薄壳山核桃CiAGL6基因的克隆、亚细胞定位及表达

王敏; 席东; 莫正海; 陈于; 赵玉强; 朱灿灿

doi:10.3969/j.issn.1000-2006.201910027

薄壳山核桃CiAGL6基因的克隆、亚细胞定位及表达

王敏, 席东, 莫正海, 陈于, 赵玉强, 朱灿灿

南京林业大学学报（自然科学版） ›› 2020, Vol. 44 ›› Issue (4) : 63-69.

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南京林业大学学报（自然科学版） ›› 2020, Vol. 44 ›› Issue (4) : 63-69. DOI: 10.3969/j.issn.1000-2006.201910027

研究论文

薄壳山核桃CiAGL6基因的克隆、亚细胞定位及表达

作者信息 +

Cloning, subcellular localization and expression analysis of CiAGL6 gene in pecan

Author information +

文章历史 +

摘要

目的

研究薄壳山核桃MADS-box转录因子CiAGL6基因的特性、亚细胞定位及在雌花不同发育时期的表达水平，为揭示薄壳山核桃成花分子机理提供理论依据。

方法

根据薄壳山核桃转录组数据，采用RT-PCR方法获得CiAGL6基因，对其进行生物信息学分析和亚细胞定位鉴定。利用实时荧光定量PCR技术（qRT-PCR）分析CiAGL6基因在雌花发育过程中的表达模式。

结果

获得薄壳山核桃CiAGL6基因的完整开放阅读框（ORF）780 bp，共编码259个氨基酸，具有典型的MADS-box结构域和K-box区。在该蛋白的二级结构中，α-螺旋、延伸链和无规则卷曲分别占该蛋白质氨基酸残基总数的54.44%、17.37%和28.19%。系统进化分析表明，薄壳山核桃CiAGL6与壳斗科栓皮栎QsAGL6的亲缘关系最近。亚细胞定位显示，CiAGL6蛋白定位在细胞核内。qRT-PCR分析结果表明，CiAGL6基因在雌花中的表达量显著高于雄花中的。CiAGL6基因在雌花发育的各个阶段均有表达，其中在雌花盛开期的表达量最高。

结论

CiAGL6基因在薄壳山核桃雌花形成过程中发挥重要调控作用。

Abstract

Objective

The aim was to study the characteristics, subcellular localization and expression level in different development stages of female flowers of the MADS?box transcription factor CiAGL6 in pecan, which provides a theoretical basis for revealing the molecular mechanism of pecan flower formation.

Method

According to the transcriptome data of pecan, the CiAGL6 gene was cloned using RT?PCR. Next, the bioinformatic analysis and subcellular localization analysis were performed. And the expression pattern of CiAGL6 gene in female flower development was analyzed using real?time fluorescence quantitative PCR (qRT?PCR).

Result

The complete open reading frame (ORF) of the CiAGL6 gene was 780 bp, which encodes 259 amino acids and contains a conserved MADS?box domain and K?domain．In the secondary structure of the protein, a?helix, extension chain, and irregular curl accounted for 54.44%, 17.37%, and 28.19% of the total amino acid residues of the protein, respectively. The phylogenetic analysis showed that CiAGL6 has the closest relationship with QsAGL6 of Quercus suber in Fagaceae. The subcellular localization assay revealed CiAGL6 was located in the nucleus. The qRT?PCR analysis showed that the expression of CiAGL6 gene in female flowers was significantly higher than that in male flowers. CiAGL6 was expressed in all stages of female flower development, and the highest expression was found in female flowers at the blooming stage.

Conclusion

The CiAGL6 gene plays an important regulatory role in pecan female flower formation.

导出引用

王敏, 席东, 莫正海, 陈于, 赵玉强, 朱灿灿. 薄壳山核桃CiAGL6基因的克隆、亚细胞定位及表达[J]. 南京林业大学学报（自然科学版）. 2020, 44(4): 63-69 https://doi.org/10.3969/j.issn.1000-2006.201910027

WANG Min, XI Dong, MO Zhenghai, CHEN Yu, ZHAO Yuqiang, ZHU Cancan. Cloning, subcellular localization and expression analysis of CiAGL6 gene in pecan[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2020, 44(4): 63-69 https://doi.org/10.3969/j.issn.1000-2006.201910027

