Characterization of Purple Acid Phosphatase Family and Functional Analysis ofGmPAP7a/7bInvolved in Extracellular ATP Utilization in Soybean
Characterization of Purple Acid Phosphatase Family and Functional Analysis ofGmPAP7a/7bInvolved in Extracellular ATP Utilization in Soybean
复制标题
大豆紫酸性磷酸酶家族特征及参与细胞外ATP利用的GmPAP7a/7b功能分析
DOI：
10.3389/fpls.2020.00661
复制
发表时间：
2020-06-24
影响因子：
5.6
通讯作者：
Tian, Jiang
中科院分区：
文献类型：
--
作者：
Zhu, Shengnan;Chen, Minhui;Tian, Jiang
低磷酸盐（Pi）有效性限制了酸性土壤中作物的生长和产量。尽管根相关酸性磷酸酶（APases）在细胞外有机磷(P)利用中发挥着重要作用，但对大豆（Glycine max）这一重要豆科作物的研究还很少。在本研究中，研究了细胞内（叶和根）和根相关APase活性在pi充足和pi缺乏条件下的动态变化。此外，还分析了大豆紫色酸性磷酸酶（PAP）家族成员的全基因组鉴定及其在Pi饥饿下的表达模式。gmpap7a7和gmpap7b的功能随后被表征，它们的表达被Pi饥饿上调。缺磷处理4 d后，叶片细胞内APase活性显著升高，1 d后，根系细胞内及相关APase活性显著升高，但缺磷处理5 ~ 16 d时，根系细胞内及相关APase活性持续升高。此外，在大豆基因组中共鉴定出38个gmpap成员。在缺磷处理16 d时，叶片中有19个gmpap成员转录本上调，根部中有17个gmpap成员转录本上调，尽管在缺磷处理2 d时，没有任何gmpap成员对缺磷反应。Pi饥饿上调gmpap7a7和gmpap7b，随后选择它们进行进一步分析。GmPAP7a和GmPAP7b均表现出较高的体外抗三磷酸腺苷（ATP）活性。此外，过表达gmpap7a7和gmpap7bin的大豆毛状根显著提高了根相关的APase活性，从而促进了细胞外ATP的利用。综上所述，GmPAP7a和GmPAP7b可能参与大豆根相关APase活性，从而在细胞外ATP利用中发挥作用。
Low phosphate (Pi) availability limits crop growth and yield in acid soils. Although root-associated acid phosphatases (APases) play an important role in extracellular organic phosphorus (P) utilization, they remain poorly studied in soybean (Glycine max), an important legume crop. In this study, dynamic changes in intracellular (leaf and root) and root-associated APase activities were investigated under both Pi-sufficient and Pi-deficient conditions. Moreover, genome-wide identification of members of thepurple acid phosphatase(PAP) family and their expression patterns in response to Pi starvation were analyzed in soybean. The functions of bothGmPAP7aandGmPAP7b, whose expression is up regulated by Pi starvation, were subsequently characterized. Phosphate starvation resulted in significant increases in intracellular APase activities in the leaves after 4 days, and in root intracellular and associated APase activities after 1 day, but constant increases were observed only for root intracellular and associated APase activities during day 5-16 of P deficiency in soybean. Moreover, a total of 38GmPAPmembers were identified in the soybean genome. The transcripts of 19GmPAPmembers in the leaves and 17 in the roots were upregulated at 16 days of P deficiency despite the lack of a response for anyGmPAPmembers to Pi starvation at 2 days. Pi starvation upregulatedGmPAP7aandGmPAP7b, and they were subsequently selected for further analysis. Both GmPAP7a and GmPAP7b exhibited relatively high activities against adenosine triphosphate (ATP)in vitro. Furthermore, overexpressingGmPAP7aandGmPAP7bin soybean hairy roots significantly increased root-associated APase activities and thus facilitated extracellular ATP utilization. Taken together, these results suggest that GmPAP7a and GmPAP7b might contribute to root-associated APase activities, thus having a function in extracellular ATP utilization in soybean.