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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (05): 642-649.doi: 10.3724/SP.J.1006.2018.00642

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Identification and Expression Analysis of Likely Orthologs of Tobacco Salicylic Acid Binding Protein 2 in Common Beans

Ren-Feng XUE1,**(), Li WANG2,**, Ming FENG1, Wei-De GE1,*()   

  1. 1 Crop Research Institute, Liaoning Academy of Agricultural Sciences, Shenyang 110161, Liaoning, China
    2 Rural Energy and Environment Agency, Ministry of Agriculture, Beijing 100125, China
  • Received:2017-09-10 Accepted:2018-01-08 Online:2018-05-20 Published:2018-01-29
  • Contact: Ren-Feng XUE,Li WANG,Wei-De GE E-mail:xuerf82@163.com;snowweide@163.com
  • Supported by:
    This work was supported by the National Natural Science Foundation of China (31401447), the Liaoning Doctor Startup Foundation (201501113), the New Varieties Breeding, High Quality and Effective Technology Integration and Demonstration of Special Grain and Oil Crop (2018416023), and the China Agriculture Research System (CARS-08-Z8).

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Abstract:

Salicylic acid can induce the systematic resistant response in plants, and salicylic acid-binding protein 2 is an important esterase in the regulation of salicylic acid content in plant cells. In this study, we used the bioinformatics method to search the orthologous genes of tobacco salicylic acid-binding protein 2 in common bean, which were designated as PvMES1-PvMES7, then analyzed the changes of MES activity and free SA accumulation in plant roots. The expression level of 7 PvMES genes in susceptible varieties BRB130 and Baidaodou, resistant varieties Heiyundou and 260205 infected by Fusarium oxysporum f. sp. phaseoli isolate FOP-DM01 was detected by Real time PCR, showing that the expression of PvMES1, PvMES3, PvMES4, PvMES5, and PvMES6 was significantly increased at three days after FOP-DM01 infection, with most significant increase of 7.6 folds and 5.6 folds for PvMES5 gene in Heiyundou and PvMES1 gene in 260205 at 0 day, respectively. The MES activity and free SA content in roots of Heiyundou and 260205 were also improved, which activated the relevant defense response reaction mediated by SA in the host. The results of this study provide a theoretical basis for the resistance molecular breeding of Fusarium wilt in common beans.

Key words: common bean, Fusarium wilt, salicylic acid binding protein 2, orthologous genes, system acquired resistance

Table 1

Primers used in the experiment"

基因名称
Gene name		 引物名称
Primer name		 引物序列
Sequence for qPCR (5°-3°)
PvMES1 QMES1F GCTCTATCAACTTTCCCCTG
QMES1R CATCGTTGGTAATCCAAAGT
PvMES2 QMES2F CTATCAACTCTCCCCAACTC
QMES2R CAAAGTAATTGCAAGGTCTAT
PvMES3 QMES3F CAGAAAATTGAAGACGTTGG
QMES3R AACATAGGAGGGCTTGCGC
PvMES4 QMES4F GAGTACCTTGGGAGAGAAT
QMES4R TAGAGAAGCTCTTCTGTTGG
PvMES5 QMES5F GCAATTCCCAGAAAAAATTCTG
QMES5R CTTGGGACCAAAGAACATCAA
PvMES6 QMES6F ACCAAGTTTCTGTCCACTGC
QMES6R TACTCTTTTGGAATTGTCAA
PvMES7 QMES7F GGAGTTGAATTTTTGAGATCTA
QMES7R CATCAAATTATCTTTTCCAGC
PvPR1 QPR1F TGCTAAAGACGCCGATACCA
QPR1R GCAACACCTCCAACTATGCT
Actin Act-F GAAGTTCTCTTCCAACCATCC
Act-R TTTCCTTGCTCATTCTGTCCG

Table 2

Nucleotide sequence analysis of PvMES family members and tobacco (Nt) SABP2"

基因名称
Gene		 基因位点
Genetic locus		 基因序列位置
Sequence location		 E值
E-value		 一致性
Identity (%)		 染色体
Chromosome
PvMES1 Phvul.003G248200.1 48561200-48564505 1.7×10-35 59.2 3
PvMES2 Phvul.003G248300.1 48567955-48569848 7.0×10-34 57.6 3
PvMES3 Phvul.003G248025.1 48530639-48532632 1.6×10-29 63.8 3
PvMES4 Phvul.003G248050.1 48539059-48541466 1.9×10-28 61.4 3
PvMES5 Phvul.003G248075.1 48545538-48547254 1.9×10-28 63.1 3
PvMES6 Phvul.002G022300.1 2389494-2391172 3.4×10-25 58.4 2
PvMES7 Phvul.010G043800.2 6682398-6684451 5.1×10-4 45.3 10

Fig. 1

Multiple sequence alignment of PvMES1-PvMES7 and tobacco SABP2 (NtSABP2) Identical residues are shaded in black and similar residues in gray; the catalytic triad residues are indicated by arrows, and residues that contact salicylic acid are indicated with black triangles."

Fig. 2

Phylogenetic tree of PvMES1-PvMES7 and NtSABP2"

Fig. 3

Expression of PvMES1-PvMES7 and PvPR1 induced by Fusarium oxysporum f. sp. phaseoli DM01 isolate (FOP-DM01) Bars represented by different letters are significantly different (P < 0.05) among four common bean varieties at the same time point."

Fig. 4

MES activity induced by Fusarium oxysporum f. sp. phaseoli DM01 isolate (FOP-DM01) Bars represented by different letters are significantly different (P < 0.05) among four common bean varieties at the same time point."

Fig. 5

SA content induced by Fusarium oxysporum f. sp. phaseoli DM01 isolate (FOP-DM01) Bars represented by different letters are significantly different (P < 0.05) among four common bean varieties at the same time point."

Fig. 6

Disease assessment of the common bean varieties after inoculating with FOP-DM01 A: Disease progress of the common bean varieties at three weeks post inoculation with FOP-DM01; B: Disease index of the common bean varieties plants at three weeks post inoculation with FOP-DM01. Bars represented by different letters are significantly different (P < 0.05) among four common bean varieties at the same time point."

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