Please wait a minute...
Journal of Integrative Agriculture  2024, Vol. 23 Issue (04): 1222-1237    DOI: 10.1016/j.jia.2024.01.011
Horticulture Advanced Online Publication | Current Issue | Archive | Adv Search |

Genome-wide identification of the CONSTANS-LIKE (COL) family and mechanism of fruit senescence regulation by PpCOL8 in sand pear (Pyrus pyrifolia)

Yue Xu1*, Shurui Song2*, Huiying Wang1*, Xilong Cao1, Xinran Zhao1, Wenli Wang1, Liyue Huo1, Yawei Li1, Misganaw Wassie2, Bin Lu1, Liang Chen2, Haiyan Shi1#

1 College of Horticulture, Hebei Agricultural University, Baoding 071001, China

2 Wuhan Botanical Garden, Chinese Academy of Sciences, Wuhan 430074, China

Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
摘要  “黄金梨”属于砂梨系统,营养品质和口感极佳。然而,由于其货架期短,黄金梨”产业化发展受到严重限制。水杨酸(Salicylic acid, SA)是一种植物激素,可以延缓果实衰老从而延长货架期。但是,SA在果实衰老过程中如何调控CONSTANS-LIKE基因(COLs)以及COL基因在砂梨果实衰老中的调控作用尚不清楚。本研究在砂梨中鉴定出22个COL基因,包括通过转录组鉴定的4个COL基因(PpCOL8PpCOL9aPpCOL9bPpCOL14)和通过全基因组分析鉴定的18个COL基因。根据COL蛋白的结构域,将这些COL基因分为三个亚家族。PpCOL8具有两个B-box基序和一个CCT结构域,属于第一亚家族。相比之下,其他三个PpCOLs,PpCOL9a,PpCOL9b和PpCOL14,具有相似的保守蛋白结构域和基因结构,属于第三亚家族。PpCOL8PpCOL9aPpCOL9bPpCOL14在梨各组织中均表现出不同的表达模式,且均在果实发育早期优势表达。此外,外源SA处理抑制了PpCOL8的表达,却上调了PpCOL9aPpCOL9b的表达。有趣的是,PpCOL8与PpMADS相互作用,而PpMADS是一个MADS-box蛋白基因,在果实中优势表达且SA上调其表达。在过表达PpCOL8基因的砂梨果实中,乙烯生成量和丙二醛(MDA)含量均增加;而且砂梨果实中抗氧化酶(POD和SOD)活性以及PpPOD1PpSOD1的表达量均下调,表明PpCOL8促进砂梨果实衰老。而在过表达PpMADS的砂梨果实中,相应的变化则相反,说明PpMADS延缓砂梨果实衰老。PpCOL8PpMADS的共转化亦延缓砂梨果实衰老。本研究揭示了PpCOL8通过SA信号通路与PpMADS相互作用在梨果实衰老中起关键作用。

