Cloning of the Self-incompatibility SFB Gene from Chinese Apricot 'Xiaobaixing and Construction of the SFB Expression Vectors


文章信息:

Liu, H. N., Feng, J. R., Liu, X. F., Li, W. H., Lv, W. J., & Luo, M. (2016). Cloning of the self-incompatibility sfb gene from chinese apricot 'xiaobaixing and construction of the sfb expression vectors. Journal of the American Society for Horticultural Science, 141(5): 407-413.

通讯作者:冯建荣





Three kinds of expression vectors of a pollen-S determinant were constructed to provide a reference for molecular breeding of self-compatible (SC) Prunus species. An S-haplotype-specific F-box (SFB) protein gene from the ‘Xiaobaixing’ apricot (Prunus armeniaca) was cloned by reverse transcription polymerase chain reaction (RTPCR) and 3′-rapid-amplification of cDNA ends (3′-RACE). A 1136-bp sequence complementary to the 3′-end of the cDNA (GenBank accession number KP938528.2) with a 912-bp complete open reading frame (ORF) was obtained. The deduced amino acid sequence contained an F-box domain, two variable regions, and two hypervariable regions with structural characteristics similar to SFB in other Rosaceae plants. Sense, antisense, and RNA interference (RNAi) vectors for SFB were constructed by enzyme restriction. The target fragment was restricted using the corresponding restriction enzyme and then directionally inserted between the 35S cauliflower mosaic virus promoter and the nopaline synthase terminator (NOS-ter) of the expression vector pCAMBIA-35S-MCS-NOS-NPTII. The intron-containing hairpin RNA (ihpRNA) was obtained by fusion PCR. The constructed vectors were transferred into Agrobacterium tumefaciens strain LBA4404 by freezing/thawing. The RNAi vector of SFB was also transformed in tobacco (Nicotiana tabacum). The successful construction of these three expression vectors provides a basis for transforming ‘Xiaobaixing’ apricot and the breeding of SC Prunus cultivars. © 2016, American Society for Horticultural Science. All rights reserved.


链接:http://journal.ashspublications.org/content/141/5/407.short

 
网站服务
CopyRight@2017 版权所有:特色果蔬栽培生理与种质资源利用重点实验室
地址:石河子大学 电话: 技术支持:石河子大学网络中心