Biotechnology Bulletin ›› 2024, Vol. 40 ›› Issue (3): 181-192.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0960
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MEI Xian-jun1,2(), SONG Hui-yang2, LI Jing-hao2, MEI Chao2,3, SONG Qian-na2,3, FENG Rui-yun2,3(), CHEN Xi-ming1,2()
Received:
2023-10-11
Online:
2024-03-26
Published:
2024-04-08
Contact:
FENG Rui-yun, CHEN Xi-ming
E-mail:1732244325@qq.com;516834898@qq.com;fengruiyun1970@163.com
MEI Xian-jun, SONG Hui-yang, LI Jing-hao, MEI Chao, SONG Qian-na, FENG Rui-yun, CHEN Xi-ming. Cloning and Expression Analysis of StDof5 Gene in Potato[J]. Biotechnology Bulletin, 2024, 40(3): 181-192.
引物名称Primer name | 引物序列Primer sequence(5'-3') |
---|---|
Actin-F | GGGATGGAGAAGTTTGGTGGTGG |
Actin-R | CTTCGACCAAGGGATGGTGTAGC |
Dof-F.2 | GCTTGACTTTGATGGAGTGGTGG |
Dof-R.2 | AAGATTGGCTATCGGAGGTGC |
ABF4-F | CTTCTGCACATACAAGGATATTCAA |
ABF4-R | TGCCTAGCCAAAGGGAAATCA |
MYB2-F | ATGACATGGACCTCCGACGA |
MYB2-R | CGTTCGCTTTAGACCAGCAC |
SnRK2s-F | TTGCTGTAGAGAAGGTGGGT |
SnRK2s-R | GTCCTACTGTCACTGCCGTC |
DREB2A-F | CACAGAAGAAAAGAGAAATGGCT |
DREB2A-R | TCCACTATAATCCAACGGCAGA |
ABI3-F | TGCTGACATGAAGTGTGGCA |
ABI3-R | TTTTGCCCTCCGACTTTGGT |
ABI5-F | TAAAACAGGCCCTGGCGGAG |
ABI5-R | TTGCGCCTTTGTTTGAGCTT |
Table 1 Primer sequences for fluorescence quantification
引物名称Primer name | 引物序列Primer sequence(5'-3') |
---|---|
Actin-F | GGGATGGAGAAGTTTGGTGGTGG |
Actin-R | CTTCGACCAAGGGATGGTGTAGC |
Dof-F.2 | GCTTGACTTTGATGGAGTGGTGG |
Dof-R.2 | AAGATTGGCTATCGGAGGTGC |
ABF4-F | CTTCTGCACATACAAGGATATTCAA |
ABF4-R | TGCCTAGCCAAAGGGAAATCA |
MYB2-F | ATGACATGGACCTCCGACGA |
MYB2-R | CGTTCGCTTTAGACCAGCAC |
SnRK2s-F | TTGCTGTAGAGAAGGTGGGT |
SnRK2s-R | GTCCTACTGTCACTGCCGTC |
DREB2A-F | CACAGAAGAAAAGAGAAATGGCT |
DREB2A-R | TCCACTATAATCCAACGGCAGA |
ABI3-F | TGCTGACATGAAGTGTGGCA |
ABI3-R | TTTTGCCCTCCGACTTTGGT |
ABI5-F | TAAAACAGGCCCTGGCGGAG |
ABI5-R | TTGCGCCTTTGTTTGAGCTT |
Fig. 1 Bioinformatics analysis of StDof5 protein A: Transmembrane domain of the StDof5 protein; B: secondary structure prediction of potato StDof5 protein; C: hydrophilic nature of potato StDof5 protein; D: tertiary structure prediction of potato StDof5 protein
Fig. 5 Construction of the potato StDof5 vector A: Agrobacterium vector construction; B: potato genetically transformed tissue culture; C: identification of overexpressed potato plants. M: Marker 2000 bp. P: Plasmid. DES: Wild type. 1-8: Positive single clone. 6-10, 14, 16, 24: Overexpression positive plants
Fig. 6 Analysis of Dof5 gene expression in different tissues of potato DES: Wild type. OP: Overexpressed positive plants. Different letters indicate a significant difference(P<0.05). The same below
测量指标Measurement index | 盐浓度Salting strength/(mol·L-1) | 品种名称Variety name | ||||||
---|---|---|---|---|---|---|---|---|
DES | OP-6 | OP-7 | OP-9 | OP-10 | OP-16 | OP-24 | ||
株高Plant height/cm | 0 | 6.87±0.38a | 7.10±0.17a | 5.57±0.40a | 5.43±0.15a | 6.53±0.85a | 7.37±0.75a | 7.83±1.01a |
50 | 3.70±010b | 4.83±0.65b | 3.70±0.36b | 4.37±0.21b | 3.97±0.35b | 6.00±0.72b | 4.13±0.45b | |
100 | 2.13±0.76c | 2.97±0.42c | 2.00±0.53c | 1.77±0.31c | 2.80±0.36c | 3.00±0.44c | 2.93±0.15c | |
鲜重Fresh weight/g | 0 | 0.21±0.01a | 0.23±0.01a | 0.19±0.01a | 0.21±0.02a | 0.24±0.02a | 0.19±0.02a | 0.35±0.01a |
