Function Identification of ZmCOR410 from Zoysia matrella in Resistance to Abiotic Stresses

doi:10.7668/hbnxb.20192992

Abstract

Abstract:

In order to explore the molecular mechanism of Zoysia grass in resistance to abiotic stresses,in the present study we reported the function of a cold responding gene from Zoysia matrella designated as ZmCOR410. The sequence information of the gene was obtained on the base of transcriptome and genomic data. The expression profile of ZmCOR410 in response to low temperature was detected by qRT-PCR,and the functions of the gene under abiotic stresses were estimated in Arabidopsis and yeast by genetic transformation.The results showed that the CDS of the gene was 927 bp in length,encoding an acid dehydrin that contains 308 amino acids. In the polygenetic tree of COR410 homologues from gramineous grasses,ZmCOR410 had a close relationship with the homologues of Cleistogenes songorica and Eragrostis curvula,two species that were high tolerant to drought stress. In the genomic DNA,there were four copies of core sequence of DRE-cis element in the 1 700 bp region upstream of the CDS of ZmCOR410,and its mRNA was accumulated in leaves exposed to cold. Compared to WT plants,Arabidopsis plants over expressing ZmCOR410 showed reduced injury in leaves after a freezing-temperature exposure,and showed higher survival rates under drought and high temperature stresses. Yeast cells harboring ZmCOR410 were also more tolerant to high temperature stress than the control cells. The results indicated that the product of ZmCOR410 could enhance cells resistance to freezing cold,high temperature and drought stresses,which would help Zoysia matrella going through adverse environments.

Key words: Zoysia matrella, COR genes, Abiotic stress, Dehydrins

ZHANG Xiaohan, CHEN Yanliang, MA Xin, ZHANG Shanshan, WEI Shanjun. Function Identification of ZmCOR410 from Zoysia matrella in Resistance to Abiotic Stresses[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(4): 53-61. doi: 10.7668/hbnxb.20192992.

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Figures/Tables 9

Fig.1 The amplification of the coding sequence of ZmCOR410 by PCR

Fig.2 The structure of ZmCOR410 in the genome DNA Exon and intron is showed as white block and black block,respectively;(+)and(-)represents sense strand and anti sense strand,respectively;1 indicates the translation initiation site.

Fig.3 Sequence characters of ZmCOR410 The box is K-segment;Underline is S-segment.

Fig.4 Hydrophobicity analysis and secondary structure prediction of ZmCOR410 protein A.Hydrophobicity analysis of COR410 protein;B.Predicted secondary structure of COR410 protein.

Fig.5 Transmembrane analysis of ZmCOR410 protein

Fig.6 Phylogenetic tree analysis of ZmCOR410 protein

Fig.7 Relative expression of ZmCOR410 gene in Z. matrella under low temperature stress Different lowercase letters indicate that there are significant differences between processing times at 0.05 level.

Fig.8 Tolerance ability of transgenic plants to abiotic stresses Different lowercase indicate significant difference(P<0.05).

Fig.9 The survival rate of yeast cells exposed to high temperature stress A.The recombinant yeast-expression vector;B.The survival rate of yeast cells.

