Cloning and Preliminary Functional Exploration of the Transposon-Derived Gene RcHTD in Rose

doi:10.16420/j.issn.0513-353x.2024-0842

Acta Horticulturae Sinica ›› 2025, Vol. 52 ›› Issue (9): 2377-2386.doi: 10.16420/j.issn.0513-353x.2024-0842

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Cloning and Preliminary Functional Exploration of the Transposon-Derived Gene RcHTD in Rose

LI Maofu¹^,²^,³, YANG Yuan¹^,²^,⁴, WANG Hua¹^,²^,³, SUN Pei¹^,²^,³, KANG Yanhui¹^,²^,³, SUN Xiangyi¹^,²^,³, LUO Jie¹^,²^,³, LIU Xin¹^,²^,³, JIN Min¹^,²^,³, JIN Wanmei¹^,²^,³^,^*()

¹ Institute of Forestry and Pomology， Beijing Academy of Agriculture and Forestry Sciences， Beijing 100093， China
² Key Laboratory of Biology and Genetic Improvement of Horticultural Crops（North China）， Ministry of Agriculture， Beijing 100093， China
³ Beijing Engineering Research Center of Functional Floriculture， Beijing 100093， China
⁴ Beijing Engineering Research Center for Deciduous Fruit Trees， Beijing 100093， China

Received:2025-01-05 Revised:2025-04-07 Online:2025-09-25 Published:2025-09-24
Contact: JIN Wanmei

Abstract

Abstract:

To investigate the regulatory role of Harbinger transposon-derived genes in anthocyanin synthesis in rose，the petals of the rose variety（Rosa chinensis‘Slater’s Crimson China’）were used as experimental material. In preliminary experiments，Rosa1 was employed as bait to screen a petal cDNA library，resulting in the identification of an interacting gene. This study successfully cloned the gene and characterized it as a transposon-derived gene，named RcHTD，which has an open reading frame of 1 560 bp and encodes 519 amino acids. Multiple sequence alignment analysis revealed that RcHTD encompasses a DDE transposon subfamily domain. Phylogenetic tree analysis indicated that RcHTD clustered closely with the genes AT5G12010，AT4G29780，AtALP1 and AtHHP1，which encode Harbinger-type transposases. Expression profilingof RcHTD in different developmental stages and tissues of Rosa chinensis‘Slater’s Crimson China’petals showed that RcHTD is expressed in all stages and tissues. Subcellular localization studies confirmed that the RcHTD protein is localized in the cell nucleus. Yeast two-hybrid experiments indicated that RcHTD physically interacts with RcMYB114. EMSA experiments further confirmed that RcHTD specifically binds to the RcMYB114 promoter，indicating its role in regulating anthocyanin synthesis，thereby consequently influencing the coloration of rose red petal.

Key words: Rosa chinensis, transposon-derived genes, RcHTD, gene function

LI Maofu, YANG Yuan, WANG Hua, SUN Pei, KANG Yanhui, SUN Xiangyi, LUO Jie, LIU Xin, JIN Min, JIN Wanmei. Cloning and Preliminary Functional Exploration of the Transposon-Derived Gene RcHTD in Rose[J]. Acta Horticulturae Sinica, 2025, 52(9): 2377-2386.

