荒漠苔藓齿肋赤藓(Syntrichia caninervis Mitt.)瞬时遗传转化方法建立

Establishment of Transient Genetic Transformation Method for Desert Moss Syntrichia caninervis Mitt.

以野生极端耐干藓类植物齿肋赤藓(S. caninervis)为转化受体,进行农杆菌浸染,对GUS基因的瞬时表达率进行组织染色和分子检测分析。结果表明:GUS基因在野生齿肋赤藓植株个体中有一定表达,农杆菌培养过程中农杆菌浓度和瞬时转化过程中共培养时间对瞬时转化率有很大的影响。其中,最优条件为用1/2 MS+100μmol/L AS+3%蔗糖+0.02%Tween20,p H=5.6调整菌液浓度至0.8,得到最高转化苗GUS瞬时表达率,为38%。本研究建立了一种齿肋赤藓外源基因瞬时遗传转化方法,为后续抗逆基因功能原位快速鉴定提供理论参考。

Using wild extreme dry-resistant moss S. caninervis as the transformation receptor to carry out agrobacterium infection, the transient expression rate of GUS gene was analyzed by histochemical staining and molecular detection. The results showed that GUS gene had a certain expression in the individuals of wild S. caninervis, and the Agrobacterium concentration in the process of Agrobacterium cultivation and the co-culture time in the transient transformation process had a great influence on the transient transformation rate. Among them, the optimal condition was to adjust concentration of the bacteria solution to 0.8 with 1/2 MS+100 μmol/L AS+3% sucrose + 0.02% Tween20, pH=5.6. The highest transient expression rate of transformed seedlings GUS was 38%. In this study, a method of transient genetic transformation of exogenous genes of Syntrichia caninervis Mitt. was established for rapid identification of the function of stress-resistant genes in situ.