The Chinese pear(Pyrus spp.)exhibits typical gametophytic self-incompatibility(GSI),which inhibits self-crossing and promotes outcrossing,similar to other fruit species in the Rosaceae family.Thus,S-compatible cultiva...The Chinese pear(Pyrus spp.)exhibits typical gametophytic self-incompatibility(GSI),which inhibits self-crossing and promotes outcrossing,similar to other fruit species in the Rosaceae family.Thus,S-compatible cultivars are required in pear orchards to ensure successful pollination and stable yields.In this study,84 native Chinese pear accessions were genotyped by allele-specific PCR using one pair allele consensus primers and 29 pairs of S-allele-specific primers that were designed in this study.After cloning and sequencing the PCR products,the S-genotypes of all 84 pear accessions,including wild and cultivated accessions,were determined.The reported 34 S-alleles and a novel S-allele were isolated from these pear accessions.These S-alleles were expressed specifically in the style.Sequence analysis identified that six pear cultivars originated in China shared the same S-RNases with P.communis(Pc).These findings supported the hypothesis that oriental and occidental Pyrus spp.may share the same pool of alleles at the S-locus.A novel S-RNase was isolated in ‘Putiandouli’,‘Daguoshanli’,‘Yunhongli1,and‘Dianli’and deposited as S67-RNase under accession number MT773568.Furthermore,the deduced amino acid sequences exhibited high similarity(99.56%)to S32-RNase in Malus.The high similarity between S-RNase in Pyrus and Malus indicated that the existence of S-RNase could have predated speciation between Pyrus and Malus.Furthermore,S-allele information was rearranged in Asian and European pears to provide information for selecting the best pollinator for widely cultivated pear cultivars in China.This information is useful for pear production,cross-breeding,and understanding the mechanism of the self-incompatibility reaction.展开更多
Based on the c DNA sequences from hyper variable(HV) regions of identified 52 S-alleles in Oriental pear cultivars, S-RNase c DNA probes were designed, and a c DNA microarray for S-RNase detections was established. Ea...Based on the c DNA sequences from hyper variable(HV) regions of identified 52 S-alleles in Oriental pear cultivars, S-RNase c DNA probes were designed, and a c DNA microarray for S-RNase detections was established. Each microarray contained 240 sites from 55 c DNA probes, including all specific c DNA sequences from the HV regions of the S-alleles. Using the c DNA of pistils of tested pear cultivars as template and Cy3 fluorescently labeling primers by PCR amplification, microarray hybridization detected the S-genotype of each pear cultivar. The genotypes inferred from the c DNA microarray hybridization signals of pear cultivars such as ‘Lijiang Huangsuanli', ‘Xiuyu', ‘Midu Yuli', ‘Baimianli', and ‘Deshengxiang' were similar to the known genotypes of all tested cultivars. The S-RNase c DNA microarrays and the oligonucleotide gene chips were then used to conduct parallel testing of 24 P. pyrifolia cultivars with unknown S-genotypes. In conclusion, the construction of c DNA microarrays has further improved the pear S-RNase detection platform.展开更多
基金financially supported by the National Natural Science Foundation of China (Grant No.32172511)A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutionsthe Key Project of New Variety Breeding of Agriculture of Jiangsu Province (Grant No.PZCZ201725)。
文摘The Chinese pear(Pyrus spp.)exhibits typical gametophytic self-incompatibility(GSI),which inhibits self-crossing and promotes outcrossing,similar to other fruit species in the Rosaceae family.Thus,S-compatible cultivars are required in pear orchards to ensure successful pollination and stable yields.In this study,84 native Chinese pear accessions were genotyped by allele-specific PCR using one pair allele consensus primers and 29 pairs of S-allele-specific primers that were designed in this study.After cloning and sequencing the PCR products,the S-genotypes of all 84 pear accessions,including wild and cultivated accessions,were determined.The reported 34 S-alleles and a novel S-allele were isolated from these pear accessions.These S-alleles were expressed specifically in the style.Sequence analysis identified that six pear cultivars originated in China shared the same S-RNases with P.communis(Pc).These findings supported the hypothesis that oriental and occidental Pyrus spp.may share the same pool of alleles at the S-locus.A novel S-RNase was isolated in ‘Putiandouli’,‘Daguoshanli’,‘Yunhongli1,and‘Dianli’and deposited as S67-RNase under accession number MT773568.Furthermore,the deduced amino acid sequences exhibited high similarity(99.56%)to S32-RNase in Malus.The high similarity between S-RNase in Pyrus and Malus indicated that the existence of S-RNase could have predated speciation between Pyrus and Malus.Furthermore,S-allele information was rearranged in Asian and European pears to provide information for selecting the best pollinator for widely cultivated pear cultivars in China.This information is useful for pear production,cross-breeding,and understanding the mechanism of the self-incompatibility reaction.
基金supported by the National Natural Science Foundation of China(31272124)
文摘Based on the c DNA sequences from hyper variable(HV) regions of identified 52 S-alleles in Oriental pear cultivars, S-RNase c DNA probes were designed, and a c DNA microarray for S-RNase detections was established. Each microarray contained 240 sites from 55 c DNA probes, including all specific c DNA sequences from the HV regions of the S-alleles. Using the c DNA of pistils of tested pear cultivars as template and Cy3 fluorescently labeling primers by PCR amplification, microarray hybridization detected the S-genotype of each pear cultivar. The genotypes inferred from the c DNA microarray hybridization signals of pear cultivars such as ‘Lijiang Huangsuanli', ‘Xiuyu', ‘Midu Yuli', ‘Baimianli', and ‘Deshengxiang' were similar to the known genotypes of all tested cultivars. The S-RNase c DNA microarrays and the oligonucleotide gene chips were then used to conduct parallel testing of 24 P. pyrifolia cultivars with unknown S-genotypes. In conclusion, the construction of c DNA microarrays has further improved the pear S-RNase detection platform.
