TSA4, Encoding an RsgA GTPase, is Required for Temperature-Dependent Chloroplast Development in Rice

doi:10.1016/j.rsci.2025.08.001

Rice Science ›› 2025, Vol. 32 ›› Issue (5): 599-602.DOI: 10.1016/j.rsci.2025.08.001

• Letters • Previous Articles Next Articles

TSA4, Encoding an RsgA GTPase, is Required for Temperature-Dependent Chloroplast Development in Rice

Xue Kexin¹^,³^,^#, Guo Liang²^,^#, Min Lizhang², Chen Zuwu², Liu Wenqiang², Li Xiaoxiang², Dong Zheng², He Pan¹, Zhang Haiwen³(), Pan Xiaowu¹^,²()

¹Longping Agricultural College, Hunan University, Changsha 410125, China
²State Key Laboratory of Hybrid Rice, Hunan Hybrid Rice Research Center, Hunan Academy of Agricultural Sciences, Changsha 410125, China
³Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China

Received:2025-01-12 Accepted:2025-05-09 Online:2025-09-28 Published:2025-10-11
Contact: Pan Xiaowu (panxiaowu@hunaas.cn); Zhang Haiwen (zhanghaiwen@caas.cn).
About author:^#These authors contributed equally to this work

1. .pdf(408KB)

Abstract

Xue Kexin, Guo Liang, Min Lizhang, Chen Zuwu, Liu Wenqiang, Li Xiaoxiang, Dong Zheng, He Pan, Zhang Haiwen, Pan Xiaowu. TSA4, Encoding an RsgA GTPase, is Required for Temperature-Dependent Chloroplast Development in Rice[J]. Rice Science, 2025, 32(5): 599-602.

Figures/Tables 2

Fig. 1. Phenotypic characterization of wild type (WT, Zhonghua 11) and tsa4 mutant plants. A, Phenotypes of seedlings of WT and tsa4 mutant at 5-day-old grown in the field. B, Phenotypes of seedlings of WT and tsa4 mutant at 2-week-old grown in the field. C, Field temperature data from 1st April to 14th April, 2024. D-G, Phenotypes of WT and tsa4 mutants grown at temperatures of 25 ºC (D and F) and 35 ºC (E and G). H and I, Pigment contents of WT and tsa4 seedlings grown at temperatures of 25 ºC (H) and 35 ºC (I). Chl a, Chlorophyll a; Chl b, Chlorophyll b; Car, Carotenoid. Data are mean ± SD (n = 3). ** indicates significant difference at the 0.01 level. J-Q, Chloroplast ultrastructure in cells of the WT and tsa4 plants at 25 ºC (J-M) and 35 ºC (N-Q). Chloroplast (J, L, N and P, Scale bars are 5 μm) and thylakoid (K, M, O and Q, Scale bars are 2 μm) are clearly detected. CP, Chloroplast; Thy, Thylakoid.

Fig. 2. Map‐based cloning of TSA4 gene and construction and characteristics of tsa4 mutant. A, TSA4 was located between InDel markers S2 and S3 on chromosome 4 (Chr. 4). The locus was further narrowed down to a 158-kb region between InDel markers S6 and S7. The mutation site of TSA4 is shown by a black arrow. Five black boxes indicate five exons of TSA4. ATG and TAG is initiation codon and termination codon of TSA4. B, Mutations of two CRISPR-Cas9 knockout lines with a GG deletion (KO#1) and an A insertion (KO#2). C and D, Phenotypes of WT and TSA4 knockout lines (KO#1 and KO#2) at the seedling stage at 25 ºC (C) and 35 ºC (D). E, Free green fluorescent protein (GFP) and TSA4‐GFP fusion protein were transiently expressed in tobacco epidermal cells. Red signal is chloroplast auto-fluorescence. Scale bars, 20 μm. F and G, Relative expression levels of TSA4 in different rice organs of WT at 25 ºC (F) and in WT and tsa4 leaves grown under different temperatures (G). H-I, Relative expression levels of chloroplast development-related genes in WT and tsa4 mutant grown at 25 ºC (H) and 35 ºC (I). Tubulin gene is served as the internal control. In F-I, Data are mean ± SD (n = 3). * and ** indicate significant difference at the 0.05 and 0.01 probability level, respectively.

