中早熟马铃薯‘陇薯20号’高效再生体系建立及抗草铵膦种质创制

doi:10.3724/SP.J.1006.2025.54024

Abstract

Abstract:

To establish an efficient regeneration system for the new early-mid maturing, vegetable-use potato cultivar ‘Longshu 20’ and to develop novel germplasm resistant to glufosinate, we systematically evaluated the embryogenic callus induction efficiency and shoot regeneration capacity of test-tube plantlet stem segments cultured on MS medium supplemented with varying concentrations and combinations of 6-benzylaminopurine (6-BA), naphthaleneacetic acid (NAA), and gibberellic acid (GA₃). The highest rates of embryogenic callus induction and shoot differentiation were observed on medium MS + 1.0 mg L^-1 6-BA + 0.5 mg L^-1 NAA + 4.5 mg L^-1 GA₃ + 3% sucrose. On day 30, the embryogenic callus induction rate reached 89.83%, while the shoot differentiation rate reached 91.81% by day 45. A Bar gene overexpression vector was constructed using recombinant DNA technology and introduced into stem segments via Agrobacterium-mediated transformation. After selection on MS medium containing 2.0 mg L^-1 phosphinothricin (PPT), six independent transgenic lines were successfully obtained. Both transgenic and non-transgenic control lines were grown and treated with commercial glufosinate-ammonium at an active ingredient content of 1271 g hm^-2 during the seedling stage. Within 9 days, all non-transgenic control plants had completely withered and died, while all transgenic lines exhibited stable resistance to glufosinate-ammonium. Phenotypic analysis at maturity revealed variation among transgenic lines in terms of tuber number per plant, yield per plant, and key quality traits. Notably, transgenic line S_20-4 showed no significant differences from the non-transgenic control in tuber number, yield, or key quality parameters such as starch, crude protein, and reducing sugar content. These results demonstrate that S_20-4 successfully integrates glufosinate resistance while concurrently maintaining the yield and quality traits of the recipient cultivar. The efficient regeneration system developed in this study provides a solid foundation for the genetic improvement of ‘Longshu 20’, and the transgenic line S_20-4 represents a promising candidate for direct application in breeding programs targeting herbicide resistance and high yield.

Key words: potato, Longshu 20, regeneration system, Bar gene, glyphosate

JIA Xiao-Xia, QI En-Fang, WEN Guo-Hong, MA Sheng, HUANG Wei, LYU He-Ping, LI Jian-Wu, QU Ya-Ying, DING Ning. Establishment of regeneration system and creation of glufosinate-resistant germplasm for early-mid maturing potato ‘Longshu 20’[J].Acta Agronomica Sinica, 2025, 51(9): 2285-2294.

Figures/Tables 8

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Table 2

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培养基编号 Medium number	基础培养基 Basic medium	6-苄基腺嘌呤 6-BA (mg L^-1)	萘乙酸 NAA (mg L^-1)	赤霉素 GA₃ (mg L^-1)
A	MS	1.00	0.25	2.00
B	MS	2.00	0.25	4.50
C	MS	3.00	0.25	7.00
D	MS	1.00	0.50	4.50
E	MS	2.00	0.50	7.00
F	MS	3.00	0.50	2.00
G	MS	1.00	0.75	7.00
H	MS	2.00	0.75	2.00
I	MS	3.00	0.75	4.50

培养基编号 Medium number	外植体数 Number of explants	培养15 d时愈伤组织诱导率 Induction rate of callus tissue after 15 days of cultivation (%)	培养30 d时胚型愈伤组织诱导率 Induction rate of embryogenic callus tissue after 30 days of cultivation (%)	培养45 d时胚性愈伤不定芽诱导率 Induction rate of adventitious buds from embryogenic callus after 45 days of cultivation (%)
A	113	45.61±1.21 d	18.16±0.73 d	0.00±0.00 d
B	117	66.56±1.14 c	9.64±0.83 e	8.96±1.23 c
C	111	25.48±0.89 f	0.00±0.00 g	—
D	115	92.50±1.03 a	89.83±0.98 a	91.81±2.14 a
E	113	43.57±1.23 d	21.23±0.87 c	0.00±0.00 d
F	114	27.14±0.97 f	0.00±0.00 g	—
G	116	32.28±1.06 e	6.64±0.96 f	0.00±0.00 d
H	118	46.83±0.89 d	17.05±0.86 d	0.00±0.00 d
I	114	77.78±1.17 b	48.45±0.95 b	16.23±1.96 b

株系名称 Name of strains	干物质含量 Content of dry matter (%)	淀粉含量 Content of starch (%)	粗蛋白含量 Content of crude protein (%)	还原糖含量 Content of reduced sugar (%)	维生素C含量Content of vitamin C (mg 100^-1 g^-1)	单株结薯数 Number of tubers per plant	单株产量 Yield of per plant (g)
S₂₀	17.80±1.24 ab	11.79±1.03 a	2.17±0.11 a	0.21±0.03 c	11.80±0.26 a	4.29±0.12 a	627.26±14.96 a
S_20-1	17.34±0.97 ab	12.03±0.98 a	2.18±0.12 a	0.24±0.05 c	10.69±0.45 a	3.12±0.09 e	475.07±12.62 b
S_20-2	15.06±1.02 b	11.96±1.32 a	2.21±0.06 a	0.48±0.02 a	11.02±0.69 a	3.98±0.15 b	509.94±21.07 b
S_20-3	18.90±1.56 a	10.89±1.01 a	2.13±0.09 a	0.36±0.03 b	10.67±1.02 a	3.69±0.03 bc	391.78±18.74 c
S_20-4	17.65±0.89 ab	11.98±0.89 a	2.23±0.10 a	0.22±0.02 c	11.96±0.98 a	4.33±0.08 a	635.12±21.04 a
S_20-5	16.89±1.96 b	12.21±0.93 a	2.21±0.09 a	0.39±0.06 b	11.06±1.32 a	3.73±0.14 b	321.46±19.64 d
S_20-6	16.07±1.47 b	11.32±0.93 a	2.15±0.07 a	0.37±0.05 b	10.78±1.23 a	3.48±0.18 cd	311.46±17.57 d

Establishment of regeneration system and creation of glufosinate-resistant germplasm for early-mid maturing potato ‘Longshu 20’

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