Analysis of the PotatoGARPTranscription Factor Family and Functional Validation of StARR14-X1

doi:10.13560/j.cnki.biotech.bull.1985.2025-0701

Abstract

Abstract:

Objective To identify key genes regulating anthocyanin accumulation in the tubers of colored potatoes (Solanum tuberosum L.), provide genetic resources and technical references for potato molecular breeding. Method GARP transcription factors were screened based on transcriptome sequencing data of purple potato tubers at different developmental stages. Bioinformatics analysis was conducted, and functional verification of StARR14-X1 gene related to anthocyanin regulation was performed. Result The 12 GARP transcription factors were screened through transcriptome data. The amino acid sequence lengths of 12 GARP transcription factors were concentrated between 148-686 aa, the molecular weight ranged from 16.66 to 75.44 kD, the pI value ranged from 5.13 to 8.05, the fatty index ranged from 71.36 to 97.36, and the instability coefficient ranged from 35.59 to 75.94, all of which belonged to hydrophilic proteins. Subcellular localization prediction showed that 12 GARP transcription factors were distributed in the nucleus, cytoplasm and chloroplasts. The StARR14-X1 gene was successfully transformed into tetraploid tobacco using the Agrobacterium-mediated leaf disc method, and transgenic tobacco plants overexpressing StARR14-X1 were obtained. The anthocyanin contents in the stems and petals of StARR14-X1-overexpressing transgenic lines increased significantly. Conclusion StARR14-X1 gene has a positive regulatory function in anthocyanin accumulation and promotes anthocyanin accumulation.

Key words: potato, anthocyanin, GARP transcription factor, positive regulation

WU Xiao-juan, ZHAO Xue-wen, WANG Pei-jie, NIE Hu-shuai, LI Nan, MA Yu, YANG En-ze, MA Jie-ru, MA Yan-hong. Analysis of the PotatoGARPTranscription Factor Family and Functional Validation of StARR14-X1[J]. Biotechnology Bulletin, 2026, 42(6): 22-30.

Figures/Tables 11

Fig. 1 Genetic transformation process diagram of StARR14-X1 transgenic tobaccoA: Infected leaves. B: Callus culture. C: Induction of shoot differentiation. D: Rooting culture

Table 1 Physicochemical properties and subcellular localization of potato GARP transcription factors

基因 Gene name	氨基酸数Number of amino acids （aa）	分子量 Molecular weight （Da）	理论等电点 PI	不稳定系数 Instability index	总平均亲水性 Gravy	脂肪族指数 Aliphatic index	亚细胞定位 Subcellular localization
StORR24-like-1	686	75 439.30	6.36	43.01	-0.627	71.36	细胞核
StORR9	226	25 635.06	5.38	63.55	-0.611	85.35	细胞核
StORR9-like	226	25 628.02	5.13	67.28	-0.620	84.91	细胞核
StARR5-like	248	27 532.26	5.29	75.94	-0.458	80.52	细胞核
StARR1-like	424	46 469.10	5.92	54.21	-0.501	74.29	细胞核
StORR10-like	163	18 701.66	6.74	40.77	-0.290	97.36	细胞核
StARR2-like	675	74 652.52	5.93	45.56	-0.564	74.64	细胞核
StARR5	202	22 405.65	5.38	58.08	-0.373	93.07	细胞核
StARR17-like	148	16 656.27	6.08	35.59	-0.345	86.89	细胞质
StORR24-like-2	669	74 026.52	6.78	48.79	-0.693	71.69	细胞核
StARR14-X1	394	42 295.94	8.05	62.75	-0.298	83.15	叶绿体、细胞核
StARR11	481	54 100.85	6.39	44.27	-0.592	72.43	细胞核

Fig. 2 Relative positions of motifs in potato (Nicotiana tabacum) GARP transcription factors

Fig. 3 Conserved motifs in potato GARP transcription factors

Table 2 Functional prediction of 12 GARP transcription factors

基因名称 Gene name	GO 注释的基因名称 Gene name of GO annotation	GO 编号 GO ID	GO 信息 GO term
StORR24-like-1	PGSC0003DMG400007823	GO：0031537	花青素代谢过程的调节
StORR9	PGSC0003DMG400029839	GO：0000156	磷酸响应调节因子活性
StORR9-like	PGSC0003DMG400029852	GO：0000156	磷酸响应调节因子活性
StARR5-like	PGSC0003DMG402022640	GO：0006351	DNA模板的转录
StARR1-like	PGSC0003DMG400023534	GO：0031537	花青素代谢过程的调节
StORR10-like	PGSC0003DMG400003084	GO：0005634	细胞核
StARR2-like	PGSC0003DMG401025624	GO：0016049	细胞生长
StARR5	PGSC0003DMG400024578	GO：0005515	蛋白质结合
StARR17-like	PGSC0003DMG400016643	GO：0006351	DNA模板的转录
StORR24-like-2	PGSC0003DMG400003196	GO：0005634	细胞核
StARR14-X1	PGSC0003DMG400029514	GO：0031537	花青素代谢过程的调节
		GO：0048364	根的发育
StARR11	PGSC0003DMG400011980	GO：0003677	DNA结合

Fig. 4 Correlation coefficient graph of transcriptome samplesThe correlation coefficient r is highly positive and negative correlation between ± 1 and ± 0.8. The redder the color, the stronger the correlation; the whiter the color, the weaker the correlation

Fig. 5 Expression patterns of 12 GARP genes in three development stages of Huasong 66The tuber formation stage is the mean value of three replicates (S11, S12, S13); the tuber expansion stage is the mean value of three replicates (S21, S22, S23); and the tuber maturation stage is the mean value of three replicates (S31, S32, S33). The Maiwei Cloud Platform was used for normalization of genes at the sample level, and the normalized values range from -2 to 2

Fig. 6 Expressions analysis of 8 GARP transcription factors in three periods of different potato varietiesS1: Tuber formation stage. S2: Tuber expansion stage. S3: Tuber maturity stage. Different lowercase letters indicate significant differences at 0.05 level (Duncan test). The same below

Fig. 8 Relative expressions of the roots, stems, leaves and flowers in wild-type and transgenic plantsA, B, C, and D are the relative expressions in the roots, stems, leaves, and flowers, respectively. WT refers to the wild-type plant; OE1, OE4, OE5, OE8 are four different lines of transgenic tobacco with StARR14-X1 gene, the same below

Fig. 9 Plant morphology of wild-type tobacco and transgenic tobacco overexpressing StARR14-X1

Fig. 10 Anthocyanin contents in wild-type and transgenic plants

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