Liu Xiaoman, Zhao Xiaolei, Zhang Lijun, Lu Wenjing, Li Xiaojuan, Xiao Kai
College of Agronomy, Agricultural University of Hebei, Baoding 071001, China.
College of Life Sciences, Agricultural University of Hebei, Baoding 071001, China.
Funct Plant Biol. 2013 May;40(4):329-341. doi: 10.1071/FP12242.
An expressed sequence tag (EST) highly similar to BdPT1-4, a phosphate transporter (PT) gene in Brachypodium distachyon, was obtained in a wheat root cDNA subtractive suppression library containing genes that respond to low-phosphate (Pi) stress. The DNA sequence covering this EST (designated as TaPht1;4) was determined based on screening a wheat DNA library. TaPht1;4 consists of two exons and one intron and encodes a 555 amino acid (aa) polypeptide with a molecular weight of 60.85kDa and an isoelectric point of 7.60. TaPht1;4 contains 12 conserved membrane-spanning domains similar to previously reported PTs in diverse plant species. Yeast complement analysis in low-Pi medium confirmed that TaPht1;4 confers the capacity to uptake Pi to MB192, a yeast strain with a defective high-affinity PT; with an apparent Km of 35.3μM. The TaPht1;4 transcripts were specifically detected in the root and were highly induced under Pi deficiency. TaPht1;4 was also expressed following a diurnal pattern, i.e. high levels during daytime and low levels during night-time. TaPht1;4 overexpression and downregulation dramatically altered the plant phenotypic features under low-Pi conditions. Samples that overexpressed TaPht1;4 had significantly improved growth traits and accumulated more Pi than the wild-type plant and those with downregulated TaPht1;4 expression. Therefore, TaPht1;4 is a high-affinity PT gene that plays a critical role in wheat Pi acquisition under Pi deprivation.
在一个包含对低磷(Pi)胁迫有响应的基因的小麦根cDNA消减抑制文库中,获得了一个与二穗短柄草中的磷酸盐转运体(PT)基因BdPT1-4高度相似的表达序列标签(EST)。基于对小麦DNA文库的筛选,确定了覆盖该EST的DNA序列(命名为TaPht1;4)。TaPht1;4由两个外显子和一个内含子组成,编码一个555个氨基酸(aa)的多肽,分子量为60.85kDa,等电点为7.60。TaPht1;4含有12个保守的跨膜结构域,与不同植物物种中先前报道的PT相似。在低磷培养基中的酵母互补分析证实,TaPht1;4赋予了高亲和力PT缺陷的酵母菌株MB192吸收Pi的能力;其表观Km为35.3μM。TaPht1;4转录本在根中被特异性检测到,并且在Pi缺乏时被高度诱导。TaPht1;4也呈现昼夜表达模式,即白天水平高而夜间水平低。在低磷条件下,TaPht1;4的过表达和下调显著改变了植物的表型特征。过表达TaPht1;4的样品比野生型植物具有显著改善的生长性状并且积累了更多的Pi,而TaPht1;4表达下调的样品则相反。因此,TaPht1;4是一个高亲和力PT基因,在缺磷条件下对小麦获取Pi起着关键作用。