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Identification of two plastid transit peptides for construction of pollen-inactivation system in rice

文献类型： 外文期刊

作者： Wang, Menglong 1 ; Peng, Xiaoqun 1 ; Wang, Changjian 3 ; Tang, Xiaoyan 2 ;

作者机构： 1.Huizhou Univ, Sch Life Sci, Huizhou 516007, Peoples R China

2.Chinese Acad Agr Sci, Agr Genom Inst Shenzhen, Shenzhen Branch, Guangdong Lab Lingnan Modern Agr, Shenzhen 518120, Peoples R China

3.South China Normal Univ, Sch Life Sci, Guangdong Prov Key Lab Biotechnol Plant Dev, Guangzhou 510631, Peoples R China

4.Shenzhen Inst Mol Crop Design, Shenzhen 518107, Peoples R China

关键词： Rice (Oryza sativa L.); Seed production technology (SPT); Amyloplast targeting signal peptide (ASP); Pollen-inactivation; Male-sterile

期刊名称：MOLECULAR BREEDING （ 影响因子：3.1； 五年影响因子：3.1 ）

ISSN： 1380-3743

年卷期： 2024 年 44 卷 5 期

页码：

收录情况： SCI

摘要： Hybrid seed production technology (SPT) is achieved through the utilization of a recessive nuclear male-sterile mutant transformed with a transgenic cassette comprising three essential components: the wild-type gene to restore the fertility of the male-sterile mutant, an alpha-amylase gene to disrupt transgenic pollen grains, and red fluorescence protein gene DsRed to distinguish the transgenic seeds from the nontransgenic male sterile seeds. In rice, we establish the pollen disruption system by introducing an amyloplast targeting signal peptide (ASP) at the N-terminus of maize alpha-amylase protein ZM-AA1(Delta SP) (ZM-AA1 with the N-terminal signal peptide removed). The ASP facilitates the transport of ZM-AA1(Delta SP) protein into amyloplast where it degrades starch, resulting in disruption of the pollen fertility. To obtain such signal peptides for rice, we searched the rice proteins homologous to the defined wheat amyloplast proteins followed by protein-protein interaction network predictions and targeting signal peptides prediction. These analyses enabled the identification of four candidate ASPs in rice, which were designated as ASP1, ASP2, ASP3, and ASP4, respectively. ASP1 and ASP2, when linked with ZM-AA1(Delta SP), exhibited the capability to disrupt transgenic pollen grains, whereas ASP3 and ASP4 did not produce this effect. Interestingly, the localization experiments showed that ASP3 and ASP4 were able to target the proteins into chloroplast. The ASP1 and ASP2 sequences provide valuable tools for genetic engineering of the rice male-sterile system, which will contribute to the hybrid rice breeding and production.

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