References 50

[1]	Brede A D, Sun S C. Diversity of turfgrass germplasm in the Asian Pacific rim countries and potential for reducing genetic vulnerability[J]. Crop Science, 1995, 35(2):317-321. doi: 10.2135/cropsci1995.0011183X003500020004x. doi: 10.2135/cropsci1995.0011183X003500020004x URL
[2]	刘建秀, 贺善安, 陈守良. 华东地区结缕草属植物形态类型及其坪用价值[J]. 草地学报, 1997, 5(1):42-47. doi: 10.11733/j.issn.1007-0435.1997.01.007. doi: 10.11733/j.issn.1007-0435.1997.01.007
	Liu J X, He S N, Chen S L. Morphologicol type and turfgrass quality of zoysiagrass in East China[J]. Acta Agrestia Sinica, 1997, 5(1):42-47.
[3]	陈莹, 何八斤, 周禾, 张剑岷. 结缕草属种质资源评述[J]. 草业科学, 2008, 25(1):106-112.
	Chen Y, He B J, Zhou H, Zhang J M. A review for germplasm resources in Zoysia genus[J]. Pratacultural Science, 2008, 25(1):106-112.
[4]	王艳, 张绵. 结缕草和早熟禾解剖结构与其抗旱性、耐践踏性和弹性关系的对比研究[J]. 辽宁大学学报(自然科学版), 2000, 27(4):371-375. doi: 10.3969/j.issn.1000-5846.2000.04.020. doi: 10.3969/j.issn.1000-5846.2000.04.020
	Wang Y, Zhang M. Comparison study of anatomy structure of Zoysia japonica and Poa pratensis related with characteristics of drought and tramp resistance and elasticity[J]. Journal of Liaoning University (Natural Sciences Edition), 2000, 27(4):371-375.
[5]	刘莉, 胡涛, 傅金民. 中国沿海地区野生结缕草属分布现状调查与耐盐性评价[J]. 草业科学, 2012, 29(8):1250-1255.
	Liu L, Hu T, Fu J M. China littoral region wild Zoysia distribution and salt tolerance evaluation[J]. Pratacultural Science, 2012, 29(8):1250-1255.
[6]	刘俊祥, 孙振元, 巨关升, 韩蕾, 钱永强. 结缕草对重金属镉的生理响应[J]. 生态学报, 2011, 31(20):6149-6156.
	Liu J X, Sun Z Y, Ju G S, Han L, Qian Y Q. Physiological response of Zoysia japonica to Cd²⁺[J]. Acta Ecologica Sinica, 2011, 31(20):6149-6156.
[7]	李西, 吴亚娇, 孙凌霞. 铅胁迫对三种暖季型草坪草生长和生理特性的影响[J]. 草业学报, 2014, 23(4):171-180. doi: 10.11686/cyxb20140421. doi: 10.11686/cyxb20140421
	Li X, Wu Y J, Sun L X. Growth and physiological responses of three warm-season turfgrasses to lead stress[J]. Acta Prataculturae Sinica, 2014, 23(4):171-180.
[8]	胡利珍, 关贤交, 杨知建, 陈乐谞. 野生结缕草坪用性状的综合评价[J]. 草业科学, 2010, 27(10):23-26. doi: 10.3969/j.issn.1001-0629.2010.10.004. doi: 10.3969/j.issn.1001-0629.2010.10.004
	Hu L Z, Guan X J, Yang Z J, Chen L X. Comprehensive evaluation on turf characteristics of Zoysia japonica[J]. Pratacultural Science, 2010, 27(10):23-26.
[9]	邵伯琴, 施桂芬, 李彦. 青岛胶州湾地区结缕草属种质资源的保护和开发利用[J]. 中国草地, 1997(6):61-64.
	Shao B Q, Shi G F, Li Y. Protection, developinent and utilization of germplasm resonrces of lawngrass in the Jiaozhou-bay area of Qingdao[J]. Grassland of China, 1997(6):61-64.
[10]	董厚德, 宫莉君. 中国结缕草生态学及其资源开发与应用[M]. 北京: 中国林业出版社, 2001.
	Dong H D, Gong L J. Ecology of Zoysia japonica and the exploitation and application of its resources in China[M]. Beijing: China Forestry Publishing House, 2001.
[11]	马进, 孟瑾, 蔡建国. 草坪草种在江南草坪上的应用和探讨[J]. 草业科学, 2003, 20(7):90-93. doi: 10.3969/j.issn.1001-0629.2003.07.027. doi: 10.3969/j.issn.1001-0629.2003.07.027
	Ma J, Meng J, Cai J G. Discussion on application of turfgrass species in lawn of south of the Yangtze River[J]. Pratacultural Science, 2003, 20(7):90-93.
