蓝莓VcTPS/VcTPP基因家族鉴定及VcTPS24对花诱导的功能分析

doi:10.3864/j.issn.0578-1752.2025.13.013

Abstract

Abstract:

【Objective】 Trehalose-6-phosphate synthase (TPS), as a key enzyme in the trehalose synthesis pathway, is involved in regulating various developmental process such as germination, plant growth, flowering, and stress resistance. In this study, genes encoding TPS and TPP in the blueberry trehalose synthesis pathway were identified. We analyzed the molecular mechanism of VcTPS24 in regulating floral induction through molecular biology methods, laying the foundation for revealing the sugar signaling pathway regulating blueberry floral induction. 【Method】 Bioinformatics methods was used to analyze the number of amino acid, gene structure, conserved motifs, promoter sequences, and subcellular localization of VcTPS/VcTPP family members in the trehalose metabolism pathway of blueberries (Vaccinium corymbosum L.). The expression characteristics of VcTPS class I genes during blueberry floral differentiation were analyzed through qRT-PCR experiments. We analyzed the expression of VcTPS24 in blueberries under hormone and low temperature stress by exogenous application of gibberellin, auxin and other hormones, and 4 ℃ low temperature treatment. The regulatory effects of VcSOC1 and VcSVP on the VcTPS24 promoter were analyzed by dual luciferase assay. We validated the function of VcTPS24 in regulating growth and floral induction by overexpressing it in tobacco. 【Result】 A total of 28 VcTPS and 19 VcTPP family members were identified from blueberry, which can be divided into two subfamilies. VcTPS family members encoding proteins ranging from 465 to 1 382 amino acids, with an average isoelectric point (pI) of 6.02. Most members have 3 exon structures. VcTPP family members encoding proteins ranging from 241 to 434 amino acids, with an average isoelectric point (pI) of 7.48. Most members have 11 exon structures. Cis acting elements such as G-Box, P-Box, LTR, and circadian, which are involved in light, hormone, stress response, and regulation of plant growth, were found in the promoter sequences of VcTPS/VcTPP family members. Members of the VcTPS/VcTPP family are mostly located in chloroplasts and vacuoles. The expression level of VcTPS class I genes significantly increased during blueberry floral induction, and the expression level of VcTPS24 was significantly higher through the entire period of flower bud differentiation than in the undifferentiated group. Subcellular localization assay showed that the expression of VcTPS24 can be significantly upregulated in blueberry seedlings by exogenous IAA, GA₃, SA, MeJA, and low temperature stress. The dual luciferase assay showed that the promoter activity of VcTPS24 was significantly increased by VcSOC1, while it was significantly decreased by VcSVP. VcTPS24 is located in the nucleus and cytoplasm. Under the condition of no exogenous sucrose supply, the germination and radicle elongation of tobacco seeds overexpressing VcTPS24 were promoted, but excessive sucrose hindered the growth of tobacco seedlings overexpressing VcTPS24. The expression level of VcTPS24 was upregulated under short day condition. Under both long and short day conditions, the relative expression levels of NtLFY, NtFUL, and NtSOC1 in overexpressing VcTPS24 tobacco lines were all increased to varying degrees, and the formation time of their inflorescences was significantly earlier than that of the wild type. This indicates that VcTPS24 is involved in the regulation of floral induction. 【Conclusion】 A total of 28 blueberry VcTPS and 19 VcTPP family members were identified, and the VcTPS class I gene played an important role during blueberry floral induction. The expression of VcTPS24 was influenced by exogenous hormones, and the overexpression of VcTPS24 can promote tobacco flowering, which may be related to the upregulated expression of flowering genes.

Key words: blueberry (Vaccinium corymbosum L.), trehalose, dual luciferase reporter assay, trehalose-6-phosphate synthase, floral induction

WU XinLiang, WU HuiLing, ZHANG SuiLin, CAO Man, FENG Xin, ZHOU BingJie, HOU ZhiXia. Identification of the VcTPS/VcTPP Gene Family in Blueberries and Functional Analysis of VcTPS24 in Floral Induction[J].Scientia Agricultura Sinica, 2025, 58(13): 2663-2681.

