[1]王春雷,高季平,赵宪坤.利用酶联免疫反应鉴定梨树S基因型[J].江苏农业学报,2015,(05):1124-1128.[doi:doi:10.3969/j.issn.1000-4440.2015.05.028]
　WANG Chun-lei,GAO Ji-ping,ZHAO Xian-kun.S genotyping in pear by PCR-ELISA[J].,2015,(05):1124-1128.[doi:doi:10.3969/j.issn.1000-4440.2015.05.028]
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利用酶联免疫反应鉴定梨树S基因型()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年05期
页码:
1124-1128
栏目:
园艺
出版日期:
2015-10-31

文章信息/Info

Title:
S genotyping in pear by PCR-ELISA
作者:
王春雷; 高季平; 赵宪坤
（扬州大学园艺与植物保护学院，江苏扬州225009）
Author(s):
WANG Chun-lei1; GAO Ji-ping; ZHAO Xian-kun
(School of Horticulture and Plant Protection, Yangzhou University, Yangzhou 225009，China)
关键词:
梨; 自交不亲和性; S基因型鉴定; 酶联免疫反应
Keywords:
pear; self-incompatibility; S genotyping; ELISA
分类号:
S661.2
DOI:
doi:10.3969/j.issn.1000-4440.2015.05.028
文献标志码:
A
摘要:
梨是蔷薇科植物，具有典型的配子体自交不亲和性。获取各品种S基因型是果树生产中一项重要内容。现有S基因型鉴定方法存在一些缺陷。利用通用引物扩增S-RNase高变区，结合单链核苷酸探针杂交、酶联免疫反应，成功鉴定8个品种的7个S基因型。PCR酶联免疫法结果显示爱宕（S2S5），爱甘水（S4S5），二十世纪（S2S4），丰水（S3S5），今村秋（S1S6），新高（S3S9）和幸水（S4S5）7个品种都能与各自对应的S基因型探针杂交结合，颜色反应明显，无明显假阳性反应。秋荣（S4smS5）S4sm-RNase基因在花柱中不表达，只在秋荣花柱中检测到S5-RNase基因信号。与目前已有的方法相比，PCR酶联免疫法操作简单，结果可靠，能够广泛用于梨树S基因型鉴定。
Abstract:
Pear, belonging to the family Rosaceae, possesses gametophytic self-incompatibility. Since SI inhibits self-fertilization, it is essential to obtain information on S genotypes of cultivars for successful planting. A method using a semi-automated sequencer is not convenient. S genotyping by dot-blot analysis requires considerable time and skill, and it is not suitable for identifying S genotypes of a small number of cultivars. A rapid and reliable new method for S genotyping is needed to be developed. To solve those problems, in this study, a PCR-ELISA using a set consensus primers to amplify the fragment covering hypervariable region of S-RNase genes, using allele-specific oligonucleotide probes to discriminate the PCR fragment, and using enzyme linked immunosorbent assays (ELISA) to detect the signal allowed us to identify 7 S haplotypes of 8 cultivars. The signals in Atago（S2S5），Aikansui（S4S5），Nijisseiki（S2S4），Housui（S3S5），Imamuraaki（S1S6），Niitaka（S3S9）, and Kousui（S4S5）were consistent with their respective S genotypes. In‘Akiba’（S4smS5），the S4sm-RNase were not expressed in the style; only S5-RNase signals were detected. Compared with the common methods, the PCR-ELISA method is easily-operated and reliable, which is generally applicable to identify the S genotype of pear. 

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期：2015-05-11 基金项目：高等学校全国优秀博士学位论文作者专项基金项目（201467）〖JP〗 作者简介：王春雷，（1981-）男，江苏南京人，博士，教授，研究方向：果树生殖生理。（Tel）18118252060;（E-mail）wangcl@yzu.edu.cn
更新日期/Last Update: 2015-10-31