[12]	郭海林, 刘建秀. 结缕草属植物育种进展概述[J]. 草业学报, 2004, 13(3):106-112. doi: 10.3321/j.issn:1004-5759.2004.03.019. doi: 10.3321/j.issn:1004-5759.2004.03.019
	Guo H L, Liu J X. Zoysia grass breeding progress summary[J]. Acta Prataculturae Sinica, 2004, 13(3):106-112.
[13]	李亚, 谢晓金, 宣继萍, 刘建秀. 中国结缕草属(Zoysia spp.)植物抗寒性评价[J]. 草地学报, 2003, 11(3):240-245. doi: 10.3969/j.issn.1007-0435.2003.03.010. doi: 10.3969/j.issn.1007-0435.2003.03.010
	Li Y, Xie X J, Xuan J P, Liu J X. Assessment of cold-tolerance of China's Zoysia spp.[J]. Acta Agrestia Sinica, 2003, 11(3):240-245.
[14]	胡化广, 刘建秀, 宣继萍, 何秋, 程晓丽, 郭爱桂. 结缕草属植物的抗旱性初步评价[J]. 草业学报, 2007, 16(1):47-51. doi: 10.3321/j.issn:1004-5759.2007.01.008. doi: 10.3321/j.issn:1004-5759.2007.01.008
	Hu H G, Liu J X, Xuan J P, He Q, Cheng X L, Guo A G. Assessment of drought resistance of Zoysia[J]. Acta Prataculturae Sinica, 2007, 16(1):47-51.
[15]	黄春琼, 刘国道, 白昌军. 41份结缕草属种质资源耐盐性评价[J]. 热带作物学报, 2017, 38(7):1260-1266. doi: 10.3969/j.issn.1000-2561.2017.07.013. doi: 10.3969/j.issn.1000-2561.2017.07.013
	Huang C Q, Liu G D, Bai C J. Evaluation of salt tolerance among 41 Zoysia accessions[J]. Chinese Journal of Tropical Crops, 2017, 38(7):1260-1266.
[16]	Tanaka H, Hirakawa H, Kosugi S, Nakayama S, Ono A, Watanabe A, Hashiguchi M, Gondo T, Ishigaki G, Muguerza M, Shimizu K, Sawamura N, Inoue T, Shigeki Y, Ohno N, Tabata S, Akashi R, Sato S. Sequencing and comparative analyses of the genomes of Zoysia grasses[J]. DNA Research, 2016, 23(2):171-180. doi: 10.1093/dnares/dsw006. doi: 10.1093/dnares/dsw006 URL
[17]	Xuan J P, Song Y F, Zhang H X, Liu J X, Guo Z R, Hua Y L. Comparative proteomic analysis of the stolon cold stress response between the C4 perennial grass species Zoysia japonica and Zoysia metrella[J]. PLoS One, 2013, 8(9):e75705. doi: 10.1371/journal.pone.0075705. doi: 10.1371/journal.pone.0075705 URL
[18]	Wei S J, Du Z L, Gao F, Ke X, Li J, Liu J X, Zhou Y J. Global transcriptome profiles of Meyer zoysiagrass in response to cold stress[J]. PLoS One, 2015, 10(6):e0131153. doi: 10.1371/journal.pone.0131153. doi: 10.1371/journal.pone.0131153 URL
[19]	Long S X, Yan F Y, Yang L, Sun Z Y, Wei S J. Responses of Manila grass(Zoysia matrella)to chilling stress:From transcriptomics to physiology[J]. PLoS One, 2020, 15(7):e0235972. doi: 10.1371/journal.pone.0235972. doi: 10.1371/journal.pone.0235972 URL
[20]	Wang R, Wang X, Liu K, Zhang X J, Zhang L Y, Fan S J. Comparative transcriptome analysis of halophyte Zoysia macrostachya in response to salinity stress[J]. Plants, 2020, 9(4):458. doi: 10.3390/plants9040458. doi: 10.3390/plants9040458 URL
[21]	Wang J J, An C, Guo H L, Yang X Y, Chen J B, Zong J Q, Li J J, Liu J X. Physiological and transcriptomic analyses reveal the mechanisms underlying the salt tolerance of Zoysia japonica Steud[J]. BMC Plant Biology, 2020, 20(1):114. doi: 10.1186/s12870-020-02330-6. doi: 10.1186/s12870-020-02330-6 URL
