源于新疆野生油菜的线粒体基因orf113b和orf146导致异源细胞质雄性不育花粉败育

doi:10.1016/j.jia.2024.09.018

Journal of Integrative Agriculture ›› 2026, Vol. 25 ›› Issue (4): 1384-1401.DOI: 10.1016/j.jia.2024.09.018

源于新疆野生油菜的线粒体基因orf113b和orf146导致异源细胞质雄性不育花粉败育

收稿日期:2024-06-09 修回日期:2024-09-24 接受日期:2024-09-11 出版日期:2026-04-20 发布日期:2026-03-10

The mitochondrial genes orf113b and orf146 from Xinjiang wild rapeseed cause pollen abortion in alloplasmic male sterility

Man Xing^{1, 2}, Bo Hong^{1, 2}, Chunyun Guan^{1, 2, 3}, Mei Guan^{1, 2, 3#}

¹College of Agronomy, Hunan Agricultural University, Changsha 410128, China
²Hunan Branch of National Oilseed Crops Improvement Center, Changsha 410128, China
³Southern Regional Collaborative Innovation Center for Grain and Oil Crops in China, Changsha 410128, China

Received:2024-06-09 Revised:2024-09-24 Accepted:2024-09-11 Online:2026-04-20 Published:2026-03-10
About author:Man Xing, E-mail: xingman18971@outlook.com; #Correspondence Mei Guan, E-mail: guanmei@hunau.edu.cn
Supported by:
This work was supported by the National Natural Science Foundation of China (3240150455), the China Agriculture Research System of MOF and MARA (CARS-13) and the National High-Tech R&D Program of China (863 Program, 2011AA10A104).

摘要/Abstract

摘要：

利用远缘杂交技术，以新疆野生油菜（野芥，Sinapis arvensis）为母本，以甘蓝型油菜湘油15号（XY15，Brassica napus L.）为父本，从杂交后代中选育出具有新疆野生油菜的细胞质雄性不育油菜Nsa CMS。其不育的因素及其潜在机制尚不清楚。本研究首次成功组装并分析了1258A (Nsa CMS)的线粒体基因组。该线粒体基因组全长263,010 bp，包含91个基因，包括33个蛋白质编码基因和36个orf。一个新的线粒体基因orf113b和一个突变的、截断的基因orf146，可能与1258A中观察到的雄性不育有关。ORF113b和ORF146抑制细胞生长，影响线粒体氧化磷酸化途径中与复合物I、III和V相关的基因表达，并触发活性氧（ROS）爆发。此外，转录组数据分析揭示了与orf113b和orf146共表达的关键核基因，表明它们的异常表达可能受到线粒体逆行信号的影响。该信号可导致绒毡层非典型程序性细胞死亡（PCD），导致花粉不育。总之，本研究不仅首次鉴定了远缘杂交而来的Nsa CMS线粒体基因组，明确orf113b和orf146对花粉不育至关重要。线粒体基因诱发的ROS可能是导致调控花粉发育关键核基因异常表达的主要触发因素，为探究Nsa CMS花粉败育的分子机制提供了新的思路。

Abstract:

Nsa CMS is a type of cytoplasmic male sterility (CMS) in rapeseed that originated from the cross between Xinjiang wild rapeseed (Sinapis arvensis) and Xiangyou 15 (Brassica napus L.). Although this CMS variant shows promising applications, the factors contributing to its sterility and their underlying mechanisms remain unclear. To the best of our knowledge, we successfully assembled and analyzed the mitochondrial genome of 1258A (Nsa CMS) for the first time. This mitochondrial genome spans 263,010 bp and contains 91 genes, including 33 protein-coding and 36 orf genes. Our analysis identified a novel mitochondrial gene, orf113b, and a mutated, truncated gene, orf146, both likely linked to the sterility observed in 1258A. ORF113b and ORF146 were found to impede cell growth, disrupt gene expression associated with complexes I, III, and V of the mitochondrial oxidative phosphorylation pathway, and trigger reactive oxygen species (ROS) production. In addition, transcriptome data analysis revealed key nuclear genes co-expressed with orf113b and orf146, suggesting that their aberrant expression may be influenced by retrograde signaling from the mitochondria. This signaling could lead to atypical programmed cell death (PCD) in the tapetum layer, resulting in pollen sterility. In conclusion, our study not only provides the first characterization of the Nsa CMS mitochondrial genome but also identifies orf113b and orf146 as crucial for pollen sterility. Furthermore, it suggests that the ROS induced by these mitochondrial genes may play a central role in the abnormal regulation of nuclear genes essential for pollen development, thus offering new insights into the molecular mechanisms underlying Nsa CMS.

