摘要
为深入探究AtbHLH059的生物学功能,本研究筛选到AtbHLH059功能缺陷突变体(atbhlh059)并构建了过表达株系(OEs).在电镜下观察各株系花粉并结合DAPI染色分析,发现atbhlh059突变体的部分花粉形态异常并且没有精细胞;亚历山大染色和花粉萌发实验证明这类花粉败育.qRT-PCR检测各株系精细胞发育标记基因表达情况,结果显示DUO1(duo pollen1)、DUO3(duo pollen3)、FBL17(f-box-like17)、CYCB1(cyclin b1)和GCS1(genera tive cell specific 1)基因在atbhlh059株系中的表达量与野生型相比明显下调,在AtbHLH059过表达株系中没有显著差异.结果表明,AtbHLH059在拟南芥花粉精细胞形成、花粉形态建成过程中是必需的.
Abstract
To further investigate the biological functions of AtbHLH059,this study identified AtbHLH059 loss-of-function mutant(atbhlh059)and constructed overexpression lines(OEs).The mature pollen from various lines were observed using an electron microscope,and found that some pollen staining with DAPI,had abnormal morphology and no sperm cells in atbhlh059 mutants.Alexander staining and pollen germina-tion experiments confirmed that these pollen grains were sterile.qRT-PCR results revealed that the expres-sion levels of DUO1(duo pollen1),DUO3(duo pollen3),FBL17(f-box-like17),CYCB1(cyclin b1)and GCS1(genera tive cell specific 1),which are sperm cell development marker genes,in atbhlh059 were significantly down-regulated while there was no significant difference in overexpression lines,compared with those in the wild-type plants.In conclusion,AtbHLH059 regulates pollen sperm cell formation and pollen morphology and fertility in Arabidopsis thaliana.
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