Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (4): 713-723.doi: 10.16420/j.issn.0513-353x.2022-0100
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KAN Liping, SHI Xiaoqian, YANG Han, JIN Yumeng, CHEN Liyan, ZHANG Lijuan, XU Yangchun, SHEN Qirong, DONG Caixia()
Received:
2022-11-30
Revised:
2023-02-02
Online:
2023-04-25
Published:
2023-04-27
Contact:
*(E-mail:cxdong@njau.edu.cn)
CLC Number:
KAN Liping, SHI Xiaoqian, YANG Han, JIN Yumeng, CHEN Liyan, ZHANG Lijuan, XU Yangchun, SHEN Qirong, DONG Caixia. Cloning and Function Identification of a Potassium Transporter Gene PbKT12 in Pear Fruit[J]. Acta Horticulturae Sinica, 2023, 50(4): 713-723.
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URL: https://www.ahs.ac.cn/EN/10.16420/j.issn.0513-353x.2022-0100
用途 Usage | 引物 Primer | 序列(5′-3′) Sequence |
---|---|---|
基因克隆Gene cloning | PbKT12-F/R | ATGTCAATAATGTCCGTGTCG TCAGACCATGTAAGTCATTCC |
亚细胞定位Subcellular localization | PbKT12N-F/R | AGAGATCTCGAGCTCAAGCTTATGTCAATAATGTCCGTGTCG CCTTGCTCACCATCAGGATCCGGACCATGTAAGTCATTCCGA |
PbKT12-2288-F/R | ACCACCCATAATACCCATAAACCGGTCAACATGTGGAGCA GCGATGGCAAACAGCTATTATGCGTCACTGGATTTTGGTTT | |
qRT-PCR | qRT-PbKT12-F/R | CGTGGTTCGCTCTTGGTGGTT CCTGAATGGCTCGCACAGAGAA |
内参基因Reference gene | Actin-F/R | CTCCCAGGGCTGTGTTTCCTA CTCCATGTCATCCCAGTTGCT |
酵母表达载体构建pYES2 expression | pYES2-PbKT12-F/R | ACTATAGGGAATATTAAGCTTATGTCAATAATGTCCGTGTCG AGTGGATCCGAGCTCGGTACCTCAGACCATGTAAGTCATTCC |
转基因载体的构建 pCAMBIA1305.1-expression | pCAMBIA1305.1- KT12-F/R | TCCTCTAGATCGACCTGCAGATGTCAATAATGTCCGTGTCG ACGACGGCCAGTGCCAAGCTTTCAGACCATGTAAGTCATTCCG |
Table 1 Primers in this study
用途 Usage | 引物 Primer | 序列(5′-3′) Sequence |
---|---|---|
基因克隆Gene cloning | PbKT12-F/R | ATGTCAATAATGTCCGTGTCG TCAGACCATGTAAGTCATTCC |
亚细胞定位Subcellular localization | PbKT12N-F/R | AGAGATCTCGAGCTCAAGCTTATGTCAATAATGTCCGTGTCG CCTTGCTCACCATCAGGATCCGGACCATGTAAGTCATTCCGA |
PbKT12-2288-F/R | ACCACCCATAATACCCATAAACCGGTCAACATGTGGAGCA GCGATGGCAAACAGCTATTATGCGTCACTGGATTTTGGTTT | |
qRT-PCR | qRT-PbKT12-F/R | CGTGGTTCGCTCTTGGTGGTT CCTGAATGGCTCGCACAGAGAA |
内参基因Reference gene | Actin-F/R | CTCCCAGGGCTGTGTTTCCTA CTCCATGTCATCCCAGTTGCT |
酵母表达载体构建pYES2 expression | pYES2-PbKT12-F/R | ACTATAGGGAATATTAAGCTTATGTCAATAATGTCCGTGTCG AGTGGATCCGAGCTCGGTACCTCAGACCATGTAAGTCATTCC |
转基因载体的构建 pCAMBIA1305.1-expression | pCAMBIA1305.1- KT12-F/R | TCCTCTAGATCGACCTGCAGATGTCAATAATGTCCGTGTCG ACGACGGCCAGTGCCAAGCTTTCAGACCATGTAAGTCATTCCG |
Fig. 6 The relative expression of PbKT12 gene in leaf and fruit of‘Huangguan’pear under different potassium levels Different lowercase letters indicate significant differences of root growth index(P < 0.05). The same below.
Fig. 8 The rooting(A)and flowering(B)phenotypes of the potassium donor(+ K)and potassium deficient(-K)Arabidopsis thaliana overexpression lines PbKT12-OE and WT
拟南芥株系 Arabidopsis | MS培养液 | 根平均直径/mm Mean root diameter | 根表面积/cm2 Root surface area | 总根长/cm Total root length | 单株开花数 Blossom number | |
---|---|---|---|---|---|---|
处理 | K+/(mmol · L-1) | |||||
野生型WT | -K | 0 | 0.18 ± 0.03 b | 7.18 ± 1.37 c | 69.28 ± 25.51 b | 5.00 ± 1.10 c |
+ K | 20 | 0.19 ± 0.02 b | 9.69 ± 1.50 b | 97.99 ± 18.47 b | 3.50 ± 1.05 c | |
PbKT12-OE | -K | 0 | 0.29 ± 0.09 a | 7.17 ± 1.60 c | 75.09 ± 28.57 b | 8.17 ± 1.33 b |
+ K | 20 | 0.18 ± 0.02 b | 12.19 ± 1.70 a | 156.63 ± 38.76 a | 10.67 ± 2.73 a |
Table 2 Effect of potassium in culture medium on root growth and flowering number of PbKT12-OE and WT
拟南芥株系 Arabidopsis | MS培养液 | 根平均直径/mm Mean root diameter | 根表面积/cm2 Root surface area | 总根长/cm Total root length | 单株开花数 Blossom number | |
---|---|---|---|---|---|---|
处理 | K+/(mmol · L-1) | |||||
野生型WT | -K | 0 | 0.18 ± 0.03 b | 7.18 ± 1.37 c | 69.28 ± 25.51 b | 5.00 ± 1.10 c |
+ K | 20 | 0.19 ± 0.02 b | 9.69 ± 1.50 b | 97.99 ± 18.47 b | 3.50 ± 1.05 c | |
PbKT12-OE | -K | 0 | 0.29 ± 0.09 a | 7.17 ± 1.60 c | 75.09 ± 28.57 b | 8.17 ± 1.33 b |
+ K | 20 | 0.18 ± 0.02 b | 12.19 ± 1.70 a | 156.63 ± 38.76 a | 10.67 ± 2.73 a |
Fig. 9 Distribution of K+ in Arabidopsis PbKT12-OE and WT plants with potassium supply(+ K)and potassium deficiency(-K)conditions Red indicates high K+ content and blue indicates low K+ content.
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