[22]	Wang Z, Zhang F, Xuan J P, Zong J Q, Liu J X. Isolation and expression profiles of the ZjDREB1 gene encoding a DRE-binding transcription factor from zoysiagrass(Zoysia japonica)[J]. The Journal of Horticultural Science and Biotechnology, 2012, 87(1):77-83. doi: 10.1080/14620316.2012.11512834. doi: 10.1080/14620316.2012.11512834 URL
[23]	冯婉倩, 马礼鹏, 蒋笑笑, 张红, 李玥, 韦善君. 结缕草转录因子基因ZjDREB1.2的克隆及胁迫下的表达分析[J]. 北方园艺, 2018(18): 75-85. doi: 10.11937/bfyy.20180246. doi: 10.11937/bfyy.20180246
	Feng W Q, Ma L P, Jiang X X, Zhang H, Li Y, Wei S J. Cloning and expression analysis of a transcription factor gene ZjDREB1.2 from Zoysia[J]. Northern Horticulture, 2018(18):75-85.
[24]	Feng W Q, Li J, Long S X, Wei S J. A DREB1 gene from zoysiagrass enhances Arabidopsis tolerance to temperature stresses without growth inhibition[J]. Plant Science, 2019, 278:20-31. doi: 10.1016/j.plantsci.2018.10.009. doi: 10.1016/j.plantsci.2018.10.009 URL
[25]	Teng K, Tan P H, Guo W E, Yue Y S, Fan X F, Wu J Y. Heterologous expression of a novel Zoysia japonica C₂ H₂ zinc finger gene, ZjZFN1,improved salt tolerance in Arabidopsis[J]. Frontiers in Plant Science, 2018, 9:1159. doi: 10.3389/fpls.2018.01159. doi: 10.3389/fpls.2018.01159 pmid: 30154810
[26]	Zuo Z F, Kang H G, Park M Y, Jeong H, Sun H J, Song P S, Lee H Y. Zoysia japonica MYC type transcription factor ZjICE1 regulates cold tolerance in transgenic Arabidopsis[J]. Plant Science:an International Journal of Experimental Plant Biology, 2019, 289:110254. doi: 10.1016/j.plantsci.2019.110254. doi: 10.1016/j.plantsci.2019.110254 URL
[27]	Zuo Z F, Kang H G, Hong Q C, Park M Y, Sun H J, Kim J, Song P S, Lee H Y. A novel basic helix-loop-helix transcription factor,ZjICE2 from Zoysia japonica confers abiotic stress tolerance to transgenic plants via activating the DREB/CBF regulon and enhancing ROS scavenging[J]. Plant Molecular Biology, 2020, 102(4/5):447-462. doi: 10.1007/s11103-019-00957-0. doi: 10.1007/s11103-019-00957-0 URL
[28]	张胤冰, 孙鑫博, 樊波, 韩烈保, 张雪, 袁建波, 许立新. 结缕草ZjNAC 基因的克隆与表达分析[J]. 草业学报, 2016, 25(4):239-245. doi: 10.11686/cyxb2015453. doi: 10.11686/cyxb2015453
	Zhang Y B, Sun X B, Fan B, Han L B, Zhang X, Yuan J B, Xu L X. Cloning and expression of ZjNAC from Zoysia japonica[J]. Acta Prataculturae Sinica, 2016, 25(4):239-245.
[29]	Chen Y, Li L L, Zong J Q, Chen J B, Guo H L, Guo A G, Liu J X. Heterologous expression of the halophyte Zoysia matrella H⁺-pyrophosphatase gene improved salt tolerance in Arabidopsis thaliana[J]. Plant Physiology and Biochemistry, 2015, 91:49-55. doi: 10.1016/j.plaphy.2015.04.004. doi: 10.1016/j.plaphy.2015.04.004 pmid: 25874657
[30]	闫逢英, 农钧绣, 锁忠花, 韦善君. 结缕草Zjlea3基因逆境响应模式及抗逆功能鉴定[J]. 华北农学报, 2020, 35(3):55-62. doi: 10.7668/hbnxb.20190692. doi: 10.7668/hbnxb.20190692
	Yan F Y, Nong J X, Suo Z H, Wei S J. Expression patterns and stress resistance function identification of Zjlea3 gene from Zoysia japonica[J]. Acta Agriculturae Boreali-Sinica, 2020, 35(3):55-62.
[31]	龙思欣, 农钧绣, 王璐, 锁忠花, 韦善君. 日本结缕草耐逆基因ZjLEA2的克隆及功能探究[J]. 基因组学与应用生物学, 2021, 40(1):349-356. doi: 10.13417/j.gab.040.000349. doi: 10.13417/j.gab.040.000349