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Figures/Tables 12

Table 1

Basic information of the VcTPS/VcTPP family in Vaccinium corymbosum"

蛋白质 Protein	基因组名称 Gene ID	分子量 Molecular weight (kDa)	理论等电点 Theoretical isoelectric point (pI)	氨基酸数量 No. of amino acid (aa)	脂肪族指数 Aliphatic index (AI)	总平均亲水性 Grand average of hydropathicity (GRAVY)	亚细胞定位预测 Prediction of subcellular localization
VcTPS1	VaccDscaff1-augustus-396.21	109711.99	6.79	973	87.65	-0.269	叶绿体Chloroplast
VcTPS2	VaccDscaff2-augustus-264.26	101361.67	6.11	896	93.28	-0.140	叶绿体Chloroplast
VcTPS3	VaccDscaff3-augustus-173.14	101383.72	6.08	896	93.72	-0.132	叶绿体Chloroplast
VcTPS4	VaccDscaff4-augustus-204.23	96344.97	5.96	857	84.47	-0.228	液泡Vacuole
VcTPS5	VaccDscaff4-augustus-305.15	96296.98	5.78	852	87.85	-0.238	细胞质Cytoplasm
VcTPS6	VaccDscaff5-snap-373.29	103449.36	6.66	921	84.14	-0.346	叶绿体Chloroplast
VcTPS7	VaccDscaff6-snap-157.32	104767.98	6.07	922	84.89	-0.281	叶绿体Chloroplast
VcTPS8	VaccDscaff6-augustus-415.25	97533.97	5.90	865	90.75	-0.164	叶绿体Chloroplast
VcTPS9	VaccDscaff8-snap-53.38	102274.98	6.57	911	84.63	-0.333	叶绿体Chloroplast
VcTPS10	VaccDscaff9-augustus-192.33	51250.97	5.47	465	76.49	-0.247	液泡Vacuole
VcTPS11	VaccDscaff9-augustus-279.19	96393.19	5.90	852	87.73	-0.232	细胞质Cytoplasm
VcTPS12	VaccDscaff10-processed-46.5	98352.52	6.59	875	82.77	-0.370	细胞质Cytoplasm
VcTPS13	VaccDscaff14-snap-228.37	97032.44	6.18	857	91.52	-0.187	叶绿体Chloroplast
VcTPS14	VaccDscaff20-augustus-148.22	96872.05	5.50	856	90.93	-0.217	细胞质Cytoplasm
VcTPS15	VaccDscaff22-processed-180.0	155153.79	6.04	1382	85.17	-0.385	液泡Vacuole
VcTPS16	VaccDscaff25-snap-274.23	97028.45	6.18	857	91.4	-0.190	叶绿体Chloroplast
VcTPS17	VaccDscaff28-augustus-174.15	96922.17	5.50	856	90.48	-0.215	细胞质Cytoplasm
VcTPS18	VaccDscaff35-augustus-209.24	96292.13	5.85	852	87.85	-0.222	叶绿体Chloroplast
VcTPS19	VaccDscaff36-processed-141.5	96279.97	5.85	852	87.73	-0.230	细胞质Cytoplasm
VcTPS20	VaccDscaff37-snap-73.33	104704.9	6.07	992	85.31	-0.276	叶绿体Chloroplast
VcTPS21	VaccDscaff37-augustus-296.26	97625.08	5.96	865	90.18	-0.172	叶绿体Chloroplast
VcTPS22	VaccDscaff38-augustus-13.19	97520.93	5.95	865	90.42	-0.174	叶绿体Chloroplast
VcTPS23	VaccDscaff38-augustus-206.34	100031.5	5.78	883	85.78	-0.229	叶绿体Chloroplast
VcTPS24	VaccDscaff38-augustus-295.59	106154.43	6.77	946	84.41	-0.346	叶绿体Chloroplast
VcTPS25	VaccDscaff39-augustus-6.34	97603.07	5.93	865	90.64	-0.169	叶绿体Chloroplast
