摘要
梨品种S基因型鉴定对梨栽培中授粉品种选择和遗传育种都具有重要意义。本研究利用梨S-RNase基因荧光标记的特异引物PCR扩增获得梨品种荧光标记的cDNA特异产物;进一步完善梨S-RNase基因cDNA芯片,以被检测梨品种cDNA特异序列与梨S-RNase基因cDNA芯片杂交检测不同梨品种S基因型,并发现新的S-RNase基因。结果表明:利用梨S-RNase基因cDNA芯片鉴定了泸定王皮梨、兴山24号、弥渡百合等35个未知S基因型梨品种,确定了各品种的S基因型。结合PCRRFLP及DNA克隆和测序等技术,发现了7个新的S-RNase基因资源,获得了新S-RNase基因序列。序列分析表明各新S-RNase基因均具有S-RNase基因特异区域序列的典型特征;进化分析显示7个新S-RNase基因主要属于蔷薇科苹果亚科S-RNase类群,且存在种间和属间比种内和属内进化关系更近的现象。7个新的S基因分别命名为:PpS_(53)(Pyrus pyrifolia S53)、PpS_(54)、PpS_(55)、PpS_(56)、PpS_(57)、PpS_(58)和PpS_(59),GenBank登录号分别为:KX581753、KX581754、KX581755、KX581756、KX581757、KX581751和KX581752。
Identification of S-genotypes of pear cuhivars is useful for allocating suitable pollination trees and pear breeding program. The fluorescently-labeled specific cDNA sequences of S-RNases were obtained by PCR am- plification using Cy3-labeled specific primers. The S-RNase cDNA microarray was further improved with which the pear S-genotype was detected and novel S-RNase genes were discovered. The S-genotypes of thirty-five pear culti- vats,in total,including Luding Wangpili＇ , ＇ Xingshan 24＇ and＇ Midu Baihe＇ etc, were identified and seven new Pyrus pyrifolla （ N. L. Burman） Nakai. S-RNase genes were discovered. Then the DNA sequences of the seven new S-RNase genes were identified by DNA cloning and sequencing combined with PCR-RFLP method. Sequence analy- sis demonstrated that the seven new genes showed typical structural features with specific domains of S-RNase. Evo- lution analysis indicated that the seven genes belonged to the S-RNase group of Maloideae, Rosaceae, and the evolu- tionary relationship among S-RNases was closer between species or genera than intra-species or intra-genus. The sev-en S-RNases were named PpS53 （ Pyruspyrifolia S53 ）, PpS54, PpS55 , PpS56 ,PpS57, PpS58 and PpS59, and had been de- posited under the GenBank database with accession numbers KX581753, KX581754, KX581755, KX581756, KX581757, KX581751 and KX581752, respectively.
出处
《植物遗传资源学报》
CAS
CSCD
北大核心
2017年第3期520-529,共10页
Journal of Plant Genetic Resources
基金
国家自然科学基金面上项目(31272124)
国家林业局林业科技成果推广项目([2014]53)
