CAJ | 학술논문

以SNAC1基因作为筛选标记基因,NaC1作为筛选剂,通过农杆菌介导法将SNAC1和GUS基因导入棉花细胞并得到胚性愈伤组织.经过PCR检测证实,外源基因已经整合到棉花基因组中,GUS染色证明GUS基因得到表达.研究了NaC1作为棉花转化细胞的筛选剂在农杆菌介导转化中的应用浓度及方法,即NaC1的筛选浓度在1.1%～1.5%(W/V)之间,在愈伤组织诱导初期适当低一点,随着愈伤组织的生长而加大筛选浓度.由于NaC1不利于胚的分化,经过2～3次继代筛选后要及时去除NaC1以促进胚的分化.
이SNAC1기인작위사선표기기인,NaC1작위사선제,통과농간균개도법장SNAC1화GUS기인도입면화세포병득도배성유상조직.경과PCR검측증실,외원기인이경정합도면화기인조중,GUS염색증명GUS기인득도표체.연구료NaC1작위면화전화세포적사선제재농간균개도전화중적응용농도급방법,즉NaC1적사선농도재1.1%～1.5%(W/V)지간,재유상조직유도초기괄당저일점,수착유상조직적생장이가대사선농도.유우NaC1불리우배적분화,경과2～3차계대사선후요급시거제NaC1이촉진배적분화.
Employing SNAC1 gene as a selection marker gene and NaC1 as a selection agent, SNAC1 and GUS genes were intro-duced into cotton genome via Agrobacterium. Transgenic calli were confirmed by PCR analysis, and expression of the GUS gene was showed with GUS staining. The concentration and method for employing NaC1 as a selection agent had been studied. The reasonable selection concentration of NaC1 should be 1.1%～1.5%(W/V). The start concentration of NaC1 in callus induction medium should be lower and the concentration increased as calli proliferated. Because NaC1 is not beneficial for embryo differ-entiation and development, NaC1 should be removed from the culture medium after the calli were subcultured for 2～3 times.