陆地棉微管蛋白基因Ghtub12转棉花研究

Study on Micro-tubulin Gene Ghtub12 to Cotton in Gossypium hirsutum

棉花(Gossypium spp.)是重要的纤维作物,棉花微管蛋白基因在棉纤维发育过程中特异优势表达,影响棉纤维品质。微管蛋白基因Ghtub12为Tubulin基因家族中的一员,在棉纤维发育中机理尚不明确。以陆地棉(Gossypium hirsutum Linn.)R15下胚轴为受体,以0.5 mg/L Basta为筛选标记,用农杆菌介导法将Ghtub12遗传转化棉花。100个胚轴段经过抗性愈伤组织诱导,得到143块愈伤组织,出愈率为71.50%;再经过3次抗性筛选获得126块愈伤组织,对愈伤组织进行PCR检测,102块检测出bar基因,即抗性愈伤组织转化率为81.00%;其中有45块愈伤组织诱导产生胚性愈伤组织,分化率为44.12%。胚性愈伤组织进行成苗培养得60株苗,对其进行PCR检测,56株扩增出条带,阳性率为93.3%。获得了转微管蛋白基因Ghtub12棉花,为进一步研究Ghtub12基因在棉花中的可能机理和作用提供了试验材料。

Cotton is the world＇s important fiber crop, cotton tubulin gene in cotton fiber development process specific advantages of expression, affecting cotton fiber quality. Tubulin gene Ghtub12 as a member of tile tubulin gene family, the mechanism of cotton fiber development is not clear. In this experiment, R15 hypocotyls were used as acceptor, and 0.5 mg/L Basra was used as a screening marker. Agrobacterium tumefaciens-mediated transformation method was used to study the genetic transformation. 100 calli were induced by resistance callus, and 143 callus were obtained after 1 month. The recovery rate was 71.50%. After 3 times, the callus was obtained and the callus was obtained. The transformation rate was 81.00%. Among them, 45 callus induced embryogenic callus, the differentiation rate was 44.12%. Embryogenic calli were divided into 60 seedlings. The positive rate was 93.3%. The transgenic tubulin gene Ghtub12 was obtained, which provided experimental material for further study on the possible mechanism and effect of Ghtub12 gene in cotton.