Key words: Brassica napus L. , cytoplasmic male sterility , orf113b , orf146 , reactive oxygen species (ROS)

Man Xing, Bo Hong, Chunyun Guan, Mei Guan. 源于新疆野生油菜的线粒体基因orf113b和orf146导致异源细胞质雄性不育花粉败育[J]. Journal of Integrative Agriculture, 2026, 25(4): 1384-1401.

Man Xing, Bo Hong, Chunyun Guan, Mei Guan. The mitochondrial genes orf113b and orf146 from Xinjiang wild rapeseed cause pollen abortion in alloplasmic male sterility[J]. Journal of Integrative Agriculture, 2026, 25(4): 1384-1401.

参考文献

Belchí-Navarro S, Almagro L, Sabater-Jara A B, Fernández-Pérez F, Bru R, Pedreño M A. 2013. Induction of trans-resveratrol and extracellular pathogenesis-related proteins in elicited suspension cultured cells of Vitis vinifera cv. Monastrell. Journal of Plant Physiology, 170, 258–264.

Chase C D. 2007. Cytoplasmic male sterility: A window to the world of plant mitochondrial-nuclear interactions. Trends in Genetics, 23, 81–90.

Chen H, Zhang S, Li R, Peng G, Chen W, Rautengarten C, Liu M, Zhu L, Xiao Y, Song F, Ni J, Huang J, Wu A, Liu Z, Zhuang C, Heazlewood J L, Xie Y, Chu Z, Zhou H. 2023. BOTRYOID POLLEN 1 regulates ROS-triggered PCD and pollen wall development by controlling UDP-sugar homeostasis in rice. The Plant Cell, 35, 3522–3543.

Chen L, Liu Y G. 2014. Male sterility and fertility restoration in crops. Annual Review of Plant Biology, 65, 579–606.

Christensen A C. 2021. Plant mitochondria are a riddle wrapped in a mystery inside an Enigma. Journal of Molecular Evolution, 89, 151–156.

Dewey R E, Selote D, Griffin H C, Dickey A N, Jantz D, Smith J J, Matthiadis A, Strable J, Kestell C, Smith W A. 2023. Cytoplasmic male sterility and abortive seed traits generated through mitochondrial genome editing coupled with allotopic expression of atp1 in tobacco. Frontiers in Plant Science, 14, 1253640.

Du K, Xiao Y, Liu Q, Wu X, Jiang J, Wu J, Fang Y, Xiang Y, Wang Y. 2019. Abnormal tapetum development and energy metabolism associated with sterility in SaNa-1A CMS of Brassica napus L. Plant Cell Reports, 38, 545–558.

Eubel H, Heazlewood J L, Millar A H. 2006. Isolation and Subfractionation of Plant Mitochondria for Proteomic Analysis. Humana Press, Methods in Molecular Biology, USA. pp. 49–62.

Feng Y, Wang Y, Lu H, Li J, Akhter D, Liu F, Zhao T, Shen X, Li X, Whelan J, Zhang T, Hu J, Pan R. 2023. Assembly and phylogenomic analysis of cotton mitochondrial genomes provide insights into the history of cotton evolution. The Crop Journal, 11, 1782–1792.

Forner J, Kleinschmidt D, Meyer E H, Gremmels J, Morbitzer R, Lahaye T, Schöttler M A, Bock R. 2023. Targeted knockout of a conserved plant mitochondrial gene by genome editing. Nature Plants, 9, 1818–1831.

