Cloning and Promotor Analysis of DfMYB3 from Dendrocalamus farinosus

doi:10.7525/j.issn.1673-5102.2021.05.011

Abstract

Abstract:

Based on the transcriptome database of Dendrocalamus farinosus， a MYB transcription factor named DfMYB3 was cloned from the leave of D. farinosus. Its open reading frame length was 1 287 bp， encoding 428 amino acids， and GenBank accession Number was KY963358. The conserved domain analysis showed that DfMYB3 had the typical SANT domains and DNA-Binding domains. Phylogenetic tree analysis showed that DfMYB3 clustered with R2R3-MYB transcription factors in Saccharum， Arabidopsis and Populus. Subcellular localization showed that DfMYB3 protein was expressed in both the nucleus and cell membrane，and was more significant in the nucleus. The sequence of 5′ flanking of DfMYB3 gene about 2 000 bp was cloned by chromosome walking method. Analysis of PlantCARE online software showed that the sequence had typical promoter characteristics， and contained hormones responsive elements such as GA， ABA， MeJA as well as stress responsive elements such as drought. The expression level of DfMYB3 gene was significantly up-regulated after 100 μmol·L^-1 GA and 100 μmol·L^-1 ABA treatment， indicating that DfMYB3 gene was involved in response to GA and ABA treatments. In order to explore the function of DfMYB3 promoter， the expression vector of GUS gene fused with DfMYB3 promoter was constructed and genetically transformed into tobacco. The results showed that GUS signal was detected in both leaf and stem of transgenic tobacco， especially in the veins.

Key words: Dendrocalamus farinosus, MYB transcription factor, promotor, GA and ABA treatments, GUS staining

CLC Number:

S795.9

Tian-Tian DU, Hui-Ping LI, Bo-Ya WANG, Qian OU, Yan HUANG, Ying CAO, Shang-Lian HU. Cloning and Promotor Analysis of DfMYB3 from Dendrocalamus farinosus[J]. Bulletin of Botanical Research, 2021, 41(5): 729-737.

Figures/Tables 8

Table 1

Fig.1

Fig.2

Fig.3

Table 2

Element analysis of DfMYB3 promoter

元件 Site name	物种 Organism	位置 Position	链 Strand	评分 Matrix score	序列 Sequence	功能 Function
ABRE	拟南芥Arabidopsis thaliana	705	+	5	ACGTG	cis-acting element involved in the abscisic acid responsiveness
ABRE	大麦Hordeum vulgare	976	_	9	CGTACGTGCA	cis-acting element involved in the abscisic acid responsiveness
AuxRR-core	烟草Nicotiana tabacum	537	-	7	GGTCCAT	cis-acting regulatory element involved in auxin responsiveness
Box 4	欧芹Petroselinum crispum	96	+	6	ATTAAT	part of a conserved DNA module involved in light responsiveness
CGTCA-motif	大麦Hordeum vulgare	191	+	5	CGTCA	cis-acting regulatory element involved in the MeJA-responsiveness
CGTCA-motif	大麦Hordeum vulgare	813	-	5	CGTCA	cis-acting regulatory element involved in the MeJA-responsiveness
CGTCA-motif	大麦Hordeum vulgare	604	-	5	CGTCA	cis-acting regulatory element involved in the MeJA-responsiveness
CGTCA-motif	大麦Hordeum vulgare	854	-	5	CGTCA	cis-acting regulatory element involved in the MeJA-responsiveness
G-Box	豌豆Pisum sativum	704	-	6	CACGTT	cis-acting regulatory element involved in light responsiveness
GARE-motif	甘草Brassica oleracea	1017	-	7	TCTGTTG	gibberellin-responsive element
GATA-motif	拟南芥Arabidopsis thaliana	1574	-	7	GATAGGA	part of a light responsive element
GT1-motif	拟南芥Arabidopsis thaliana	673	-	6	GGTTAA	light responsive elemen
MBS	拟南芥Arabidopsis thaliana	841	+	6	CAACTG	MYB binding site involved in drought-inducibility
MBS	拟南芥Arabidopsis thaliana	1045	+	6	CAACTG	MYB binding site involved in drought-inducibility
P-box	水稻Oryza sativa	775	-	7	CCTTTTG	gibberellin-responsive element
TCA-element	烟草Nicotiana tabacum	1108	-	9	CCATCTTTTT	cis-acting element involved in salicylic acid responsiveness
TGACG-motif	大麦Hordeum vulgare	191	-	5	TGACG	cis-acting regulatory element involved in the MeJA-responsiveness
TGACG-motif	Hordeum vulgare	813	+	5	TGACG	cis-acting regulatory element involved in the MeJA-responsiveness
TGACG-motif	Hordeum vulgare	604	+	5	TGACG	cis-acting regulatory element involved in the MeJA-responsiveness
TGACG-motif	Hordeum vulgare	854	+	5	TGACG	cis-acting regulatory element involved in the MeJA-responsiveness

Table 2

Fig.4

Fig.5

Fig.6

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引物名称 Primer name	引物序列（5′→3′） Primer sequences（5′→3′）
DfMYB3-F	ATGGGGAGGCATTCTTGTTG
DfMYB3-R	CTAGATATGCTCAAAAGATA
RT-DfMYB3-F	AAGCAACGGTATGGTATTCGAGCCA
RT-DfMYB3-R	GCACATATCCCTTCCATGTTGAATT
Tublin-F	GCCGTGAATCTCATCCCCTT
Tublin-R	TTGTTCTTGGCATCCCACAT
DfMYB3-F（KpnⅠ）	GGGGTACCCCATGGGGAGGCATTCTTGTTG
DfMYB3-R（HindⅢ）	CCCAAGCTTGGGGATATGCTCAAAAGATA
P3	TTGCTTGTAGCAACAAGAATGC
P2	TGTTCCCAAGGACAGCATGGAG
P1	TTTCAGGGCGATTCTTATCATA
pDfMYB3-F（HindⅢ）	CCAAGCTTGGTATCGAGTTTCGTGTTCCCAAG
pDfMYB3-R（XbaI）	GCTCTAGAGCTTCGGTGGAATGCACAACCTGA

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