Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (10): 164-172.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0055
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HAN Lei1,2(), LI Jun-lin1,2, GAO Ai-ping3, HUANG Jian-feng3, LI Jian-zhao1(), SONG Zhi-zhong1,3,4()
Received:
2022-01-11
Online:
2022-10-26
Published:
2022-11-11
Contact:
LI Jian-zhao,SONG Zhi-zhong
E-mail:hanlei1610@qq.com;jianzhaoli95@163.com;3614@ldu.edu.cn
HAN Lei, LI Jun-lin, GAO Ai-ping, HUANG Jian-feng, LI Jian-zhao, SONG Zhi-zhong. Cloning,Expression and Functional Analysis of Potassium Channel Gene MiSPIK in Mangifera indica[J]. Biotechnology Bulletin, 2022, 38(10): 164-172.
引物名称 Primer name | 引物序列 Primer sequence(5'-3') |
---|---|
GP1-F | TCTTCACCCCTCTGGA(A/G)TT(C/T)GG |
GP1-R | GAGGGGACCGGCC(A/G)TC(A/G)TA(A/C)TC |
GP2-out | CTAATGGTTAATACCAGCAATC |
GP2-in | CAGAAGGAAGTGTTCCTCTATG |
GP3-out | GCTGGTGCAGCCCTTCTGAATCAG |
GP3-in | GACGTAACATACTTGACCCAC |
MiSPIK-F | CCAGTCTAGGCAACAAACCAAG |
MiSPIK-R | CGCCTCCATTGGCTATCAGATCTG |
pHB-MiSPIK-F | GCCGAGCTCATGAAAATAACGTGGCTGAGAA |
pHB-MiSPIK-R | GCGTCTAGATTACAACCCACCATGAGTTTTAC |
MiActin-F | GAGGAGAAACAGAAGCAAGT |
MiActin-R | AATCATCTTGCTTCTCACCC |
Table 1 Primers used in study
引物名称 Primer name | 引物序列 Primer sequence(5'-3') |
---|---|
GP1-F | TCTTCACCCCTCTGGA(A/G)TT(C/T)GG |
GP1-R | GAGGGGACCGGCC(A/G)TC(A/G)TA(A/C)TC |
GP2-out | CTAATGGTTAATACCAGCAATC |
GP2-in | CAGAAGGAAGTGTTCCTCTATG |
GP3-out | GCTGGTGCAGCCCTTCTGAATCAG |
GP3-in | GACGTAACATACTTGACCCAC |
MiSPIK-F | CCAGTCTAGGCAACAAACCAAG |
MiSPIK-R | CGCCTCCATTGGCTATCAGATCTG |
pHB-MiSPIK-F | GCCGAGCTCATGAAAATAACGTGGCTGAGAA |
pHB-MiSPIK-R | GCGTCTAGATTACAACCCACCATGAGTTTTAC |
MiActin-F | GAGGAGAAACAGAAGCAAGT |
MiActin-R | AATCATCTTGCTTCTCACCC |
Fig. 1 RACE cloning process of MiSPIK A:Amplification of conserved domain(M:DL2000 marker. 1:PCR product of conserved domain). B:RACE PCR amplification of both ends(M:DL2000 marker. 3:RACE PCR product of 3'-end. 4:RACE PCR product of 5'-end). C:Amplification of full length CDS(M:DL2000 marker. 1:PCR product of CDS)
Fig. 8 Generation of heterologous over-expression transg-enic seedlings A:Schematic diagram of recombinant expression vector construction. B:PCR identification of T0 transgenic lines
指标Indicator | 对照Control | T1转基因株系T1 transgenic lines |
---|---|---|
总鲜重Total fresh weight/g | 11.73±1.34 | 16.86±1.38** |
地上部鲜重Fresh weight of shoots/g | 10.53±1.12 | 15.59±1.09** |
根部鲜重Fresh weight of roots/g | 1.20±0.09 | 1.27±0.13 |
总根长Total root length/cm | 19.26±0.21 | 19.34±0.23 |
地上部钾含量K+ concentration of shoots/(g·kg-1 DW) | 33.47±2.89 | 42.56±3.32* |
根部钾含量K+ concentration of roots/(g·kg-1 DW) | 22.18±1.65 | 29.26±2.14** |
Table 2 Physiological indicator analysis of T1 transgenic lines
指标Indicator | 对照Control | T1转基因株系T1 transgenic lines |
---|---|---|
总鲜重Total fresh weight/g | 11.73±1.34 | 16.86±1.38** |
地上部鲜重Fresh weight of shoots/g | 10.53±1.12 | 15.59±1.09** |
根部鲜重Fresh weight of roots/g | 1.20±0.09 | 1.27±0.13 |
总根长Total root length/cm | 19.26±0.21 | 19.34±0.23 |
地上部钾含量K+ concentration of shoots/(g·kg-1 DW) | 33.47±2.89 | 42.56±3.32* |
根部钾含量K+ concentration of roots/(g·kg-1 DW) | 22.18±1.65 | 29.26±2.14** |
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