高温胁迫下结薯因子StSP6A对马铃薯试管块茎形成的影响

doi:10.3864/j.issn.0578-1752.2026.18.002

Abstract

Abstract:

【Objective】High temperatures, as a major abiotic stress factor, affect potato tuber formation. Exploring the mechanisms underlying potato tuber formation under high-temperature stress holds significant theoretical value for elucidating the intrinsic mechanisms by which environmental factors regulate tuber development.【Method】Using in vitro seedlings from 103 potato germplasm accessions as experimental material, the seedlings were subjected to either optimal temperature (17 ℃) or high temperature (28 ℃) in plant growth chamber for 150 days. After collecting and analyzing the phenotypic traits of the seedlings, representative germplasm accessions were selected for transcriptomic analysis, which was validated using real-time quantitative PCR (RT-qPCR). Furthermore, following the construction of an SP6A overexpression vector, genetic transformation was performed using Longshu 10. After subjecting the transgenic material to high-temperature treatment, phenotypic evaluation and molecular-level analysis were conducted.【Result】After treatment at 17 ℃ and 28 ℃, 103 accessions were classified into four temperature response patterns: widespread tuber-forming type (GroupⅠ, 46 accessions; tubers formed at both temperatures), heat-sensitive type (GroupⅡ, 33 accessions; tubers formed only at 17 ℃), high temperatures /high sugar induced sensitive type (GroupⅢ, 11 accessions; tubers formed only at 28 ℃), and non-tuber-forming (GroupⅣ, 13 accessions; no tubers formed at either temperature). Furthermore, a variance analysis indicated that high-temperature stress shortened the number of days to tuber formation, reduced plant height, and significantly decreased the weight and number of test-tube tubers. At the same time, the coefficients of variation for those indicators all increased significantly. Transcriptome analysis revealed that Longshu 11 (which tuberizes only at 17 ℃) and Lily (which tuberizes only at 28 ℃) had 3 398 and 2 197 differentially expressed genes, respectively. with the differentially expressed genes in Longshu 11 focused on molecular functions such as catalytic activity, as well as metabolic processes and pathways, while those in Lily were enriched in cell wall remodeling and carbohydrate metabolism pathways. A combined transcriptomic analysis revealed that the two groups shared 730 differentially expressed genes, which were classified into eight categories based on their expression patterns. Among the 14 key differentially expressed genes, the tuber-specific gene SP6A is the primary factor responsible for the differences in tuber formation in in vitro tubers under high-temperature stress. An increased number of microtubers was observed in the overexpression plants (SP6A-OE) under 17 ℃ treatment. Under 28 ℃ heat treatment, although the formation of in vitro tubers was inhibited in both the SP6A-OE and control, the SP6A-OE line still produced more in vitro tubers. Western blot results indicated that the expression level of the SP6A protein in SP6A-OE plants was lower at 28 ℃ than at 17 ℃. Meanwhile, RT-qPCR results showed that SP6A expression was suppressed at high temperatures, whereas the expression levels of both SP5G and HSFA6b were significantly elevated at 28 ℃.【Conclusion】Under high-temperature stress, different potato germplasm exhibit varying tuber-forming habits. Overexpression of StSP6A alleviates the inhibitory effect caused by heat stress on axillary tuber formation, thereby enhancing the ability of in-vitro seedlings to form tubers under high-temperature conditions.

Key words: potato, high temperature stress, SELF-PRUNING 6A (StSP6A), genetic modification, in vitro tubers

DING Ning, LI QingQuan, XU Jing, VLADIMIR N. Obraztsov, JIA XiaoXia, HUANG Wei, QI EnFang. The Effect of the Tuber-Specific Protein StSP6A on in vitro Potato Tuber Formation Under High-Temperature Stress[J].Scientia Agricultura Sinica, 2026, 59(18): 3973-3988.

