Determination Of S-genotype Of Huiyang Hongli And The S Genes Cloning Of Polle

Self-incompatibility, a common biological phenomenon in angiosperm, is an important protective mechanism of preventing close breeding and species retrogression that formed during long period evolution. Pear is an kind of ro-saceae plants. It is a typical gametophytic self-incompatibility (GSI) fruit tree. When nature pollination, it has low outputs. So it need arrange varieties for pollination or to perform artificial pollination to adept to overcome. The Huiyang Hongli is a fine cultivated variety and the breeding material.which is with thin peel and crisp pulp, big fruit shape and small core. Because there has no correlative reports about Huiyang Hongli, we research the Huiyang Hongli's S gene by ascertaining its self-incompatibility, isolating its S gene, identifying its S genotype and polle S gene cloning. The mai n results are shown as follows:1.The research validates Huiyang Hongli's self-incompatibility by culturing style in vitro with farm selfed-pollination.2.Using a pair primer of PF:5'-TGCCTCGCTCTTGAACAA-3'and PR:5-A C(A/G)TTCGGCCAAATAATT-3'specific to pear S-RNases, we carried out the PCR amplification from genomic DNAs of Huiyang Hongli.S46-RNase was isolated from Huiyang Hongli. The number of S46-RNase amino acid in signal peptide, HV region, conserved region 1(C1) and conserved region 2 (C2) was 27,15,12 and 11.There is an ami no acid substitutions in conserved region 1 of S46.S46 shows the highest simil ar to Sm in the amino acid sequences. But there are five different amino acids in the hyper variable regions (HV) between S46and Sm. The S-genotype of Huiyang Hongli pear was identified as S46S473.Basing on the highest similar pear Sm and S29 full sequence characteristicthat reported, we designed and synthesized the primer PSF:5'-TGCCTCGC(T/C)CTTG AACAAAT-3:å’ŒPSR:5'-AGCCGC ACTCTAATATAGAAT-3, and using them PCR the Huiyang Hongli's genomic DNA.S46-RNase were amplified isolated from Huiyang Hongli. Which with a 874bp sequence and 146bp intron in it. 4.Using three pairs of specific primer, that of AF1:5'-AGGGACTCGTAGTT TGATTTAGTTA-3'and AR1:5'-ATTACAAAATGCTGAAAGTCCAAAA-3'; BF1: 5'-ACATGAGAAGATTTCCCGTGGAACA-3'and BR1:5'-CCCTCAACTCACTT GA (C/T) TGAAACAA-3'; RF1:5'-GTGTGTAATTCATGTGCATGG-3'and RR1:5'-TGGAACGTTTCCCTCAACTC-3', we carried out the PCR amplification from genomic DNA of Huiyang Hongli.and separatied and isolated three pairs of pollen S gene, such as SFBB47-alpha and SFBB46-alpha (1179bp and 1209bp), SFBB47-beta and SFBB46-beta (1189bp and 1192bp), SFBB47-gamma and SFBB46-gamma (1206bp and 1245bp), the length this gene is as follow:1179bp and 1209bp,1189bp and 1192bp,1206bp and 1245bp.