	Long S X, Nong J X, Wang L, Suo Z H, Wei S J. Cloning and preliminary function and analysis of ZjLEA2 from Zoysia japonica[J]. Genomics and Applied Biology, 2021, 40(1):349-356.
[32]	Teng K, Tan P H, Xiao G Z, Han L B, Chang Z H, Chao Y H. Heterologous expression of a novel Zoysia japonica salt-induced Glycine-rich RNA-binding protein gene,ZjGRP, caused salt sensitivity in Arabidopsis[J]. Plant Cell Reports, 2017, 36(1):179-191. doi: 10.1007/s00299-016-2068-x. doi: 10.1007/s00299-016-2068-x pmid: 27796490
[33]	张雪, 孙鑫博, 樊波, 张胤冰, 韩烈保, 许立新. 结缕草ZjCSD基因的克隆及表达分析[J]. 草业学报, 2017, 26(2):102-110. doi: 10.11686/cyxb2016260. doi: 10.11686/cyxb2016260
	Zhang X, Sun X B, Fan B, Zhang Y B, Han L B, Xu L X. Molecular cloning and expression analysis of ZjCSD from Zoysia japonica[J]. Acta Prataculturae Sinica, 2017, 26(2):102-110.
[34]	樊波, 孙鑫博, 张胤冰, 张雪, 袁建波, 张海兰, 肖维阳, 韩烈保, 许立新. 结缕草ZjCCS基因的克隆与表达分析[J]. 草地学报, 2016, 24(2):447-452. doi: 10.11733/j.issn.1007-0435.2016.02.030. doi: 10.11733/j.issn.1007-0435.2016.02.030
	Fan B, Sun X B, Zhang Y B, Zhang X, Yuan J B, Zhang H L, Xiao W Y, Han L B, Xu L X. Cloning and expression analysis of ZjCCS from Zoysia japonica[J]. Acta Agrestia Sinica, 2016, 24(2):447-452.
[35]	Thomashow M F. Role of cold-responsive genes in plant freezing tolerance[J]. Plant Physiology, 1998, 118(1):1-8. doi: 10.1104/pp.118.1.1. doi: 10.1104/pp.118.1.1 pmid: 9733520
[36]	耿晓东, 周英, 于明华, 汪成忠, 钱剑林. NaCl胁迫对小黄花菜生长及相关生理指标的影响[J]. 广西植物, 2021, 41(6):930-936. doi: 10.11931/guihaia.gxzw202007053. doi: 10.11931/guihaia.gxzw202007053
	Geng X D, Zhou Y, Yu M H, Wang C Z, Qian J L. Effects of NaCl stress on growth and related physiological indexes in Hemerocallis minor[J]. Guihaia, 2021, 41(6):930-936.
[37]	张杰, 侯小歌, 王俊英, 胡炳义. 高温大曲中酵母菌的分离、鉴定及耐高温性[J]. 中国酿造, 2011, 30(11):112-114. doi: 10.3969/j.issn.0254-5071.2011.11.031. doi: 10.3969/j.issn.0254-5071.2011.11.031
	Zhang J, Hou X G, Wang J Y, Hu B Y. Isolation,identification and thermotolerance of yeast from high temperature Daqu[J]. China Brewing, 2011, 30(11):112-114.
[38]	Danyluk J, Houde M, Rassart É, Sarhan F. Differential expression of a gene encoding an acidic dehydrin in chilling sensitive and freezing tolerant Gramineae species[J]. FEBS Letters, 1994, 344(1): 20-24. doi: 10.1016/0014-5793(94)00353-X. doi: 10.1016/0014-5793(94)00353-X pmid: 7910142
[39]	Graether S P, Boddington K F. Disorder and function:A review of the dehydrin protein family[J]. Frontiers in Plant Science, 2014, 5:576. doi: 10.3389/fpls.2014.00576. doi: 10.3389/fpls.2014.00576 pmid: 25400646
[40]	Zhu B, Choi D W, Fenton R, Close T J. Expression of the barley dehydrin multigene family and the development of freezing tolerance[J]. Molecular & General Genetics, 2000, 264(1/2):145-153. doi: 10.1007/s004380000299. doi: 10.1007/s004380000299
[41]	Lee S C, Lee M Y, Kim S J, Jun S H, An G, Kim S R. Characterization of an abiotic stress-inducible dehydrin gene, OsDhn1,in rice(Oryza sativa L.)[J]. Molecules and Cells, 2005, 19(2):212-218.
