A β-Carotene Ketolase Gene NfcrtO from Subaerial Cyanobacteria Confers Drought Tolerance in Rice
Received date: 2023-06-12
Accepted date: 2023-10-10
Online published: 2024-02-06
Nostoc flagelliforme is a terrestrial cyanobacterium that can resist many types of stressors, including drought, ultraviolet radiation, and extreme temperatures. In this study, we identified the drought tolerance gene NfcrtO, which encodes a β-carotene ketolase, through screening the transcriptome of N. flagelliforme under water loss stress. Prokaryotic expression of NfcrtO under 0.6 mol/L sorbitol or under 0.3 mol/L NaCl stress significantly increased the growth rate of Escherichia coli. When NfcrtO was heterologously expressed in rice, the seedling height and root length of NfcrtO-overexpressing rice plants were significantly higher than those of the wild type (WT) plants grown on ½ Murashige and Skoog solid medium with 120 mmol/L mannitol at the seedling stage. Transcriptome analysis revealed that NfcrtO was involved in osmotic stress, antioxidant, and other stress-related pathways. Additionally, the survival rate of the NfcrtO-overexpression lines was significantly higher than that of the WT line under both hydroponic stress (24% PEG and 100 mmol/L H2O2) and soil drought treatment at the seedling stage. Physiological traits, including the activity levels of superoxide dismutase, peroxidase, catalase, total antioxidant capacity, and the contents of proline, trehalose, and soluble sugar, were significantly improved in the NfcrtO-overexpression lines relative to those in the WT line under 20% PEG treatment. Furthermore, when water was withheld at the booting stage, the grain yield per plant of NfcrtO-overexpression lines was significantly higher than that of the WT line. Yeast two-hybrid analysis identified interactions between NfcrtO and Dna J protein, E3 ubiquitin-protein ligase, and pyrophosphate-energized vacuolar membrane proton pump. Thus, heterologous expression of NfcrtO in rice could significantly improve the tolerance of rice to osmotic stress, potentially facilitating the development of new rice varieties.
Gao Ningning, Ye Shuifeng, Zhang Yu, Zhou Liguo, Ma Xiaosong, Yu Hanxi, Li Tianfei, Han Jing, Liu Zaochang, Luo Lijun . A β-Carotene Ketolase Gene NfcrtO from Subaerial Cyanobacteria Confers Drought Tolerance in Rice[J]. Rice Science, 2024 , 31(1) : 62 -76 . DOI: 10.1016/j.rsci.2023.10.002
| [1] | Ai Y F, Yang Y W, Gao X, Qiu B S. 2014a. Heterologous expression of three stress-responsive genes from Nostoc flagelliforme confers tolerance to abiotic stresses in Escherichia coli. J Appl Phycol, 26: 123-129. |
| [2] | Ai Y F, Yang Y W, Qiu B S, Gao X. 2014b. Unique WSPA protein from terrestrial macroscopic cyanobacteria can confer resistance to osmotic stress in transgenic plants. World J Microbiol Biotechnol, 30(9): 2361-2369. |
| [3] | Arnon D I. 1949. Copper enzymes in isolated chloroplasts. polyphenoloxidase in Beta vulgaris. Plant Physiol, 24(1): 1-15. |
| [4] | Bar Eyal L, Ranjbar Choubeh R, Cohen E, Eisenberg I, Tamburu C, Dorogi M, Ünnep R, Appavou M S, Nevo R, Raviv U, Reich Z, Garab G, van Amerongen H, Paltiel Y, Keren N. 2017. Changes in aggregation states of light-harvesting complexes as a mechanism for modulating energy transfer in desert crust cyanobacteria. Proc Natl Acad Sci USA, 114(35): 9481-9486. |