VcTPS26	VaccDscaff39-augustus-233.28	100186.69	5.78	883	85.66	-0.225	叶绿体Chloroplast
VcTPS27	VaccDscaff44-augustus-176.31	96872.05	5.50	856	90.93	-0.217	细胞质Cytoplasm
VcTPS28	VaccDscaff46-augustus-60.18	97060.12	5.93	863	91.32	-0.169	液泡Vacuole
VcTPP1	VaccDscaff13-augustus-12.21	42711.95	8.84	386	84.40	-0.257	液泡Vacuole
VcTPP2	VaccDscaff19-snap-26.29	48525.61	6.21	434	85.32	-0.257	叶绿体Chloroplast
VcTPP3	VaccDscaff20-snap-26.33	42818.92	6.61	383	81.67	-0.327	叶绿体Chloroplast
VcTPP4	VaccDscaff21-augustus-85.22	43152.42	8.81	384	79.95	-0.351	叶绿体Chloroplast
VcTPP5	VaccDscaff22-augustus-142.24	44825.27	6.73	396	88.13	-0.395	叶绿体Chloroplast
VcTPP6	VaccDscaff23-augustus-237.19	42680.54	9.08	387	72.82	-0.445	液泡Vacuole
VcTPP7	VaccDscaff26-augustus-82.30	42608.65	5.69	380	82.32	-0.292	叶绿体Chloroplast
VcTPP8	VaccDscaff26-augustus-83.16	43024.25	8.50	382	80.86	-0.334	叶绿体Chloroplast
VcTPP9	VaccDscaff28-snap-330.33	42814.92	6.84	383	82.19	-0.329	液泡Vacuole
VcTPP10	VaccDscaff29-augustus-42.42	39118.42	7.12	346	77.51	-0.496	液泡Vacuole
VcTPP11	VaccDscaff29-augustus-84.21	44352.86	8.88	394	81.88	-0.338	叶绿体Chloroplast
VcTPP12	VaccDscaff33-augustus-254.24	43199.47	8.90	384	79.69	-0.364	叶绿体Chloroplast
VcTPP13	VaccDscaff35-snap-140.24	26586.25	4.61	241	80.08	-0.033	叶绿体Chloroplast
VcTPP14	VaccDscaff37-augustus-226.21	42763.88	8.64	387	81.37	-0.281	叶绿体Chloroplast
VcTPP15	VaccDscaff38-snap-84.29	42550.6	8.65	385	80.78	-0.285	液泡Vacuole
VcTPP16	VaccDscaff39-snap-90.9	42535.59	8.64	385	80.78	-0.291	叶绿体Chloroplast
VcTPP17	VaccDscaff43-augustus-226.19	44804.23	6.53	396	87.88	-0.394	液泡Vacuole
VcTPP18	VaccDscaff46-augustus-22.31	44840.24	6.29	396	88.13	-0.394	液泡Vacuole
VcTPP19	VaccDscaff48-snap-89.34	42832.87	6.61	383	81.93	-0.337	叶绿体Chloroplast

Table 1

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Fig. 5

Fig. 6

Fig. 7

Fig. 8

Table 2

Fig. 9

Fig. 10

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处理 Treatment	叶长 Length of leaf (cm)	叶宽 Width of leaf (cm)	株高 Plant height (cm)	节间长 Internode length (cm)
OE-长日照 LD	16.0±1.5	10.5±1.0	28.6±2.5	2.4±0.3
OE-短日照 SD	19.5±1.0	10.0±1.0	16.5±1.0	2.0±0.1
YS-长日照 LD	20.0±1.5	11.5±1.5	17.3±2.0	1.8±0.1
YS-短日照 SD	15.5±1.5	11.0±1.0	9.5±1.5	1.5±0.5

Identification of the VcTPS/VcTPP Gene Family in Blueberries and Functional Analysis of VcTPS24 in Floral Induction

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