A novel dominant selection system for plant transgenics based on phosphite metabolism catalyzed by bacterial alkaline phosphatase.
A novel dominant selection system for plant transgenics based on phosphite metabolism catalyzed by bacterial alkaline phosphatase.
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基于细菌碱性磷酸酶催化亚磷酸盐代谢的新型植物转基因显性选择系统
DOI：
10.1371/journal.pone.0259600
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Zou Z
中科院分区：
文献类型：
--
作者：
Yuan H;Wang Y;Liu Y;Zhang M;Zou Z
选择标记是植物遗传转化中必不可少的手段，常用的选择标记是抗生素或除草剂抗性标记。然而，随着人们对转基因生物安全性的日益关注，许多新的、安全的选择性标记不断涌现，其中一个突出的代表就是亚磷酸盐（Phi）与其脱氢酶（PTDH，如PtxD）的结合。由于细菌碱性磷酸酶（BAP）可以类似于PtxD将毒性Phi氧化转化为可代谢的磷酸盐（Pi），因此我们利用它作为PtxD的替代物来开发替代的基于Phi的选择系统。我们首先验证了大肠杆菌BAP（EcBAP）确实具有氧化Phi为Pi的额外酶活性。EcBAP可作为农杆菌介导的烟草遗传转化的显性选择标记。尽管转化烟草细胞的所涉及的Phi选择令人惊讶地需要Pi的存在，但与常规卡那霉素选择和众所周知的PtxD/Phi系统相比，它显示出相当大的转化效率和显著加速的转化过程。此外，EcBAP转基因烟草可以代谢有毒的Phi作为磷（P）肥料，从而潜在的Phi抗性，并在Phi胁迫下竞争性地拥有相对于野生型烟草和杂草的优势生长。因此，这种新型的BAP/Phi偶联系统，集成了生物安全显性选择标记，植物磷利用和杂草管理的多重优势，可以提供一个PTDH旁路技术选择工程植物物种，特别是那些对可持续农业非常重要的。
Selective markers are generally indispensable in plant genetic transformation, of which the frequently used are of antibiotic or herbicide resistance. However, the increasing concerns on transgenic biosafety have encouraged many new and safe selective markers emerging, with an eminent representative as phosphite (Phi) in combination to its dehydrogenase (PTDH, e.g. PtxD). As bacterial alkaline phosphatase (BAP) can resemble PtxD to oxidatively convert toxic Phi into metabolizable phosphate (Pi), herein we harnessed it as the substitute of PtxD to develop an alternative Phi-based selection system. We first validated the Escherichia coli BAP (EcBAP) did own an extra enzymatic activity of oxidizing Phi to Pi. We further revealed EcBAP could be used as a dominant selective marker for Agrobacterium-mediated tobacco transformation. Although the involved Phi selection for transformed tobacco cells surprisingly required the presence of Pi, it showed a considerable transformation efficiency and dramatically accelerated transformation procedure, as compared to the routine kanamycin selection and the well-known PtxD/Phi system. Moreover, the EcBAP transgenic tobaccos could metabolize toxic Phi as a phosphorus (P) fertilizer thus underlying Phi-resistance, and competitively possess a dominant growth over wild-type tobacco and weeds under Phi stress. Therefore, this novel BAP/Phi-coupled system, integrating multiple advantages covering biosafe dominant selective marker, plant P utilization and weed management, can provide a PTDH-bypass technological choice to engineer transgenic plant species, especially those of great importance for sustainable agriculture.
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影响因子：
13.8
作者：
Loera-Quezada MM;Leyva-González MA;Velázquez-Juárez G;Sanchez-Calderón L;Do Nascimento M;López-Arredondo D;Herrera-Estrella L
通讯作者：
Herrera-Estrella L
影响因子：
13.8
作者：
Benedetti M;Barera S;Longoni P;Guardini Z;Herrero Garcia N;Bolzonella D;Lopez-Arredondo D;Herrera-Estrella L;Goldschmidt-Clermont M;Bassi R;Dall'Osto L
通讯作者：
Dall'Osto L
影响因子：
4.6
作者：
Manna M;Achary VM;Islam T;Agrawal PK;Reddy MK
通讯作者：
Reddy MK
影响因子：
2.7
作者：
Dormatey R;Sun C;Ali K;Fiaz S;Xu D;Calderón-Urrea A;Bi Z;Zhang J;Bai J
通讯作者：
Bai J
影响因子：
46.9
作者：
Lizbeth Lopez-Arredondo, Damar;Herrera-Estrella, Luis
通讯作者：
Herrera-Estrella, Luis