Figures/Tables 9

Table 1

Table 2

Phenotypic characteristics of 103 potato germplasms under high-temperature stress"

品种名称 Variety name	品种编号 Serial	结薯天数 Days to tuber formation (d)		株高 Plant height (cm)		块茎鲜重 Weight (g)		试管薯数目 No. of tubers per plant
品种名称 Variety name	品种编号 Serial	17 ℃	28 ℃	17 ℃	28 ℃	17 ℃	28 ℃	17 ℃	28 ℃
S. demissum	S1	42.11	70.85	6.05	3.60	1.17	0.29	1.33	2.00
Сюрприз	S2	48.24	61.58	13.73	8.10	2.43	2.36	3.33	1.67
Лад	S3	50.58	97.57	4.80	9.73	0.07	0.73	0.33	1.00
云薯701 Yunshu 701	S4	45.42	87.52	9.87	5.60	3.58	0.64	3.00	1.00
冀张薯12号Jizhangshu 12	S5	115.23	98.25	13.45	12.12	4.00	3.36	2.00	2.67
大西洋Atlantic	S6	42.24	45.62	13.20	10.32	2.28	1.62	3.33	1.33
费乌瑞它Favorita	S7	76.08	133.95	15.67	11.73	1.76	0.79	3.00	1.00
东农310 Dongnong 310	S8	91.24	133.43	12.47	5.10	5.13	1.33	2.67	0.33
Десятка	S9	107.86	133.52	16.17	7.30	2.11	0.27	3.00	0.67
天薯14号Tianshu 14	S10	42.48	70.11	12.80	7.27	1.31	0.79	2.67	0.67
S.stenotomum	S11	87.54	76.51	12.05	10.42	1.38	1.77	3.00	2.33
Мастак	S12	104.46	45.57	14.48	6.28	8.26	1.05	3.33	2.67
Волат	S13	61.13	45.10	8.35	10.87	1.48	3.48	1.67	1.67
Лель	S14	83.27	42.52	7.83	12.57	2.20	2.17	2.67	4.00
Бриз	S15	42.58	42.78	9.90	7.60	2.10	3.65	3.33	2.33
Вилия	S16	45.75	42.62	6.30	10.35	2.80	1.76	2.67	1.67
Фальварак	S17	61.24	42.24	8.47	10.40	2.99	1.84	3.33	1.00
陇薯10号Longshu 10	S18	87.33	50.11	9.60	14.53	3.08	2.84	2.67	3.00
Палац	S19	87.31	48.82	9.37	11.53	1.27	0.94	1.33	0.33
Jenь	S20	87.43	52.23	13.57	9.47	1.34	1.32	2.33	1.67
Уладар	S21	52.00	45.75	6.17	10.37	3.02	2.00	3.00	3.00
Максимум	S22	70.45	45.21	9.70	11.57	2.59	4.26	2.33	2.67
Гарантия	S23	45.23	45.50	8.43	10.13	2.54	1.77	3.00	2.00
Водар	S24	61.12	50.85	13.20	13.60	3.62	5.24	2.67	3.00
中薯7号Zhongshu 7	S25	70.24	45.12	11.50	8.80	2.05	0.53	2.00	0.33
云薯103 Yunshu 103	S26	87.85	45.24	11.63	8.60	0.60	1.41	1.67	1.33
云薯901 Yunshu 901	S27	70.28	42.12	14.08	8.57	1.97	1.22	0.67	2.00
云薯505 Yunshu 505	S28	87.33	45.24	10.73	12.80	4.03	0.83	2.33	1.33