References 17

[1]	Janowski M, Zoschke R, Scharff L B, et al. 2018. AtRsgA from Arabidopsis thaliana is important for maturation of the small subunit of the chloroplast ribosome. Plant J, 96(2): 404-420.
[2]	Jarvis P, López-Juez E. 2013. Biogenesis and homeostasis of chloroplasts and other plastids. Nat Rev Mol Cell Biol, 14(12): 787-802.
[3]	Kurata N, Miyoshi K, Nonomura K I, et al. 2005. Rice mutants and genes related to organ development, morphogenesis and physiological traits. Plant Cell Physiol, 46(1): 48-62.
[4]	Liere K, Maliga P. 1999. In vitro characterization of the tobacco rpoB promoter reveals a core sequence motif conserved between phage-type plastid and plant mitochondrial promoters. EMBO J, 18(1): 249-257.
[5]	Lim C, Kang K, Shim Y, et al. 2020. Rice ETHYLENE RESPONSE FACTOR 101 promotes leaf senescence through jasmonic acid- mediated regulation of OsNAP and OsMYC2. Front Plant Sci, 11: 1096.
[6]	Lin D Z, Jiang Q, Ma X J, et al. 2018. Rice TSV3 encoding Obg-like GTPase protein is essential for chloroplast development during the early leaf stage under cold stress. G3-Genes Genomes Genet, 8(1): 253-263.
[7]	Manuell A L, Quispe J, Mayfield S P. 2007. Structure of the chloroplast ribosome: Novel domains for translation regulation. PLoS Biol, 5(8): e209.
[8]	Schmitz A J, Glynn J M, Olson B J S C, et al. 2009. Arabidopsis FtsZ2-1 and FtsZ2-2 are functionally redundant, but FtsZ-based plastid division is not essential for chloroplast partitioning or plant growth and development. Mol Plant, 2(6): 1211-1222.
[9]	Seo D H, Jang J, Park D, et al. 2024. PEP-ASSOCIATED PROTEIN 3 regulates rice tiller formation and grain yield by controlling chloroplast biogenesis. Plant Physiol, 194(2): 805-818.
[10]	Shajani Z, Sykes M T, Williamson J R. 2011. Assembly of bacterial ribosomes. Annu Rev Biochem, 80: 501-526.
[11]	Sharma M R, Wilson D N, Datta P P, et al. 2007. Cryo-EM study of the spinach chloroplast ribosome reveals the structural and functional roles of plastid-specific ribosomal proteins. Proc Natl Acad Sci USA, 104(49): 19315-19320.
[12]	Shin D H, Lou Y, Jancarik J, et al. 2004. Crystal structure of YjeQ from Thermotoga maritima contains a circularly permuted GTPase domain. Proc Natl Acad Sci USA, 101(36): 13198-13203.
[13]	Song J, Wei X J, Shao G N, et al. 2014. The rice nuclear gene WLP1 encoding a chloroplast ribosome L13 protein is needed for chloroplast development in rice grown under low temperature conditions. Plant Mol Biol, 84(3): 301-314.
[14]	Swiatecka-Hagenbruch M, Liere K, Börner T. 2007. High diversity of plastidial promoters in Arabidopsis thaliana. Mol Genet Genomics, 277(6): 725-734.
[15]	Tiller N, Weingartner M, Thiele W, et al. 2012. The plastid-specific ribosomal proteins of Arabidopsis thaliana can be divided into non-essential proteins and genuine ribosomal proteins. Plant J, 69(2): 302-316.
[16]	Vothknecht U C, Westhoff P. 2001. Biogenesis and origin of thylakoid membranes. BBA Mol Cell Res, 1541: 91-101.
[17]	Zheng K L, Zhao J, Lin D Z, et al. 2016. The rice TCM5 gene encoding a novel Deg protease protein is essential for chloroplast development under high temperatures. Rice, 9(1): 13.

TSA4, Encoding an RsgA GTPase, is Required for Temperature-Dependent Chloroplast Development in Rice

RichHTML

PDF

Supplement

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 2

References 17

Related Articles 0

Recommended Articles

Metrics