RESUMO
Papaya ringspot virus (PRSV) severely affects the global papaya industry. Transgenic papaya has been proven to have effective resistance to PRSV isolates from Hawaii, Thailand, Taiwan, and other countries. However, those transgenic cultivars failed to show resistance to Hainan Island isolates. Some 76 PRSV samples, representative of all traditional papaya planting areas across five cities (Wen Chang, n = 13; Cheng Mai, n = 14; Chang Jiang, n = 11; Le Dong, n = 25; and San Ya, n = 13) within Hainan Province, were investigated. Results revealed three genetic diversity groups (Hainan I, II, and III) that correlated with geographical distribution. Frequent mutations among PRSV isolates from Hainan were also observed. The high genetic divergence in PRSV isolates from Hainan is likely to be the cause of the failure of genetically modified papaya that targets sequence-specific virus.
Assuntos
Carica/virologia , Variação Genética , Doenças das Plantas/virologia , Potyvirus/genética , China , FilogeniaRESUMO
Plant thioglucosidases are the only known S-glycosidases in the large superfamily of glycosidases. These enzymes evolved more recently and are distributed mainly in Brassicales. Thioglucosidase research has focused mainly on the cruciferous crops due to their economic importance and cancer preventive benefits. In this study, we cloned a novel myrosinase gene, CpTGG1, from Carica papaya Linnaeus. and showed that it was expressed in the aboveground tissues in planta. The recombinant CpTGG1 expressed in Pichia pastoris catalyzed the hydrolysis of both sinigrin and glucotropaeolin (the only thioglucoside present in papaya), showing that CpTGG1 was indeed a functional myrosinase gene. Sequence alignment analysis indicated that CpTGG1 contained all the motifs conserved in functional myrosinases from crucifers, except for two aglycon-binding motifs, suggesting substrate priority variation of the non-cruciferous myrosinases. Using sinigrin as substrate, the apparent K(m) and V(max) values of recombinant CpTGG1 were 2.82 mM and 59.9 µmol min⻹ mg protein⻹ , respectively. The K(cat) /K(m) value was 23 s⻹ mM⻹ . O-ß-glucosidase activity towards a variety of substrates were tested, CpTGG1 displayed substrate-dependent and ascorbic acid-independent O-ß-glucosidase activity towards 2-nitrophenyl-ß-D-glucopyranoside and 4-nitrophenyl-ß-D-glucopyranoside, but was inactive towards glucovanillin and n-octyl-ß-D-glucopyranoside. Phylogenetic analysis indicated CpTGG1 belongs to the MYR II subfamily of myrosinases.
Assuntos
Ácido Ascórbico/metabolismo , Carica/enzimologia , Glicosídeo Hidrolases/metabolismo , beta-Glucosidase/metabolismo , Sequência de Aminoácidos , Carica/metabolismo , Glicosídeo Hidrolases/química , Glicosídeo Hidrolases/classificação , Dados de Sequência Molecular , Filogenia , Homologia de Sequência de Aminoácidos , beta-Glucosidase/química , beta-Glucosidase/classificaçãoRESUMO
The proteins in the food are the source of common allergic components to certain patients. Current lists of plant endogenous allergens were based on the medical/clinical reports as well as laboratory results. Plant genome sequences made it possible to predict and characterize the genome-wide of putative endogenous allergens in rice (Oryza sativa L.). In this work, we identified and characterized 122 candidate rice allergens including the 22 allergens in present databases. Conserved domain analysis also revealed 37 domains among rice allergens including one novel domain (histidine kinase-, DNA gyrase B-, and HSP90-like ATPase, PF13589) adding to the allergen protein database. Phylogenetic analysis of the allergens revealed the diversity among the Prolamin superfamily and DnaK protein family, respectively. Additionally, some allergens proteins clustered on the rice chromosome might suggest the molecular function during the evolution.