二穗短柄草成熟胚再生体系建立及农杆菌介导转化研究

Establishment of Brachypodium distachyon Regeneration System from Mature Embryos and the Study of Agrobacterium-mediated Transformation

为建立二穗短柄草组织培养及遗传转化体系,以二穗短柄草BD21-3成熟胚为外植体,对成熟胚愈伤诱导、分化以及农杆菌侵染条件进行了研究。结果表明,在含有2.5mg·L^(-1) 2,4-D的培养基上,愈伤组织出愈率最高为93.83%;在含有0.2mg·L^(-1) KT的分化培养基上,分化率最高为38.18%;对二穗短柄草胚性愈伤组织农杆菌转化和GUS染色结果表明,侵染的过程中农杆菌菌液浓度OD_(600)为0.6、侵染时间为5min时转化率最高。

To establish the regeneration and genetic transformation system of Brachypodium dis- tachyon,the mature embryos of Brachypodium distachyon BD21-3 were used as explant,and the fac- tors affecting the callus induction and the Agrobacterium infection were studied in this research. The results showed that the frequency of callus induction was 93. 83% on the basic media with 2. 5 mg · L^-1 2,4-D. The maximum regeneration frequency （38. 18%） was observed on the differential media containing 0.2 mg ·L^-1 KT. The GUS staining showed that the highest GUS activity was ob- tained when the concentration of Agrobacterium was OD600 = 0.6 and infection incubation duration was 5 min.