云薯104 Yunshu 104	S29	76.45	45.45	12.50	9.30	1.45	1.97	0.33	0.33
云薯601 Yunshu 601	S30	87.87	52.42	10.60	8.03	0.67	1.18	2.33	1.33
甘农薯7号Gannongshu 7	S31	87.52	50.20	10.97	8.50	3.22	2.96	1.33	2.00
甘农薯9号Gannongshu 9	S32	70.47	45.30	10.80	9.83	3.21	4.98	2.00	1.33
红美Hongmei	S33	87.86	42.12	13.27	11.27	3.06	3.70	2.67	2.00
维拉斯Velas	S34	91.59	45.25	15.20	11.10	1.13	2.03	2.00	1.33
青薯7号Qingshu7	S35	91.42	42.11	14.07	8.75	2.74	1.99	2.67	0.67
京张薯1号Jingzhangshu 1	S36	97.27	57.57	14.28	9.07	1.67	4.80	2.33	2.00
陇薯4号Longshu 4	S37	70.21	42.85	14.03	13.13	3.88	3.12	3.33	0.67
陇薯12号Longshu 12	S38	76.35	76.82	12.83	8.73	2.91	0.40	2.00	0.67
陇薯14号Longshu 14	S39	70.24	50.41	11.10	10.20	3.79	2.73	2.00	2.67
陇薯17号Longshu 17	S40	70.01	42.57	9.60	6.05	1.04	0.71	1.33	0.67
陇薯19号Longshu 19	S41	87.25	42.89	13.07	6.93	4.24	0.15	2.67	0.67
陇薯23号Longshu 23	S42	83.36	45.12	17.55	7.50	0.62	0.19	2.67	0.33
中薯3号Zhongshu 3	S43	70.24	42.35	10.20	10.03	1.71	1.16	3.33	0.67
天薯10号Tianshu 10	S44	133.04	70.15	13.33	8.51	1.54	2.03	3.67	1.67
新大坪Xindaping	S45	87.85	50.54	12.13	11.52	2.59	4.77	3.33	2.33
底西芮Desiree	S46	83.12	44.74	8.40	15.03	3.42	0.19	3.67	0.33
陇薯3号Longshu 3	A1	133.58	-	8.75	8.95	0.45	-	0.45	-
陇薯6号Longshu 6	A2	104.48	-	11.63	9.63	2.77	-	1.67	-
陇薯11号Longshu 11	A3	52.59	-	14.81	9.81	4.56	-	3.33	-
陇薯18号Longshu 18	A4	98.62	-	14.10	9.67	1.07	-	0.33	-
陇薯20号Longshu 20	A5	87.51	-	8.60	7.19	0.76	-	1.33	-
天薯11号Tianshu 11	A6	70.63	-	8.20	6.90	2.31	-	2.00	-
天薯12号Tianshu 12	A7	91.44	-	7.53	6.07	2.81	-	3.00	-
天薯17号Tianshu 17	A8	45.54	-	11.37	6.35	2.50	-	2.33	-
S.pinnatisecta	A9	45.85	-	15.85	13.95	0.07	-	0.67	-
Красавик	A10	76.24	-	8.73	15.87	3.03	-	2.67	-
云薯102 Yunshu102	A11	42.55	-	17.13	9.50	2.96	-	3.67	-
云薯801 Yunshu801	A12	91.47	-	12.10	8.74	2.37	-	3.00	-
冀张薯3号Jizhangshu 3	A13	42.59	-	10.20	9.14	2.67	-	3.00	-
冀张薯14号Jizhangshu 14	A14	133.25	-	13.01	5.82	0.76	-	1.33	-
黑金刚Heimeiren	A15	115.28	-	4.70	1.37	0.17	-	2.00	-
天薯19号Tianshu 19	A16	50.89	-	11.10	1.47	0.36	-	1.00	-
