紫薇花器官发育调控基因LiFUL1的分离与表达分析

doi:10.3969/j.issn.1004-1524.2020.07.09

浙江农业学报 ›› 2020, Vol. 32 ›› Issue (7): 1206-1214.DOI: 10.3969/j.issn.1004-1524.2020.07.09

紫薇花器官发育调控基因LiFUL1的分离与表达分析

黄小珍^1,2, 乔中全^1,*, 曾慧杰¹, 李永欣^1,3, 何钢², 王晓明¹

1.湖南省林业科学院林木无性系育种湖南省重点实验室,湖南长沙410004;
2.中南林业科技大学生命科学与技术学院,湖南长沙 410004;
3.长沙市木本花卉工程技术研究中心,湖南长沙410004

收稿日期:2020-02-01 出版日期:2020-07-25 发布日期:2020-07-28
通讯作者: *乔中全,E-mail:qzq110@hnlky.cn
作者简介:黄小珍(1994—),女,广西贵港人,硕士研究生,主要从事紫薇花器官发育相关分子机理的研究。E-mail:hxzyoyo@163.com
基金资助:
长沙市科技计划(kq1801026); 国家重点研发计划(2019YFD1001000)

Isolation and expression of floral organ development regulating gene LiFUL1 in Lagerstroemia indica L.

HUANG Xiaozhen^1,2, QIAO Zhongquan^1,*, ZENG Huijie¹, LI Yongxin^1,3, HE Gang², WANG Xiaoming¹

1. Hunan Key Laboratory for Breeding of Clonally Propagated Forest Trees, Hunan Academy of Forestry, Changsha 410004, China;
2. College of Life Science of Biotechnology, Central South University of Forestry & Technology, Changsha 410004, China;
3. Changsha Woody Flower Engineering Technology Research Center, Changsha 410004, China

Received:2020-02-01 Online:2020-07-25 Published:2020-07-28

摘要/Abstract

摘要： 紫薇湘韵具有只开花不结实的特性,表现为花药不开裂、花粉败育、胚珠发育异常。紫薇红叶为普通紫薇,可正常开花结实。以紫薇湘韵为试验材料,采用RT-PCR方法,从花芽中分离得到1个FRUITFULL(FUL)同源基因,命名为LiFUL1(GenBank登录号为MN894547)。序列分析结果表明,其cDNA开放阅读框长度为753 bp,编码251个氨基酸,分子质量为28.453 13 ku。序列比对和保守结构域分析结果表明,LiFUL1基因编码蛋白具有典型的MADS-MEF2和K-box 结构域,C末端含有一个保守性高的基序euFUL MOTIF,因此,LiFUL1属于MADS家族的FUL/AP1亚家族。进化分析表明,紫薇的LiFUL1基因编码的蛋白氨基酸序列与木本植物蓝桉的FUL/AP1氨基酸序列亲缘关系最近。qRT-PCR分析结果表明,LiFUL1基因在紫薇湘韵花器官分化阶段的花萼分化时期、花瓣与雄蕊分化时期、雄蕊与雌蕊分化时期的表达量显著高于紫薇红叶。另外,利用原核表达系统在大肠埃希菌中成功表达了LiFUL1蛋白,为进一步研究LiFUL1基因的功能奠定了基础。

关键词: 紫薇, LiFUL1, 亲缘关系, 原核表达, RT-qPCR

Abstract: Flowers of crape myrtle Xiangyun are not strong enough to seeding, the manifestations of Xiangyun are indehiscent anther, pollen abortion and abnormal ovule. Redleaf is common crape myrtle with normal flowering and bearing. Crape myrtle Xiangyun was selected as the experimental material, RT-PCR was employed to isolate a homologous gene of FRUITFULL (FUL) from Xiangyun, named as LiFUL1(GenBank accession No. MN894547). Sequence analysis indicated that length of the open reading frame of LiFUL1 cDNA was 753 bp, which encoded a protein with 251 amino acids, its molecular weight was 28.453 13 ku. Analysis of sequence alignment and conserved structural domain showed that LiFUL1 protein belonged to FUL/AP1, a subfamily of the MADS, for it had both typical MADS-MEF2 and K-box domain, and the highly conserved motif euFUL MOTIF at C terminus. Phylogenetic tree analysis revealed that LiFUL1 of crape myrtle was most closely related to Eucalyptus globulus’s FUL/AP1 sequence. qRT-PCR test demonstrated that expressions of LiFUL1 in Xiangyun were significantly higher than those in crape myrtle Redleaf at calyx differentiation, petals and stamens differentiation, stamens and pistils differentiation stages. Successful expression of LiFUL1 in Escherichia coli using the prokaryotic expression system provided a basis for further study of LiFUL1 gene function.

