Expression Characteristic Analysis of the BcPG17,a Gene Related to Inflorescence Stem Development of Brassica campestris

doi:10.16420/j.issn.0513-353x.2020-0520

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (3): 505-517.doi: 10.16420/j.issn.0513-353x.2020-0520

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Expression Characteristic Analysis of the BcPG17,a Gene Related to Inflorescence Stem Development of Brassica campestris

LÜ Meiling¹, CAO Jiashu²^,^*()

¹College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou 350002,China
²Laboratory of Cell & Molecular Biology,Institute of Vegetable Science,Zhejiang University,Hangzhou 310058,China

Received:2020-07-08 Online:2021-03-25 Published:2021-04-02
Contact: CAO Jiashu E-mail:jshcao@ziu.edu.cn

Abstract

Abstract:

In the present study,a PG gene BcPG17 and its promoter were cloned from Brassica campestris L. ssp. chinensis Makino‘Aijiaohuang’inbred-line Bcajh97-01. Gene expression and promoter activity were explored by qRT-PCR,in situ hybridization and transient transformation of promoter-GUS fusion. The genomic DNA fragment of BcPG17 was 2 015 bp in length,containing nine exons and eight introns. The ORF of BcPG17 was 1 344 bp in length,encoding 447 amino acids. The predicted molecular weight and the theoretical pI of BcPG17 were 48.61 kD and 8.39,respectively. The protein contained a transmembrane domain with a signal peptide sequence at the N-terminus,indicating that the protein may be associated with the cell membrane. Analysis of the amino acid sequence indicated the presence of four typical domains of PG protein,closely relating to other species of Brassica. The highest expression level of BcPG17 was found in the inflorescence stem during flowering,then to a less level in the tetrad stage and uninucleate microspore stage of pollen development. The results of in situ hybridization showed that BcPG17 had a strong signal in all tissues of inflorescence stems during flowering. The 1 442 bp promoter contained at least one cis-acting element related to meristem expression, multiple motifs or elements related to plant hormone response,and four anther-specific motifs,as well as two tapetum-degeneration- retardation（TDR）binding sites. Moreover,transient expression of the promoter-GUS showed strong signal in the second internode and node of the inflorescence stem during flowering,and there was also a significant GUS signal in the anthers in the middle development stage of flower. The above results illustrated that the BcPG17 may regulate the elongation of the inflorescence stem by interacting with other proteins,presumably related to plant hormone metabolism,and it may be involved in the development of the pollen wall by the regulation of TDR protein.

Key words: Brassica campestris, BcPG17, polygalacturonase, inflorescence stem, expression analysis

CLC Number:

S634.3

LÜ Meiling, CAO Jiashu. Expression Characteristic Analysis of the BcPG17,a Gene Related to Inflorescence Stem Development of Brassica campestris[J]. Acta Horticulturae Sinica, 2021, 48(3): 505-517.

Figures/Tables 11

References 56

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物种（基因名称） Species（gene name）			GenBank登录号 GenBank No.
拟南芥Arabidopsis thaliana（PGA3）			NP_187439.1
拟南芥Arabidopsis thaliana（PGA2）			CAA51692.1
白菜Brassica campestris（BcMF2）			ABW24665.1
白菜Brassica campestris（BcMF6）			ACP74159.1
白菜Brassica campestris（BcMF9）			ABN13878.1
白菜Brassica campestris（BcMF16）			ADJ68232.1
油菜Brassica napus（RDPG）			CAA65072.1
油菜Brassica napus（PGAZ）			CAC05658.1
甜瓜Cucumis melo（MPG2）			AAC26511.1
陆地棉Gossypium hirsutum（G9）			AAA82167.1
荔枝Litchi chinensisv（LcPG1）			AFW04075.1
番茄Solanum lycopersicum（TAPG1）			AAC28903.1
番茄Solanum lycopersicum（TAPG2）			AAC28904.1
烟草Nicotiana tabacum（PG1）			Q05967.1
桃Prunus persica（PRF5）			CAA54150.1

物种（基因名称） Species（gene name）			GenBank登录号 GenBank No.
拟南芥Arabidopsis thaliana（PGA3）			NP_187439.1
拟南芥Arabidopsis thaliana（PGA2）			CAA51692.1
白菜Brassica campestris（BcMF2）			ABW24665.1
白菜Brassica campestris（BcMF6）			ACP74159.1
白菜Brassica campestris（BcMF9）			ABN13878.1
白菜Brassica campestris（BcMF16）			ADJ68232.1
油菜Brassica napus（RDPG）			CAA65072.1
油菜Brassica napus（PGAZ）			CAC05658.1
甜瓜Cucumis melo（MPG2）			AAC26511.1
陆地棉Gossypium hirsutum（G9）			AAA82167.1
荔枝Litchi chinensisv（LcPG1）			AFW04075.1
番茄Solanum lycopersicum（TAPG1）			AAC28903.1
番茄Solanum lycopersicum（TAPG2）			AAC28904.1
烟草Nicotiana tabacum（PG1）			Q05967.1
桃Prunus persica（PRF5）			CAA54150.1

