摘要
烟草绒毡层特异启动子pTA29(TA29 promoter)已用于多种植物雄性不育和花粉发育的研究。作为外源特异表达启动子pTA29在棉花中的表达特性尚不清楚,为了研究该启动子在棉花中的表达特异性,本文构建了pTA29:gus融合基因,并将其转入棉花。研究发现在GUS染液中添加10%的乙醇可以抑制棉花花药内源GUS活性。用改良的GUS染液进行组织化学染色表明,pTA29:gus转化植株的GUS活性主要存在于花药中,并在花蕾长为6mm和15mm的花药中有2个活性高峰,而在根、茎、胚珠、花瓣、苞叶中未被检测到。与烟草不同,在pTA29:gus棉花转化植株的叶片表皮毛和花粉中也能检测到GUS活性。定量RT-PCR分析表明转化子中gus基因的转录水平与GUS活性一致。上述结果表明,烟草绒毡层特异启动子pTA29可以控制下游基因在棉花花药中优势表达;在利用该启动子进行棉花基因功能以及雄性不育研究时,应注意该启动子在棉花中的表达范围和组织特异性。
Tobacco tapetum-specific promoter pTA29 is widely used to create male sterile lines and to study pollen development in various plants. However, the pTA29 expression characteristics in cotton is still not elucidated. To determine the expression characteristics of pTA29 promoter in cotton, gus gene was fused downstream to pTA29, transformed into cotton. It was demonstrated that 10% alcohol suppressed the intrinsic GUS-like activity in cotton anther. GUS staining with modified solution showed that the GUS activity in the pTA29：gus transformants mainly existed in anthers with two peaks in the floral buds of 6 and 15 mm, while no GUS activity was detected in roots, stems, ovules, petals and bracts. Inconsistent with that in tobacco, the GUS activity was also detected in the leaf trichomes and pollens in pTA29：gus transformants. Real-time RT-PCR analysis indicated that the gus transcripts accumulated in a same pattern with the GUS activity. These results suggest that the tobacco tapetum-specific promoter pTA29 can drive the downstream genes to express preferentially in anthers of cotton. The application of pTA29 in cotton male sterility engineering was discussed.
出处
《作物学报》
CAS
CSCD
北大核心
2008年第12期2092-2098,共7页
Acta Agronomica Sinica
基金
国家高技术研究发展计划(863计划)项目(2007AA10Z134)