中图分类号： S664.1

参考文献

1	COEN E S, MEYEROWITZ E M. The war of the whorls: genetic interactions controlling flower development [J]. Nature, 1991, 353(6339): 31-37. DOI: 10.1038/353031a0.
2	唐璐璐, 宋云澎, 李交昆, 等. 花器官发育的分子机制研究进展[J]. 西北植物学报, 2013, 33(5): 1063-1070.
2	TANG L L, SONG Y P, LI J K, et al. Advances on the molecular mechanism of floral organ development [J]. Acta BotBoreal?OccidentSin, 2013, 33(5): 1063-1070. DOI: 1000-4025 (2013)05-1063-08.
3	THEISSEN G. Development of floral organ identity: stories from the MADS house [J]. Current Opinion in Plant Biology, 2001, 4(1): 75?85. DOI: 10.1016/s1369-5266(00)00139-4.
4	隋娟娟, 李晓昕, 杨秋燕, 等. 重瓣百合LiSEP3基因克隆与表达分析[J]. 南京林业大学学报(自然科学版), 2017, 41(1): 42-48.
4	SUI J J, LI X X, YANG Q Y, et al. Cloning and expression analysis of gene LiSEP3 in double lily [J]. Journal of Nanjing Forestry University (Natural Sciences Edition), 2017, 41(1): 42-48. DOI: 10.3969/j.issn.1000-2006.2017.01.007.
5	GRAMZOW L, RITZ M S, Thei?en G. On the origin of MADS-domain transcription factors [J]. Trends in Genetics, 2010, 26(4): 149-153. DOI: 10.1016/j.tig.2010.01.004.
6	IMMINK R G H, KAUFMANN K, ANGENENT G C. The ‘ABC’ of MADS domain protein behaviour and interactions [J]. Semin Cell Dev Biol, 2010, 21（1）, 87-93. DOI: 10.1016/j.semcdb. 2009.10.004.
7	牟村, 魏琦琦, 何潇, 等. 枣AGL6基因的克隆及表达分析[J]. 经济林研究, 2017, 35(4): 29 -35.
7	MU C, WEI Q Q, He X, et al. Cloning and expression analysis of AGL6 gene in Ziziphus jujube [J]. Nonwood Forest Research, 2017, 35(4): 29-35. DOI:10.14067/j.cnki.1003-8981.2017.04.005
8	隋娟娟, 李晓昕, 吴健, 等. 重瓣百合LiAGL6基因的克隆与表达分析[J]. 华北农学报, 2018, 33(2): 87-94.
8	SUI J J, LI X X, WU J, et al. Cloning and expression analysis of LiAGL6 in Double Lily[J]. Acta Agriculturae Boreali-Sinica, 2018, 33(2): 87-94. DOI: 10.7668/hbnxb.2018.02.013.
9	孙迎坤, 李纪元, 殷恒福, 等. 重瓣山茶花‘金盘荔枝’C功能基因CjAGL6的全长克隆与表达分析[J]. 植物研究, 2013, 33(3): 330-338.
9	SUN Y K, LI J Y, YIN H F, et al. Cloning and expression analysis of C function CjAGL6 gene cDNA from Camellia japonica ‘Jinpanlizhi’[J]. Bulletin of Botanical Research, 2013, 33(3): 330-338. DOI: 10.7525/j.issn.1673 -5102.2013．03．013.
10	唐文龙, 赵鹏飞, 李永华, 等. 牡丹PsAGL6基因克隆与表达分析[J]. 西北植物学报, 2018, 38 (3): 439-444.
10	TANG W L, ZHAO P F, LI Y H, et al. Cloning and expression analysis of transcription factor gene PsAGL6 in tree peony (Paeonia suffruticosa L.) [J]. Acta Botanica Boreali?Occidentalia Sinica, 2018, 38(3):439-444. DOI: CNKI: SUN: DNYX. 0. 2018-03-008.
11	韩明慧, 彭方仁, 邓秋菊,等. 薄壳山核桃雌雄花芽分化外部形态与内部结构的关系[J]. 南京林业大学学报(自然科学版), 2017, 41 （6）：1-7.
11	HAN M H, PENG F R, DENG Q J, et al. Observation of morphological and anatomical characters on staminate and pistillate flower differentiation in Carya illinoensis (Wangenh.) C. Koch [J]. Journal of Nanjing Forestry University (Natural Sciences Edition), 2017, 41 （6）：1-7.DOI: 10.3969/j.issn.1000-2006.201703020.
12	刘雨, 常君, 任华东,等. 薄壳山核桃雌花发育的解剖学研究[J]. 西南大学学报(自然科学版), 2018, 40 （6):32-37.
12	LIU Y, CHANG J, REN H D, et al. A Study on anatomy of development of female flower in Carya illinoinensis [J]. Journal of Southwest University (Natural Science), 2018, 40 (6):32-37. DOI: 10. 13718/j.cnki. xdzk.2018.06.005.