| [5] | Chen L J, Wuriyanghan H, Zhang Y Q, Duan K X, Chen H W, Li Q T, Lu X, He S J, Ma B, Zhang W K, Lin Q, Chen S Y, Zhang J S. 2013. An S-domain receptor-like kinase, OsSIK2, confers abiotic stress tolerance and delays dark-induced leaf senescence in rice. Plant Physiol, 163(4): 1752-1765. |
| [6] | Chen M Y, Teng W K, Zhao L, Hu C X, Zhou Y K, Han B P, Song L R, Shu W S. 2021. Comparative genomics reveals insights into cyanobacterial evolution and habitat adaptation. ISME J, 15(1): 211-227. |
| [7] | Chen S J, Xu K, Kong D Y, Wu L Y, Chen Q, Ma X S, Ma S Q, Li T F, Xie Q, Liu H Y, Luo L J. 2022. Ubiquitin ligase OsRINGzf1 regulates drought resistance by controlling the turnover of OsPIP2;1. Plant Biotechnol J, 20(9): 1743-1755. |
| [8] | Cui L J, Xu H Y, Zhu Z X, Gao X. 2017. The effects of the exopolysaccharide and growth rate on the morphogenesis of the terrestrial filamentous Cyanobacterium Nostoc flagelliforme. Biol Open, 6(9): 1329-1335. |
| [9] | Cui L J, Liu Y H, Yang Y W, Ye S F, Luo H Y, Qiu B S, Gao X. 2018. The drnf1 gene from the drought-adapted Cyanobacterium Nostoc flagelliforme improved salt tolerance in transgenic Synechocystis and Arabidopsis plant. Genes, 9(9): 441. |
| [10] | Dong J F, Zhou L, Feng A Q, Zhang S H, Fu H, Chen L, Zhao J L, Yang T F, Yang W, Ma Y M, Wang J, Zhu X Y, Liu Q, Liu B. 2021. The OsOXO2, OsOXO3 and OsOXO4 positively regulate panicle blast resistance in rice. Rice, 14(1): 51. |
| [11] | Fernández-González B, Sandmann G, Vioque A. 1997. A new type of asymmetrically acting β-carotene ketolase is required for the synthesis of echinenone in the Cyanobacterium Synechocystis sp. PCC 6803. J Biol Chem, 272(15): 9728-9733. |
| [12] | Gao K S. 1998. Chinese studies on the edible blue-green Alga, Nostoc flagelliforme: A review. J Appl Phycol, 10(1): 37-49. |
| [13] | Gao X. 2017. Scytonemin plays a potential role in stabilizing the exopolysaccharidic matrix in terrestrial cyanobacteria. Microb Ecol, 73(2): 255-258. |
| [14] | Gao X, Xu H Y, Ye S F, Liang W Y. 2016. A proposal on the restoration of Nostoc flagelliforme for sustainable improvement in the ecology of arid steppes in China. Environments, 3(2): 14. |
| [15] | Gao X, Liu B, Ji B Y. 2019. Profiling of small molecular metabolites in Nostoc flagelliforme during periodic desiccation. Mar Drugs, 17(5): 298. |
| [16] | Gao X, Xu H Y, Zhu Z X, She Y, Ye S F. 2020. Improved production of echinenone and canthaxanthin in transgenic Nostoc sp. PCC 7120 overexpressing a heterologous crtO gene from Nostoc flagelliforme. Microbiol Res, 236: 126455. |
| [17] | Gao X, Zhu Z X, Xu H Y, Liu L T, An J, Ji B Y, Ye S F. 2021. Cold adaptation in drylands: Transcriptomic insights into cold-stressed Nostoc flagelliforme and characterization of a hypothetical gene with cold and nitrogen stress tolerance. Environ Microbiol, 23(2): 713-727. |
| [18] | Gong M Y, Bassi A. 2016. Carotenoids from microalgae: A review of recent developments. Biotechnol Adv, 34(8): 1396-1412. |
| [19] | Gong Z Z, Xiong L M, Shi H Z, Yang S H, Herrera-Estrella L R, Xu G H, Chao D Y, Li J R, Wang P Y, Qin F, Li J J, Ding Y L, Shi Y T, Wang Y, Yang Y Q, Guo Y, Zhu J K. 2020. Plant abiotic stress response and nutrient use efficiency. Sci China Life Sci, 63(5): 635-674. |
| [20] | Graus D, Konrad K R, Bemm F, Patir Nebioglu M G, Lorey C, Duscha K, Güthoff T, Herrmann J, Ferjani A, Cuin T A, Roelfsema M R G, Schumacher K, Neuhaus H E, Marten I, Hedrich R. 2018. High V-PPase activity is beneficial under high salt loads, but detrimental without salinity. New Phytol, 219(4): 1421-1432. |
