摘要
利用农杆菌介导的花蕾浸泡转化法将拟南芥生长素反应因子8(Auxin Response Factor8,AtARF8)基因导入油菜花序后获得转化种子。在转化种子的抗性筛选中会产生种子内生菌污染现象。通过对内生菌的细胞学观察及PCR分子鉴定,确定种子内生菌来自转染的农杆菌LBA 4404;选择抗生素头孢曲松钠和硫酸庆大霉素进行抑菌试验,50 mg/L头孢曲松钠可以抑制农杆菌生长,同时对植株真叶数量、根系数量、株高、叶片颜色等影响较小。因此,头孢曲松钠可作为该转基因材料抗菌素抗性筛选中种子内生菌的抑制剂。
Arabidopsis Auxin Response Factor 8 （AtARF 8） gene can weaken the apical dominance and improve side sprouts growing when it is overexpressed. The rote-dip transformation method was used to transfer AtARF8 gene into rape （Brassica napus L. ） flowers and 8 143 candidate seeds were harvested from transformed plants. During screening for the transgenic plant, however,250 mg/L kanamycin was not able to effectively control contamination of endogenous microbe from seeds. Through cytological identification and PCR technique, endogenous microbe were determined to be agrobacterium LBA 4404. Different antibacterial substances such as gentamicin sulphate and ceftriaxone sodium were employed to eliminate contamination resulted from the endogenous bacteria. It was found that 50 mg/L ceftriaxone sodium could not only control the contamination but also had litde effects on the growth and development of the plant. So 50 mg/L ceftriaxone sodium can be used as the appropriate antibiotic in anti-antibiotic screening of transgenic Brassica napus seeds.
出处
《种子》
CSCD
北大核心
2009年第4期38-40,共3页
Seed
基金
广东省科技计划项目(2005B2090101015)
关键词
转基因油菜种子
内生菌
农杆菌
抗生素
transgenic Brassica napus seed
endogenous microbe
agrobacterium
antibiotics