Figures/Tables 8

References 31

[1]	Bourque G, Burns K H, Gehring M, Gorbunova V, Seluanov A, Hammell M, Imbeault M, Izsvák Z, Levin H L, Macfarlan T S, Mager D L, Feschotte C. 2018. Ten things you should know about transposable elements. Genome Biology, 19 (1):199. doi: 10.1186/s13059-018-1577-z pmid: 30454069
[2]	Bundock P, Hooykaas P. 2005. An Arabidopsis hAT-like transposase is essential for plant development. Nature, 436 (7048):282-284.
[3]	Cam H P, Noma K, Ebina H, Levin H L, Grewal S I. 2008. Host genome surveillance for retrotransposons by transposon-derived proteins. Nature, 451 (7177):431-436.
[4]	Duan C G, Wang X, Xie S, Pan L, Miki D, Tang K, Hsu C C, Lei M, Zhong Y, Hou Y J, Wang Z, Zhang Z, Mangrauthia S K, Xu H, Zhang H, Dilkes B, Tao W A, Zhu J K. 2017. A pair of transposon-derived proteins function in a histone acetyltransferase complex for active DNA demethylation. Cell Research, 27 (2):226-240.
[5]	Fan F, Cheng M, Yuan H, Li N, Liu M, Cai M, Luo X, Ahmad A, Li N, Li S. 2023. A transposon-derived gene family regulates heading date in rice. Plant Science,337:111871.
[6]	Feschotte C, Jiang N, Wessler S R. 2002. Plant transposable elements:where genetics meets genomics. Nature Reviews Genetics, 3 (5):329-341. doi: 10.1038/nrg793 pmid: 11988759
[7]	Gao Y, Xu Z, Zhang L, Li S, Wang S, Yang H, Liu X, Zeng D, Liu Q, Qian Q, Zhang B, Zhou Y. 2020. MYB61 is regulated by GRF4 and promotes nitrogen utilization and biomass production in rice. Nature Communications, 11 (1):5219.
[8]	Hassan A H, Mokhtar M M, El Allali A. 2024. Transposable elements:multifunctional players in the plant genome. Frontiers in Plant Science,14:1330127.
[9]	Jiang N, Bao Z, Zhang X, Eddy S R, Wessler S R. 2004. Pack-MULE transposable elements mediate gene evolution in plants. Nature, 431 (7008):569-573.
[10]	Kapitonov V V, Jurka J. 2004. Harbinger transposons and an ancient HARBI1 gene derived from a transposase. DNA Cell Biology, 23 (5):311-324.
[11]	Krasileva K V. 2019. The role of transposable elements and DNA damage repair mechanisms in gene duplications and gene fusions in plant genomes. Current Opinion in Plant Biology,48:18-25.
[12]	Li M, Zhang H, Yang Y, Wang H, Xue Z, Fan Y, Sun P, Zhang H, Zhang X, Jin W. 2022. Rosa1,a transposable element-like insertion,produces red petal coloration in rose through altering RcMYB114 transcription. Frontiers in Plant Science,13:857684.
[13]	Li Maofu, Yang Yuan, Wang Hua, Liu Jiashen, Jin Wanmei. 2017. Cloning and expression analysis on RcbHLH gene from Rosa chinensis and interaction analysis between MYB and WD 40 transcription factors. Acta Horticulturae Sinica, 44 (10):1949-1958. (in Chinese)
	李茂福, 杨媛, 王华, 刘佳棽, 金万梅. 2017. 月季bHLH基因的克隆、表达及其与MYB和WD40的互作分析. 园艺学报, 44 (10):1949-1958. doi: 10.16420/j.issn.0513-353x.2017-0183
[14]	Liu Keke, Tang Dingqin, Zhou Mingbing. 2021. The evolution pathway of transposon-derived genes and its effect on biological growth and development. Chinese Journal of Cell Biology, 43 (1):152-160. (in Chinese)
	刘珂珂, 汤定钦, 周明兵. 2021. 转座子衍生基因的演变途径及对生物生长发育的影响. 中国细胞生物学学报, 43 (1):152-160
[15]	Mao D, Tao S, Li X, Gao D, Tang M, Liu C, Wu D, Bai L, He Z, Wang X, Yang L, Zhu Y, Zhang D, Zhang W. 2022. The Harbinger transposon-derived gene PANDA epigenetically coordinates panicle number and grain size in rice. Plant Biotechnology Journal, 20 (6):1154-1166.
[16]	Mikina W, Hałakuc P, Milanowski R. 2024. Transposon-derived introns as an element shaping the structure of eukaryotic genomes. Journal of Experimental Botany, 15 (1):15.
[17]	Naito K, Zhang F, Tsukiyama T, Saito H, Hancock C N, Richardson A O, Okumoto Y, Tanisaka T, Wessler S R. 2009. Unexpected consequences of a sudden and massive transposon amplification on rice gene expression. Nature, 461 (7267):1130-1134.
[18]	Niu X M, Xu Y C, Li Z W, Bian Y T, Hou X H, Chen J F, Zou Y P, Jiang J, Wu Q, Ge S, Balasubramanian S, Guo Y L. 2019. Transposable elements drive rapid phenotypic variation in Capsella rubella. Proceedings of the National Academy of Sciences of the United States of America, 116 (14):6908-6913.
[19]	Sinzelle L, Kapitonov V V, Grzela D P, Jursch T, Jurka J, Izsvák Z, Ivics Z. 2008. Transposition of a reconstructed Harbinger element in human cells and functional homology with two transposon-derived cellular genes. Proceedings of the National Academy of Sciences of the United States of America, 105 (12):4715-4720.
[20]	Stefanov B A, Nowacki M. 2024. The roles of transposable elements in transgenerational inheritance and genome evolution. In:Witzany G ed. Epigenetics in Biological Communication. Cham: Springer Nature Switzerland,369-385.
[21]	Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGA5:molecular evolutionary genetics analysis using maximum likelihood,evolutionary distance,and maximum parsimony methods. Molecular Biology and Evolution, 28 (10):2731-2739.
[22]	Tang W, Ji Q, Huang Y, Jiang Z, Bao M, Wang H, Lin R. 2013. FAR-RED ELONGATED HYPOCOTYL3 and FAR-RED IMPAIRED RESPONSE1 transcription factors integrate light and abscisic acid signaling in Arabidopsis. Plant Physiology, 163 (2):857-866.
[23]	Wang D, Su Y, Wang X, Lei H, Yu J. 2012. Transposon-derived and satellite-derived repetitive sequences play distinct functional roles in Mammalian intron size expansion. Evolutionary Bioinformatics,8:301-319.
[24]	Wang L, Dou K, Moon S, Tan F J, Zhang Z Z. 2018. Hijacking oogenesis enables massive propagation of LINE and retroviral transposons. Cell, 174 (5):1082. doi: S0092-8674(18)30843-2 pmid: 30057117
[25]	Wang L, Tracy L, Su W, Yang F, Feng Y, Silverman N, Zhang Z Z Z. 2022. Retrotransposon activation during Drosophila metamorphosis conditions adult antiviral responses. Nature Genetics, 54 (12):1933-1945. doi: 10.1038/s41588-022-01214-9 pmid: 36396707
[26]	Xiang Yuanping, Wang Yidan, He Hongjun, Xu Qijiang. 2020. Plant transposable elements and the effects of insertion mutations on flower development in horticultural plants. Acta Horticulturae Sinica, 47 (11):2247-2266. (in Chinese) doi: 10.16420/j.issn.0513-353x.2020-0361
	相元萍, 王一丹, 贺洪军, 徐启江. 2020. 植物转座子类型及其插入突变对园艺植物花发育影响的研究进展. 园艺学报, 47 (11):2247-2266.
[27]	Yaakov B, Kashkush K. 2011. Methylation,transcription,and rearrangements of transposable elements in synthetic allopolyploids. Int J Plant Genomics,569826.
[28]	Yang H, Zhang J T, Zhang H W, Cao R X, Tang D L, Wang L J. 2024. 5-Aminolevulinic acid against strawberry Fusarium wilt:Bidirectional regulation of biocontrol agents and pathogens. Horticultural Plant Journal, 10 (6):1349-1361.
[29]	Yao M C, Mao D. 2022. The Harbinger transposon-derived gene PANDA epigenetically coordinates panicle number and grain size in rice. Genes Development, 20 (6):1154-1166.
[30]	Zhang L, Wang R, Xing Y, Xu Y, Xiong D, Wang Y, Yao S. 2021. Separable regulation of POW1 in grain size and leaf angle development in rice. Plant Biotechnol Journal, 19 (12):2517-2531.
[31]	Zhou X, He J, Velanis C N, Zhu Y, He Y, Tang K, Zhu M, Graser L, de Leau E, Wang X, Zhang L, Andy Tao W, Goodrich J, Zhu J, Zhang C. 2021. 3 deacetylation at genes but not TEs in Arabidopsis. Journal of Integrative Plant Biology, 63 (8):1462-1474.