青薯2号Qingshu 2	A17	70.52	-	10.60	7.35	2.43	-	1.67	-
青薯9号Qingshu 9	A18	45.63	-	14.77	8.81	1.82	-	2.67	-
陇薯25号Longshu 25	A19	45.41	-	16.20	10.23	1.43	-	2.33	-
陇薯29号Longshu 29	A20	42.05	-	9.40	5.82	3.12	-	3.33	-
露辛达v7	A21	133.82	-	12.07	9.82	1.68	-	1.00	-
S.microdonatum	A22	115.00	-	11.08	15.75	0.21	-	0.33	-
早大白Zaodabai	A23	87.28	-	12.07	9.58	2.44	-	3.33	-
陇薯27号Longshu 27	A24	87.24	-	12.63	8.99	3.90	-	3.00	-
甘农薯2号Gannongshu 2	A25	61.85	-	13.84	12.69	2.97	-	2.33	-
希森6号Xisen 6	A26	52.95	-	7.06	6.12	5.38	-	3.00	-
青薯168 Qingshu 168	A27	45.20	-	16.77	13.95	5.83	-	2.67	-
陇薯5号Longshu 5	A28	133.25	-	13.47	8.43	1.40	-	2.33	-
陇薯7号Longshu 7	A29	70.65	-	11.90	10.80	5.91	-	3.00	-
陇薯15号Longshu 15	A30	87.74	-	12.13	8.73	1.64	-	2.33	-
陇薯16号Longshu 16	A31	50.72	-	12.53	9.63	2.79	-	1.67	-
陇薯28号Longshu 28	A32	87.58	-	12.80	6.71	3.17	-	3.67	-
陇薯紫2号Longshuzi 2	A33	70.62	-	7.40	12.30	1.53	-	3.33	-
庄薯3号Zhuangshu 3	B1	-	52.87	15.33	5.83	-	1.33	-	1.33
S.stoloniferum	B2	-	42.22	13.86	14.30	-	1.85	-	2.00
丽薯1号Lishu 1	B3	-	83.10	14.70	7.67	-	1.36	-	1.00
Фиолетовый	B4	-	83.45	13.24	5.17	-	2.28	-	1.00
云薯304 Yunshu304	B5	-	45.01	12.30	9.92	-	0.05	-	0.33
甘农薯4号Gannongshu 4	B6	-	70.56	11.03	11.80	-	0.85	-	1.00
闵薯1号Minshu 1	B7	-	91.45	13.90	7.87	-	3.24	-	3.33
青薯6号Qingshu 6	B8	-	42.05	11.30	6.18	-	2.08	-	1.33
青薯10号Qingshu 10	B9	-	42.48	7.57	5.65	-	2.07	-	0.67
Белы	B10	-	104.15	8.92	7.78	-	0.52	-	0.67
Лилея	B11	-	85.68	13.89	6.86	-	2.11	-	1.00
陇薯9号Longshu 10	C1	-	-	13.76	8.12	-	-	-	-
天薯9号Tianshu 9	C2	-	-	2.69	6.53	-	-	-	-
中薯5号Zhongshu 5	C3	-	-	12.57	8.25	-	-	-	-
夏波帝Shepody	C4	-	-	14.13	6.98	-	-	-	-
合作88 Cooperation-88	C5	-	-	12.37	7.67	-	-	-	-
S.chacoense	C6	-	-	9.12	14.47	-	-	-	-
陇薯21号Longshu 21	C7	-	-	12.17	7.39	-	-	-	-
Здабытак	C8	-	-	8.60	11.89	-	-	-	-
云薯902 Yunshu 902	C9	-	-	5.50	6.82	-	-	-	-
陇薯红1号Longshuhong 1	C10	-	-	13.56	10.81	-	-	-	-
天薯15号Tianshu 15	C11	-	-	11.73	7.47	-	-	-	-
克新6号Kexin 6	C12	-	-	16.57	13.89	-	-	-	-
陇薯紫1号Longshuzi 1	C13	-	-	10.79	5.46	-	-	-	-

