摘要
以‘砀山酥梨’为材料,采用酶联免疫分析法(ELISA),测定叶片中内源IAA的含量。依据已构建的缺铁叶片SSH文库中生长素抑制蛋白(ARP)基因片段的序列信息,应用RACE技术克隆其cDNA全长,通过实时荧光定量(qRT-PCR)技术,分析ARP基因的相对表达量。结果表明:(1)ARP基因cDNA全长为707bp,其中开放阅读框为351bp,编码116个氨基酸,推测的蛋白质分子量为12.82kD;该蛋白可能定位于微体,属于非分泌型、非跨膜蛋白类,并具有ARP基因家族的保守结构域。(2)在不同程度缺铁叶片中ARP基因的表达量存在差异,随着缺铁程度的增加,表达量显著升高,同时叶片中内源IAA含量逐渐降低。据此推测,ARP基因可能负反馈调节缺铁黄化叶片中IAA的水平,从而调控叶片的生长发育。
According to the ARP（Auxin-repressed protein） gene fragment sequence,which was obtained from the SSH（suppression subtractive hybridization） library constructed by iron-deficiency and normal leaves.The full length of ARP was cloned by the method of RACE（repeat amplification of cDNA end）.The ARP expression levels were analyzed by qRT-PCR（real-time quantitative PCR） technology.The results showed that the IAA content in leaf was decreased gradually with the increasing of iron-deficiency degree;the full-length cDNA of ARP was 707 bp,its coding region covered 351 bp and encoded a polypeptide containing 116 amino acids.It was predicted that the ARP might positioned on the microbody in cell,was a nontransmembrane protein with non secretory type,had a conserved domains with auxin gene family;and its molecular weight was 12.82 kD;There were positive significant differences between ARP relative expressions in leaves with different iron-deficiency degrees.The ARP expression increased with the increasing of iron-deficiency degree.It was speculated that the endogenous IAA level was negatively regulated by the ARP in iron-deficiency leaves,to control the leaf growth and development.
出处
《西北植物学报》
CAS
CSCD
北大核心
2013年第2期240-246,共7页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家梨产业技术体系项目(nycytx-29-12)
