摘要
海带孢子体的培育是海带育种育苗技术中的关键步骤,通过转基因技术与海带孢子体育苗相结合来实施有目的的海带品种改良是一项亟待解决的关键技术。本研究利用基因枪法将抗除草剂Bar(bialaphos resistance)基因转入海带雌配子体中,再将转化获得的雌配子体细胞与雄配子体细胞进行杂交,获得了含有抗除草剂基因的海带孢子体幼苗,为海带的转基因技术条件下的育种应用开拓了新的途径。经PCR扩增证明,利用基因枪法转化海带雌配子体细胞,Bar基因已整合到海带配子体细胞的核基因组中,目标基因的转化率可达16.7%。雌配子体的转化体细胞与雄配子体细胞在15℃下共培养10天,就会有大量的合子形成,共培养20天以后许多合子形成孢子体,大约80%左右的合子细胞发育成孢子体,到了30天就会有肉眼可见的2mm左右的孢子体幼苗。
Cultivation of kelp sporophyte has been used as key step in breeding and propagation technology, recently the combination of transgenic technology with kelp sporophyte cultivation is considered as a poten- tial technique to be solved in kelp breeding. The Bar（bialaphos resistance） gene was transformed into the female gametophytes of L. japonica by means of micro--particle bombardment and the transgenic female gametophytes were hybridized with the male gametophyte. In this study, we obtained the kelp sporophyte seedlings containing Bar gene for the transgenic breeding. The PCR amplification results showed that Bar gene had been integrated into the kelp gametophyte genome and the transgenic rate could reach to 16.7G. Under 15 ℃ temperature, the transgenic gametophyte cells were co--cultured with male gametophyte. A number of zygotic cells began to appear after 10 days co--culture, and within more sporophytes were ob- served through the microscope after 20 days. It was found that about 80% zygotic cells developed into young sporophytes, the sporophyte seedlings were visible around 2mm after 30 days.
出处
《青岛农业大学学报（自然科学版）》
2017年第2期133-136,共4页
Journal of Qingdao Agricultural University(Natural Science)
基金
烟台市科技攻关项目(2013LGS010)