Ge X Y, Chen J L, Li O Q, Zou M, Tao B L, Zhao L, Wen J, Yi B, Tu J X, Shen J X. 2025. ORF138 causes abnormal lipid metabolism in the tapetum leading to Ogu cytoplasmic male sterility in Brassica napus. Journal of Integrative Agriculture, 24, 2080–2095.

Guan C. 1996. Comparative studies of inheritable character in Xinjiang wild rape and Sinapis arvensis L. Acta Agronomica Sinica, 22, 214–219. (in Chinese)

Guan C, Yin M Z. 2014. Breeding of YeYou cytoplasmic male sterile line 1193A. In: Proceedings of the Academic Annual Meeting of the Crop Science Society of China in 2014. Nanjing, China. pp. 50–52. (in Chinese)

Handa H, Gualberto J M, Grienenberger J M. 1995. Characterization of the mitochondrial orfB gene and its derivative, orf224, a chimeric open reading frame specific to one mitochondrial genome of the “Polima” male-sterile cytoplasm in rapeseed (Brassica napus L.). Current Genetics, 28, 546–552.

Heng S, Gao J, Wei C, Chen F, Li X, Wen J, Yi B, Ma C, Tu J, Fu T, Shen J. 2018. Transcript levels of orf288 are associated with the hau cytoplasmic male sterility system and altered nuclear gene expression in Brassica juncea. Journal of Experimental Botany, 69, 455–466.

Heng S, Wei C, Jing B, Wan Z, Wen J, Yi B, Ma C, Tu J, Fu T, Shen J. 2014. Comparative analysis of mitochondrial genomes between the hau cytoplasmic male sterility (CMS) line and its iso-nuclear maintainer line in Brassica juncea to reveal the origin of the CMS-associated gene orf288. BMC Genomics, 15, 322.

Hu Q, Andersen S, Dixelius C, Hansen L. 2002. Production of fertile intergeneric somatic hybrids between Brassica napus and Sinapis arvensis for the enrichment of the rapeseed gene pool. Plant Cell Reports, 21, 147–152.

Iwabuchi M, Koizuka N, Fujimoto H, Sakai T, Imamura J. 1999. Identification and expression of the kosena radish (Raphanus sativus cv. Kosena) homologue of the ogura radish CMS-associated gene, orf138. Plant Molecular Biology, 39, 183–188.

Kazama T, Itabashi E, Fujii S, Nakamura T, Toriyama K. 2016. Mitochondrial ORF79 levels determine pollen abortion in cytoplasmic male sterile rice. Plant Journal, 85, 707–716.

Khan K, Tran H C, Mansuroglu B, Önsell P, Buratti S, Schwarzländer M, Costa A, Rasmusson A G, Van Aken O. 2024. Mitochondria-derived reactive oxygen species are the likely primary trigger of mitochondrial retrograde signaling in Arabidopsis. Current Biology, 34, 327–342.e4.

Kim D H, Kang J G, Kim B D. 2007. Isolation and characterization of the cytoplasmic male sterility-associated orf456 gene of chili pepper (Capsicum annuum L.). Plant Molecular Biology, 63, 519–532.

Köhler R H, Horn R, Lössl A, Zetsche K. 1991. Cytoplasmic male sterility in sunflower is correlated with the co-transcription of a new open reading frame with the atpA gene. Molecular and General Genetics, 227, 369–376.

Kuwabara K, Arimura S I, Shirasawa K, Ariizumi T. 2022. orf137 triggers cytoplasmic male sterility in tomato. Plant Physiology, 189, 465–468.

Lanciano P, Khalfaoui-Hassani B, Selamoglu N, Ghelli A, Rugolo M, Daldal F. 2013. Molecular mechanisms of superoxide production by complex III: A bacterial versus human mitochondrial comparative case study. Biochimica et Biophysica Acta, 1827, 1332–1339.

Landgren M, Zetterstrand M, Sundberg E, Glimelius K. 1996. Alloplasmic male-sterile Brassica lines containing B. tournefortii mitochondria express an ORF 3´ of the atp6 gene and a 32 kDa protein. Plant Molecular Biology, 32, 879–890.