目的Purpose	引物名称Primer name	引物序列（5′-3′）Primer sequence
基因克隆 Gene cloning	RcHTD	F：ATGGAAGTCACCCCTTTCACATTTCT
基因克隆 Gene cloning	RcHTD	R：CTAATGAAATCCAGTGCCTGCAAGGCC
RT-PCR	RcHTD	F：GCGGAGGAAGGTCGATGAGGACGATACC
	RcHTD	R：CCTGTAATTCGCGTCAAGAGTCTCC
	Actin	F：GGCTGTTCTCTCTCTGTATGC
	Actin	R：TTCTGGGCACCTGAATCTC
亚细胞定位 Subcellular localization	RcHTD	F： CGACTCTAGTCTAGAAAGCTATGGAAGTCACCCCTTTCACATTTCT
亚细胞定位 Subcellular localization	RcHTD	R： CATGGTACCGGATCCACTAGTATGAAATCCAGTGCCTGCAAGGCC
酵母双杂交 Yeast two hybrid	RcHTD	F： TGGCCATGGAGGCCAGTGAATTCATGGAAGTCACCCCTTTCACATTTCT
酵母双杂交 Yeast two hybrid	RcHTD	R： CTACGATTCATCTGCAGCTCGAGCTAATGAAATCCAGTGCCTGCAAGGCC
蛋白诱导表达 Protein induced expression EMAS 探针 EMAS probe	RcHTD Rosa1	F： GATCTGGTTCCGCGTGGATCCATGGAAGTCACCCCTTTCACATTTCT
蛋白诱导表达 Protein induced expression EMAS 探针 EMAS probe	RcHTD Rosa1	R： GATGCGGCCGCTCGAGTCGAATGAAATCCAGTGCCTGCAAGGCC AAATGAATGCTTACTTTCGGAAGTCCTTGTTC AAAAAAAAAAAAAAAATCGGAAGTCCTTGTTC

