCATG AAATAAACCC TTTATCTGAA GATGATTGTT GGTCTGTGTT TCAGAAGCAT421 GCATTTGAAC ATAGAAATAT CAATGATCAT CCGAACCTGG AGTCCATTGG TAGGAAAATT481 GTTGCCAAAT GTGGTGGATT GCCACTAGCG TTG麦黄葡萄旬阳-8(3)1 GGGGTGGGTA AGACTACGCT CGCACGGCTG GTTTACAATG ATGATTTGGC CCAAAATTTT61 GACCTAAAAG CGTGGGTTTG TGTGACTGAT GTGTTTGATC TGGAGAATAT AACAAAAGCA121 ATTCTCAATT CGGTGTTAAA TTCTGATGCA AGTGGGTCTT TGGACTTCCA ACAAGTTCAA181 AGGAAATTGA CAGACACGTT AGCAGGGAAA ACTCTTTTAC TCATTCTAGA TGACGTGTGG241 AACGGAAATT ATTGTAATTG GGATCGTTTG CGGGCTCCTT TGAGTGTGGT GGCAAAAGGA301 AGCAAAGTCA TAGTTACAAC ACGCAATAAA AATGTTGCAT TGATGATGGG AGCTCCCCAG361 AATGTTCATG AAATAAACCC TTTATCTGAA GATGATTGTT GGTCTGTGTT TCAGAAGCAT421 GCATTTGAAC ATAGAAATAT CAATGATCAT CCGAACCTGG AGTCCATTGG TAGGAAAATT481 GTTGCCAAAT GTGGTGGATT GCCACTAGCG TTG麦黄葡萄旬阳-8(5)1 GGTGTTGGTA AGACAACTCT CGCACGGCAG GTTTACAAGT ATGATTTGGC CAAAATTTTG61 AACTAAAAGC GTGGGTTTGT GTGACTGATG TGTTTGATGT GGAGAATATA ACAAAAGCAA121 TTCTCAATTC GGTATTAGAG TCTGATGCAA GTGGCTCTTT GGACTTCCAA CAAGTTCAAA181 AGAAATTGAC AGACACGTTA GCAGGGAAAA CTTTTTTACT CGTTATAGAC GACGTGTGGA241 ACGAAAATTG TGGTCATTGG GATCTTTTGC GGGCTCCTTT TAGTGTAGGG TCCAAAGGAA301 GCAAAGTCAT AGTTACAACA CGCAATAAAA ATGTCGCATT GATGATGGGA GCTGCCAAGA361 ATGTTCATGA ACTAAACCCT TTATCTGAAG ATGCTTGTTG GTCTGTGTTT CAGAAGCATG421 CATTTGAACA TAGAAATATC AATGATCATC CGAACCTGGT GTCCATTGGC AGGAAAATTG481 TTGGCAAATG TGGTGGATTG CCACTAGCGT TG麦黄葡萄旬阳-8(7)1 GGGGTTGGGA AGACTACTCT GGCCAAAAAA GTTTACGACA ACCAGAGAGT GGTGGGACAC61 TTCGATTGCC ATGCTTGGGT CACTGTGTCT CAATCATTCA AGATGGAGGA GCTACTACGG121 AACATGACAA AGAAATTCTA TCAGGGAAGA AAGGAAGCAG TTCCTGAGGG TATCGATACA181 ATAGACGAGA TGTCACTGAT TTCCTTAATC AGGCAGTATT TACAAGATAA AAGGTATGTG241 GTTGTTTTTG ATGATGTATG GAAACTAGAT TTCTGGGGAT TTATTAAATA TGTTTTACCA301 GAAAATGGAA ACGGCAGTAG GATAATAATC ACTACACGTA ATGATGAGGT AGCCTCTTCC361 TGCAAAGAGT CGTCATTTGA TTACATCCAC AAGCTGCAAC CCCTGTCCCC AAAGAGTTCC421 TGGGAACTCT TTTGCAAGAA GACATTCCAG GGTGGATGTC CTCCAGAATT GGAGAAACTG481 TCTCTTCATA TTGTCAAAAG ATGTGGAGGA TTGCCACTAG CGTTG麦黄葡萄旬阳-8(8)1 GGGGTGGGGA AGACAACCCT TTTGACCCAA ATCAACAATG AGTTCCTCAA AACAACTCAT61 GATTTTGATG TTGTGATCTG GGCTGTGGTG TCACGAGATC CAGACTTTCC CAAGGTTCAA121 GATGAAATTG GGAAAAAGGT TGGGTTTTGT GATGGTATAT GGAGAAATAA AAGCAAGGAT181 GAAAAAGCCA TAGACATCTT TAGAGCCTTG CGCAAAAAGA GGTTTGTGTT GTTGTTAGAT241 GACGTATGGG AGCCGGTAAA TCTATCAGTC TTGGGAGTTC CAGTTCCAAA TGAAGAAAAT301 AAGTCCAAGC TAGTATTCAC AACCCGATCC GAGGATGTAT GCCGACAAAT GGAAGCTCAA361 AAGAATATCA AAGTGGAGTG CTTGGCATGG CAGGAATCCT GGGATTTGTT TCAGAAGAAG421 GTTGGACAAG ACACACTAGA CTCCCATGCA GAAATCCCCA TGCTAGCTGA AATGGTTGCC481 AAAGAATGTT GCGGATTGCC ACTAGCGTTG