一个编码P型PPR蛋白的基因SiTCD1调控谷子低温下早期叶绿体发育

doi:10.1016/j.jia.2024.12.022

Journal of Integrative Agriculture ›› 2026, Vol. 25 ›› Issue (7): 2739-2754.DOI: 10.1016/j.jia.2024.12.022

一个编码P型PPR蛋白的基因SiTCD1调控谷子低温下早期叶绿体发育

收稿日期:2024-08-20 修回日期:2024-12-19 接受日期:2024-11-11 出版日期:2026-07-20 发布日期:2026-06-09

SiTCD1 encodes a P-type PPR protein that affects early chloroplast development at low temperatures in foxtail millet

Zhilan Wang^{1, 2*}, Xiaofen Du^{1, 2*}, Kangni Han², Miao Li^{1, 2}, Shichao Lian², Yuxin Li², Yanfang Li^{1, 2}, Linyi Zhang², Xingchun Wang^{1, 3#}, Jun Wang^{1, 2#}

¹College of Agriculture, Shanxi Agricultural University, Taigu 030800, China
²Houji Laboratory in Shanxi Province/Millet Research Institute, Shanxi Agricultural University, Changzhi 046011, China
³College of Life Sciences, Shanxi Agricultural University, Taigu 030800, China

Received:2024-08-20 Revised:2024-12-19 Accepted:2024-11-11 Online:2026-07-20 Published:2026-06-09
About author:Zhilan Wang, Tel: +86-355-2204016, E-mail: wangyan11111ai@163.com; Xiaofeng Du, Tel: +86-355-2204016, E-mail: dxf6285210@126.com; #Correspondence Jun Wang, Tel/Fax: +86-355-2204158, E-mail: 128wan@163.com; Xingchun Wang, Tel/Fax: +86-354-6286908, E-mail: wxingchun@sxau.edu.cn * These authors contributed equally to this study.
Supported by:
This research was supported by the Central Guidance on Local Science and Technology Development Fund of Shanxi Province, China (YDZJSX2022A043), the Minor Crop Molecular Breeding Platform Special Project of Shanxi Academy of Agricultural Sciences, China (YGC2019FZ3), the Key R&D Projects of Shanxi Province, China (202102140601003), the Modern Agriculture Industry Technology System Construction Project of Shanxi Province, China (2024CYJSTX04-01), the Shanxi Province Agricultural Key Core Technology Research, China (NYGG19), and the Shanxi Province Science and Technology Innovation Team Project, China (2015013001-09).

摘要/Abstract

摘要：

叶绿体基因的表达依赖于细胞核编码因子对其进行RNA代谢加工，但在冷胁迫下该机制仍不明确。在本研究中，我们分离鉴定了一个谷子(Setaria italica)温度敏感叶绿素缺乏突变体sitcd1 (temperature-sensitive chlorophyll-deficient 1)，在低温条件（光照20°C黑暗18°C，L20/D18）下，sitcd1早期发育叶片叶绿素含量降低，叶绿体结构异常，呈现白化表型。图位克隆结果表明，SiTCD1编码一个定位于叶绿体中的P型PPR蛋白。同时发现该基因受低温诱导表达且在早期叶片中高表达。在sitcd1遗传背景下，过表达SiTCD1的转基因株系在低温下出现近乎正常的绿叶表型。此外，REMSAs和RT-PCR结果表明，SiTCD1在体外直接与质体基因atpF结合，可能参与了低温下质体基因atpF的剪接。RNA-seq分析表明，在低温条件下，sitcd1中一些需要ATP提供能量的代谢（如卟啉、叶绿素和谷胱甘肽代谢）相关基因表达水平下调，导致其叶绿素含量、还原性谷胱甘肽（GSH）降低，谷胱甘肽氧化还原对（GSH/GSSG）比值下降。相对于萌发期和苗期，sitcd1在芽期对冷胁迫更敏感，利用195份种质芽期耐冷表型，发现携带SiTCD1^Hap2种质比携带SiTCD1^Hap1种质具有更强的芽期耐冷性。以上结果表明，SiTCD1在谷子低温早期叶绿体发育中发挥重要作用。

Abstract:

Chloroplast gene expression relies on nucleus-encoded factors for RNA metabolism processing, but the mechanisms under cold stress remain poorly understood. In this study, we isolated and characterized a foxtail millet (Setaria italica) mutant, temperature-sensitive chlorophyll-deficient (sitcd1), which exhibited reduced chlorophyll content and abnormal chloroplasts, resulting in an albino phenotype during early leaf development at low temperatures (20°C during the day and 18°C at night). Map-based cloning revealed that SiTCD1 encoded a P-type PPR protein localized in chloroplasts. In sitcd1 background, transgenic lines of SiTCD1 overexpression appeared nearly normal green leaves under L20/D18 condition. SiTCD1 was especially expressed in earlier development of leaves under low temperature. Additionally, SiTCD1 directly bound to the plastid gene atpF in vitro, which might participate in the splicing of plastid gene atpF under low temperature. RNA-seq indicated that the expression of genes related to metabolism (such as porphyrin, chlorophyll and glutathione metabolism), which required ATP for energy, was down-regulated in sitcd1, resulting in decreased chlorophyll content, GSH, and its redox couple (GSH/GSSG) at low temperature. As sitcd1 exhibited more sensitive at the bud bursting stage than germination and seedling stage under cold stress, we identified two haplotypes of SiTCD1 (SiTCD1^Hap1 and SiTCD1^Hap2) in 195 accessions, and found that accessions carrying the SiTCD1^Hap2 allele were more tolerant to cold stress than those with the SiTCD1^Hap1 allele at the bud bursting stage. In summary, our results suggest that SiTCD1 is essential for early chloroplast development under low temperature in foxtail millet.

Key words: foxtail millet (Setaria italica) , chloroplast development , SiTCD1 , plastid gene atpF , low temperature

Zhilan Wang, Xiaofen Du, Kangni Han, Miao Li, Shichao Lian, Yuxin Li, Yanfang Li, Linyi Zhang, Xingchun Wang, Jun Wang. 一个编码P型PPR蛋白的基因SiTCD1调控谷子低温下早期叶绿体发育[J]. Journal of Integrative Agriculture, 2026, 25(7): 2739-2754.

Zhilan Wang, Xiaofen Du, Kangni Han, Miao Li, Shichao Lian, Yuxin Li, Yanfang Li, Linyi Zhang, Xingchun Wang, Jun Wang. SiTCD1 encodes a P-type PPR protein that affects early chloroplast development at low temperatures in foxtail millet[J]. Journal of Integrative Agriculture, 2026, 25(7): 2739-2754.

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一个编码P型PPR蛋白的基因SiTCD1调控谷子低温下早期叶绿体发育

SiTCD1 encodes a P-type PPR protein that affects early chloroplast development at low temperatures in foxtail millet

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