Establishment of a Genetic Transformation System for Potato ‘Xinjiang No. 2’

doi:10.13560/j.cnki.biotech.bull.1985.2025-1016

Abstract

Abstract:

Objective To establish an efficient regeneration and genetic transformation system for potatoes (Solanum tuberosum L.) variety ‘Xinjiang No. 2’, and to provide technical support for virus-resistant breeding. Method Using ‘Xinjiang No. 2’ potato as the experimental material, Agrobacterium-mediated transformation was employed to introduce the plant interference vector plasmid pCAMBIA2300-CP-RNAi into potato stem explants. The effects of pre-culture duration, plant hormone combinations, and types and concentrations of antibiotics on callus induction and shoot differentiation were investigated. By real-time quantitative PCR (RT-qPCR) technology, with the Actin gene as an internal reference, the mRNA expression of the CP gene was detected. The actual content of PVY virus in plants was quantitatively measured using enzyme-linked immunosorbent assay (ELISA) to validate the RNAi silencing effect and virus suppression. Result The optimal hormone and antibiotic combination for callus induction was 2.0 mg/L 6-BA + 0.8 mg/L 2,4-D + 50 mg/L Kan + 200 mg/L TMT when pre-culture and co-culture durations were both 2 days, achieving an induction rate of 85%. The callus showed a compact morphology, fresh green color, and a dumbbell-shaped swelling at both ends of the stem segments. For shoot differentiation, the optimal hormone and antibiotic ratio was 2 mg/L 6-BA + 2 mg/L ZT + 0.5 mg/L GA₃ + 50 mg/L Kan + 200 mg/L TMT, resulting in the highest germination and transformation rates, with well-developed seedlings. Transgenic seedlings regenerated into plants on 1/2 MS medium supplemented with 0.6 mg/L IAA + 75 mg/L Kan + 200 mg/L TMT, demonstrating robust root systems. Compared to non-transformed control plants, the relative expression of the CP gene in transgenic plants was significantly downregulated, and PVY virus content was markedly reduced, confirming that the constructed RNAi vector successfully triggered specific gene silencing and effectively inhibited virus accumulation in plants. Conclusion This study successfully establishes a stable and efficient genetic transformation system for the potato variety ‘Xinjiang No. 2’. The constructed RNAi interference vector effectively suppresses Y virus accumulation in plants.

Key words: Solanum tuberosum L., plant hormone ratio, genetic transformation system, transgenic plants

ZHAO Wen-juan, LI Hui, YANG Xue-ying, WANG Zhao-lu, ZHU Jian-bo. Establishment of a Genetic Transformation System for Potato ‘Xinjiang No. 2’[J]. Biotechnology Bulletin, 2026, 42(6): 87-97.

Figures/Tables 16

References 22

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试验编号 Test number	6-苄氨基嘌呤 6-Benzylaminopurine（6-BA）	2，4-二氯苯氧乙酸 2，4-Dichlorophenoxyacetic acid （2，4-D）	萘乙酸 1-Naphthaleneacetic acid （NAA）	3-吲哚乙酸 Indole-3-acetic acid （IAA）	6-糠氨基嘌呤 Kinetin （KT）	玉米素核苷 Zeatin riboside （ZR）
C1	2.0	0	0.6	0	0	0
C2	2.0	0.8	0	0	0	0
C3	0	0	0	0.2	0	2.0
C4	2.0	0.6	0	0	0.4	0

试验编号 Test number	6-苄氨基嘌呤 6-Benzylaminopurine（6-BA）	2，4-二氯苯氧乙酸 2，4-Dichlorophenoxyacetic acid （2，4-D）	萘乙酸 1-Naphthaleneacetic acid （NAA）	3-吲哚乙酸 Indole-3-acetic acid （IAA）	6-糠氨基嘌呤 Kinetin （KT）	玉米素核苷 Zeatin riboside （ZR）
C1	2.0	0	0.6	0	0	0
C2	2.0	0.8	0	0	0	0
C3	0	0	0	0.2	0	2.0
C4	2.0	0.6	0	0	0.4	0

预培养时间 Pre-cultivation time （d）	外植体数 Number of explants	愈伤数 Number of callus	愈伤形成率 Callus formation rate （%）	再生芽数 Number of regenerated buds	出苗率 Seedling emergence rate （%）	转化频率 Conversion frequency （%）
0	60	2	3.3	0	0	0
1	60	21	35.0	4	19.0	0
2	60	49	81.7	23	46.9	21.73
3	60	39	65.0	11	28.2	9.1
4	60	11	18.3	1	9.1	0

预培养时间 Pre-cultivation time （d）	外植体数 Number of explants	愈伤数 Number of callus	愈伤形成率 Callus formation rate （%）	再生芽数 Number of regenerated buds	出苗率 Seedling emergence rate （%）	转化频率 Conversion frequency （%）
0	60	2	3.3	0	0	0
1	60	21	35.0	4	19.0	0
2	60	49	81.7	23	46.9	21.73
3	60	39	65.0	11	28.2	9.1
4	60	11	18.3	1	9.1	0

处理 Treatment	外植体数 Number of explants	愈伤数 Number of callus	愈伤形成率 Callus formation rate （%）	愈伤组织形态 Morphology of callus tissue
C1	60	27	45	白色须状物
C2	60	51	85	鲜绿色、结构紧密
C3	60	40	66.7	黄绿色，结构松散
C4	60	30	50	白色须状物、结构松散