13	WoodBRUCE W. Pollination characteristics of pecan trees and orchards [J]. Horttechnology, 2000, 10(1):120-126. DOI:10.21273/HORTTECH.10.1.120.
14	张计育, 莫正海, 李永荣, 等. 薄壳山核桃MADS?box基因CiMADS9的克隆与功能分析[J]. 园艺学报, 2015, 42(6):1049-1056.
14	ZHANG J Y, MO Z H, LI Y Y, et al. Cloning and functional analysis of MADS?box CiMADS9 gene from Carya illinoinensis [J]. Acta Horticulturae Sinica, 2015, 42(6): 1049-1056. DOI: 10.16420/j.issn.0513-353x.2014-1094.
15	ZHANG J Y, WANG M, MO Z H, et al. Molecular characterization and functional analysis of an AGAMOUS?like gene CiAG from Pecan [J]. Hort Science, 2016, 51(6):664-668. DOI: 10.21273/HORTSCI.51.6.664.
16	WANG M, XI D, CHEN Y, et al. Morphological characterization and transcriptome analysis of pistillate flowering in pecan (Carya illinoinensis)[J]. Scientia Horticulturae, 2019, 257：108674. DOI: 10.1016/j.scienta.2019.108674.
17	REN X L, XIN G L, DU X M, et al. Abnormal tapetum development in hermaphrodites of an androdioecious tree, Tapiscia sinensis [J]. Tree physiology, 2020, 41（1）： 108-118. DOI: 10.1093/treephys/tpz080.
18	LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real?time quantitative PCR and the 2^-△△Ct method [J]. Methods, 2001, 25(4):402-408. DOI: 10.1006/meth.2001.1262.
19	YU X H, CHEN G P, GUO X H, et al. Silencing SlAGL6, a tomato AGAMOUS?LIKE6 lineage gene, generates fused sepal and green petal [J]. Plant Cell Reports, 2017, 36(6): 959-969. DOI: 10.1007/s00299-017-2129-9.
20	LI H F, LIANG W Q, JIA R D, et al. The AGL6?like gene OsMADS6 regulates floral organ and meristem identities in rice [J]. Cell Research, 2010, 20(3):299-313. DOI: 10.1038/cr.2009.143.
21	RIJPKEMA A S, ZETHOF J, GERATS T, et al. The petunia AGL6 gene has a SEPALLATA?like function in ?oral patterning [J]. The Plant Journal, 2009, 60(1): 1-9. DOI:10.1111/j.1365-313X.2009.03917.x.
22	HSU H F, HUANG C H, CHOU L T, et al. Ectopic expression of an orchid (Oncidium Gower Ramsey) AGL6?like gene promotes flowering by activating flowering time genes in Arabidopsis thaliana [J]. Plant and Cell Physiology, 2003, 44(8):783-794. DOI: 10.1093/pcp/pcg099.
23	WANG B G, ZHANG Q, WANG L G, et al. The AGL6?like gene CpAGL6, a potential regulator of floral time and organ Identity in Wintersweet (Chimonanthus praecox) [J]. Journal of Plant Growth Regulation, 2011, 30(3):343-352. DOI: 10.1007/s00344-011-9196-x.
24	SU Y L, LIU J X, LIANG W Q, et al. Wheat AGAMOUS LIKE 6 transcription factors function in stamen development by regulating the expression of Ta APETALA3[J]. Development, 2019, 146(20):dev. 177527. DOI: 10.1242/dev.177527.
25	FAN J H, LI WEN Q, DONG X C, et al. Ectopic expression of a hyacinth AGL6 homolog caused earlier flowering and homeotic conversion in Arabidopsis [J]. Science in China Series C: Life Sciences, 2007, 50(5): 676–689. DOI: 10.1007/s11427-007-0083-4.
26	廖望, 闫晓雪, 吴军, 等. 麻疯树雌雄花中MADS?box基因的表达分析[J]. 广西植物, 2018, 38(2): 180-187.
26	LIAO W, YAN X X, WU J, et al. Expression analysis on MADS?BOX genes in male and female flowers of Jatropha curca [J]. Guihaia, 2018, 38(2):180-187. DOI:10.11931/guihaia.gxzw201707001.