Cloning and Expression Analysis of the Cytokinin Response Regulator VlRR5 in Kyoho Grapevine

doi:10.16420/j.issn.0513-353x.2020-0668

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (8): 1437-1445.doi: 10.16420/j.issn.0513-353x.2020-0668

• Research Papers • Next Articles

Cloning and Expression Analysis of the Cytokinin Response Regulator VlRR5 in Kyoho Grapevine

BIAN Lu, GUO Dalong, YU Keke, WEI Tonglu, PEI Maosong, LIU Hainan, YU Yihe()

College of Horticulture and Plant Protection,Henan University of Science and Technology,Luoyang,Henan 471023,China

Received:2021-02-23 Revised:2021-06-07 Online:2021-08-25 Published:2021-09-06
Contact: YU Yihe E-mail:yuyihe@haust.edu.cn

Abstract

Abstract:

There are three fruit-dropping peaks in grape growth and development. Grape fruit-dropping phenomenon is more serious in Kyoho grapevine. In this study,a cytokinin response regulator VlRR5 was cloned from Kyoho grapevine,which has a phosphoric acid receptor domain（REC）and a MYB-like DNA binding domain. Sequence analysis revealed that cDNA of VlRR5 was 2 597 bp in length,encoding 693 amino acids,and located on chromosome 4 of grape. Subcellular localization assay showed that VlRR5 was localized in the nucleus. The yeast transformed into pGBKT7-VlRR5 could grow normally and turn blue on SD/-Trp/-Ade/-His/+X-α-Gal deficient medium,which proved that VlRR5 was a transcription factor with transcriptional activation activity. By quantitative real-time PCR analysis,VlRR5 was characterized by tissue specific expression and showed high expression level in leaf and stem,which was increased and then decreased in the development stage of young fruit. The expression level increased after treatment with exogenous cytokinin,and decreased after treatment with the cytokinin synthesis inhibitor lovastatin. These results indicated that VlRR5 can respond to cytokinin and plays an important role in participating in cytokinin to improve the fruiting rate of grape.

Key words: grapevine, cytokinin, CPPU, cytokinin response regulators, VlRR5

CLC Number:

S663.1

BIAN Lu, GUO Dalong, YU Keke, WEI Tonglu, PEI Maosong, LIU Hainan, YU Yihe. Cloning and Expression Analysis of the Cytokinin Response Regulator VlRR5 in Kyoho Grapevine[J]. Acta Horticulturae Sinica, 2021, 48(8): 1437-1445.

Figures/Tables 9

References 19

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引物名称基因 Primer	碱基序列（5'-3'） Primer sequence
QT-VlRR5	F：TGGGTATGCATTGCCTTTCT;R：GAAGAAGGAAGCTCTGCGTTT
VlRR5	F：ATGACAGTTGAACCGAGGGT;R：TTAATGGAAAAGAACTTGAT
Y1H-RR5	F：ATGGCCATGGAGGCCGAATTCATGACAGTTGAACCGAGGGT;
Y1H-RR5	R：ATGCGGCCGCTGCAGGAATTCTTAATGGAAAAGAACTTGAT
LS-RR5	F：AGAGAACACGGGGGACTCTAGAATGACAGTTGAACCGAGGGT;
LS-RR5	R：CTTCTCCCTTACCCATGGTACCTTAATGGAAAAGAACTTGAT

引物名称基因 Primer	碱基序列（5'-3'） Primer sequence
QT-VlRR5	F：TGGGTATGCATTGCCTTTCT;R：GAAGAAGGAAGCTCTGCGTTT
VlRR5	F：ATGACAGTTGAACCGAGGGT;R：TTAATGGAAAAGAACTTGAT
Y1H-RR5	F：ATGGCCATGGAGGCCGAATTCATGACAGTTGAACCGAGGGT;
Y1H-RR5	R：ATGCGGCCGCTGCAGGAATTCTTAATGGAAAAGAACTTGAT
LS-RR5	F：AGAGAACACGGGGGACTCTAGAATGACAGTTGAACCGAGGGT;
LS-RR5	R：CTTCTCCCTTACCCATGGTACCTTAATGGAAAAGAACTTGAT

顺式作用元件 cis-acting element	功能 Function	序列 Sequence	数量 Amount
gGTAAAGAAA	脱落酸响应Abscisic acid responsiveness	ACGTG	1
ARE	厌氧诱导Anaerobic induction	AAACCA	3
AuxRR-core	生长素响应Auxin responsiveness	GGTCCAT	1
CAT-box	分生组织表达Meristem expression	GCCACT	1
CGTCA-motif	茉莉酸甲酯响应MeJA-responsiveness	CGTCA	2
G-box	光响应Light responsiveness	TACGTG	2
MBS	参与干旱诱导的MYB结合位点MYB binding site involved in drought-inducibility	CAACTG	9
Pc-CMA2a	光响应元件的一部分Part of a light responsive element	CAGCCAATCACAG	1
TATA-box	在转录开始-30左右的核心启动子元件 Core promoter element around-30 of transcription start	TATA	5
TCA-element	水杨酸响应Salicylic acid responsiveness	TCAGAAGAGG	1
TGACG-motif	茉莉酸甲酯响应MeJA-responsiveness	TGACG	2
CAAT-box	启动子和增强子区域的顺式作用元件 Common cis-acting element in promoter and enhancer regions	CAAAT	49

顺式作用元件 cis-acting element	功能 Function	序列 Sequence	数量 Amount
gGTAAAGAAA	脱落酸响应Abscisic acid responsiveness	ACGTG	1
ARE	厌氧诱导Anaerobic induction	AAACCA	3
AuxRR-core	生长素响应Auxin responsiveness	GGTCCAT	1
CAT-box	分生组织表达Meristem expression	GCCACT	1
CGTCA-motif	茉莉酸甲酯响应MeJA-responsiveness	CGTCA	2
G-box	光响应Light responsiveness	TACGTG	2
MBS	参与干旱诱导的MYB结合位点MYB binding site involved in drought-inducibility	CAACTG	9
Pc-CMA2a	光响应元件的一部分Part of a light responsive element	CAGCCAATCACAG	1
TATA-box	在转录开始-30左右的核心启动子元件 Core promoter element around-30 of transcription start	TATA	5
TCA-element	水杨酸响应Salicylic acid responsiveness	TCAGAAGAGG	1
TGACG-motif	茉莉酸甲酯响应MeJA-responsiveness	TGACG	2
CAAT-box	启动子和增强子区域的顺式作用元件 Common cis-acting element in promoter and enhancer regions	CAAAT	49

Cloning and Expression Analysis of the Cytokinin Response Regulator VlRR5 in Kyoho Grapevine

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