Establishment of an Efficient Genetic Transformation System Mediated by Agrobacterium rhizogenes in Turnip

doi:10.16420/j.issn.0513-353x.2025-0140

Acta Horticulturae Sinica ›› 2025, Vol. 52 ›› Issue (5): 1389-1398.doi: 10.16420/j.issn.0513-353x.2025-0140

• New Technology and New Method • Previous Articles

Establishment of an Efficient Genetic Transformation System Mediated by Agrobacterium rhizogenes in Turnip

MA Xi¹, ZHANG Jinrui¹, ZHUANG Hongmei², CHEN Wei³, ZHANG Mi¹, YUAN Xiaoqi¹, YI Xiaofang¹, WANG Congcong¹, WANG Haiyun¹, WANG Yan¹^,^**()

¹ State Key Laboratory of Crop Genetics and Germplasm Enhancement and Utilization，Key Laboratory of Horticultural Crop Biology and Genetic Improvement（East China），College of Horticulture，Nanjing Agricultural University，Nanjing 211800，China
² Xinjiang Vegetable Engineering Technology Research Center，Institute of Fruit and Vegetable Research，Xinjiang Uyghur Autonomous Region Academy of Agricultural Sciences（Luntai Fruit Tree Resource Garden of the National Fruit Tree Germplasm Resources），Urumqi 830091，China
³ Jiangsu Engineering and Technology Center for Modern Horticulture，Jiangsu Polytechnic College of Agriculture and Forestry，Zhenjiang，Jiangsu 212400，China

Received:2025-02-22 Revised:2025-04-28 Online:2025-05-23 Published:2025-05-21
Contact: WANG Yan

Abstract

Abstract:

Turnip（Brassica rapa var. rapa L.）has problems such as poor regeneration and low transformation efficiency. The influenced factors including explants type，bacterial infection concentration，infection time，co-cultivation time and acetosyringone（AS）were firstly screened to establish the genetic transformed hairy roots system by Agrobacterium rhizogenes with a white skin turnip variety from Xinjiang Keping. The results showed that 6 d rootless seedlings were selected as explant and the suspension added 200 μmol · L^-1 AS with the bacterial concentration OD₆₀₀ of 0.4 was infected for 12 min. After 36 h co-cultivation，hairy roots were exhibited the highest transformation efficiency in MS bacteriostatic medium with 400 mg · L^-1 carbenicillin，as well as the visual marker RUBY was applied to efficiently screening for transgenic positive hairy roots. Furthermore，based on this system combined with developmental regulators，the stable positive transgenic plants were successfully obtained with a transformation efficiency of 10.61%.The whole process from seed germination，shoot regeneration to rooting could be completed within 3 months.

Key words: turnip, Agrobacterium rhizogenes, hairy root, RUBY, developmental regulator

MA Xi, ZHANG Jinrui, ZHUANG Hongmei, CHEN Wei, ZHANG Mi, YUAN Xiaoqi, YI Xiaofang, WANG Congcong, WANG Haiyun, WANG Yan. Establishment of an Efficient Genetic Transformation System Mediated by Agrobacterium rhizogenes in Turnip[J]. Acta Horticulturae Sinica, 2025, 52(5): 1389-1398.

Figures/Tables 11

References 16

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外植体类型 Explant type	诱导率/% Inductivity	平均毛状根数 Average number of hairy roots	阳性率/% Positive rate
带柄子叶 Cotyledon with petiole	89.66 ± 4.78 a	12.83 ± 1.49 a	2.15 ± 1.60 b
下胚轴 Hypocotyl	16.67 ± 9.43 b	0.20 ± 0.14 c	8.33 ± 5.89 ab
生长点 Growing point	35.00 ± 14.72 b	3.77 ± 2.29 b	3.98 ± 1.50 ab
无根苗 Rootless seedling	92.06 ± 4.88 a	12.98 ± 0.80 a	10.64 ± 1.82 a