Key words: crape myrtle, LiFUL1, genetic relationship, prokaryotic expression, RT-qPCR

中图分类号:

S685.99

黄小珍, 乔中全, 曾慧杰, 李永欣, 何钢, 王晓明. 紫薇花器官发育调控基因LiFUL1的分离与表达分析[J]. 浙江农业学报, 2020, 32(7): 1206-1214.

HUANG Xiaozhen, QIAO Zhongquan, ZENG Huijie, LI Yongxin, HE Gang, WANG Xiaoming. Isolation and expression of floral organ development regulating gene LiFUL1 in Lagerstroemia indica L.[J]. , 2020, 32(7): 1206-1214.

参考文献

[1] 孙宜, 孙猛, 石青松. 紫薇新品种扦插繁殖试验[J]. 浙江农业科学, 2019, 60(10):1729-1731.
SUN Y, SUN M, SHI Q S.Cutting propagation of new cultivars of Lagerstroemia[J]. Journal of Zhejiang Agricultural Sciences, 2019, 60(10):1729-1731. (in Chinese)
[2] 郑丽梅. 紫薇种质资源收集过程中硬枝扦插繁殖技术[J]. 福建林业, 2019(4):34-38.
ZHENG L M.The hard cutting propagation technique for the germplasm collection of Lagerstroemia indica[J]. Fujian Forestry, 2019(4):34-38.(in Chinese with English abstract)
[3] 王莹, 李敏, 谈峰, 等. 紫叶紫薇新品系叶色变化理化因素研究[J]. 江苏农业科学, 2018, 46(24):150-153.
WANG Y, LI M, TAN F, et al.Study on physiological and biochemical factors of leaf color changes of a new crape myrtle (Lagerstroemia indica) cultivar[J]. Jiangsu Agricultural Sciences, 2018, 46(24):150-153.(in Chinese)
[4] 吴际洋, 焦垚, 叶远俊, 等. 大花紫薇与紫薇杂交F₁群体表型评价及分子标记连锁分析[J]. 园艺学报, 2018, 45(11):2153-2163.
WU J Y, JIAO Y, YE Y J, et al.Phenotypic assessment and linkage analysis of main ornamental traits of Lagerstroemia speciosa × L. indica to SSR markers[J]. Acta Horticulturae Sinica, 2018, 45(11):2153-2163.(in Chinese with English abstract)
[5] 高德良, 宋化稳, 吕娟, 等. 几种新颖杀虫剂对紫薇长斑蚜的室内毒力测定及田间药效评价[J]. 北方园艺, 2019(23):68-73.
GAO D L, SONG H W, LYU J, et al.Bioactivity and field efficacy of several novel insecticides against Tinocallis kahawaluokalani[J]. Northern Horticulture, 2019(23):68-73.(in Chinese with English abstract)
[6] 乔中全, 王晓明, 蔡能, 等. 紫薇新品种‘丹霞’[J]. 园艺学报, 2019, 46(10):2069-2070.
QIAO Z Q, WANG X M, CAI N, et al.A new Lagerstroemia indica cultivar ‘Danxia’[J]. Acta Horticulturae Sinica, 2019, 46(10):2069-2070.(in Chinese with English abstract)
[7] 曾慧杰, 王晓明, 乔中全, 等. 紫薇新品种‘紫韵’[J]. 园艺学报, 2019, 46(增刊2): 2906-2907.
ZENG H J, WANG X M, QIAO Z Q, et al.A new Lagerstroemia indica cultivar ‘Ziyun ’[J]. Acta Horticulturae Sinica, 2019, 46(Suppl. 2): 2906-2907. (in Chinese with English abstract)
[8] 马水晶, 方淦民, 周斌, 等. 矮紫薇扦插繁殖[J]. 中国花卉园艺, 2019(18):40-42.
MA S J, FANG G M, ZHOU B, et al.Dwarf crape myrtle cuttage propagation[J]. China Flowers & Horticulture, 2019(18):40-42.(in Chinese)
[9] NG M, YANOFSKY M F.Function and evolution of the plant MADS-box gene family[J]. Nature Reviews. Genetics, 2001, 2(3):186-195.
[10] MESSENGUY F, DUBOIS E.Role of MADS box proteins and their cofactors in combinatorial control of gene expression and cell development[J]. Gene, 2003, 316:1-21.
[11] LILJEGREN S J, DITTA G S, ESHED Y, et al.SHATTERPROOF MADS-box genes control seed dispersal in Arabidopsis[J]. Nature, 2000, 404(6779):766-770.
[12] WANG L, YIN X J, CHENG C X, et al.Evolutionary and expression analysis of a MADS-box gene superfamily involved in ovule development of seeded and seedless grapevines[J]. Molecular Genetics and Genomics, 2015, 290(3):825-846.
[13] LITT A, IRISH V F.Duplication and diversification in the APETALA1/FRUITFULL floral homeotic gene lineage: implications for the evolution of floral development[J]. Genetics, 2003, 165(2):821-833.
[14] FERRÁNDIZ C, GU Q, MARTIENSSEN R, et al. Redundant regulation of meristem identity and plant architecture by FRUITFULL, APETALA1 and CAULIFLOWER[J]. Development (Cambridge, England), 2000, 127(4):725-734.