目的Purpose	引物名称Primer name	引物序列（5′-3′）Primer sequence
基因克隆 Gene cloning	RcHTD	F：ATGGAAGTCACCCCTTTCACATTTCT
基因克隆 Gene cloning	RcHTD	R：CTAATGAAATCCAGTGCCTGCAAGGCC
RT-PCR	RcHTD	F：GCGGAGGAAGGTCGATGAGGACGATACC
	RcHTD	R：CCTGTAATTCGCGTCAAGAGTCTCC
	Actin	F：GGCTGTTCTCTCTCTGTATGC
	Actin	R：TTCTGGGCACCTGAATCTC
亚细胞定位 Subcellular localization	RcHTD	F： CGACTCTAGTCTAGAAAGCTATGGAAGTCACCCCTTTCACATTTCT
亚细胞定位 Subcellular localization	RcHTD	R： CATGGTACCGGATCCACTAGTATGAAATCCAGTGCCTGCAAGGCC
酵母双杂交 Yeast two hybrid	RcHTD	F： TGGCCATGGAGGCCAGTGAATTCATGGAAGTCACCCCTTTCACATTTCT
酵母双杂交 Yeast two hybrid	RcHTD	R： CTACGATTCATCTGCAGCTCGAGCTAATGAAATCCAGTGCCTGCAAGGCC
蛋白诱导表达 Protein induced expression EMAS 探针 EMAS probe	RcHTD Rosa1	F： GATCTGGTTCCGCGTGGATCCATGGAAGTCACCCCTTTCACATTTCT
蛋白诱导表达 Protein induced expression EMAS 探针 EMAS probe	RcHTD Rosa1	R： GATGCGGCCGCTCGAGTCGAATGAAATCCAGTGCCTGCAAGGCC AAATGAATGCTTACTTTCGGAAGTCCTTGTTC AAAAAAAAAAAAAAAATCGGAAGTCCTTGTTC

Cloning and Preliminary Functional Exploration of the Transposon-Derived Gene RcHTD in Rose

