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

doi:10.3969/j.issn.1000-2006.201910027

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2020, Vol. 44 ›› Issue (4): 63-69.doi: 10.3969/j.issn.1000-2006.201910027

Previous Articles Next Articles

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

WANG Min¹(), XI Dong², MO Zhenghai¹, CHEN Yu¹, ZHAO Yuqiang¹, ZHU Cancan¹()

^1.Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing 210095, China
^2.Shanghai Qingpu District Agricultural Technology Extension Service Center, Shanghai 201700, China

Received:2019-10-25 Revised:2020-03-19 Online:2020-07-22 Published:2020-08-13
Contact: ZHU Cancan E-mail:wm120624@163.com;zhucancan858@163.com

Abstract

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.

Key words: pecan, CiAGL6 gene, phylogenetic analysis, subcellular localization, gene expression

CLC Number:

S664.1

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.

Figures/Tables 4

References 26

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.
	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.
	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, Theißen 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.
	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.
	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.
	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.
	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.
	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.
	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.
	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 ﬂoral 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.
	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.

Related Articles 15

[1]	HUANG Biyun, ZHUO Renying, QIAO Guirong. Comparative analysis of different types of callus in moso bamboo [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2023, 47(6): 141-149.
[2]	WANG Jianchao, QIU Wenmin, JIN Kangming, LU Zhuchou, HAN Xiaojiao, ZHUO Renying, LIU Xiaoguang, HE Zhengquan. Comprehensive analysis of WRKY gene family in Sedum plumbizincicola responding to cadmium stress [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2023, 47(2): 49-60.
[3]	ZHANG Ting, BAI Yang, QI Xiwu, YU Xu, FANG Hailing, LI Li, LIU Dongmei, LIANG Chengyuan, LI Weilin. Cloning and expression analyses of MhWRKY57 response to jasmonic acid in Mentha canadensis [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2022, 46(6): 279-287.
[4]	JIA Zhanhui, JIA Xiaodong, XU Mengyang, MO Zhenghai, ZHAI Min, XUAN Jiping, ZHANG Jiyu, WANG Gang, WANG Tao, GUO Zhongren. Cloning and expression analyses of the key enzyme gene of procyanidins biosynthesis in pecan (Carya illinoinensis) [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2022, 46(5): 49-57.
[5]	TIAN Mengyang, DOU Quanqin, XIE Yinfeng, TANG Wenhua, JI Yanhong. Study on photosynthetic characteristics of four pecan cultivars [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2022, 46(5): 67-74.
[6]	WANG Zhanjun, WU Ziqi, WANG Zhaoxia, OU Zulan, LI Jie, CAI Qianwen, XU Zhongdong, ZHANG Zhaoliang. A comparative study of the evolution and codon usage bias in WOX gene family of three Camellia sinensis cultivars [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2022, 46(2): 71-80.
[7]	NA Xiaoying, LIU Gang, LIU Guifeng, WANG Xiuwei. Effects of the four genes expressions and photosynthetic parameters on seedlings growth of different birch clones [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2022, 46(2): 88-94.
[8]	LIN Lili, HU Anqi, CHEN Gang, ZHANG Jiyue, CAO Guangqiu, CAO Shijiang. Cloning and expression characteristics of ClWRKY44 gene from Cunninghamia lanceolata [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2022, 46(1): 203-209.
[9]	WANG Peilong, YANG Ni, ZHANG Aoran, Tangnver•Sailike , LI Shuang, GAO Caiqiu. Cloning ThPCS1 gene of Tamarix hispida to improve cadmium tolerance [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2021, 45(3): 71-78.
[10]	LI Mengjuan, ZHU Liming, HUO Junnan, ZHANG Jingbo, SHI Jisen, CHENG Tielong. Cloning and expression analyses of NtCBL1,NtCBL2 gene of Nitraria tangutorum [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2021, 45(3): 93-99.
[11]	CHEN Wei, CHENG Tielong, JI Jing, WU Yanyan, XIE Tiantian, JIANG Zeping, SHI Shengqing. Identification of three gene families in the GABA shunt and their expression analysis in poplar [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2020, 44(5): 67-77.
[12]	ZHOU Xiaofeng, MO Zhenghai, SHANG Yangjuan, LUO Min, SU Wenchuan, TAN Pengpeng, HAN Minghui, DENG Qiuju, TIAN Zhaoxia, PENG Fangren. Cloning and expression analysis of the CiDGAT1 from Carya illinoinensis [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2020, 44(4): 55-62.
[13]	TIAN Xueyao, ZHOU Jie, WANG Baosong, HE Kaiyue, HE Xudong. Cloning and expression pattern analysis of NAC genes in Salix [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2020, 44(1): 119-124.
[14]	ZHENG Xiaoqin, CHEN Wenjing, CAO Fan, FENG Gang, LI Yongrong, PENG Fangren. Effects of the proportional fertilization on the nutrient content and photosynthesis of young grafted pecan seedlings [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2019, 43(5): 169-174.
[15]	ZHANG Chunhong, XIONG Zhenhao, WU Wenlong, LI Weilin. CAD enzyme activity and gene expression in connection with lignin synthesis during fruit development and ripening process of blackberry [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2019, 43(01): 141-148.

Metrics

Comments

www.nldxb.njfu.edu.cn

Edited ＆ Published by Editorial Department of Nanjing Forestry University(Natural Sciences Edition)
Address： No.159 Longpan Road,Nanjing 210037 Jiangsu,P.R.China
E-mail ：xuebao@njfu.edu.cn , xuebao@njfu.com.cn
Telephone +86-25-85428247,85427076
Distributed by China International Book Trading Corporation P.O. Box 399, Beijing ,P.R. China

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

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 4

References 26

Related Articles 15

Recommended Articles

Metrics

Comments

Author

Reader

Reviewer