不同性别类型黄瓜ACC合酶基因的单核苷酸多态性标记和酶切扩增长度多态性标记  

Development of SNP Marker and CAPS Marker Linked to ACC Synthase Gene in Different Sexual Phenotypes of Cucumber

作　　者：向太和[1] 王利琳[1] 庞基良[1] 胡江琴[1] 申屠连峰[1] 吴锴[1] 

机构地区：[1]杭州师范学院生命科学学院,杭州310036

出　　处：《生物化学与生物物理进展》2006年第4期362-367,共6页Progress In Biochemistry and Biophysics

基　　金：浙江省自然科学基金资助项目(Y304083).

摘　　要：黄瓜的性别分化与乙烯密切相关,1-氨基环丙烷-1-羧酸(ACC)合酶是乙烯生物合成过程中的关键酶.根据ACC合酶基因家族的保守序列设计PCR引物,从8个不同性别类型(雌雄同株、强雌性和全雌性)黄瓜品种中克隆了长度为1188bp的ACC合酶基因(CS-ACS2)片段(GenBank登记号为:DQ115884～DQ115886和DQ115875～DQ115879).经测序分析,3个雌雄同株性别类型品种的序列完全相同.与之相比,5个强雌性和全雌性品种中存在8个单核苷酸多态性(SNPs)标记,SNPs标记为4个A←→G和4个T←→C之间的转换.在8个SNPs中,有1个SNP位于内含子区域,其余7个SNPs都位于外显子区域.在7个位于外显子区域的SNPs中,有3个为非编码区的SNPs,4个为cSNPs.而在4个cSNPs中,有3个导致了编码的氨基酸序列改变.研究结果表明,与雌雄同株性别类型相比,雌性系中均存在单核苷酸的变异,这提示ACC合酶基因CS-ACS2的单核苷酸变异可能与黄瓜雌性系的发生形成有关.另一方面,根据SNP多态性还发展了一个酶切扩增长度多态性(CAPS)标记C-MT700.利用CAPS标记C-MT700能将强雌性优良品种MT-705与其他黄瓜品种相区别,该标记在黄瓜育种生产上具有一定的应用价值.此外,研究获得的SNPs标记和CAPS标记丰富了黄瓜的分子标记种类.Ethylene regulates sex expression in cucumber plants （Cucumis sativus L.）. ACC synthase is a key factor during the ethylene biosynthesis. Pairs of PCR primers were synthesized corresponding to the conserved sequences of ACC synthase gene family. The 1 188 bp DNA fragment of ACC synthase gene （CS-A CS2） was amplified from genomic DNA of 8 different sexual phenotypes of cucumber respectively （Gerd3ank accession number is DQl15884 ~DQl15886 and DQl15875 -DQ115879）. 8 SNPs have been identified by sequences analysis between 3 monoecious lines and 5 subgynoecious lines and gynoecious lines, which including 4 A←→G and 4 T←→C transition. Of these 8 SNPs, one locus is in intron and 7 loci in exons. Of the 7 SNPs located in exons, 3 SNPs are non-coding SNPs and 4 SNPs are coding SNPs （cSNPs） of which 3 induced changes of encoding amino acid of ACC synthase. The results of SNPs from subgynoecious lines and gynoecious lines suggest that single nucleotide mutation events of CS-A CS2 might be correlated with the development of subgynoecious lines and gynoecious lines in cucumber. Furthermore, CAPS marker C-MT705 was developed for identifying elite subgynoecious cultivar MT-705, which could be valuable in cucumber breeding. Besides, the SNPs and CAPS markers obtained in the study enriched molecular markers of cucumber.

关 键 词：黄瓜 性别类型 1-氨基环丙烷-1-羧酸(ACC)合酶基因 单核苷酸多态性(SNP) 酶切扩增长度多态性(CAPS) 

分 类 号：Q943.2[生物学—植物学] S642.2[农业科学—蔬菜学]