Table 2

Table 3

Fig. 1

Fig. 2

Fig. 3

Table 4

Fig. 4

Fig. 5

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引物Primer	正向引物Forward primer (5′-3′)	反向引物Reverse primer (5′-3′)
StSP6A-pART-CAM-FLAG-GFP	TCATTTGGAGAGGACACGCATGCCTAGA GTTGATCCATTGATAGTTGGTCG	TCGTCATCCTTGTAGTCGAATGCGCGAC GTCCTCCAGT
CDI	AGATTCATTCCTTTATCTGGAGGT	TCCCGTCTCCAACAACATCG
SBT5.6	GCACCTGTCATGGCACATTTT	CGCTCCACGCTGCTAGTATG
NFD4	TCAAGGTTCCGGCTTCCTTC	TCCTCCCCAATCATTTTCCCA
HSP22.7	TCTGCCGCTTTCTATTGCGA	TCCGGCAAACGAAACTGTCT
HSFA6b	GGAACAGAGAGGAAGCAGCA	GGAATCCGTTAGCCTCTCCG
SP6A	CCCAAGCGACCCTAACTTGA	CCAACCTGGGGCATACACTG
PYL4	AATCACTACAGCGCCACACA	CTACGTTACCGTCTCCGTCG
LEA	GCTAGACCACATCAACGTGC	TCGCGATACCAATCGTGACA
MAP3Ka	TGGATGGCTCCAGAGGTAGT	GCTGAACAGCTGTCAAACGG
SP5G	GGTGTGTAGACTTTGGTGTGGTTT	GGCCTCAAGGCACATCCAT
EF1a	GATGAAATCGTGAAGGAAGTTTCTTC	CAGTCAAGGTTGGTAGACCT

指标 Index	处理 Treatments	最大值 Maximum	最小值 Minimum	极差 Range	平均值 Mean	标准差 Standard deviation	变异系数 Coefficient of variation (%)
结薯天数 Days to tuber formation (d)	17 ℃	133.04	42.11	90.93	75.57	20.90	27.65
结薯天数 Days to tuber formation (d)	28 ℃	133.95	42.11	91.84	58.15	23.01	39.58
株高 Plant height (cm)	17 ℃	17.55	4.80	12.75	11.47	2.85	24.85
株高 Plant height (cm)	28 ℃	15.03	3.60	11.43	9.63	2.44	25.34
块茎鲜重 Weight (g)	17 ℃	8.26	0.07	8.19	2.48	1.42	57.23
块茎鲜重 Weight (g)	28 ℃	5.24	0.15	5.09	1.94	1.40	72.31
试管薯数目 No. of tubers per plant	17 ℃	3.67	0.33	3.34	2.44	0.83	34.07
试管薯数目 No. of tubers per plant	28 ℃	4.00	0.33	3.67	1.53	0.91	59.34

基因ID Gene ID	基因 Gene	陇薯11号中表达量 Longshu 11 FPKM	Lily中表达量 Lily FPKM	描述 Description
DM8C02G15320	SBT5.6	5.85	-9.37	假想蛋白Hypothetical protein
DM8C03G24430	CDI	9.61	-7.20	假想蛋白Hypothetical protein
DM8C03G24450	CDI	9.61	-7.20	假想蛋白Hypothetical protein
DM8C08G19980	NFD4	9.32	-10.28	核融合缺陷蛋白Nuclear fusion defective 4-like protein
DM8C11G12130	HSP22.7	5.64	-14.18	小分子热激蛋白Heat shock protein
MSTRG.26382	TAP	5.14	-13.43	转运衔接蛋白TAP1-like protein
DM8C01G29290	PECS-2.1	-5.76	5.02	果胶酯酶Pectinesterase 2
DM8C10G19610	PYL4	-6.40	4.42	脱落酸受体Abscisic acid receptor PYL4-like
DM8C08G28600	LEA	-8.06	4.43	晚期胚胎富集蛋白Late embryogenesis abundant (LEA) hydroxyproline-rich
DM8C04G26070	MtN21	-12.91	4.39	细胞壁合成相关蛋白WAT1-related protein
DM8C05G26810	SP6A	-11.01	3.46	开花因子Flowering locus T protein
DM8C09G20490	HSFA6b	-10.50	3.85	热激转录因子Heat stress transcription factor
DM8C09G20500	HSFA6b	-9.46	3.30	热激转录因子Heat stress transcription factor A-7a-like isoform X2
DM8C04G24740	MAP3Ka	-4.02	11.27	丝裂原活化蛋白激酶激酶激酶3 Mitogen-activated protein kinase kinase kinase 3

The Effect of the Tuber-Specific Protein StSP6A on in vitro Potato Tuber Formation Under High-Temperature Stress

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