L’Homme Y, Stahl R J, Li X Q, Hameed A, Brown G G. 1997. Brassica nap cytoplasmic male sterility is associated with expression of a mtDNA region containing a chimeric gene similar to the pol CMS-associated orf224 gene. Current Genetics, 31, 325–335.

Li P, Zhang D, Su T, Wang W, Yu Y, Zhao X, Li Z, Yu S, Zhang F. 2020. Genome-wide analysis of mRNA and lncRNA expression and mitochondrial genome sequencing provide insights into the mechanisms underlying a novel cytoplasmic male sterility system, BVRC-CMS96, in Brassica rapa. Theoretical and Applied Genetics, 133, 2157–2170.

Liang Z M, Li J, Feng J Y, Li Z, Nangia V, Mo F, Liu Y. 2025. Brassinosteroids improve the redox state of wheat florets under low-nitrogen stress and alleviate degeneration. Journal of Integrative Agriculture, 24, 2920–2939.

Liu G, Tian H, Huang Y Q, Hu J, Ji Y X, Li S Q, Feng Y Q, Guo L, Zhu Y G. 2012. Alterations of mitochondrial protein assembly and jasmonic acid biosynthesis pathway in Honglian (HL)-type cytoplasmic male sterility rice. Journal of Biological Chemistry, 287, 40051–40060.

Liu J, Xiang R, Wang W, Mei D, Li Y, Mason A S, Fu L, Hu Q. 2015. Cytological and molecular analysis of Nsa CMS in Brassica napus L. Euphytica, 206, 279–286.

Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods, 25, 402–408.

Luo D, Xu H, Liu Z, Guo J, Li H, Chen L, Fang C, Zhang Q, Bai M, Yao N, Wu H, Wu H, Ji C, Zheng H, Chen Y, Ye S, Li X, Zhao X, Li R, Liu Y G. 2013. A detrimental mitochondrial-nuclear interaction causes cytoplasmic male sterility in rice. Nature Genetics, 45, 573–577.

Melonek J, Duarte J, Martin J, Beuf L, Murigneux A, Varenne P, Comadran J, Specel S, Levadoux S, Bernath-Levin K, Torney F, Pichon J P, Perez P, Small I. 2021. The genetic basis of cytoplasmic male sterility and fertility restoration in wheat. Nature Communications, 12, 1036.

Menassa R, L’Homme Y, Brown G G. 1999. Post-transcriptional and developmental regulation of a CMS-associated mitochondrial gene region by a nuclear restorer gene. Plant Journal, 17, 491–499.

Mittler R, Zandalinas S I, Fichman Y, Van Breusegem F. 2022. Reactive oxygen species signalling in plant stress responses. Nature Reviews Molecular Cell Biology, 23, 663–679.

Molendijk A J, Ruperti B, Singh M K, Dovzhenko A, Ditengou F A, Milia M, Westphal L, Rosahl S, Soellick T, Uhrig J, Weingarten L, Huber M, Palme K. 2007. A cysteine-rich receptor-like kinase NCRK and a pathogen-induced protein kinase RBK1 are Rop GTPase interactors. Plant Journal, 53, 909–923.

Ng S, De Clercq I, Van Aken O, Law S R, Ivanova A, Willems P, Giraud E, Van Breusegem F, Whelan J. 2014. Anterograde and retrograde regulation of nuclear genes encoding mitochondrial proteins during growth, development, and stress. Molecular Plant, 7, 1075–1093.

Nikornpun M, Sukwiwat K, Wongsing K, Kumchai J. 2023. Development of male sterile lines of CMS chilies (Capsicum annuum L.) from F1 hybrids. Breeding Science, 73, 158–167.

Park J Y, Lee Y P, Lee J, Choi B S, Kim S, Yang T J. 2013. Complete mitochondrial genome sequence and identification of a candidate gene responsible for cytoplasmic male sterility in radish (Raphanus sativus L.) containing DCGMS cytoplasm. Theoretical and Applied Genetics, 126, 1763–1774.

