Kong Youbin, Li Xihuan, Ma Jun, Li Wenlong, Yan Guijun, Zhang Caiying
North China Key Laboratory for Germplasm Resources of Education Ministry, Department of Plant Genetics and Breeding, Hebei Agricultural University, Baoding, 071001, People's Republic of China.
Plant Cell Rep. 2014 Apr;33(4):655-67. doi: 10.1007/s00299-014-1588-5. Epub 2014 Mar 5.
GmPAP4 , a novel plant PAP gene in soybean, has phytase activity. Over-expressing GmPAP4 can enhance Arabidopsis growth when phytate is the sole P source in culture. Phosphorus (P) is an important macronutrient for plant growth and development. However, most of the total P in soils is fixed into organic phosphate (Po). Purple acid phosphatase (PAP) can hydrolyze Po in the soil to liberate inorganic phosphate and enhance plant P utilization. We isolated a novel PAP gene, GmPAP4, from soybean (Glycine max). It had an open reading frame of 1,329 bp, encoding 442 amino acid residues. Sequence alignment and phylogenetics analysis indicated that GmPAP4 was similar to other plant PAPs with large molecular masses. Quantitative real-time PCR analysis showed that the induced expression of GmPAP4 was greater in P-efficient genotype Zhonghuang15 (ZH15) than in P-inefficient genotype Niumaohuang (NMH) during the periods of flowering (28-35 days post phytate stress; DPP) and pod formation (49-63 DPP). Moreover, peak expression, at 63 DPP, was about 3-fold higher in 'ZH15' than in 'NMH'. Sub-cellular localization showed that GmPAP4 might be on plasma membrane or in cytoplasm. Over-expressing GmPAP4 in Arabidopsis resulted in significant rises in P acquisition and utilization compared with the wild-type (WT). Under phytate condition, transgenic Arabidopsis plants showed increases of approximately 132.7 % in dry weight and 162.6 % in shoot P content compared with the WT. Furthermore, when phytate was added as the sole P source in cultures, the activity of acid phosphatase was significantly higher in transgenic plants. Therefore, GmPAP4 is a novel PAP gene that functions in plant's utilization of organic phosphate especially under phytate condition.
GmPAP4是大豆中一个新的植物紫色酸性磷酸酶(PAP)基因,具有植酸酶活性。当植酸是培养基中唯一的磷源时,过表达GmPAP4能促进拟南芥生长。磷(P)是植物生长发育所需的一种重要大量元素。然而,土壤中大部分的总磷被固定为有机磷(Po)。紫色酸性磷酸酶(PAP)可以水解土壤中的有机磷以释放无机磷,从而提高植物对磷的利用率。我们从大豆(Glycine max)中分离出一个新的PAP基因GmPAP4。它有一个1329 bp的开放阅读框,编码442个氨基酸残基。序列比对和系统发育分析表明,GmPAP4与其他大分子质量的植物PAP相似。实时定量PCR分析表明,在开花期(植酸胁迫后28 - 35天;DPP)和结荚期(49 - 63 DPP),磷高效基因型中黄15(ZH15)中GmPAP4的诱导表达高于磷低效基因型牛毛黄(NMH)。此外,在63 DPP时,“ZH15”中的峰值表达比“NMH”高约3倍。亚细胞定位显示,GmPAP4可能位于质膜上或细胞质中。与野生型(WT)相比,在拟南芥中过表达GmPAP4导致磷吸收和利用率显著提高。在植酸条件下,转基因拟南芥植株的干重比WT增加了约132.7%,地上部磷含量增加了162.6%。此外,当在培养基中添加植酸作为唯一的磷源时,转基因植物中酸性磷酸酶的活性显著更高。因此,GmPAP4是一个新的PAP基因,在植物对有机磷的利用中发挥作用,尤其是在植酸条件下。