Scientia Agricultura Sinica

Figure/Table detail

Function of Copper-Resistant Gene copA of Ralstonia solanacearum

WANG XiaoNing,LIANG Huan,WANG Shuai,FANG WenSheng,XU JingSheng,FENG Jie,XU Jin,CAO AoCheng

Scientia Agricultura Sinica, 2019, 52(5): 837-848. DOI: 10.3864/j.issn.0578-1752.2019.05.006

引物名称 Primer name	引物序列 Primer sequence (5′-3′)	用途Purpose
759f	GTCGCCGTCAACTCACTTTCC	青枯菌特异性验证 Specificity verification of R. solanacearum
760r	GTCGCCGTCAGCAATGCGGAATCG	青枯菌特异性验证 Specificity verification of R. solanacearum
copA Upf	GGTCTTAAUCCCGTGAGGTTGGGAGGTGA	扩增copA 上游片段 Amplification of upstream of copA
copA Upr	GGCATTAAUTGCTCGACTTTCACAGCGGA	扩增copA 上游片段 Amplification of upstream of copA
copA Downf	GGACTTAAUGATCGAGTTTTCCTTACGTAA	扩增copA 下游片段 Amplification of downstream of copA
copA Downr	GGGTTTAAUTCACGCAGCGCCGTGTAGTCC	扩增copA 下游片段 Amplification of downstream of copA
copAf	ATATGGCGGCCTCATCATCG	基因缺失菌株验证 Verification of gene deletion strain
copAr	CGAGAAAAGCCCTGTCCAGT	基因缺失菌株验证 Verification of gene deletion strain
pBRRf	CATTAGGCACCCCAGGCTTTAC	质粒线性化 Plasmid linearization
pBRRr	CCTCTTCGCTATTACGCCAGC	质粒线性化 Plasmid linearization
copA Hf	CCTCGAGGTCGACGGTATCGATATGCGCAGCAATCGTGCATCCCGCC	copA 扩增 Amplification of copA
copA Hr	GAACTAGTGGATCCCCCGGGCTTCAGGCCACCACCACTTCGCGGAAC	copA 扩增 Amplification of copA
copA Testf	GCCGTTTGTGATGGCTTCC	基因互补菌株验证 Verification of gene complementary strain
copA Testr	CTTATTCAGGCGTAGCACCAGG	基因互补菌株验证 Verification of gene complementary strain
gyrBf	GACCTTCCAGGGGTTGATCG	RT-qPCR
gyrBr	TCTCCCCCAGCCCCTTATAC
copAf	ATATGGCGGCCTCATCATCG
copAr	CGAGAAAAGCCCTGTCCAGT
hrpBf	GAAGTGGCCGCCCATATC
hrpBr	GCTTGCGGTAGCCCTTGA
hrpGf	GGACACATTCCACGTTCTGCA
hrpGr	CCATGAAATTCGCCGTATTGA
popAf	TTCAGGAGCTTCACCAGGTCT
popAr	CAACACCAATGGCAACTCCAA

Table 2 Primer sequences used in this study

Other figure/table from this article

Fig. 3 PCR detection of copA gene deletion and complementary strains
Fig. 8 Biolog metabolic microarray detection
Purple pore is defined as positive and pores shallower than purple are defined as boundary values. The picture indicates the metabolic results of Po82, ΔPo82copA and pBBR-ΔPo82copA strains in 96 h, respectively. The OD₅₉₀of α -D-glucose (micropore plate C1) in the three strains is 0.632, 0.521 and 0.372, respectively. The OD₅₉₀of D-trehalose (micropore plate A4) in the three strains is 0.620, 0.480 and 0.383, respectively. The OD₅₉₀of L-alanine (micropore plate E3) in the three strains is 0.786, 0.370 and 0.436, respectively. The OD₅₉₀of glucuronamide (micropore plate F6) in the three strains is 0.566, 0.388 and 0.380, respectively
Table 1 Strains and plasmids used in this study
Fig. 1 Constituent components of GEN Ⅲ MicroPlant
Fig. 2 Phylogenetic tree based on copA sequence
Fig. 4 Induced expression of copA
The asterisk on the column indicates significant difference in gene expression under different inductive concentrations (P <0.05)
Fig. 5 Growth curve test of the wild-type Po82, ΔPo82copA and pBBR-ΔPo82copA strains
* indicates significant difference between the mutant strain and wild-type strain under the same treatment (0.01<P ≤0.05); ** indicates extremely significant difference between the mutant strain and wild-type strain under the same treatment (P <0.01). The same as below
Fig. 6 Pathogenicity test of the wild-type Po82, ΔPo82copA and pBBR-ΔPo82copA strains on tomato
Fig. 7 Expression analysis of pathogenic genes in the wild-type Po82 and ΔPo82copA strains
Fig. 9 The mobility of the wild-type Po82, ΔPo82copA and pBBR-ΔPo82copA strains

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