光周期对棉花愈伤组织发生及体胚发生相关基因表达的影响  

Effect of Light Cycles on Cotton Callus Induction and the Expression of Somatic Embryogenesis Related Gene

作　　者：米贯勋 袁伦[1] 樊可心 张玉祥[1] 崔百明[1] 

机构地区：[1]石河子大学生命科学学院,新疆石河子832003

出　　处：《新疆农业科学》2014年第8期1373-1380,共8页Xinjiang Agricultural Sciences

基　　金：转基因生物新品种培育科技重大专项(2011ZX08005-002)

摘　　要：【目的】研究光周期对棉花愈伤组织发生及体胚发生相关基因表达的影响。【方法】利用石蜡切片对不同培养时期下胚轴细胞形态进行观察;RT-PCR技术对体胚发生相关基因表达量进行分析。【结果】石蜡切片观察发现,光处理后继代培养3 d的下胚轴在b处理(暗处理3 d后光处理3 d)下形成层细胞增殖较快;继代培养7 d时,新陆早33号、45号、50号和珂字棉312号在皮层部位都观察到次生分生细胞团的出现;其中,新陆早45号在b处理和c处理(暗处理12 h后光处理12 h,培养6 d)下以及珂字棉312在四个光周期处理下形成层部位也观测到明显的次生分生细胞团。实时荧光定量PCR表明,光周期b处理下的下胚轴材料GhLEC1、GhSERK1基因相对表达量较高。【结论】光周期b处理更有利于愈伤组织发生,并且该处理下体胚发生相关基因的表达量相对最高。【Objective】To study the effect of light cycles on cotton callus induction and the expression of somatic embryogenesis related gene.【Method】Paraffin section was used to observe cell morphology and RT-PCR techniques was employed to study the expression of somatic embryogenesis related gene.【Result】Paraffin section of hypocotyls after light cycles treatment was cultured 3 d,their primary mertstem were quicker proliferated under b treatment（dark treatment for 3 d and then light treatment for 3 d）; sub-meristem cell masses in outer-cortex were observed in four cotton hypocotyls and cultutred for 7 d;hypocotyls of Xinluzao 45 under b（dark treatment for 3 d and then light treatment for 3 d） and c treatment（dark treatment for 12 h and then light treatment for 12 h,cultured for 6 d） and coker 312 under four light cycles treatment sub-meristem cell masses in procambium were also observed. RT-PCR result showed that the expression of GhSERK1,GhLec1 was relatively higher under b treatment（dark treatment for 3 d and then light treatment for 3 d）,compared with other treatments.【Conclusion】These results indicated that dark treatment for 3 d and then light treatment for 3 d is more advantageous for cotton callus induction and the expression of gene GhSERK1 and GhLecl.

关 键 词：棉花 光周期 石蜡切片 荧光定量PCR GhLEC1基因 GhSERK1基因 

分 类 号：S562[农业科学—作物学] S503