Abstract  Pyrus pyrifolia Nakai ‘Whangkeumbae’ is a sand pear fruit with excellent nutritional quality and taste.  However, the industrial development of pear fruit is significantly limited by its short shelf life.  Salicylic acid (SA), a well-known phytohormone, can delay fruit senescence and improve shelf life.  However, the mechanism by which SA regulates CONSTANS-LIKE genes (COLs) during fruit senescence and the role of COL genes in mediating fruit senescence in sand pear are poorly understood.  In this study, 22 COL genes were identified in sand pear, including four COLs (PpCOL8, PpCOL9a, PpCOL9b, and PpCOL14) identified via transcriptome analysis and 18 COLs through genome-wide analysis.  These COL genes were divided into three subgroups according to the structural domains of the COL protein.  PpCOL8, with two B-box motifs and one CCT domain, belonged to the first subgroup.  In contrast, the other three PpCOLs, PpCOL9a, PpCOL9b, and PpCOL14, with similar conserved protein domains and gene structures, were assigned to the third subgroup.  The four COLs showed different expression patterns in pear tissues and were preferentially expressed at the early stage of fruit development.  Moreover, the expression of PpCOL8 was inhibited by exogenous SA treatment, while SA up-regulated the expression of PpCOL9a and PpCOL9b.  Interestingly, PpCOL8 interacts with PpMADS, a MADS-box protein preferentially expressed in fruit, and SA up-regulated its expression.  While the production of ethylene and the content of malondialdehyde (MDA) were increased in PpCOL8-overexpression sand pear fruit, the antioxidant enzyme (POD and SOD) activity and the expression of PpPOD1 and PpSOD1 in the sand pear fruits were down-regulated, which showed that PpCOL8 promoted sand pear fruit senescence.  In contrast, the corresponding changes were the opposite in PpMADS-overexpression sand pear fruits, suggesting that PpMADS delayed sand pear fruit senescence.  The co-transformation of PpCOL8 and PpMADS also delayed sand pear fruit senescence.  The results of this study revealed that PpCOL8 can play a key role in pear fruit senescence by interacting with PpMADS through the SA signaling pathway.
Keywords:  Pyrus pyrifolia        CONSTANS-LIKE gene        salicylic acid        fruit senescence        MADS   
Received: 12 January 2023   Accepted: 26 December 2023
Fund: This work was supported by the National Natural Science Foundation of China (32272654), the Natural Science Foundation of Hebei Province, China (C2023204016), the Hebei Province Introduced Overseas-Scholar Fund, China (C20220361), the S&T Program of Hebei, China (20326330D), and the Hebei Province Outstanding Youth Fund, China (2016, 2019).

About author:  #Correspondence Haiyan Shi, Tel/Fax: +86-312-7528300, E-mail: yyshhy@hebau.edu.cn * These authors contributed equally to this study.

Cite this article: 

Yue Xu, Shurui Song, Huiying Wang, Xilong Cao, Xinran Zhao, Wenli Wang, Liyue Huo, Yawei Li, Misganaw Wassie, Bin Lu, Liang Chen, Haiyan Shi. 2024.

Genome-wide identification of the CONSTANS-LIKE (COL) family and mechanism of fruit senescence regulation by PpCOL8 in sand pear (Pyrus pyrifolia) . Journal of Integrative Agriculture, 23(04): 1222-1237.

Barry C S, Giovannoni J J. 2006. Ripening in the tomato Green-ripe mutant is inhibited by ectopic expression of a protein that disrupts ethylene signaling. Proceedings of the National Academy of Sciences of the United States of America, 103, 7923–7928.

Chaurasia A K, Patil H B, Azeez A, Subramaniam V R, Krishna B, Sane A P, Sane P V. 2016. Molecular characterization of CONSTANS-Like (COL) genes in banana (Musa acuminata L. AAA Group, cv. Grand Nain). Physiology and Molecular Biology of Plants, 22, 1–15.

Chen C J, Chen H, Zhang Y, Thomas H R, Frank M H, He Y H, Xia R. 2020. TBtools: An integrative toolkit developed for interactive analyses of big biological data. Molecular Plant, 13, 1194–1202.

Chen J, Chen J Y, Wang J N, Kuang J F, Shan W, Lu W J. 2012. Molecular characterization and expression profiles of MaCOL1, a CONSTANS-like gene in banana fruit. Gene, 496, 110–117.

Chen J, Li F F, Li Y X, Wang Y S, Wang C Z, Yuan D B, Jiang Y M. 2019. Exogenous procyanidin treatment delays senescence of harvested banana fruit by enhancing antioxidant responses and in vivo procyanidin content. Postharvest Biology and Technology, 158, 110999.

Cheng X F, Wang Z Y. 2005. Overexpression of COL9, a CONSTANS-LIKE gene, delays flowering by reducing expression of CO and FT in Arabidopsis thaliana. The Plant Journal, 43, 758–768.

Crocco C D, Botto J F. 2013. BBX proteins in green plants: Insights into their evolution, structure, feature and functional diversification. Gene, 531, 44–52.

