Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (10): 175-183.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0369
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TAO Na1,2(), LI Mao-xing1,2, GUO Hua-chun1,2()
Received:
2023-04-19
Online:
2023-10-26
Published:
2023-11-28
Contact:
GUO Hua-chun
E-mail:2019157697@qq.com;ynghc@126.com
TAO Na, LI Mao-xing, GUO Hua-chun. Optimization of Sweet Potato Genetic Transformation System Mediated by Agrobacterium rhizogenes[J]. Biotechnology Bulletin, 2023, 39(10): 175-183.
发根农杆菌菌株 Agrobacterium strain | 冠瘿碱类型 Crown gallin type | 抗性 Resistance |
---|---|---|
Ar.1193 | 农杆碱型 Agropine | 链霉素 Streptomycin |
K599 | 黄瓜碱型 Cucumopine | 链霉素 Streptomycin |
C58C1 | 农杆碱型 Agropine | 链霉素 Streptomycin |
ArQual | 农杆碱型 Agropine | 链霉素 Streptomycin |
MSU440 | 农杆碱型 Agropine | 链霉素 Streptomycin |
Table 1 A. rhizogenes strains and their resistances
发根农杆菌菌株 Agrobacterium strain | 冠瘿碱类型 Crown gallin type | 抗性 Resistance |
---|---|---|
Ar.1193 | 农杆碱型 Agropine | 链霉素 Streptomycin |
K599 | 黄瓜碱型 Cucumopine | 链霉素 Streptomycin |
C58C1 | 农杆碱型 Agropine | 链霉素 Streptomycin |
ArQual | 农杆碱型 Agropine | 链霉素 Streptomycin |
MSU440 | 农杆碱型 Agropine | 链霉素 Streptomycin |
引物名称 Primer name | 引物序列 Primer sequences(5'-3') | 产物长度 Product length/bp |
---|---|---|
rolB-F | AAGTGCTGAAGGAACAATC | 585 |
rolB-R | CAAGTGAATGAACAAGGAAC | |
HBFD1-F | ATGGCTCCTGTTCTCCTTGG | 1 224 |
HBFD1-R | CGCTCCTTGTGATGCTCG |
Table 2 List of primers
引物名称 Primer name | 引物序列 Primer sequences(5'-3') | 产物长度 Product length/bp |
---|---|---|
rolB-F | AAGTGCTGAAGGAACAATC | 585 |
rolB-R | CAAGTGAATGAACAAGGAAC | |
HBFD1-F | ATGGCTCCTGTTCTCCTTGG | 1 224 |
HBFD1-R | CGCTCCTTGTGATGCTCG |
Fig. 2 Effects of different strains and infection times on hairy roots Different lowercase letters indicate a significant difference between treatments (P <0.05). The same below
Fig. 3 Hairy roots induction rates of different sweet potato varieties(lines) by different strains A-E shows the hairy root induction of ‘Xushu 22’ ‘Taizhong No. 6’ ‘1610’ ‘Huishu’ and ‘YS’. The same below
发根农杆菌菌株 Agrobacterium rhizogenes strain | 甘薯品种(系)Sweet potato varieties(lines) | |||||
---|---|---|---|---|---|---|
泰中6号 Taizhong No. 6 | 1610 1610 | 徐薯22 Xushu22 | YS YS | 灰薯 Huishu | 平均诱导率 Mean induction rate | |
Ar.1193 | 56.85±4.38B | 25.93±2.26EF | 43.34±0.68CD | 10.39±0.46HI | 12.81±2.0GH | 29.86 |
K599 | 59.19±4.51B | 48.73±3.09BC | 75.24±5.24A | 25.43±1.70EF | 15.46±1.67FG | 44.81 |
C58C1 | 58.89±4.84B | 57.16±4.76B | 11.80±1.28HI | 19.31±1.60FG | 20.0±0.00FG | 33.43 |
ArQual | 42.94±3.84CD | 22.26±1.73FG | 79.40±1.29A | 19.0±2.58FG | 14.02±2.04FG | 35.52 |
MSU440 | 36.53±4.28DE | 24.60±2.75FG | 79.63±1.85A | 5.69±0.29I | 18.38±2.38FG | 32.97 |
平均诱导率 Average induction rate | 50.88 | 35.74 | 57.89 | 15.97 | 16.13 |
Table 3 Type of Agrobacterium rhizogenes and sweet potato varieties(lines)
发根农杆菌菌株 Agrobacterium rhizogenes strain | 甘薯品种(系)Sweet potato varieties(lines) | |||||
---|---|---|---|---|---|---|
泰中6号 Taizhong No. 6 | 1610 1610 | 徐薯22 Xushu22 | YS YS | 灰薯 Huishu | 平均诱导率 Mean induction rate | |
Ar.1193 | 56.85±4.38B | 25.93±2.26EF | 43.34±0.68CD | 10.39±0.46HI | 12.81±2.0GH | 29.86 |
K599 | 59.19±4.51B | 48.73±3.09BC | 75.24±5.24A | 25.43±1.70EF | 15.46±1.67FG | 44.81 |
C58C1 | 58.89±4.84B | 57.16±4.76B | 11.80±1.28HI | 19.31±1.60FG | 20.0±0.00FG | 33.43 |
ArQual | 42.94±3.84CD | 22.26±1.73FG | 79.40±1.29A | 19.0±2.58FG | 14.02±2.04FG | 35.52 |
MSU440 | 36.53±4.28DE | 24.60±2.75FG | 79.63±1.85A | 5.69±0.29I | 18.38±2.38FG | 32.97 |
平均诱导率 Average induction rate | 50.88 | 35.74 | 57.89 | 15.97 | 16.13 |
Fig. 5 PCR detection of rolB gene in sweet potato hairyroots 1 and 2 are untransformed roots of sweet potato, as negative controls. 3-7 are the hairy roots induced by the stem segments of sweet potato infected by Ar. 1193, K599, C58C1, ArQual, and MSU440. The same below
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