• Title/Summary/Keyword: polygalacturonase-inhibiting protein

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Characterization of an Apple Polygalacturonase-Inhibiting Protein (PGIP) That Specifically Inhibits an Endopolygalacturonase (PG) Purified from Apple Fruits Infected with Botryosphaeria dothidea

  • Lee Dong-Hoon;Bae Han-Hong;Kang In-Kyu;Byun Jae-Kyun;Kang Sang-Gu
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1192-1200
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    • 2006
  • An apple polygalacturonase-inhibiting protein (PGIP), which specifically inhibits endopolygalacturonase (PG, EC 3.2.1.15) from Botryosphaeria dothidea, was purified from Botryosphaeria dothidea-infected apple (Malus domestica cv. Fuji) fruits. The purified apple PGIP had a molecular mass of 40 kDa. The N-terminal amino acid sequence of the purified protein showed high homologies to those of PGIP from pear (100%), tomato (70%), and bean (65%). We also purified polygalacturonase (PG) from B. dothidea. The PG hydrolyzes pectic components of plant cell walls. When the extracted apple pectic cell wall material was treated with purified apple PGIP and B. dothidea PG, the amount of uronic acid released was lower than that treated with B. dothidea PG alone. This result demonstrates that PGIP functions specifically by inhibiting cell wall maceration of B. dothidea PG Furthermore, we characterized the de novo function of the PGIP against PG on the solubilization and depolymerization of polyuronides from cell wall of apple fruits inoculated with B. dothidea. This result demonstrated that the PGIP of plants exhibits one of the direct defense mechanisms against pathogen attack by inhibiting PGs that are released from pathogens for hydrolysis of cell wall components of plants.

Characterization of an Apple Polygalacturonase-inhibiting Protein (PGIP) from Apple Fruits (사과 과실로부터 분리한 polygalacturonase-inhibiting protein(PGIP)의 생화학적 특성)

  • Lee, Dong-Hoon;Kang, Sang-Gu;Kang, In-Kyu;Lee, Yoon-Kyeong;Choi, Cheol;Byun, Jae-Kyun
    • Journal of Life Science
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    • v.16 no.4
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    • pp.653-658
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    • 2006
  • An apple polygalacturonase-inhibiting protein (PGIP), that specifically inhibited endopolygalacturonase (PG, EC 3.2.1.15) from Botryosphaeria dothidea, was purified from B. dothidea infected apple (Malus domestica cv. Fuji) fruits. The apple PGIP was a mixed-type inhibitor of PG from B. dothidea. Optimal temperature for the maximum enzyme activity was $40^{\circ}C$, and optimum pH of the purified PGIP was pH 5.0. PGIP was stable up to temperature of $60^{\circ}C$ and was completely suppressed after heating at $70^{\circ}C$ for 10 min, PGIP was stable at pH between 4 and 8. Inhibition of PG by PGIP was reduced by $K^+$, $Cu^{2+}$, $Mg^{2+}$, $Ca^{2+}$ and $Zn^{2+}$ metal ion, sodium dodecyl sulfate (SDS) and 1,2-diaminocyclohexane tetra acetate (CDTA).

Characterization of the Gene Encoding Radish (Raphanus sativus L.) PG-inhibiting Protein

  • Hwang, Byung-Ho;Kim, Hun;Lim, Sooyeon;Han, NaRae;Kim, Jongkee
    • Horticultural Science & Technology
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    • v.31 no.3
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    • pp.299-307
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    • 2013
  • A radish (Raphanus sativus L.) polygalacturonase-inhibiting protein (PGIP) gene was cloned and compared to the PGIP gene (BrPGIP2) from Chinese cabbage (Brassica rapa ssp. pekinensis) in order to gain more information on controlling a disease and improving produce quality. To clone the radish PGIP gene, primers were designed based on conserved sequences of two PGIP genes (BnPGIP1 and BnPGIP2) from rape (B. napus L. ssp. oleifera), Chinese cabbage and Arabidopsis thaliana. PCR cloning was performed with cDNA from the stigma of radish 'Daejinyeoreum' as a template to confirm DNA fragments which were about 600 base pair in size. Sequence analysis revealed 84.1% homology with BrPGIP2 and 70.1% with BnPGIP1. DNA walking was conducted to confirm the open reading frame of 972 bp, and the gene was named RsPGIP1. RsPGIP1 consisting with 323 amino acids (aa) has a high leucine content (54/323) and contains 10 leucine-rich repeat domains, as do most BrPGIPs of Chinese cabbage. The gene expression of RsPGIP1 was induced by abiotic stresses and methyl jasmonate. It showed enrichment in the stigma and the primary root than a leaf. Cloning RsPGIP1 will aid to further apply practices on postharvest quality maintenance and disease control of the root.

Screening Differential Expressions of Defense-related Responses in Cold-treated 'Kyoho' and 'Campbell Early' Grapevines

  • Ahn, Soon Young;Kim, Seon Ae;Han, Jae Hyun;Kim, Seung Heui;Yun, Hae Keun
    • Horticultural Science & Technology
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    • v.31 no.3
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    • pp.275-281
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    • 2013
  • Low temperature is one of the major environmental factors that affect productivity including reduced growth and budding of vines, and changes of metabolic processes in grape (Vitis spp.). To screen the specific expression of abiotic stress-related genes against cold treatment in 'Kyoho' and 'Campbell Early' grapevines, expression of various defense-related genes was investigated by RT-PCR and real-time PCR. Among the 67 genes analyzed by RT-PCR and real-time PCR, 17 and 16 types of cDNA were up-regulated, while 5 and 6 types were down-regulated in cold-treated 'Kyoho' and 'Campbell Early' grapevines, respectively. Genes encoding carotene (Cart3564 and Cart4472), chalcone isomerase (CHI), cytochrome P450 (CYP), flavonol synthase (FLS), endo-${\beta}$-glucanase precursor (Glu), glutathione peroxidase (GPX), glutathione-S-transferase (GST), leucine-rich repeats (LRR), manganese superoxide dismutase (Mn-SOD), phenylalanine ammonia lyase (PAL), polygalacturonase-inhibiting protein (PGIP), proline rich protein 2 (PRP2), small heat shock protein (sHSP), temperature induced lipocalin (TIL), and thaumatin-like protein (TLP) were up-regulated, while those encoding CBF like transcription factor (CBF1), chitinase-like protein (CLP), cold induced protein (CIP), glycerol-3-phosphate acyltransferase (GPAT), and mitogen-activated protein kinase (MAPK) were down-regulated by low temperature treatment in both in 'Kyoho' and 'Campbell Early'.