‘Shinseiki’ pear, a superior cultivars of Pyrus pyrifolia, was bred from a cross of ‘Nijisseiki’× ‘Chojuro’ in Japan. In China, many excellent pear cultivars have been released using ‘Shinseiki’ as a pare...‘Shinseiki’ pear, a superior cultivars of Pyrus pyrifolia, was bred from a cross of ‘Nijisseiki’× ‘Chojuro’ in Japan. In China, many excellent pear cultivars have been released using ‘Shinseiki’ as a parent. In this study, the S-genotypes of 13 progenies of ‘Shinseiki’ were identified by PCR-RFLP analysis, DNA sequencing and bioinformatic method. As a control, ‘Shinseiki’ was also included. PCR amplification showed that a common fragment of about 370 bp was generated in all the 13 cultivars. Restriction digestion analysis showed that PCR product amplified from ten cultivars ‘Huangguan’, ‘Yaqing’, ‘Bishanerhao’, ‘Xinya’, ‘Cuilv’, ‘Xizilv’, ‘Zhonglierhao’, ‘Zhongliyihao’, ‘Qingxiang’ and ‘Qinghua’ could be cut by S4-allele-specific endonuclease, and that from ‘Xinhang’, ‘Cuiguan’ and ‘Zaomeisu’ could be cut by S3-allele-specific endonuclease. DNA sequencing and bioinformatic analysis revealed that the undigested PCR fragment of ‘Huangguan’, ‘Cuiguan’, ‘Zaomeisu’, ‘Yaqing’, ‘Bishanerhao’, ‘Xinhang’, ‘Cuilv’, ‘Xizilv’, ‘Zhonglierhao’, ‘Zhongliyihao’, ‘Qingxiang’, ‘Xinya’ and ‘Qinghua’ was identical to S 16 , S5, S 35 , S 17 , S 16 , S1, S3, S1, S 31 , S 35 , S7, S 17 and S1, respectively. Consequently, the S-genotypes of 13 cultivars were as follows: ‘Huangguan’ （S4S 16 ）, ‘Yaqing’ （S4S 17 ）, ‘Xinhang’ （S1S3）, ‘Bishanerhao’ （S4S 16 ）, ‘Xinya’ （S4S 17 ）, ‘Cuilv’ （S3S4）, ‘Xizilv’ （S1S4）, ‘Zhonglierhao’ （S4S 31 ）, ‘Zhongliyihao’ （S4S 35 ）, ‘Zaomeisu’ （S3S 35 ）, ‘Cuiguan’ （S3S5）, ‘Qingxiang’ （S4S7） and ‘Qinghua’ （S1S4）.展开更多
A new selfincompatibility gene was isolated and identified from Pyrus bretschneideri cultivars of Yingzhiqing and Daaoao via PCR amplification, DNA sequence analysis and cross pollination tests. DNA sequence analysis ...A new selfincompatibility gene was isolated and identified from Pyrus bretschneideri cultivars of Yingzhiqing and Daaoao via PCR amplification, DNA sequence analysis and cross pollination tests. DNA sequence analysis revealed that the isolated fragment displayed a high homology with S 1 ～S 11 allele, and the identity to S 1 ～S 11 allele at the deduced amino level ranged from 56% to 72%; the high degree of variances in the hypervariable （HV） region resulted from the presence of substitution, deletion and insertion of 9 to 15 amino acids. The new Sallele was named S 12 RNase and its accession number was AY250987 in GeneBank. The sizes of HV region, intron, signal peptide, C1 region, C2 region were 39 AA, 341 bp, 27 AA, 11 AA and 10 AA, respectively. The cross pollination tests were carried out using Pyrus pyrifolia cultivars that contained S 1 ～S9RNase genes as female parents, and the cultivars of Daaoao and Yingzhiqing as male parents, respectively. The results showed that all of {%P.pyrifolia%} cultivars were compatible with Daaoao and Yingzhiqing, whereas the cross pollination between Daaoao and Yingzhiqing were incompatible, further confirming that the DNA fragment isolated was a new Sgene.展开更多
文摘‘Shinseiki’ pear, a superior cultivars of Pyrus pyrifolia, was bred from a cross of ‘Nijisseiki’× ‘Chojuro’ in Japan. In China, many excellent pear cultivars have been released using ‘Shinseiki’ as a parent. In this study, the S-genotypes of 13 progenies of ‘Shinseiki’ were identified by PCR-RFLP analysis, DNA sequencing and bioinformatic method. As a control, ‘Shinseiki’ was also included. PCR amplification showed that a common fragment of about 370 bp was generated in all the 13 cultivars. Restriction digestion analysis showed that PCR product amplified from ten cultivars ‘Huangguan’, ‘Yaqing’, ‘Bishanerhao’, ‘Xinya’, ‘Cuilv’, ‘Xizilv’, ‘Zhonglierhao’, ‘Zhongliyihao’, ‘Qingxiang’ and ‘Qinghua’ could be cut by S4-allele-specific endonuclease, and that from ‘Xinhang’, ‘Cuiguan’ and ‘Zaomeisu’ could be cut by S3-allele-specific endonuclease. DNA sequencing and bioinformatic analysis revealed that the undigested PCR fragment of ‘Huangguan’, ‘Cuiguan’, ‘Zaomeisu’, ‘Yaqing’, ‘Bishanerhao’, ‘Xinhang’, ‘Cuilv’, ‘Xizilv’, ‘Zhonglierhao’, ‘Zhongliyihao’, ‘Qingxiang’, ‘Xinya’ and ‘Qinghua’ was identical to S 16 , S5, S 35 , S 17 , S 16 , S1, S3, S1, S 31 , S 35 , S7, S 17 and S1, respectively. Consequently, the S-genotypes of 13 cultivars were as follows: ‘Huangguan’ （S4S 16 ）, ‘Yaqing’ （S4S 17 ）, ‘Xinhang’ （S1S3）, ‘Bishanerhao’ （S4S 16 ）, ‘Xinya’ （S4S 17 ）, ‘Cuilv’ （S3S4）, ‘Xizilv’ （S1S4）, ‘Zhonglierhao’ （S4S 31 ）, ‘Zhongliyihao’ （S4S 35 ）, ‘Zaomeisu’ （S3S 35 ）, ‘Cuiguan’ （S3S5）, ‘Qingxiang’ （S4S7） and ‘Qinghua’ （S1S4）.
文摘A new selfincompatibility gene was isolated and identified from Pyrus bretschneideri cultivars of Yingzhiqing and Daaoao via PCR amplification, DNA sequence analysis and cross pollination tests. DNA sequence analysis revealed that the isolated fragment displayed a high homology with S 1 ～S 11 allele, and the identity to S 1 ～S 11 allele at the deduced amino level ranged from 56% to 72%; the high degree of variances in the hypervariable （HV） region resulted from the presence of substitution, deletion and insertion of 9 to 15 amino acids. The new Sallele was named S 12 RNase and its accession number was AY250987 in GeneBank. The sizes of HV region, intron, signal peptide, C1 region, C2 region were 39 AA, 341 bp, 27 AA, 11 AA and 10 AA, respectively. The cross pollination tests were carried out using Pyrus pyrifolia cultivars that contained S 1 ～S9RNase genes as female parents, and the cultivars of Daaoao and Yingzhiqing as male parents, respectively. The results showed that all of {%P.pyrifolia%} cultivars were compatible with Daaoao and Yingzhiqing, whereas the cross pollination between Daaoao and Yingzhiqing were incompatible, further confirming that the DNA fragment isolated was a new Sgene.
