Identification of a novel eIF4E1 allele in tobacco and the development and application of its codominant molecular marker

doi:10.3969/j.issn.1004-1524.20250419

Acta Agriculturae Zhejiangensis ›› 2026, Vol. 38 ›› Issue (7): 1327-1335.DOI: 10.3969/j.issn.1004-1524.20250419

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Identification of a novel eIF4E1 allele in tobacco and the development and application of its codominant molecular marker

WANG Rengang¹(), LIN Shifeng¹^,^*(), WANG Zili¹, YU Shizhou¹, ZHANG Jie¹, YU Jing¹, LONG Mingjin¹, LI Li²

¹ Key Laboratory of Molecular Genetics of CNTC, Guizhou Academy of Tobacco Science, Guiyang 550081, China
² School of Life Sciences, Guizhou Normal University, Guiyang 550025, China

Received:2025-02-18 Online:2026-07-25 Published:2026-08-20

Abstract

Abstract:

To fully explore and effectively utilize the excellent alleles of the recessive resistance gene eIF4E1 against Potato virus Y (PVY) in tobacco, taking the PVY resistant sun-cured tobacco variety, Panxiandaliuye, as the research object, allelism test and sequence analysis were performed to finalize the allelic relationship of its resistance gene with eIF4E1 and the gene mutation type. A co-dominant molecular marker based on bi-directional PCR amplification of specific alleles (Bi-PASA) was developed for detecting the mutant eIF4E1 gene in Panxiandaliuye. The results showed that the resistance to PVY in Panxiandaliuye was referable to the mutation of eIF4E1 gene. There was a 2-bp deletion in position 267-268 in the coding region cDNA sequence of eIF4E1 gene in Panxiandaliuye, which caused a frame-shift mutation and resulted in a premature stop codon in exon 2. A codominant molecular marker was developed for the novel eIF4E1 mutant allele in Panxiandaliuye, which could effectively distinguish the genotypes of resistant and susceptible parents and their hybrid offsprings. In summary, a novel eIF4E1 allele with a two base pair deletion was discovered for the first time in the sun-cured tobacco Panxiandaliuye, and an ideal codominant molecular marker been developed for molecular marker-assisted selection for PVY resistance in tobacco.

Key words: sun-cured tobacco, Potato virus Y, eIF4E1, novel allele, codominant molecular marker

CLC Number:

S572

WANG Rengang, LIN Shifeng, WANG Zili, YU Shizhou, ZHANG Jie, YU Jing, LONG Mingjin, LI Li. Identification of a novel eIF4E1 allele in tobacco and the development and application of its codominant molecular marker[J]. Acta Agriculturae Zhejiangensis, 2026, 38(7): 1327-1335.

Figures/Tables 10

Table 1 Primers used in the test

用途 Usage	引物名称 Primer name	引物序列(5'→3') Primer sequence(5'→3')
eIF4E1基因5'端显性分子标记	CF2	TTTGGTTTGATAATCCTATGGCT
Dominant molecular marker at the 5'-end of eIF4E1 gene	GR11	GAAGGCAAGATATTCAGGAGCT
eIF4E1基因3'端显性分子标记	3'-F	AATTCGTGCCACGTTAGGGC
Dominant molecular marker at the 3'-end of eIF4E1 gene	3'-R	TTGAGTCGTCCTGCAATTTGC
eIF4E1基因全长序列扩增	eIF4E1-F	CAAGTACCCTTTTCCTACTAAAATCTATAACTAAG
Full-length amplification of eIF4E1 gene	eIF4E1-R	GGCATTAATTCAAAACCAAACGAG
盘县大柳叶eIF4E1新等位基因共显性分子标记	A	TCTGCAAACAGTGTTTACAATAATAgAC
Codominant molecular marker for the novel eIF4E1 allele in Panxiandaliuye	B	AACTAACTTGCTTGGGTGGcTG
	P	TGTCTGCCTTAGATGCATGTTG
	Q	CGGACGCTAACTACTGCTCCAC

Fig.1 Resistance identification of tobacco to PVY

Fig.2 Molecular marker detection of eIF4E1 gene in Panxiandaliuye A, Detection results of molecular marker at 5' end; b, Detection results of molecular marker at 3' end; M, DL 2 000 DNA marker; 1, TN90; 2, Honghuadajinyuan; 3, Panxiandaliuye.

Fig.3 PCR amplification products of eIF4E1 gene in Panxiandaliuye M1, DL 2 000 DNA marker; M2, DL 10 000 DNA marker; 1, Amplification products using cDNA as template; 2, Amplification products using genomic DNA as template.

Fig.4 Comparison of eIF4E1 gene of Panxiandaliuye and Honghuadajinyuan and genomic DNA sequence The premature termination codon (TAA) in the eIF4E1.Pan gene is marked with red box.

Fig.5 Primer design for bidirectional PCR amplification of specific alleles

Fig.6 Amplification products of genomic DNA of resistant and susceptible materials by Bi-PASA genetic marker primers A, PCR amplification with primers P and B; B, PCR amplification with primers A and Q; C, PCR amplification with primers P, B, A and Q; M, DL 2 000 marker; 1, Panxiandaliuye; 2, Honghuadajinyuan×Panxiandaliuye; 3, K326×Panxiandaliuye; 4, Guiyan 5×Panxiandaliuye; 5, Honghuadajinyuan; 6, K326; 7, Guiyan 5.

Fig.7 Amplification of Bi-PASA genetic marker in F2 population and parents M, DL 2 000 marker; P1, Honghuadajinyuan; P2, Panxiandaliuye; 1-22, F2 individuals; R, Resistant individual; S, Susceptible individual.

Fig.8 Amplification of Bi-PASA genetic markers in some individuals of backcross progenies M, DL 2 000 marker; P1, Honghuadajinyuan; P2, Panxiandaliuye; 1-22, Backcross population.

Fig.9 Schematic comparison between dominant molecular marker assisted selection breeding (A) and co-dominant molecular marker assisted selection breeding (B) for transferring single recessive genes

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Identification of a novel eIF4E1 allele in tobacco and the development and application of its codominant molecular marker

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