摘要
砂梨是重要的经济树种,表现出典型的配子体自交不亲和性,在生产和育种上需鉴定品种的S基因型以确定品种间的亲和性。选取7个砂梨品种为试验材料,使用梨S-RNase(S基因)通用引物进行基因组PCR扩增,产物通过1.8%的琼脂糖凝胶电泳分析。结果表明:‘楚比香’等4个品种中产生了预期的2条电泳条带,而其他3个品种都只产生1条带产物,通过6%的聚丙烯酰胺电泳对该3个品种的PCR产物进一步分析,结果产物被成功分离。将7个品种中分离到的14个条带分别回收、克隆、测序及序列分析,从中鉴别出10个具有梨S-RNase基因序列特征的S基因,其中‘政和大雪梨’中494bp的基因片段被鉴定为新的S基因,暂命名为S43-RNase(GenBank接受号EF566873)。RT-PCR试验证明S43-RNase仅在花柱中特异表达,符合S-RNase的表达特征。通过比对S43-RNase的基因组序列和cDNA序列,确定其内含子大小为294bp。在推导氨基酸水平上,S43-RNase与苹果亚科其他S-RNase表现出65%～92%的相似性。
Pyrus pyrifolia is a commercially important fruit tree which exhibits gametophytic seLf-incompatibility （GSI）. It is necessary to identify S-genotypes of cultivars for determination of cross-compatible combination prior to performing pear plantation and breeding programs. In this study, seven cuhivars of P. pyrifolia were used for S-genotype analysis by PCR-based molecular method with primers designed from conserved sequences of known pear S-RNases. Amplified products were analyzed by 1.8 % agarose gel electrophoresis. In each of the four cultivars ＇ Chubixiang＇ , ＇ Huangpishui, Zhenghedaxueli＇ and ＇ Hongtaiyang＇ , two expected bands were generated. The amplified products in the other three cultivars did not show length polymorphism, and therefore were further separated by 6% polyacrylamide gel electrophoresis （PAGE）. A total of 14 purified fragments from the seven cultivars were cloned and sequenced. Sequence analysis revealed that 10 alleles with typical structural features of pear SRNase were identified, of which one from ＇Zhenghedaxueli＇, with 494 bp, was determined as a new S-RNase allele that was tentatively denominated as S43-RNase （GenBank accession No. EF566873）. RT-PCR revealed that the S43-RNase was expressed specifically in the styles, which is consistent with the expression pattern of S-RNases. Comparison of genomic and cDNA sequences revealed there was an intron of 294 bp in the S43-RNase gene. The deduced amino acid sequence shared 65% to 92% similarity with other Maloideae S-RNases. This study will be helpful in pear production and breeding programs.
出处
《林业科学》
EI
CAS
CSCD
北大核心
2008年第10期42-48,共7页
Scientia Silvae Sinicae
基金
the Key Project of State Forestry Administration (2006-12)
Scientific Research Fund of Central South University of Forestry & Technology (2008007B)