Sabater-Jara A B, Almagro L, Pedreño M A. 2014. Induction of extracellular defense-related proteins in suspension cultured-cells of Daucus carota elicited with cyclodextrins and methyl jasmonate. Plant Physiology and Biochemistry, 77, 133–139.

Sang S, Cheng H, Hao M, Ding B, Mei D, Wang H, Wang W, Liu J, Fu L, Liu K, Hu Q. 2021. Mitochondrial localization of ORF346 causes pollen abortion in alloplasmic male sterility. The Crop Journal, 9, 1320–1329.

Sang S F, Mei D S. Liu J, Zaman Q U, Zhang H Y, Hao M Y, Fu L, Wang H, Cheng H T, Hu Q. 2019. Organelle genome composition and candidate gene identification for Nsa cytoplasmic male sterility in Brassica napus. BMC Genomics, 20, 813.

Shinada T, Kikuchi Y, Fujimoto R, Kishitani S. 2006. An alloplasmic male-sterile line of Brassica oleracea harboring the mitochondria from Diplotaxis muralis expresses a novel chimeric open reading frame, orf72. Plant & Cell Physiology, 47, 549–553.

Singh S, Bhatia R, Kumar R, Behera T K, Kumari K, Pramanik A, Ghemeray H, Sharma K, Bhattacharya R C, Dey S S. 2021. Elucidating mitochondrial DNA markers of ogura-based CMS lines in Indian cauliflowers (Brassica oleracea var. botrytis L.) and their floral abnormalities due to diversity in cytonuclear interactions. Frontiers in Plant Science, 12, 631489.

Takatsuka A, Kazama T, Arimura S, Toriyama K. 2022. TALEN-mediated depletion of the mitochondrial gene orf312 proves that it is a Tadukan-type cytoplasmic male sterility-causative gene in rice. Plant Journal, 110, 994–1004.

Takatsuka A, Kazama T, Toriyama K. 2021. Cytoplasmic male sterility-associated mitochondrial gene orf312 derived from rice (Oryza sativa L.) cultivar Tadukan. Rice, 14, 46.

Tanaka Y, Tsuda M, Yasumoto K, Yamagishi H, Terachi T. 2012. A complete mitochondrial genome sequence of Ogura-type male-sterile cytoplasm and its comparative analysis with that of normal cytoplasm in radish (Raphanus sativus L.). BMC Genomics, 13, 352.

Tang H, Zheng X, Li C, Xie X, Chen Y, Chen L, Zhao X, Zheng H, Zhou J, Ye S, Guo J, Liu Y G. 2016. Multi-step formation, evolution, and functionalization of new cytoplasmic male sterility genes in the plant mitochondrial genomes. Cell Research, 27, 130–146.

Vercellino I, Sazanov L A. 2021. The assembly, regulation and function of the mitochondrial respiratory chain. Nature Reviews. Molecular Cell Biology, 23, 141–161.

Wang C, Lezhneva L, Arnal N, Quadrado M, Mireau H. 2021. The radish Ogura fertility restorer impedes translation elongation along its cognate CMS-causing mRNA. Proceedings of the National Academy of Sciences of the United States of America, 118, 35.

Wang H M, Ketela T, Keller W A, Gleddie S C, Brown G G. 1995. Genetic correlation of the orf224/atp6 gene region with Polima CMS in Brassica somatic hybrids. Plant Molecular Biology, 27, 801–807.

Wang J, Kan S, Liao X, Zhou J, Tembrock L R, Daniell, H, Jin S, Wu Z. 2024. Plant organellar genomes: Much done, much more to do. Trends in Plant Science, 29, 754–769.

Wang K, Gao F, Ji Y, Liu Y, Dan Z, Yang P, Zhu Y, Li S. 2013. ORFH79 impairs mitochondrial function via interaction with a subunit of electron transport chain complex III in Honglian cytoplasmic male sterile rice. New Phytologist, 198, 408–418.

Wang R, Ba Q, Zhang L, Wang W, Zhang P, Li G. 2022. Comparative analysis of mitochondrial genomes provides insights into the mechanisms underlying an S-type cytoplasmic male sterility (CMS) system in wheat (Triticum aestivum L.). Functional & Integrative Genomics, 22, 951–964.