| [21] | Hagemann M. 2011. Molecular biology of cyanobacterial salt acclimation. FEMS Microbiol Rev, 35(1): 87-123. |
| [22] | Hu H H, Dai M Q, Yao J L, Xiao B Z, Li X H, Zhang Q F, Xiong L Z. 2006. Overexpressing a NAM, ATAF, and CUC (NAC) transcription factor enhances drought resistance and salt tolerance in rice. Proc Natl Acad Sci USA, 103(35): 12987-12992. |
| [23] | Karmakar S, Ali Molla K, Chanda P K, Sarkar S N, Datta S K, Datta K. 2016. Green tissue-specific co-expression of chitinase and oxalate oxidase 4 genes in rice for enhanced resistance against sheath blight. Planta, 243(1): 115-130. |
| [24] | Latif A, Sun Y, Pu C X, Ali N. 2023. Rice curled its leaves either adaxially or abaxially to combat drought stress. Rice Sci, 30(5): 405-416. |
| [25] | Li X X, Ding M M, Wang M, Yang S J, Ma X R, Hu J H, Song F, Wang L X, Liang W Y. 2022. Proteome profiling reveals changes in energy metabolism, transport and antioxidation during drought stress in Nostoc flagelliforme. BMC Plant Biol, 22(1): 162. |
| [26] | Liang W Y, Zhou Y W, Wang L X, You X R, Zhang Y P, Cheng C L, Chen W. 2012. Ultrastructural, physiological and proteomic analysis of Nostoc flagelliforme in response to dehydration and rehydration. J Proteomics, 75(18): 5604-5627. |
| [27] | Liu W, Cui L J, Xu H Y, Zhu Z X, Gao X. 2017. Flexibility-rigidity coordination of the dense exopolysaccharide matrix in terrestrial cyanobacteria acclimated to periodic desiccation. Appl Environ Microbiol, 83(22): e01619-17. |
| [28] | 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(4): 402-408. |
| [29] | Llewellyn C A, Airs R L, Farnham G, Greig C. 2020. Synthesis, regulation and degradation of carotenoids under low level UV-B radiation in the filamentous cyanobacterium Chlorogloeopsis fritschii PCC 6912. Front Microbiol, 11: 163. |
| [30] | Luo L J. 2010. Breeding for water-saving and drought-resistance rice (WDR) in China. J Exp Bot, 61(13): 3509-3517. |
| [31] | Luo L J, Mei H W, Yu X Q, Xia H, Chen L, Liu H Y, Zhang A N, Xu K, Wei H B, Liu G L, Wang F M, Liu Y, Ma X S, Lou Q J, Feng F J, Zhou L G, Chen S J, Yan M, Liu Z C, Bi J G, Li T F, Li M S. 2019. Water-saving and drought-resistance rice: From the concept to practice and theory. Mol Breed, 39: 145. |
| [32] | Peng Z, Huwanixi A, Wan C H. 2023. Identification of novel smORFs and microprotein acting in response to rehydration of Nostoc flagelliforme. Proteomics, 23(12): e2200473. |
| [33] | Sand-Jensen K. 2014. Ecophysiology of gelatinous Nostoc colonies: Unprecedented slow growth and survival in resource-poor and harsh environments. Ann Bot, 114(1): 17-33. |
| [34] | Scherer S, Ernst A, Chen T W, Böger P. 1984. Rewetting of drought-resistant blue-green algae: Time course of water uptake and reappearance of respiration, photosynthesis, and nitrogen fixation. Oecologia, 62(3): 418-423. |
| [35] | Shang J L, Chen M, Hou S W, Li T, Yang Y W, Li Q, Jiang H B, Dai G Z, Zhang Z C, Hess W R, Qiu B S. 2019. Genomic and transcriptomic insights into the survival of the subaerial cyanobacterium Nostoc flagelliforme in arid and exposed habitats. Environ Microbiol, 21(2): 845-863. |
| [36] | Sinha R P, Klisch M, Walter Helbling E, Häder D P. 2001. Induction of mycosporine-like amino acids (MAAs) in cyanobacteria by solar ultraviolet-B radiation. J Photochem Photobiol B, 60(2/3): 129-135. |