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 8

References 31

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	MO Jinxia, LI Fang, XIONG Xinting, ZHONG Zaofa, PENG Ting. Characterization of PtSARD1 and Its Preliminary Role in Drought Tolerance [J]. Acta Horticulturae Sinica, 2025, 52(6): 1399-1411.
[2]	MA Yuwan, LIU Ao, LIU Xiangdong, ZHANG Yajing, DONG Xuanke, LI Yufan, CHEN Jiren. Cloning of the RcRAP2.7 Gene and its Expression Analysis Under Abiotic Stresses in Rosa chinensis‘Yueyuehong’ [J]. Acta Horticulturae Sinica, 2025, 52(4): 921-932.
[3]	REN Hengze, LI Danying, YU Yating, LÜ Wuyun, HAO Xinyuan, WANG Xinchao, WANG Yuchun. Research Advances of Strategies to Engineer VIGS Vectors and Its Application in Plants [J]. Acta Horticulturae Sinica, 2024, 51(7): 1455-1473.
[4]	LU Jing, WEI Suyun, YIN Tongming, CHEN Yingnan. Research Progress on the Cytokinin Type-A Response Regulator Gene Family [J]. Acta Horticulturae Sinica, 2023, 50(9): 1867-1888.
[5]	GUO Jing, LIAO Manyu, JIN Yan, MA Xiaochuan, ZHANG Fen, LU Xiaopeng, DENG Ziniu, SHENG Ling. Functional Analysis of Transcription Factor CsbHLH3 in Regulating Citric Acid Metabolism of Citrus Fruit [J]. Acta Horticulturae Sinica, 2023, 50(5): 947-958.
[6]	SHEN Yuxiao, ZOU Jinyu, LUO Ping, SHANG Wenqian, LI Yonghua, HE Songlin, WANG Zheng, SHI Liyun. Genome-wide Identification and Abiotic Stress Response Analysis of PP2C Family Genes in Rosa chinensis‘Old Blush’ [J]. Acta Horticulturae Sinica, 2023, 50(10): 2139-2156.
[7]	HUANG Ling, HU Xianmei, LIANG Zehui, WANG Yanping, CHAN Zhulong, XIANG Lin. Cloning and Function Identification of Anthocyanidin Synthase Gene TgANS in Tulipa gesneriana [J]. Acta Horticulturae Sinica, 2022, 49(9): 1935-1944.
[8]	YANG Yuyan, DUAN Xinyuan, HE Zhilin, BING Qihao, CHEN Suoying, LIU Xiaoman, ZENG Ming, LIU Xiaogang. Cloning and Function Characterization of UDP-L-rhamnose Synthase from Fortunella crassifolia [J]. Acta Horticulturae Sinica, 2022, 49(8): 1663-1672.
[9]	TAO Xin, ZHU Rongxiang, GONG Xin, WU Lei, ZHANG Shaoling, ZHAO Jianrong, ZHANG Huping. Fructokinase Gene PpyFRK5 Plays an Important Role in Sucrose Accumulation of Pear Fruit [J]. Acta Horticulturae Sinica, 2022, 49(7): 1429-1440.
[10]	MA Mingying, HAO Chenxing, ZHANG Kai, XIAO Guihua, SU Hanying, WEN Kang, DENG Ziniu, MA Xianfeng. CsSWEET2a Promotes the Infection of Xanthomonas citri subsp. citri [J]. Acta Horticulturae Sinica, 2022, 49(6): 1247-1260.
[11]	LI Chonghui, YANG Guangsui, ZHANG Zhiqun, YIN Junmei. A Novel R2R3-MYB Transcription Factor Gene AaMYB6 Involved in Anthocyanin Biosynthesis in Anthurium andraeanum [J]. Acta Horticulturae Sinica, 2021, 48(10): 1859-1872.
[12]	ZHENG Hao, ZHANG Fen, JIAN Yue, HUANG Wenli, LIANG Sha, JIANG Min, YUAN Qiao, WANG Qiaomei, SUN Bo. Cloning and Function Identification of Dihydroflavonol 4-Reductase Gene BoaDFR in Chinese Kale [J]. Acta Horticulturae Sinica, 2021, 48(1): 73-82.
[13]	WU Jinghua，YANG Chao，and WU Shaohua*. Expression and Function Analysis of AGL6 Homologous Genes in Cymbidium ensifolium [J]. ACTA HORTICULTURAE SINICA, 2019, 46(6): 1123-1133.
[14]	LIU Shengyu1，ZHANG Shikui2，LU Juanfang1，LI Wenhui2，and XI Wanpeng1,3,*. Molecular Cloning and Function Analysis of Carotenoid Cleavage Dioxygenase 1（CCD1）in Apricot Fruit [J]. ACTA HORTICULTURAE SINICA, 2018, 45(3): 471-482.
[15]	ZHANG Shi-Zhong, FU Ying, ZHOU Bo, JIANG Ze-Sheng, XU Rui-Rui, SHU Huai-Rui. The Construction and Instruction Manual of Apple Functional Genomic Database [J]. ACTA HORTICULTURAE SINICA, 2012, 39(11): 2245-2250.