Bioinformatics and expression analysis of flowering regulation gene BrFT in Brassica rapa L.

doi:10.3969/j.issn.1004-1524.2023.05.03

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (5): 992-1000.DOI: 10.3969/j.issn.1004-1524.2023.05.03

• Crop Science • Previous Articles Next Articles

Bioinformatics and expression analysis of flowering regulation gene BrFT in Brassica rapa L.

YAO Yanlin¹^,²(), MA Li², LIU Lijun², PU Yuanyuan¹^,², LI Xuecai¹^,², WANG Wangtian³, FANG Yan¹^,², SUN Wancang¹^,², WU Junyan¹^,²^,^*()

1. Rapeseed Engineering Research Center of Gansu Province/College of Agronomy, Gansu Agricultural University, Lanzhou 730070, China
2. Gansu Provincial Key Laboratory of Aridland Crop Science, Lanzhou 730070, China
3. College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China

Received:2022-03-10 Online:2023-05-25 Published:2023-06-01

Abstract

Abstract:

FT gene is an important gene associated with flowering in plants. In order to explore the role of FT gene in regulating flowering in photoperiod pathway of Brassica rapa L. In this study, 3 BrFT genes were identified from the winter rapeseed genome using Arabidopsis FT genes as queries,bioinformatics methods were used to analyze the gene sequence characteristics, and the organ specific expression of the gene was studied by qRT-PCR. Screening genes were cloned from different Brassica rapa L., and the expression patterns of genes under 4 ℃, 16 h light/8 h darkness conditions were detected. The results showed that 3 of which contained 4 exons and 3 introns, and distributed on chromosomes A02 and A07, among which BrFT1 and BrFT2 had obvious collinear relationship, and all the three BrFT proteins contained PEBP conserved motif. The 2 000 bp upstream promoter region of the gene all contained light response elements,some regulatory elements involved in anaerobic induction, defense and stress response,and BrFT1 and BrFT2 contain action elements involved in plant growth and development and hormone induction. qRT-PCR detection showed that BrFT could be expressed in roots, stems, leaves, flower buds and flowers,and the expression level in leaves was higher than that in other organs, the expression level of BrFT1 was higher than that of the other two members. So the BrFT1 gene in Longyou 6, Longyou 17, Tianyou 2 were cloned and the sequence alignment similarity was 99.5%. BrFT1 in Brassica rapa L. showed an up-regulation trend at 4 ℃, 16 h light/8 h darkness, but there were differences among varieties. Combined with the observation of flowering in different types of Brassica rapa L. with different temperature, it showed that excessive expression of BrFT1 gene could promote the early flowering of Brassica rapa L.

Key words: Brassica rapa L., FT gene, bioinformatics analysis, gene clone, expression analysis

CLC Number:

S565.4

YAO Yanlin, MA Li, LIU Lijun, PU Yuanyuan, LI Xuecai, WANG Wangtian, FANG Yan, SUN Wancang, WU Junyan. Bioinformatics and expression analysis of flowering regulation gene BrFT in Brassica rapa L.[J]. Acta Agriculturae Zhejiangensis, 2023, 35(5): 992-1000.

Figures/Tables 11

References 31

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引物名称	引物序列	引物用途
The name of the primer	Primer sequence (5'→3')	Use of primers
BrFT1-F	ATGTCTTTAAGTAATAGAGATCCTCTTGTGGT	基因克隆
BrFT1-R	CTAACTTCTTCGTCCTCCGCAG	Gene clone
BrFT1-F	ACGAGAGTCCAAGGCCCAAC	实时荧光定量PCR
BrFT1-R	TGGCGCCATCCTGGTTCATA	Real-time fluorescence quantitative PCR
BrFT2-F	TCCCTGCGACAACTGGAACAAAC	实时荧光定量PCR
BrFT2-R	CGGAACAATACCAGCACGAGACG	Real-time fluorescence quantitative PCR
BrFT3-F	AGGTTCAACACTCGTGAGTTCGC	实时荧光定量PCR
BrFT3-R	CTTCTTCCTCCGCAGCCATTCTC	Real-time fluorescence quantitative PCR
L17-BrFT1-F	ACGAGAGTCCAAGGCCCAAC	实时荧光定量PCR
L17-BrFT1-R	TGGCGCCATCCTGGTTCATA	Real-time fluorescence quantitative PCR
Actin-F	TGTGCCAATCTACGAGGGTTT	实时荧光定量PCR
Actin-R	TTTCCCGCTCGGCTGTTGT	Real-time fluorescence quantitative PCR