| [37] | Sun Z T, Shen S C, Tian B, Wang H, Xu Z J, Wang L Y, Hua Y J. 2009. Functional analysis of γ-carotene ketolase involved in the carotenoid biosynthesis of Deinococcus radiodurans. FEMS Microbiol Lett, 301(1): 21-27. |
| [38] | Takenaka H, Yamaguchi Y, Sakaki S, Watarai K, Tanaka N, Hori M, Seki H, Tsuchida M, Yamada A, Nishimori T, Morinaga T. 1998. Safety evaluation of Nostoc flagelliforme (Nostocales, Cyanophyceae) as a potential food. Food Chem Toxicol, 36(12): 1073-1077. |
| [39] | Tamadaddi C, Sagar V, Verma A K, Afsal F, Sahi C. 2021. Expansion of the evolutionarily conserved network of J-domain proteins in the Arabidopsis mitochondrial import complex. Plant Mol Biol, 105(4/5): 385-403. |
| [40] | Tamadaddi C, Verma A K, Zambare V, Vairagkar A, Diwan D, Sahi C. 2022. J-like protein family of Arabidopsis thaliana: The enigmatic cousins of J-domain proteins. Plant Cell Rep, 41(6): 1343-1355. |
| [41] | Wada N, Sakamoto T, Matsugo S. 2013. Multiple roles of photosynthetic and sunscreen pigments in cyanobacteria focusing on the oxidative stress. Metabolites, 3(2): 463-483. |
| [42] | Wang B, Yang L, Zhang Y Q, Chen S L, Gao X, Wan C H. 2019. Investigation of the dynamical expression of Nostoc flagelliforme proteome in response to rehydration. J Proteomics, 192: 160-168. |
| [43] | Wang L K, Feng Z X, Wang X, Wang X W, Zhang X G. 2010. DEGseq: An R package for identifying differentially expressed genes from RNA-seq data. Bioinformatics, 26(1): 136-138. |
| [44] | Wang L X, Lei X T, Yang J, Wang S P, Liu Y, Liang W Y. 2018. Comparative transcriptome analysis reveals that photosynthesis contributes to drought tolerance of Nostoc flagelliforme (Nostocales, Cyanobacteria). Phycologia, 57(1): 113-120. |
| [45] | Wang L X, Wang S P, Li X X, Wang M, Zhang Z, Shi J, Xu T T, Liang W Y. 2020. Overexpression of cphA gene from Nostoc flagelliforme improves the drought tolerance of Arabidopsis thaliana. S Afr J Bot, 132: 127-131. |
| [46] | Wang L X, Li X X, Wang M, Ma X R, Song F, Hu J H, Liang W L, Liang W Y. 2022. Carbon metabolism and the ROS scavenging system participate in Nostoc flagelliforme’s adaptive response to dehydration conditions through protein acetylation. J Proteome Res, 21(2): 482-493. |
| [47] | Wang M, Zhu Q, Li X X, Hu J H, Song F, Liang W L, Ma X R, Wang L X, Liang W Y. 2022. Effect of drought stress on degradation and remodeling of membrane lipids in Nostoc flagelliforme. Foods, 11(12): 1798. |
| [48] | Wu S J, Yu K Q, Li L, Wang L X, Liang W Y. 2021. Enhancement of exopolysaccharides production and reactive oxygen species level of Nostoc flagelliforme in response to dehydration. Environ Sci Pollut Res, 28(26): 34300-34308. |
| [49] | Xia H, Zhang X X, Liu Y, Bi J G, Ma X S, Zhang A N, Liu H Y, Chen L, Zhou S, Gao H, Xu K, Wei H B, Liu G L, Wang F M, Zhao H Y, Luo X X, Hou D P, Lou Q J, Feng F J, Zhou L G, Chen S J, Yan M, Li T F, Li M S, Wang L, Liu Z C, Yu X Q, Mei H W, Luo L J. 2022. Blue revolution for food security under carbon neutrality: A case from the water-saving and drought- resistance rice. Mol Plant, 15(9): 1401-1404. |
| [50] | Xiang Y, Tang N, Du H, Ye H Y, Xiong L Z. 2008. Characterization of OsbZIP23 as a key player of the basic leucine zipper transcription factor family for conferring abscisic acid sensitivity and salinity and drought tolerance in rice. Plant Physiol, 148(4): 1938-1952. |