Cui L, Zheng F Y, Wang J F, Zhang C L, Zhang D D, Gao S N, Zhang C H, Ye J, Zhang Y Y, Ouyang B, Wang T T, Hong Z L, Ye Z B, Zhang J H. 2022. The tomato CONSTANS-LIKE protein SlCOL1 regulates fruit yield by repressing SFT gene expression. BMC Plant Biology, 22, 429.

Eddy S R. 2011. Accelerated profile HMM searches. PLoS Computational Biology, 7, e1002195.

Elitzur T, Yakir E, Quansah L, Fei Z J, Vrebalov J, Khayat E, Giovannoni J J, Friedman H. 2016. Banana MaMADS transcription factors are necessary for fruit ripening and molecular tools to promote shelf-life and food security. Plant Physiology, 171, 380–391.

Fan H, He C S. 1998. Inhibition of ethylene generation of post-harvest apple fruit by salicylic acid. Plant Physiology Communications, 34, 248–251.

Fu J X, Yang L W, Dai S L. 2015. Identification and characterization of the CONSTANS-like gene family in the short-day plant Chrysanthemum lavandulifolium. Molecular Genetics and Genomics, 290, 1039–1054.

Gangappa S N, Botto J F. 2014. The BBX family of plant transcription factors. Trends in Plant Science, 19, 460–470.

Gao Y H, Yang Q S, Yan X H, Wu X Y, Yang F, Li J Z, Wei J, Ni J B, Ahmad M, Bai S L, Teng Y W. 2021. High-quality genome assembly of ‘Cuiguan’ pear (Pyrus pyrifolia) as a reference genome for identifying regulatory genes and epigenetic modifications responsible for bud dormancy. Horticulture Research, 8, 197–212.

Griffiths S, Dunford R P, Coupland G, Laurie D A. 2003. The evolution of CONSTANS-like gene families in barley, rice, and Arabidopsis. Plant Physiology, 131, 1855–1867.

Hassan I, Zhang Y X, Du G Q, Wang G Y, Zhang J H. 2007. Effect of salicylic acid (SA) on delaying fruit senescence of Huang Kum pear. Frontiers of Agriculture in China, 1, 456–459.

Hassidim M, Harir Y, Yakir E, Kron I, Green R M. 2009. Over-expression of CONSTANS-LIKE 5 can induce flowering in short-day grown Arabidopsis. Planta, 230, 481–491.

Kumar S, Stecher G, Tamura K. 2016. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33, 1870–1874.

Leslie C A, Romani R J. 1988. Inhibition of ethylene biosynthesis by salicylic acid. Plant Physiology, 88, 833–837.

Li T, Tan D M, Liu Z, Jiang Z Y, Wei Y, Zhang L C, Li X Y, Yuan H, Wang A D. 2015. Apple MdACS6 regulates ethylene biosynthesis during fruit development involving ethylene-responsive factor. Plant and Cell Physiology, 56, 1909–1917.

Li Y F, Zhao Y Q, Zhang M, Jia G X, Zaccai M. 2018. Functional and evolutionary characterization of the CONSTANS-like family in Lilium×formolongi. Plant and Cell Physiology, 59, 1874–1888.

Liang R Z, Luo C, Liu Y, Hu W L, Guo Y H, Yu H X, Lu T T, Chen S Q, Zhang X J, He X H. 2023. Overexpression of two CONSTANS-like 2 (MiCOL2) genes from mango delays flowering and enhances tolerance to abiotic stress in transgenic Arabidopsis. Plant Science, 327, 111541.

Liu H, Dong S Y, Sun D Y, Liu W, Gu F W, Liu Y Z, Guo T, Wang H, Wang J F, Chen Z Q. 2016a. CONSTANS-like 9 (OsCOL9) interacts with receptor for activated C-kinase 1(OsRACK1) to regulate blast resistance through salicylic acid and ethylene signaling pathways. PLoS ONE, 11, e0166249.

Liu H, Gu F W, Dong S Y, Liu W, Wang H, Chen Z Q, Wang J F. 2016b. CONSTANS-like 9 (COL9) delays the flowering time in Oryza sativa by repressing the Ehd1 pathway. Biochemical and Biophysical Research Communications, 479, 173–178.

