摘要
豌豆是研究植物发育遗传的经典模式植物.虽然已经克隆到了一些与豌豆花发育有关的基因,但是由于豌豆基因组大、序列信息少以及缺少有效的遗传转化方法,豌豆花发育研究的发展受到了限制.病毒诱导的基因沉默(virus induced gene silencing,VIGS)技术是近年来发展起来的一种反向遗传学快速研究基因功能的方法.本文利用基于豌豆早褐病毒(pea early browning virus,PEBV)的VIGS体系研究了豌豆中AGAMOUS同源基因(Pisum sativum AGAMOUS homologous genes,PsAGs)的功能.在PsAGs沉默之后,豌豆花表现为雄蕊花瓣化,心皮开裂,内生出一朵不完整的花.半定量RT-PCR分析结果显示,在沉默植株中PsAGs的mRNA转录水平显著下降.mRNA原位杂交结果显示,花发育早期,PsAG基因在花原基中央表达,后期在第三、四轮花器官中表达,表明在豌豆有多个AGAMOUS同源基因.实验结果表明,豌豆中可能存在多个AGAMOUS同源基因,彼此间功能冗余且相对保守,同时暗示着在对基因家族成员进行功能研究时,VIGS是有效手段之一.
Pisum sativum L. is a classic model plant for study on plant developmental genetics. Although some genes related to pea floral development have been cloned, the study on pea floral development is restricted because of lacking sequence informations and effective genetic transformation methods, and the pea＇s genome is too big. Virus induced gene silencing （VIGS） comes forth in recent years, and it is a rapid method to study gene functions in reverse genetics way. In this article, we study on the functions of Pisum sativum AGAMOUS homologous genes（PsAGs） by VIGS based on pea early browning virus （PEBV）. After silencing PsAGs, stamen become petaloid, carpel dehisce, an incomplete flower grows from the pea flower center. We find PsAGs mRNA transcriptional level declines dramatically in silencing plant through semi-quantitative RT-PCR, In mRNA in situ hybridization, it shows that PsAG gene expresses in the floral center in floral early development, later, it expresses in the third and fourth whirls. All experiments show that there are many AGAMOUS homologous genes in pea, and their functions are redundant and conservative relatively. It also shows that VIGS is an effective way to study on the functions of gene family members.
出处
《科学通报》
EI
CAS
CSCD
北大核心
2009年第20期3207-3212,共6页
Chinese Science Bulletin
基金
国家自然科学基金资助项目(批准号:30671059)