50 | 0.14±0.01b | 0.19±0.02b | 0.12±0.02b | 0.16±0.02b | 0.16±0.02b | 0.16±0.01a | 0.28±0.02b | |
100 | 0.09±0.01c | 0.12±0.02c | 0.09±0.01c | 0.09±0.01c | 0.12±0.01c | 0.12±0.02b | 0.15±0.02c | |
根长Root length/cm | 0 | 12.13±0.25a | 12.20±0.46a | 12.23±2.03a | 14.70±0.62a | 17.63±0.35a | 12.90±1.49a | 12.03±0.72a |
50 | 9.00±0.26b | 9.73±0.12b | 10.93±1.79ab | 13.07±0.31b | 13.73±0.31b | 10.53±0.61b | 11.67±0.31a | |
100 | 7.53±0.25c | 8.93±060b | 6.70±2.74b | 4.00±0.72c | 10.1±0.20c | 9.73±0.31b | 8.60±0.53b | |
根数Root | 0 | 6.33±1.15a | 7.33 ±2.31a | 8.00±0.00a | 7.00±1.00a | 8.67±1.53a | 6.00±0.00a | 9.67±2.08a |
50 | 3.67±2.08a | 5.67±2.31a | 4.00±1.00b | 5.33±2.31a | 4.67±1.53b | 6.67±2.89a | 5.67±1.15b | |
100 | 3.33±1.53a | 5.00±1.00a | 3.33±2.08b | 1.00±0.00b | 4.33±2.31b | 1.67±0.58b | 3.00±1.73b | |
叶片数Number of leaves | 0 | 7.33±0.58a | 8.33±0.58a | 7.67±0.58a | 6.67±0.58a | 7.00±0.00a | 7.00±1.00a | 6.67±0.58a |
50 | 6.33±0.58a | 7.00±0.00a | 6.33±1.15a | 6.00±1.00ab | 7.00±0.00a | 7.00±1.00a | 6.66±0.58a | |
100 | 7.00±1.00a | 7.00±1.00a | 6.67±0.58a | 5.00±0.00b | 6.33±1.15a | 6.00±1.00a | 6.67±0.58a |
Table 2 Determination of the growth indicators of salt stress
测量指标Measurement index | 盐浓度Salting strength/(mol·L-1) | 品种名称Variety name | ||||||
---|---|---|---|---|---|---|---|---|
DES | OP-6 | OP-7 | OP-9 | OP-10 | OP-16 | OP-24 | ||
株高Plant height/cm | 0 | 6.87±0.38a | 7.10±0.17a | 5.57±0.40a | 5.43±0.15a | 6.53±0.85a | 7.37±0.75a | 7.83±1.01a |
50 | 3.70±010b | 4.83±0.65b | 3.70±0.36b | 4.37±0.21b | 3.97±0.35b | 6.00±0.72b | 4.13±0.45b | |
100 | 2.13±0.76c | 2.97±0.42c | 2.00±0.53c | 1.77±0.31c | 2.80±0.36c | 3.00±0.44c | 2.93±0.15c | |
鲜重Fresh weight/g | 0 | 0.21±0.01a | 0.23±0.01a | 0.19±0.01a | 0.21±0.02a | 0.24±0.02a | 0.19±0.02a | 0.35±0.01a |
50 | 0.14±0.01b | 0.19±0.02b | 0.12±0.02b | 0.16±0.02b | 0.16±0.02b | 0.16±0.01a | 0.28±0.02b | |
100 | 0.09±0.01c | 0.12±0.02c | 0.09±0.01c | 0.09±0.01c | 0.12±0.01c | 0.12±0.02b | 0.15±0.02c | |
根长Root length/cm | 0 | 12.13±0.25a | 12.20±0.46a | 12.23±2.03a | 14.70±0.62a | 17.63±0.35a | 12.90±1.49a | 12.03±0.72a |
50 | 9.00±0.26b | 9.73±0.12b | 10.93±1.79ab | 13.07±0.31b | 13.73±0.31b | 10.53±0.61b | 11.67±0.31a | |
100 | 7.53±0.25c | 8.93±060b | 6.70±2.74b | 4.00±0.72c | 10.1±0.20c | 9.73±0.31b | 8.60±0.53b | |
根数Root | 0 | 6.33±1.15a | 7.33 ±2.31a | 8.00±0.00a | 7.00±1.00a | 8.67±1.53a | 6.00±0.00a | 9.67±2.08a |
50 | 3.67±2.08a | 5.67±2.31a | 4.00±1.00b | 5.33±2.31a | 4.67±1.53b | 6.67±2.89a | 5.67±1.15b | |
100 | 3.33±1.53a | 5.00±1.00a | 3.33±2.08b | 1.00±0.00b | 4.33±2.31b | 1.67±0.58b | 3.00±1.73b | |
叶片数Number of leaves | 0 | 7.33±0.58a | 8.33±0.58a | 7.67±0.58a | 6.67±0.58a | 7.00±0.00a | 7.00±1.00a | 6.67±0.58a |
50 | 6.33±0.58a | 7.00±0.00a | 6.33±1.15a | 6.00±1.00ab | 7.00±0.00a | 7.00±1.00a | 6.66±0.58a | |
100 | 7.00±1.00a | 7.00±1.00a | 6.67±0.58a | 5.00±0.00b | 6.33±1.15a | 6.00±1.00a | 6.67±0.58a |
Fig. 9 Growth status of tissue culture seedlings after 25 d of salt stress From top to bottom, the pictures are: DES, OP-6, OP-7, OP-9, OP-10, OP-16, OP-24, with the salt stress of 0, 50 and 100 mmol/L NaCl, respectively
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