摘要
以多年生黑麦草(Lolium perenne L.)种子成熟胚为外植体,在MS附加5mg/L2,4-D、500mg/L的L-脯氨酸、130mg/L天门冬酰氨、10mg/LAgNO3的诱导培养基(MSJ)上诱导胚性愈伤组织。用基因枪法将ubiqutin启动子驱动的海藻糖合酶基因(TPS基因)转入多年生黑麦草的胚性愈伤。在含有5mg/LBialaphos的选择培养基上经过2个月的抗性筛选,抗性愈伤组织在分化培养基上生成了898株再生植株,其中的220株检测为阳性株,检测的阳性率占供检株数的24.5%。PCR分析及PCR-Southern杂交鉴定表明,TPS基因已整合到黑麦草的基因组中。抗旱性鉴定表明,转基因黑麦草在干旱胁迫条件下的保水能力增强,电解质渗出率明显低于对照,耐旱性提高。
Mature seeds of Lolium perenne L. were used as explants to initiate callus on MS induction medium supplemented with 5.0 mg/L 2,4-D ＋500 mg/L proline ＋130 mg/L L-Aspargine ＋ 10 mg/L AgNO3 （MSJ）. Trehalose synthase gene （TPS gene） driven by ubiqutin promoter was transferred into embry calluses of perennial ryegrass by microprojectile bombardent. No. of 898 regenerative plants were obtained from resistant calli which had been subcultured on selecting medium containing 5 mg/L Bialaphos for 2 months. 220 transformed plants was detected through PCR and PCR-southem analysis. The positive frequency was 24.5% （No. of positive plants/No. of regenerative plants）. Experiments of drought tolerance showed that transgenic perennial ryegrass demonstrated their improvement in water-holding power and electrolyte leakage under drought stress.
出处
《分子植物育种》
CAS
CSCD
2007年第1期27-31,共5页
Molecular Plant Breeding
基金
国家863项目草类转基因技术研究(2001AA212161)
林草生物技术育种(2002AA2122)资助。
关键词
黑麦草
海藻糖合酶基因(触因)
转基因
再生植株
耐旱性
Perennial ryegrass （Lolium perenne L.）, Trehalose synthase gene （TPS gene）, Transgenic, Plant regeneration, Drought tolerance
