FRUCTOKINASE-LIKE PROTEIN 1 interacts with TRXz to regulate chloroplast development in rice  

作　　者：Lei He Sen Zhang Zhennan Qiu Juan Zhao Wendan Nie Haiyan Lin Zhengge Zhu Dali Zeng Qian Qian Li Zhu 

机构地区：[1]State Key Laboratory of Rice Biology, China National Rice Research Institute [2]Hebei Key Laboratory of Molecular and Cellular Biology, Hebei Normal University

出　　处：《Journal of Integrative Plant Biology》2018年第2期94-111,共18页植物学报（英文版）

基　　金：supported by National Natural Science Foundation of China (31371606, 31601284, 31661143006);The Transgenic Plant Research and Commercialization Project of the Ministry of Agriculture of China (2016ZX08001003-002);Zhejiang Province Outstanding Youth Fund (LR16C130001);The Collaborative Innovation Project of the Chinese Academy of Agricultural Sciences (Y2016XT05);State Key Laboratory of Rice Biology Research Project (2017ZZKT10103)

摘　　要：Chloroplast genes are transcribed by the plastidencoded RNA polymerase（PEP） or nucleus-encoded RNA polymerase. FRUCTOKINASE-LIKE PROTEINS（FLNs） are phosphofructokinase-B（Pfk B）-type carbohydrate kinases that act as part of the PEP complex; however, the molecular mechanisms underlying FLN activity in rice remain elusive.Previously, we identified and characterized a heat-stress sensitive albino（hsa_1） mutant in rice. Map-based cloning revealed that HSA_1 encodes a putative OsFLN_2. Here, we further demonstrated that knockdown or knockout of the OsFLN_1, a close homolog of HSA_1/OsFLN_2, considerably inhibits chloroplast biogenesis and the fln_1 knockout mutants, created by clustered regularly interspaced short palindromic repeats（CRISPR） and CRISPR-associate protein_9, exhibit severe albino phenotype and seedling lethality. Moreover, OsFLN_1 localizes to the chloroplast.Yeast two-hybrid, pull-down and bimolecular fluorescencecomplementation experiments revealed that OsFLN_1 and HSA_1/OsFLN_2 interact with THIOREDOXINZ（OsTRXz） to regulate chloroplast development. In agreement with this,knockout of OsTRXz resulted in a similar albino and seedling lethality phenotype to that of the fln_1 mutants. Quantitative reverse transcription polymerase chain reaction and immunoblot analysis revealed that the transcription and translation of PEP-dependent genes were strongly inhibited in fln_1 and trxz mutants, indicating that loss of OsFLN_1, HSA_1/OsFLN_2, or OsTRXz function perturbs the stability of the transcriptionally active chromosome complex and PEP activity. These results show that OsFLN_1 and HSA_1/OsFLN_2 contribute to chloroplast biogenesis and plant growth.Chloroplast genes are transcribed by the plastidencoded RNA polymerase（PEP） or nucleus-encoded RNA polymerase. FRUCTOKINASE-LIKE PROTEINS（FLNs） are phosphofructokinase-B（Pfk B）-type carbohydrate kinases that act as part of the PEP complex; however, the molecular mechanisms underlying FLN activity in rice remain elusive.Previously, we identified and characterized a heat-stress sensitive albino（hsa_1） mutant in rice. Map-based cloning revealed that HSA_1 encodes a putative OsFLN_2. Here, we further demonstrated that knockdown or knockout of the OsFLN_1, a close homolog of HSA_1/OsFLN_2, considerably inhibits chloroplast biogenesis and the fln_1 knockout mutants, created by clustered regularly interspaced short palindromic repeats（CRISPR） and CRISPR-associate protein_9, exhibit severe albino phenotype and seedling lethality. Moreover, OsFLN_1 localizes to the chloroplast.Yeast two-hybrid, pull-down and bimolecular fluorescencecomplementation experiments revealed that OsFLN_1 and HSA_1/OsFLN_2 interact with THIOREDOXINZ（OsTRXz） to regulate chloroplast development. In agreement with this,knockout of OsTRXz resulted in a similar albino and seedling lethality phenotype to that of the fln_1 mutants. Quantitative reverse transcription polymerase chain reaction and immunoblot analysis revealed that the transcription and translation of PEP-dependent genes were strongly inhibited in fln_1 and trxz mutants, indicating that loss of OsFLN_1, HSA_1/OsFLN_2, or OsTRXz function perturbs the stability of the transcriptionally active chromosome complex and PEP activity. These results show that OsFLN_1 and HSA_1/OsFLN_2 contribute to chloroplast biogenesis and plant growth.

关 键 词：FLN FRUCTOKINASE-LIKE PROTEIN 1 interacts with TRXz to regulate chloroplast development in rice HSA PEP 

分 类 号：S511[农业科学—作物学]