Cloning and Preliminary Functional Analysis of NtTGAL11,a Transcription Factor Gene Involved in Stamen Development of Narcissus tazetta var.chinensis

doi:10.7668/hbnxb.20196617

Abstract

Abstract:

It aimed to elucidate the molecular regulatory mechanism underlying stamen development in Chinese narcissus.Based on our group's previous transcriptome data from stamens of the Chinese narcissus cultivar Jinzhanyintai from the Narcissus Research Group led by Ma Yanjun,a TGAL11 homologous sequence that is specifically and highly expressed in stamens was identified.This gene was cloned and designated NtTGAL11.Sequence analysis showed that the full-length coding sequence of NtTGAL11 was 1 020 bp,encoding a protein of 339 amino acids with a molecular mass of 38.072 31 ku.NtTGAL11 was a basic,hydrophilic protein predicted to be predominantly composed of αhelices and random coils,and it contained both a bZIP domain and a DOG1 domain.Phylogenetic analysis revealed that NtTGAL11 was most closely related to AtTGA9 and AtTGA10.Subcellular localization assays confirmed that NtTGAL11 was targeted to the nucleus.RT-qPCR analysis demonstrated that NtTGAL11 was highly expressed in the stamens of Jinzhanyintai but was expressed at low levels in other floral organs,as well as in the petaloid stamens of the double-flowered cultivar Yulinglong,consistent with the transcriptomic data.Bimolecular fluorescence complementation(BiFC)assays further showed that NtTGAL11 physically interacted with the AGAMOUS-class MADS-box protein NtMADS6.Taken together,these findings suggest that NtTGAL11 might promote stamen development by modulating the spatiotemporal expression of AG-like genes,and its expression pattern underscores a key regulatory role in stamen morphogenesis in Chinese narcissus.

Key words: Narcissus tazetta var.chinensis, Stamen development, TGAL, Bioinformatics, BiFC

CLC Number:

S682.2+
1

ZHAO Pengda, HU Tao, ZHANG Na, YANG Yuting, CHANG Yanting, JIANG Zehui, MA Yanjun. Cloning and Preliminary Functional Analysis of NtTGAL11,a Transcription Factor Gene Involved in Stamen Development of Narcissus tazetta var.chinensis[J]. Acta Agriculturae Boreali-Sinica, 2026, 41(5): 74-82. doi: 10.7668/hbnxb.20196617.

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Figures/Tables 10

Tab.1 Primer sequence

引物 Primers	序列(5'—3') Sequences(5'—3')
NtTGAL11	F:ATGGCCACACAAAATCATGAGCTC
	R:GAAGGCTGAAAATTGGCCGTG
PHG-NtTGAL11-EGFP	F:TCTCTCTCTCAAGCTATGGCCACACAAAATCATGAGCT
	R:CGGGTCATGAGCTCCGAAGGCTGAAAATTGGCCGTG
YNE-NtTGAL11	F:CTAGATCGGGAATTCATGGCCACACAAAATCATGAGCT
	R:ATGCAAGCTAAGCTTGAAGGCTGAAAATTGGCCGTG
YCE-NtMADS6	F:CTAGATCGGGAATTCATGGGTAGGGGGAAGATAGAGATCAAA
	R:ATGCAAGCTAAGCTTTCCCAGTTGAAGGGTAGTCTGC

Fig.1 Agarose gel electrophoresis of NtTGAL11 gene amplification products M.2 000 bp DNA Marker;1,2. NtTGAL11 PCR products.

Fig.2 Prediction result of hydrophilicity of NtTGAL11 protein

Fig.3 Prediction of secondary structure(A)and tertiary structure(B)of NtTGAL11 protein

Fig.4 Prediction result of phosphorylation sites in NtTGAL11 protein

Fig.5 Conserved domain analysis of the NtTGAL11 protein

Fig.6 Phylogenetic tree(A)and multiple sequence alignment(B)of NtTGAL11 and 10 TGA proteins from A.thaliana

Fig.7 Subcellular localization of NtTGAL11 protein in tobacco cells

Fig.8 BiFC assay for detecting the interaction of NtTGAL11 with NtMADS6

Fig.9 The relative expression levels of NtTGAL11 in different tissues Data are presented as means±s(n=3 biological replicates).Different lowercase letters indicate significant differences(P<0.05).

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