引物名称	引物序列	引物用途
The name of the primer	Primer sequence (5'→3')	Use of primers
BrFT1-F	ATGTCTTTAAGTAATAGAGATCCTCTTGTGGT	基因克隆
BrFT1-R	CTAACTTCTTCGTCCTCCGCAG	Gene clone
BrFT1-F	ACGAGAGTCCAAGGCCCAAC	实时荧光定量PCR
BrFT1-R	TGGCGCCATCCTGGTTCATA	Real-time fluorescence quantitative PCR
BrFT2-F	TCCCTGCGACAACTGGAACAAAC	实时荧光定量PCR
BrFT2-R	CGGAACAATACCAGCACGAGACG	Real-time fluorescence quantitative PCR
BrFT3-F	AGGTTCAACACTCGTGAGTTCGC	实时荧光定量PCR
BrFT3-R	CTTCTTCCTCCGCAGCCATTCTC	Real-time fluorescence quantitative PCR
L17-BrFT1-F	ACGAGAGTCCAAGGCCCAAC	实时荧光定量PCR
L17-BrFT1-R	TGGCGCCATCCTGGTTCATA	Real-time fluorescence quantitative PCR
Actin-F	TGTGCCAATCTACGAGGGTTT	实时荧光定量PCR
Actin-R	TTTCCCGCTCGGCTGTTGT	Real-time fluorescence quantitative PCR

基因	基因ID	开放阅读框	氨基酸长度	分子量	等电点	不稳定系数	亲水性	α-螺旋	β-折叠	延伸链	无规则卷曲
Gene	Gene ID	Open reading frame/bp	Amino acid length/aa	Molecular weight/ku	pI	Unstable coefficient	Hydro- philicity	Alpha helix/%	Beta turn/%	Extended strand/%	Random coil/%
BrFT1	Brapa02T001728	528	175	19.81	7.75	46.66	-0.34	14.86	3.43	22.86	58.85
BrFT2	Brapa02T003364	528	175	19.77	7.82	38.39	-0.34	17.14	4.00	21.71	57.15
BrFT3	Brapa02T004252	531	176	19.93	7.72	38.47	-0.29	13.64	3.98	25.57	56.81

基因	基因ID	开放阅读框	氨基酸长度	分子量	等电点	不稳定系数	亲水性	α-螺旋	β-折叠	延伸链	无规则卷曲
Gene	Gene ID	Open reading frame/bp	Amino acid length/aa	Molecular weight/ku	pI	Unstable coefficient	Hydro- philicity	Alpha helix/%	Beta turn/%	Extended strand/%	Random coil/%
BrFT1	Brapa02T001728	528	175	19.81	7.75	46.66	-0.34	14.86	3.43	22.86	58.85
BrFT2	Brapa02T003364	528	175	19.77	7.82	38.39	-0.34	17.14	4.00	21.71	57.15
BrFT3	Brapa02T004252	531	176	19.93	7.72	38.47	-0.29	13.64	3.98	25.57	56.81

时间 Time/d	开花株率 Flowering plant rate/%
时间 Time/d	天油2号 Tianyou 2	陇油6号 Longyou 6	陇油17号 Longyou 17	陇油7号 Longyou 7
0	0	0	0	0
20	46.8	15.6	23.2	13.3
30	88.9	26.6	46.7	39.6
40	98.3	40.0	73.3	46.6
50	100	73.3	93.3	71.6
60	100	86.1	93.3	86.6
70	100	95.4	97.1	94.3

Bioinformatics and expression analysis of flowering regulation gene BrFT in Brassica rapa L.

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