Wang R, Cai X, Hu S, Li Y, Fan Y, Tan S, Liu Q, Zhou W. 2020. Comparative analysis of the mitochondrial genomes of Nicotiana tabacum: Hints toward the key factors closely related to the cytoplasmic male sterility mechanism. Frontiers in Genetics, 11, 257.

Welchen E, Canal M V, Gras D E, Gonzalez D H. 2020. Cross-talk between mitochondrial function, growth, and stress signalling pathways in plants. Journal of Experimental Botany, 72, 4102–4118.

Wen J F, Zhao K, Lv J H, Huo J L, Wang Z R, Wan H J, Zhu H S, Zhang Z Q, Shao G F, Wang J, Zhang S, Yang T Y, Zhang J R, Zou X X, Deng M H. 2021. Orf165 is associated with cytoplasmic male sterility in pepper. Genetics and Molecular Biology, 44, e20210030.

Wu S Y, Hou L L, Zhu J, Wang Y C, Zheng Y L, Hou J Q, Yang Z N, Lou Y. 2023. Ascorbic acid-mediated reactive oxygen species homeostasis modulates the switch from tapetal cell division to cell differentiation in Arabidopsis. The Plant Cell, 35, 1474–1495.

Xia X J, Zhou Y H, Shi K, Zhou J, Foyer C H, Yu J Q. 2015. Interplay between reactive oxygen species and hormones in the control of plant development and stress tolerance. Journal of Experimental Botany, 66, 2839–2856.

Xiao S, Zang J, Pei Y, Liu J, Liu J, Song W, Shi Z, Su A, Zhao J, Chen H. 2020. Activation of mitochondrial orf355 gene expression by a nuclear-encoded DREB transcription factor causes cytoplasmic male sterility in maize. Molecular Plant, 13, 1270–1283.

Xing M, Guan C, Guan M. 2022. Comparative cytological and transcriptome analyses of anther development in Nsa cytoplasmic male sterile (1258A) and maintainer lines in Brassica napus produced by distant hybridization. International Journal of Molecular Sciences, 23, 2004.

Yang H, Xue Y, Li B, Lin Y, Li H, Guo Z, Li W, Fu Z, Ding D, Tang J. 2022. The chimeric gene atp6c confers cytoplasmic male sterility in maize by impairing the assembly of the mitochondrial ATP synthase complex. Molecular Plant, 15, 872–886.

Yang J, Liu X, Yang X, Zhang M. 2010. Mitochondrially-targeted expression of a cytoplasmic male sterility-associated orf220 gene causes male sterility in Brassica juncea. BMC Plant Biology, 10, 231.

You J, Li M, Li H, Bai Y, Zhu X, Kong X, Chen X, Zhou R. 2022. Integrated methylome and transcriptome analysis widen the knowledge of cytoplasmic male sterility in cotton (Gossypium barbadense L.). Frontiers in Plant Science, 13, 770098.

Zhang J, Zhang L, Liang D, Yang Y, Geng B, Jing P, Qu Y, Huang J. 2023. ROS accumulation-induced tapetal PCD timing changes leads to microspore abortion in cotton CMS lines. BMC Plant Biology, 23, 311.

Zhang Y, Han Y, Zhang M, Zhang X, Guo L, Qi T, Li Y, Feng J, Wang H, Tang H, Qiao X, Chen L, Song X, Xing C, Wu J. 2022. The cotton mitochondrial chimeric gene orf610a causes male sterility by disturbing the dynamic balance of ATP synthesis and ROS burst. The Crop Journal, 10, 1683–1694.

Zorov D B, Juhaszova M, Sollott S J. 2014. Mitochondrial reactive oxygen species (ROS) and ROS-Induced ROS release. Physiological Reviews, 94, 909–950.

源于新疆野生油菜的线粒体基因orf113b和orf146导致异源细胞质雄性不育花粉败育

The mitochondrial genes orf113b and orf146 from Xinjiang wild rapeseed cause pollen abortion in alloplasmic male sterility

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 0

编辑推荐

Metrics