[15] GU Q, FERRÁNDIZ C, YANOFSKY M F, et al. The FRUITFULL MADS-box gene mediates cell differentiation during Arabidopsis fruit development[J]. Development(Cambridge, England), 1998, 125(8):1509-1517.
[16] MÜLLER B M, SAEDLER H, ZACHGO S. The MADS-box gene DEFH28 from Antirrhinum is involved in the regulation of floral meristem identity and fruit development[J]. The Plant Journal: for Cell and Molecular Biology, 2001, 28(2):169-179.
[17] PELUCCHI N, FORNARA F, FAVALLI C, et al.Comparative analysis of rice MADS-box genes expressed during flower development[J]. Sexual Plant Reproduction, 2002, 15(3):113-122.
[18] SEYMOUR G B, RYDER C D, CEVIK V, et al.A SEPALLATA gene is involved in the development and ripening of strawberry (Fragaria×ananassa Duch.) fruit, a non-climacteric tissue[J]. Journal of Experimental Botany, 2011, 62(3):1179-1188.
[19] CALONJE M.Floral meristem identity genes are expressed during tendril development in grapevine[J]. Plant Physiology, 2004, 135(3):1491-1501.
[20] 位伟强, 刘伟, 段亚宾, 等. 牡丹开花调控转录因子基因PsFUL1的克隆与表达分析[J]. 植物生理学报, 2017, 53(4):536-544.
WEI W Q, LIU W, DUAN Y B, et al.Molecular cloning and expression analysis of flowering-regulating transcription factor gene PsFUL1 in tree peony(Paeonia suffruticosa)[J]. Plant Physiology Journal, 2017, 53(4):536-544.(in Chinese with English abstract)
[21] HONG J K, KIM S Y, KIM K S, et al.Overexpression of a Brassica rapa MADS-box gene, BrAGL20, induces early flowering time phenotypes in Brassica napus[J]. Plant Biotechnology Reports, 2013, 7(3):231-237.
[22] 徐勇, 张林, 马荣才. 桃(Prunus persica)中2个MADS box基因功能的初步鉴定和遗传作图[J]. 科学通报, 2008, 53(5):537-543.
XU Y, ZHANG L, MA R C.Preliminary identification and genetic mapping of two MADS box genes in Prunus persica[J]. Chinese Science Bulletin, 2008, 53(5):537-543.(in Chinese)
[23] 甘德芳, 吴娇, 庄丹, 等. 黄瓜AP1-FUL类MADS-box基因的克隆及表达分析[J]. 核农学报, 2014, 28(9):1585-1590.
GAN D F, WU J, ZHUANG D, et al.Cloning and expression analysis of AP1-FUL type MADS-box gene in cucumber (Cucumis sativus L.)[J]. Journal of Nuclear Agricultural Sciences, 2014, 28(9):1585-1590.(in Chinese with English abstract)
[24] 王晓明. 紫薇不育新品种“湘韵”选育及繁殖技术研究成果简介[J]. 湖南林业科技, 2014(1):133-134.
WANG X M.Brief introduction of breeding and breeding technology of a new sterile variety of crape myrtle ‘xiangyun’[J]. Hunan Forestry Science & Technology, 2014, 41(1): 133-134. (in Chinese)
[25] JIANG W, WEI D Y, ZHOU W W, et al.HDA9 interacts with the promoters of SOC1 and AGL24 involved in flowering time control in Brassica juncea[J]. Biochemical and Biophysical Research Communications, 2018, 499(3):519-523.
[26] LESEBERG C H, EISSLER C L, WANG X, et al.Interaction study of MADS-domain proteins in tomato[J]. Journal of Experimental Botany, 2008, 59(8):2253-2265.
[27] 许欢. 结实与不结实紫薇生物学特性比较研究[D]. 长沙: 中南林业科技大学, 2015.
XU H.Biological characteristic comparison research between fructiferous and acarpous Lagerstroemia indica[D]. Changsha: Central South University of Forestry & Technology, 2015.(in Chinese with English abstract)
[28] ZHOU W, WANG X M, CHEN J H, et al.Abortion categories and characteristics of acarpous crape myrtle floral organs[J]. Journal of the American Society for Horticultural Science, 2019, 144(6):387-393.

紫薇花器官发育调控基因LiFUL1的分离与表达分析

Isolation and expression of floral organ development regulating gene LiFUL1 in Lagerstroemia indica L.

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