Analysis of the B3 transcription factor alternative splicer in foxtail millet (Setaria italica L.)

doi:10.3969/j.issn.1004-1524.20231216

Acta Agriculturae Zhejiangensis ›› 2024, Vol. 36 ›› Issue (9): 1969-1976.DOI: 10.3969/j.issn.1004-1524.20231216

• Crop Seience • Previous Articles Next Articles

Analysis of the B3 transcription factor alternative splicer in foxtail millet (Setaria italica L.)

LIU Hui¹(), WANG Xiaomeng¹, YAN Liuyan¹, WANG Yongfang², YANG Pengjuan¹, GONG Keke¹, LI Xingjie¹, DONG Zhiping², JIA Xiaoping¹^,^*()

1. College of Agriculture, Henan University of Science and Technology, Luoyang 471023, Henan, China
2. National Millet Improvement Center, Institute of Millet, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050035, China

Received:2023-10-31 Online:2024-09-25 Published:2024-09-30

Abstract

Abstract:

The alternative splicing types of B3 transcriptional factor gene in different millet varieties were analyzed to identify the splicing variants with functional activity. Furthermore, the correlation between these splicing variants and heading stage was analyzed, aiming at providing evidence for further revealing the regulatory effects of B3 transcription factor on growth and development of millet by alternative splicing. Totally 12 millet varieties with different heading stages were selected to clone B3 transcriptional factor gene belonging to REM family by reverse transcription PCR (RT-PCR). The alternative splicing types of this gene were identified by multiple sequence alignment, and the activity of the splicing variants, their association with heading stage were analyzed. The results showed that a total of 8 splicing variants of B3 transcriptional factor gene were produced among the 12 millet varieties, of which splicing variant Ⅴ contained the most varieties (4), followed by splicing variant Ⅳ (2), and the remaining 6 splicing variants each contained only 1 variety. By alternative splicing, the B3 transcriptional factor gene of 12 varieties encoded 10 types of proteins, of which 5 types lacked the B3 DNA binding domain, and type Ⅱ protein shortened the heading stage of millet, while type Ⅵ protein prolonged the heading stage of millet, which was not affected by geographical environments. The above results showed that B3 transcription factor participated in the regulation of heading stage through alternative splicing in millet.

Key words: foxtail millet, B3 transcription factor, alternative splicing, heading stage, geographic environment

CLC Number:

S515

LIU Hui, WANG Xiaomeng, YAN Liuyan, WANG Yongfang, YANG Pengjuan, GONG Keke, LI Xingjie, DONG Zhiping, JIA Xiaoping. Analysis of the B3 transcription factor alternative splicer in foxtail millet (Setaria italica L.)[J]. Acta Agriculturae Zhejiangensis, 2024, 36(9): 1969-1976.

Figures/Tables 6

Table 1 Information of specific primers

引物名称 Primer name	正向引物(5’→3’) Forward primer (5’→3’)	反向引物(5’→3’) Reverse primer (5’→3’)	片段大小 Fragment size/bp
引物1 Primer 1	GTGAGGGAGGGAGAAATCG	GCTAAGTAAGTCAGGCTGAAACA	907
引物2 Primer 2	GAGAAATCGAGGGAGAAAAT	CTACCTTCCTGCTAAGACTGC	707

Fig.1 Electrophoresis results of total RNA from different foxtail millet varieties M, DL 2 000 DNA marker; 1-12 were Zheng 8041, Anfeng, Shawan Guzi, Gulyu Zao 1, Datou Nuo, Xiaozao Gu, Jinde, Da Kingdom, Hohhot Damao Gu, An 5424, Yugu 13, and Zaopiyibaqi, respectively. The same as below.

Fig.2 Electrophoresis results of PCR amplification products of B3 transcription factor genes from different foxtail millet varieties

Fig.3 cDNA sequences alignment result of B3 transcriptional factor gene in 12 millet varieties Ⅰ-Ⅶ represented different types of variable spliceosomes. The numbers 1-12 on the right correspond to the millet varieties Zheng 8041, Anfeng, Shawanguzi, Gulyuzao 1, Datounuo, Xiaozaogu, Jinde, Da Kingdom, Huhehaotedamaogu, An 5424, Yugu 13, and Zaopiyibaqi.

Fig.4 Protein sequences alignment result of B3 transcriptional factor in 12 millet varieties The numbers 1-12 on the left correspond to the millet varieties Zheng 8041, Anfeng, Shawanguzi, Gulyuzao 1, Datounuo, Xiaozaogu, Jinde, Da Kingdom, Huhehaotedamaogu, An 5424, Yugu 13, and Zaopiyibaqi.

Fig.5 Heading days among millet varities with different B3 protein types

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Analysis of the B3 transcription factor alternative splicer in foxtail millet (Setaria italica L.)

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