[42]	Lin C T, Guo W W, Everson E, Thomashow M F. Cold acclimation in Arabidopsis and wheat A response associated with expression of related genes encoding Boiling-stable polypeptides[J]. Plant Physiology, 1990, 94(3):1078-1083. doi: 10.1104/pp.94.3.1078. doi: 10.1104/pp.94.3.1078 pmid: 16667799
[43]	Zhang J Y, Wu F, Yan Q, John U P, Cao M S, Xu P, Zhang Z S, Ma T T, Zong X F, Li J, Liu R J, Zhang Y F, Zhao Y F, Kanzana G, Lü Y Y, Nan Z B, Spangenberg G, Wang Y R. The genome of Cleistogenes songorica provides a blueprint for functional dissection of dimorphic flower differentiation and drought adaptability[J]. Plant Biotechnology Journal, 2021, 19(3):532-547. doi: 10.1111/pbi.13483. doi: 10.1111/pbi.13483 URL
[44]	胥晓刚, 王锦平, 杨冬升, 胡庭兴, 陈金蓉. 弯叶画眉草在风化岩石边坡种植的适应性研究[J]. 公路, 2003, 48(11):106-108. doi: 10.3969/j.issn.0451-0712.2003.11.028. doi: 10.3969/j.issn.0451-0712.2003.11.028
	Xu X G, Wang J P, Yang D S, Hu T X, Chen J R. Study on adaptability of Eragrostis grass planting on effloresce rock slope[J]. Highway, 2003, 48(11):106-108.
[45]	Li M Y, Liu J X, Hao J N, Feng K, Duan A Q, Yang Q Q, Xu Z S, Xiong A S. Genomic identification of AP2/ERF transcription factors and functional characterization of two cold resistance-related AP2/ERF genes in celery(Apium graveolens L.)[J]. Planta, 2019, 250(4):1265-1280. doi: 10.1007/s00425-019-03222-2. doi: 10.1007/s00425-019-03222-2 URL
[46]	Ganeshan S, Vitamvas P, Fowler D B, Chibbar R N. Quantitative expression analysis of selected COR genes reveals their differential expression in leaf and crown tissues of wheat(Triticum aestivum L.)during an extended low temperature acclimation regimen[J]. Journal of Experimental Botany, 2008, 59(9):2393-2402. doi: 10.1093/jxb/ern112. doi: 10.1093/jxb/ern112 URL
[47]	Yang Y F, Al-Baidhani H H J, Harris J, Riboni M, Li Y, Mazonka I, Bazanova N, Chirkova L, Sarfraz Hussain S, Hrmova M, Haefele S, Lopato S, Kovalchuk N. DREB/CBF expression in wheat and barley using the stress-inducible promoters of HD-Zip I genes:Impact on plant development,stress tolerance and yield[J]. Plant Biotechnology Journal, 2020, 18(3):829-844. doi: 10.1111/pbi.13252. doi: 10.1111/pbi.13252 URL
[48]	Soltész A, Smedley M, Vashegyi I, Galiba G, Harwood W, Vágújfalvi A. Transgenic barley lines prove the involvement of TaCBF14 and TaCBF15 in the cold acclimation process and in frost tolerance[J]. Journal of Experimental Botany, 2013, 64(7):1849-1862. doi: 10.1093/jxb/ert050. doi: 10.1093/jxb/ert050 URL
[49]	Houde M, Dallaire S, N'Dong D, Sarhan F. Overexpression of the acidic dehydrin WCOR410 improves freezing tolerance in transgenic strawberry leaves[J]. Plant Biotechnology Journal, 2004, 2(5):381-387. doi: 10.1111/j.1467-7652.2004.00082.x. doi: 10.1111/j.1467-7652.2004.00082.x URL
[50]	Danyluk J, Perron A, Houde M, Limin A, Fowler B, Benhamou N, Sarhan F. Accumulation of an acidic dehydrin in the vicinity of the plasma membrane during cold acclimation of wheat[J]. The Plant Cell, 1998, 10(4):623-638. doi: 10.1105/tpc.10.4.623. doi: 10.1105/tpc.10.4.623 pmid: 9548987

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