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Acta Agronomica Sinica ›› 2021, Vol. 47 ›› Issue (4): 761-769.doi: 10.3724/SP.J.1006.2021.01046

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Transcription characteristics of wheat glutamine synthetase isoforms and the sequence analysis of their promoters

WANG Xiao-Chun1,2(), WANG Lu-Lu1, ZHANG Zhi-Yong1, QIN Bu-Tan1, YU Mei-Qin2, WEI Yi-Hao1, MA Xin-Ming1,*()   

  1. 1Collaborative Innovation Center of Henan Grain Crops, Henan Agricultural University, Zhengzhou 450002, Henan, China
    2College of Life Science, Henan Agricultural University, Zhengzhou 450002, Henan, China
  • Received:2020-06-03 Accepted:2020-10-17 Online:2021-04-12 Published:2020-11-02
  • Contact: MA Xin-Ming E-mail:xiaochun.w@163.com;xinmingma@126.com
  • Supported by:
    National Key Research and Development Program of China(2016YFD0300205);Modern Agricultural Technology System in Henan Province(S2010-01-G04)

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Abstract:

As a key enzyme for nitrogen assimilation in wheat, glutamine synthetase is grouped into two classes: cytosolic GS and chloroplastic GS (TaGS2), and cytosolic GS includes TaGS1, TaGSr, and TaGSe. In order to study the expression characteristics and regulatory mechanisms of GS isozymes in chromosome A, B, and D of heterohexaploid wheat, transcripts of TaGS isoforms were analyzed based on the third-generation sequencing technology transcriptome analysis, and 12 promoters of TaGS isozymes of Yumai 49 were cloned based on Chinese Spring genome, and the sequence of the promoters were analyzed. The results showed that TaGS1 was mainly transcribed on chromosome 6B, TaGSe and TaGSr on chromosome 4D, and TaGS2 on chromosome 2D. Furthermore, the distance from initiation codon ATG to initiation site of transcript for each promoter of TaGS was distinct. Promoter element analysis showed that the promoter of TaGS1 in 6B had more W-box, AC-I, ABRE, as-1, and methyl jasmonic response elements, the promoter of TaGSe in 4D had more stress response elements (MYB, MBS, LTR, etc.) and auxin response element, the promoter of TaGSr in 4D had more WRE3 and transcript factor response elements, the promoter of TaGS2 in 2D had more A-box, WRE3, ARE, and an AT enrichment region. In summary, the number, type and order of cis-elements of different promoters of TaGS isozymes were distinct, which provided the foundation for further study on the regulation mechanism of TaGS isozymes.

Key words: wheat, GS isoforms, transcription, promoter, cis-element

Fig. 1

Expression of TaGS isozymes on different chromosomes"

Table 1

Primers of TaGS isoenzymes for promoter amplification"

引物名称
Primer name		 引物序列
Primer sequence (5′-3′)		 退火温度
Annealing temperature (℃)		 PCR长度
PCR fragment (bp)		 5′端非编码区 5′NCS (bp)
GS1-6AL S CACTGCCTTCTCAGGCTTGTTA 58 2000 1594
GS1-6AL A GCCCCATGAATTTAGCATCG
GS1-6BL S TCTTAGTCCAACGTGGCATCATAGG 62 2033 1548
GS1-6BL A AGACGCCAAACTACGACTACG
GS1-6DL S CACATTCACACGTGGTTTCCTCT 58 2261 1470
GS1-6DL A GCAAAAACCTCTTCTCGTCGC
GSe-4AL S TCACCCATAATCATGCTGTGCAAAT 63 2538 1557
GSe-4AL A TACCATATGTACTCGGCGATGATCTTG
GSe-4BL A ATGGGAGGTTGGGACAGTGC 58 2508 1565
GSe-4BL S GATGCGAAGCATACGCGC
GSe-4DS S TCTGAAGATGCTCATGAGTTGCTCAAT 62 2536 1557
GSe-4DS A TACCATATGTACTCGGCGATGATCTTG
GSr-4AS S GTAATGTGGCTACGGTGTGAGTCTGTT 61 2155 1548
GSr-4AS A GGGTGCAGAATGCAAGTAGC
GSr-4BL S TCATCTCCGTGAGGAGGTCTTGA 62 2308 1615
GSr-4BL A TACTCGACGATGATCTTGTCGGTG
GSr-4DL S GCCCGTTGTACCACTGTTATCGAT 62 1624 1467
GSr-4DL A TACTCGACGATGATCTTGTCGGTG
GS2-2AL S GCATTCCTGGCTTGCATGTCTAGA 58 2008 1499
GS2-2AL A ATGATCTTGTCGGTGAAGGGC
GS2-2BL S ATTTCGCCGAGTTCAGGTCGAG 63 2153 1419
GS2-2BL A ATGATCTTGTCGGTGAAGGGC
GS2-2DL S CAGCGATCCTAGCCTTCGAAAAG 60 1878 1097
GS2-2DL A ATGATCTTGTCGGTGAAGGGC

Fig. 2

Identify PCR amplification of TaGS isoenzyme promoters on different chromosomes The arrows are the pointed to the target segment. M: marker."

Fig. 3

Identify recombinant plasmids of TaGS isoenzyme promoters by PCR"

Fig. 4

Types and distribution of cis-elements in different TaGS isozyme promoters Different colors represent different types of response elements; color and shape represent a specific response element. Red: hormone response element; Green: unknown function response element; Blue: basic element; Black: transcription factor binding site; Purple: stress response element; Pink: tissue specific response element; Yellow: light response element."

Fig. 5

Prediction of transcription initiation site of different TaGS isozyme TATA box is the RNA polymerase II recognition site."

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