顺式作用元件 cis-acting element	序列 Sequence	功能 Function	数量 Number
CAAT-box	CAAAT	启动子和增强子区域常见的顺式作用元件 Common cis-acting element in promoter and enhancer regions	8
TATA-box	TATAAAT/TATA	转录起始位点周围-30左右的核心启动子元件 Core promoter element around-30 of transcription start	44
TCA-element	CCATCTTTTT	水杨酸响应相关的顺式作用元件 cis-acting element involved in salicylic acid responsiveness	2
ABRE	ACGTG	脱落酸响应相关的顺式作用元件 cis-acting element involved in the abscisic acid responsiveness	1
TGACG/CGTCA-motif	TGACG/CGTCA	茉莉酸甲酯响应相关的顺式作用调节元件 cis-acting regulatory element involved in the MeJA-responsiveness	4
GARE-motif	TCTGTTG	赤霉素响应元件 Gibberellin-responsive element	1
CAT-box	GCCACT	与分生组织表达有关的顺式调控元件 cis-acting regulatory element related to meristem expression	1
ARE	AAACCA	厌氧诱导的关键顺式作用调节元件 cis-acting regulatory element essential for the anaerobic induction	2
G-Box	CACGTT	参与光响应的顺式作用调节元件 cis-acting regulatory element involved in light responsiveness	1
Box 4	ATTAAT	光响应相关的保守DNA模块的一部分 Part of a conserved DNA module involved in light responsiveness	4
Gap-box	CAAATGAA(A/G)A	光响应元件的一部分 Part of a light responsive element	1
TCT-motif	TCTTAC	光响应元件的一部分 Part of a light responsive element	1
AT1-motif	AATTATTTTTTATT	光响应元件的一部分 Part of a light responsive element	1
AE-box	AGAAACAA	光响应元件的一部分 Part of a light responsive element	1
GTGA-motif	GTGA	花药特异基序 Anther specific motif	4
E-box	CATTG	绒毡层降解延迟蛋白（TDR）结合位点 Tapetum Degeneration Retardation（TDR）binding site	2

顺式作用元件 cis-acting element	序列 Sequence	功能 Function	数量 Number
CAAT-box	CAAAT	启动子和增强子区域常见的顺式作用元件 Common cis-acting element in promoter and enhancer regions	8
TATA-box	TATAAAT/TATA	转录起始位点周围-30左右的核心启动子元件 Core promoter element around-30 of transcription start	44
TCA-element	CCATCTTTTT	水杨酸响应相关的顺式作用元件 cis-acting element involved in salicylic acid responsiveness	2
ABRE	ACGTG	脱落酸响应相关的顺式作用元件 cis-acting element involved in the abscisic acid responsiveness	1
TGACG/CGTCA-motif	TGACG/CGTCA	茉莉酸甲酯响应相关的顺式作用调节元件 cis-acting regulatory element involved in the MeJA-responsiveness	4
GARE-motif	TCTGTTG	赤霉素响应元件 Gibberellin-responsive element	1
CAT-box	GCCACT	与分生组织表达有关的顺式调控元件 cis-acting regulatory element related to meristem expression	1
ARE	AAACCA	厌氧诱导的关键顺式作用调节元件 cis-acting regulatory element essential for the anaerobic induction	2
G-Box	CACGTT	参与光响应的顺式作用调节元件 cis-acting regulatory element involved in light responsiveness	1
Box 4	ATTAAT	光响应相关的保守DNA模块的一部分 Part of a conserved DNA module involved in light responsiveness	4
Gap-box	CAAATGAA(A/G)A	光响应元件的一部分 Part of a light responsive element	1
TCT-motif	TCTTAC	光响应元件的一部分 Part of a light responsive element	1
AT1-motif	AATTATTTTTTATT	光响应元件的一部分 Part of a light responsive element	1
AE-box	AGAAACAA	光响应元件的一部分 Part of a light responsive element	1
GTGA-motif	GTGA	花药特异基序 Anther specific motif	4
E-box	CATTG	绒毡层降解延迟蛋白（TDR）结合位点 Tapetum Degeneration Retardation（TDR）binding site	2

Expression Characteristic Analysis of the BcPG17,a Gene Related to Inflorescence Stem Development of Brassica campestris

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