Liu L D, Ding Q Y, Liu J, Yang C L, Chen H, Zhang S F, Zhu J, Wang D J. 2020. Brassica napus COL transcription factor BnCOL2 negatively affects the tolerance of transgenic Arabidopsis to drought stress. Environmental and Experimental Botany, 178, 104171.

Liu Y, Luo C, Guo Y H, Liang R Z, Yu H X, Chen S Q, Mo X, Yang X Z, He X H. 2022. Isolation and functional characterization of two CONSTANS-like 16 (MiCOL16) genes from Mango. International Journal of Molecular Sciences, 23, 3075.

Liu Y X, Li C Y, Yang B, Jiang Q Q, Mao R Y, Liu Z T, Huang Y, Zhang M, Prusky D B. 2021. Induction of defense response against Alternaria rot in Zaosu pear fruit by exogenous L-lysine through regulating ROS metabolism and activating defense-related proteins. Postharvest Biology and Technology, 179, 111567.

Meng X L, Song Z Q. 2003. ‘Whangkeumbae’ an excellent fruit tree variety. Bulletin of Biology, 38, 61. (in Chinese)

Qi X L, Liu C G, Song L L, Li M. 2020. PaMADS7, a MADS-box transcription factor, regulates sweet cherry fruit ripening and softening. Plant Science, 301, 110634.

Robson F, Costa M M, Hepworth S R, Vizir I, Pineiro M, Reeves P H. 2001. Functional importance of conserved domains in the flowering-time gene CONSTANS demonstrated by analysis of mutant alleles and transgenic plants. The Plant Journal, 28, 619–631.

Shi H Y, Cao L W, Xu Y, Yang X, Liu S L, Liang Z S, Li G C, Yang Y P, Zhang Y X, Chen L. 2021. Transcriptional pro-files underlying the effects of salicylic acid on fruit ripening and senescence in pear (Pyrus pyrifolia Nakai). Journal of Integrative Agriculture, 20, 2424–2437.

Shi H Y, Wang Y H, Qi A Y, Zhang Y F, Xu J F, Wang A B, Zhang Y X. 2013. PpACS1b, a pear gene encoding ACC synthase, is regulated during fruit late development and involved in response to salicylic acid. Scientia Horticulturae, 164, 602–609.

Shi H Y, Zhang Y X. 2012. Pear ACO genes encoding putative 1-aminocyclopropane-1-carboxylate oxidase homologs are functionally expressed during fruit ripening and involved in response to salicylic acid. Molecular Biology Reports, 39, 9509–9519.

Shi H Y, Zhang Y X, Chen L. 2019. Expression and regulation of PpEIN3b during fruit ripening and senescence via integrating SA, glucose, and ACC signaling in pear (Pyrus pyrifolia Nakai. Whangkeumbae). Genes (Basel), 10, 476.

Takase T, Kakikubo Y, Nakasone A, Nishiyama Y, Yasuhara M, Tokioka-Ono Y, Kiyosue T. 2011. Characterization and transgenic study of CONSTANS-LIKE8 (COL8) gene in Arabidopsis thaliana: Expression of 35S:COL8 delays flowering under long-day conditions. Plant Biotechnology, 28, 439–446.

Voinnet O, Rivas S, Mestre P, Baulcombe D. 2003. Retracted: An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus. The Plant Journal, 33, 949–956.

Vrebalov J, Ruezinsky D, Padmanabhan V, White R, Medrano D, Drake R, Schuch W, Giovannoni J. 2002. A MADS box gene necessary for fruit ripening at the tomato ripening-inhibitor (rin) locus. Science, 296, 343–346.

Wang W H, Wang Z H, Tong W, Jiang X C, Sun X S. 2009. Effects of 1-MCP application on postharvest physiology and fresh-keeping of ‘Whangkeumbae’ pear fruits. Storage and Process, 1, 30–34. (in Chinese)

Wu F Q, Price B W, Haider W, Seufferheld G, Nelson R, Hanzawa Y. 2014. Functional and evolutionary characterization of the CONSTANS gene family in short-day photoperiodic flowering in soybean. PLoS ONE, 9, e85754.