| [51] | Xiao L H, Yang G, Zhang L C, Yang X H, Zhao S, Ji Z Z, Zhou Q, Hu M, Wang Y, Chen M, Xu Y, Jin H J, Xiao X, Hu G P, Bao F, Hu Y, Wan P, Li L G, Deng X, Kuang T Y, Xiang C B, Zhu J K, Oliver M J, He Y K. 2015. The resurrection genome of Boea hygrometrica: A blueprint for survival of dehydration. Proc Natl Acad Sci USA, 112(18): 5833-5837. |
| [52] | Xu H F, Dai G Z, Ye D M, Shang J L, Song W Y, Shi H Z, Qiu B S. 2020. Dehydration-induced DnaK2 chaperone is involved in PSII repair of a desiccation-tolerant cyanobacterium. Plant Physiol, 182(4): 1991-2005. |
| [53] | Xu H F, Raanan H, Dai G Z, Oren N, Berkowicz S, Murik O, Kaplan A, Qiu B S. 2021. Reading and surviving the harsh conditions in desert biological soil crust: The cyanobacterial viewpoint. FEMS Microbiol Rev, 45(6): fuab036. |
| [54] | Xue Y, He Q F. 2015. Cyanobacteria as cell factories to produce plant secondary metabolites. Front Bioeng Biotechnol, 3: 57. |
| [55] | Yan L, Gong Y Y, Luo Q, Dai G X, Teng Z N, He Y, Wu X X, Liu C, Tang D Y, Ye N H, Deng G F, Lin J Z, Liu X M. 2021. Heterologous expression of fungal AcGDH alleviates ammonium toxicity and suppresses photorespiration, thereby improving drought tolerance in rice. Plant Sci, 305: 110769. |
| [56] | Yang Y W, Yin Y C, Li Z K, Huang D, Shang J L, Chen M, Qiu B S. 2019. Orange and red carotenoid proteins are involved in the adaptation of the terrestrial cyanobacterium Nostoc flagelliforme to desiccation. Photosynth Res, 140(1): 103-113. |
| [57] | Ye S F, Gao X. 2015. Excavating abiotic stress-related gene resources of terrestrial macroscopic cyanobacteria for crop genetic engineering: Dawn and challenge. Bioengineered, 6(6): 313-315. |
| [58] | Yu S B, Ali J, Zhou S C, Ren G J, Xie H A, Xu J L, Yu X Q, Zhou F S, Peng S B, Ma L Y, Yuan D Y, Li Z F, Chen D Z, Zheng R F, Zhao Z G, Chu C C, You A Q, Wei Y, Zhu S S, Gu Q Y, He G C, Li S G, Liu G F, Liu C H, Zhang C P, Xiao J H, Luo L J, Li Z K, Zhang Q F. 2022. From Green Super Rice to green agriculture: Reaping the promise of functional genomics research. Mol Plant, 15(1): 9-26. |
| [59] | Yuan X L, An J, Zheng T, Liu W J. 2022. Exogenous melatonin improves salt tolerance mainly by regulating the antioxidant system in cyanobacterium Nostoc flagelliforme. PeerJ, 10: e14479. |
| [60] | Zhang H, Li Y Y, Zhu J K. 2018. Developing naturally stress- resistant crops for a sustainable agriculture. Nat Plants, 4(12): 989-996. |
| [61] | Zhang H, Zhao Y, Zhu J K. 2020. Thriving under stress: How plants balance growth and the stress response. Dev Cell, 55(5): 529-543. |
| [62] | Zhang H M, Zhu J H, Gong Z Z, Zhu J K. 2022. Abiotic stress responses in plants. Nat Rev Genet, 23(2): 104-119. |
| [63] | Zhang Q F. 2007. Strategies for developing green super rice. Proc Natl Acad Sci USA, 104(42): 16402-16409. |
| [64] | Zhang Z C, Wang K, Hao F H, Shang J L, Tang H R, Qiu B S. 2021. New types of ATP-grasp ligase are associated with the novel pathway for complicated mycosporine-like amino acid production in desiccation-tolerant cyanobacteria. Environ Microbiol, 23(11): 6420-6432. |
| [65] | Zhu J K. 2016. Abiotic stress signaling and responses in plants. Cell, 167(2): 313-324. |
| [66] | Zhu X Y, Xiong L Z. 2013. Putative megaenzyme DWA1 plays essential roles in drought resistance by regulating stress-induced wax deposition in rice. Proc Natl Acad Sci USA, 110(44): 17790-17795. |
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