外植体类型 Explant type	诱导率/% Inductivity	平均毛状根数 Average number of hairy roots	阳性率/% Positive rate
带柄子叶 Cotyledon with petiole	89.66 ± 4.78 a	12.83 ± 1.49 a	2.15 ± 1.60 b
下胚轴 Hypocotyl	16.67 ± 9.43 b	0.20 ± 0.14 c	8.33 ± 5.89 ab
生长点 Growing point	35.00 ± 14.72 b	3.77 ± 2.29 b	3.98 ± 1.50 ab
无根苗 Rootless seedling	92.06 ± 4.88 a	12.98 ± 0.80 a	10.64 ± 1.82 a

OD₆₀₀值 OD₆₀₀ value	诱导率/% Inductivity	平均毛状根数 Average number of hairy roots	溢菌数 Number of Agrobacterium overflow
0.2	81.03 ± 6.52 b	1.79 ± 1.09 b	少 Little
0.3	85.96 ± 4.97 ab	2.74 ± 0.71 b	无 None
0.4	92.06 ± 4.88 ab	12.98 ± 0.80 a	无 None
0.5 0.6	96.61 ± 2.42 a 63.33 ± 8.50 c	15.08 ± 0.86 a 3.97 ± 0.65 b	较少 Less 多 Much

OD₆₀₀值 OD₆₀₀ value	诱导率/% Inductivity	平均毛状根数 Average number of hairy roots	溢菌数 Number of Agrobacterium overflow
0.2	81.03 ± 6.52 b	1.79 ± 1.09 b	少 Little
0.3	85.96 ± 4.97 ab	2.74 ± 0.71 b	无 None
0.4	92.06 ± 4.88 ab	12.98 ± 0.80 a	无 None
0.5 0.6	96.61 ± 2.42 a 63.33 ± 8.50 c	15.08 ± 0.86 a 3.97 ± 0.65 b	较少 Less 多 Much

侵染时间/min Infection time	诱导率/% Inductivity	平均毛状根数 Average number of hairy roots	阳性率/% Positive rate	毛状根长势 Hairy root growing condition
4	90.00 ± 4.89 a	4.25 ± 0.37 b	6.67 ± 7.37 a	细长，稀疏 Slender and sparse
8	95.00 ± 7.44 a	4.47 ± 0.81 b	1.49 ± 1.47 b	细长，稀疏 Slender and sparse
12	92.06 ± 4.88 a	12.98 ± 0.80 a	10.64 ± 1.82 a	粗长，茂密 Thick，long，and dense
16	93.33 ± 6.24 a	10.98 ± 2.81 a	1.37 ± 1.92 b	细长，茂密 Slender and dense

Establishment of an Efficient Genetic Transformation System Mediated by Agrobacterium rhizogenes in Turnip

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共培养时间/h Co-cultivation time	诱导率/% Inductivity	平均毛状根数 Average number of hairy roots	阳性率/% Positive rate	侵染后发根时间/d Rooting time after infection
12	96.61 ± 2.50 a	10.58 ± 0.89 b	1.44 ± 0.68 b	9 ~ 10
24	96.67 ± 4.71 a	8.43 ± 1.05 c	3.56 ± 1.71 b	7 ~ 8
36	92.06 ± 4.88 a	12.98 ± 0.80 a	10.64 ± 1.82 a	7 ~ 8
48	94.92 ± 4.08 a	8.25 ± 0.76 c	3.08 ± 0.27 b	4 ~ 5

AS/（μmol · L^-1）	诱导率/% Inductivity	平均毛状根数 Average number of hairy roots	阳性率/% Positive rate
0	100.00 ± 0 a	9.15 ± 1.57 b	1.64 ± 0.48 c
100	100.00 ± 0 a	9.05 ± 0.78 b	8.84 ± 1.50 a
200	92.06 ± 4.88 b	12.98 ± 0.80 a	10.64 ± 1.82 a
300	95.00 ± 4.08 ab	9.90 ± 0.74 b	5.39 ± 0.82 b
400	98.33 ± 2.36 a	13.33 ± 0.66 a	1.88 ± 0.90 c
500	100.00 ± 0 a	13.33 ± 0.91 a	1.65 ± 0.40 c

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