Xie Q L, Hu Z L, Zhu Z G, Dong T T, Zhao Z P, Cui B L, Chen G P. 2014. Overexpression of a novel MADS-box gene SlFYFL delays senescence, fruit ripening and abscission in tomato. Scientific Reports, 4, 4367.

Xu D, Lu Z C, Jin K M, Qiu W M, Qiao G R, Han X J, Zhou R Y. 2021. SPDE: A multi-functional software for sequence processing and data extraction. Bioinformatics, 37, 3686–3687.

Xu Y, Huo L Y, Zhao K K, Li Y W, Zhao X R, Wang H Y, Wang W L, Shi H Y. 2023. Salicylic acid delays pear fruit senescence by playing an antagonistic role toward ethylene, auxin, and glucose in regulating the expression of PpEIN3a. Frontiers in Plant Science, 13, 1096645.

Ye L X, Zhang J X, Hou X J, Qiu M Q, Wang W F, Zhang J X, Hu C G, Zhang J Z. 2021. A MADS-Box gene CiMADS43 is involved in citrus flowering and leaf development through interaction with CiAGL9. International Journal of Molecular Sciences, 22, 5205.

Yin W C, Hu Z L, Cui B L, Guo X H, Hu J T, Zhu Z G, Chen G. 2017. Suppression of the MADS-box gene SlMBP8 accelerates fruit ripening of tomato (Solanum lycopersicum). Plant Physiology and Biochemistry, 118, 235–244.

Yue P T, Lu Q, Liu Z, Lv T X, Li X Y, Bu H D, Liu W T, Xu Y X, Yuan H, Wang A D. 2020. Auxin-activated MdARF5 induces the expression of ethylene biosynthetic genes to initiate apple fruit ripening. The New Phytologist, 226, 1781–1795.

Zhai R, Liu J L, Liu F X, Zhao Y X, Liu L L, Fang C, Wang H B, Li X Y, Wang Z G, Ma F W, Xu L F. 2018. Melatonin limited ethylene production, softening and reduced physiology disorder in pear (Pyrus communis L.) fruit during senescence. Postharvest Biology and Technology, 139, 38–46.

Zhang J L, Hu Z L, Yao Q Y, Guo X, Nguyen V, Li F F, Chen G. 2018. A tomato MADS-box protein, SlCMB1, regulates ethylene biosynthesis and carotenoid accumulation during fruit ripening. Scientific Reports, 8, 3413.

Zhang R, Ding J, Liu C X, Cai C P, Zhou B L, Zhang T Z, Guo W Z. 2015. Molecular evolution and phylogenetic analysis of eight COL superfamily genes in group I related to photoperiodic regulation of flowering time in wild and domesticated cotton (Gossypium) species. PLoS ONE, 10, e0118669.

Zhang S L, Yao J, Wang L, Wu N, van Nocker S, Li Z, Gao M, Wang X P. 2022. Role of grapevine SEPALLATA-related MADS-box gene VvMADS39 in flower and ovule development. The Plant Journal, 111, 1565–1579.

Zhang Y F, Shi H Y, Zhang Y X. 2013. Expression and regulation of the ethylene receptor PpERS gene during pear fruit development and following salicylic acid treatment. Plant Cell Tissue and Organ Culture, 114, 385–394.

Zhao Y, Tan D X, L Q, Chen H, Wang L, Li Q T, Gao Y, Kong J. 2013. Melatonin and its potential biological functions in the fruits of sweet cherry. Journal of Pineal Research, 55, 79–88.

Zhu Q G, Gong Z Y, Wang M M, Li X, Grierson D, Yin X R, Chen K S. 2018. A transcription factor network responsive to high CO2/hypoxia is involved in deastringency in persimmon fruit. Journal of Experimental Botany, 69, 2061–2070.

No related articles found!
No Suggested Reading articles found!