Effects of a Novel Pathogenesis-Related Class 10 (PR-10) Protein from Crotalaria pallida Roots with Papain Inhibitory Activity against Root-Knot Nematode Meloidogyne incognita

被引：41
作者：
Andrade, Lucia Betania da S. [2 ,3 ]
Oliveira, Adeliana S. [1 ,3 ]
Ribeiro, Jannison K. C. [1 ]
Kiyota, Sumika [4 ]
Vasconcelos, Ilka Maria [3 ]
Abreu de Oliveira, Jose Tadeu [3 ]
de Sales, Mauricio P. [1 ]
机构：
[1] Univ Fed Rio Grande do Norte, Dept Bioquim, Ctr Biociencias, Lab Quim & Funcao Prot Bioat, BR-59072970 Natal, RN, Brazil
[2] Univ Estadual Vale Acarau UVA, Ctr Ciencias Agr & Biol, Sobral, Ceara, Brazil
[3] Univ Fed Ceara, Dept Bioquim & Biol Mol, Fortaleza, Ceara, Brazil
[4] Inst Biol, LBPP, Sao Paulo, Brazil
来源：
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY | 2010年 / 58卷 / 07期
关键词：
Papain inhibitor; PR-10; nematode; Crotalaria pallida; Meloidogyne incognita; CYSTEINE PROTEINASE; MOLECULAR-CLONING; DIFFERENTIAL EXPRESSION; GLOBODERA-PALLIDA; METHYL JASMONATE; ROTYLENCHULUS-RENIFORMIS; TRANSGENIC RESISTANCE; ARABIDOPSIS-THALIANA; CONFERS RESISTANCE; PLANT CYSTATIN;
D O I：
10.1021/jf9044556
中图分类号：
S [农业科学];
学科分类号：
09 ;
摘要：
A novel pathogenesis-related class 10 (PR-10) protein with papain inhibitory activity, named CpPRI, was purified from Crotalaria pallida roots by ammonium sulfate precipitation followed by three reverse-phase high-performance liquid chromatographies (HPLCs). CpPRI is made up of a single polypeptide chain with a M-r of 15 kDa, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). This protein exhibited a K-i value of 1.8 X 10(-9) M and operates via a noncompetitive inhibition mechanism. The alignment of the N-terminal amino acid sequence of CpPRI with other proteins revealed its identity with PR-10 proteins. CpPRI acts against digestive proteinase from root-knot nematode Meloidogyne incognita and demonstrated nematostatic and nematicide effects on this parasite in bioassays. In a localization study, fluorescein-5-isothiocyanate (FITC) CpPRI was observed to internalize and diffuse over the entire J2 body after 6 h of incubation. This fact could explain the natural tolerance of this plant species to nematodes.
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页码：4145 / 4152
页数：8
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