剑麻四倍体诱导与倍性鉴定

Tetraploid Induction and Identification in Sisal

以剑麻H.11648愈伤组织为材料,分别采用0、2、4、6、8 g/L浓度的秋水仙碱对愈伤组织进行不同时间(1、2、3、4、5、6 d)的诱导处理,观察处理后的再生植株气孔大小、保卫细胞叶绿体数目、叶长、叶宽、叶形指数、叶片厚度以及苗头大小等,采用流式细胞仪倍性鉴定方法对处理后的再生植株体细胞DNA含量进行了检测。结果表明:用4 g/L浓度的秋水仙碱处理2 d诱变率最高,诱变率为(57.69±6.66)%,不定芽分化率为(63.33±2.89)%,分化系数为(3.55±1.24),加倍后的再生植株叶片下表皮气孔变大,单位叶面积的气孔数减少,叶片变宽变厚,叶形指数变小,植株变粗,突变体再生植株体细胞DNA含量是二倍体的2倍。

Callus derived from H.11648 were treated with 0, 2, 4, 6, 8 g/L of colehicine for 1, 2 , 3, 4, 5, 6 d, respectively. The size and density of leaf stomata, leaf length, leaf width, leaf thickness, leaf index and stem size from regenerated plants were examined, and ploidy level was determined by an analysis of flow cytometry. The results showed that among the tested combinations, the most effective treatment was the 4 g/L colchicine for 2 d which resulted in about （57.69 ~6.66）% polyploidy. The colchieine -induced autotetraploids developed leaves with significantly increased width, thickness, guard cell size when compared with the corresponding control diploids, but significantly reduced stomata density and leaf index, and the stem diameter significantly increased in colchicine- induced tetraploids compared to corresponding controls. The nuclear DNA content of putative tetraploids was indeed doubled relative to that of control diploid plants.