The cleavage of biglycan by aggrecanases

SummaryObjectiveAggrecanase-1 [a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4] and aggrecanase-2 (ADAMTS-5) have been named for their ability to degrade the proteoglycan aggrecan. While this may be the preferred substrate for these enzymes, they are also able to degrade other proteins. The aim of this work was to determine whether the aggrecanases could degrade biglycan and decorin.MethodsBiglycan, decorin and aggrecan were purified from human and bovine cartilage and subjected to degradation by recombinant aggrecanase-1 or aggrecanase-2. In vitro degradation was assessed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE) and immunoblotting, and the cleavage site in biglycan was determined by N-terminal amino acid sequencing. SDS/PAGE and immunoblotting were also used to assess in situ degradation in both normal and arthritic human articular cartilage.ResultsBoth aggrecanase-1 and aggrecanase-2 are able to cleave bovine and human biglycan at a site within their central leucine-rich repeat regions. Cleavage occurs at an asparagine–cysteine bond within the fifth leucine-rich repeat. In contrast, the closely related proteoglycan decorin is not a substrate for the aggrecanases. Analysis of human articular cartilage from osteoarthritic (OA) and rheumatoid arthritic (RA) joints showed that a biglycan degradation product of equivalent size is present in the extracellular matrix. No equivalent degradation product was, however, detectable in normal adult human articular cartilage.ConclusionBiglycan, which is structurally unrelated to aggrecan, can act as a substrate for aggrecanase-1 and aggrecanase-2, and these proteinases may account for at least part of the biglycan degradation that is present in arthritic cartilage

Avaliação da resistencia a ferrugem tropical em linhagens de milho Tropical rust evaluation in maize inbred lines

Avaliaram-se 440 linhagens endogâmicas de milho do Instituto Agronômico de Campinas, obtidas a partir de diversas populações, quanto à severidade da ferrugem tropical, causada por Physopella zeae, em condições naturais de infecção. Foram instalados 22 experimentos em blocos ao acaso com duas repetições, em 1993/94, em Ribeirão Preto (SP) e utilizados como testemunhas resistente e suscetível os híbridos Z 8568 e P 3069 respectivamente, incluídos de forma intercalar. A avaliação foi realizada aos 30 dias após o florescimento, mediante uma escala de notas de 1 a 9, correspondendo a 0%; 1%; 2,5%; 5%; 10%; 25%; 50%; 75% e >75% de área foliar afetada. Todas as populações estudadas apresentaram variabilidade quanto à resistência a P. zeae, indicando que a utilização de métodos de seleção recorrente pode ser eficaz no melhoramento de populações visando à resistência ao patógeno.<br>Four hundred and forty maize inbred lines, developed at Instituto Agronômico de Campinas, State of São Paulo, Brazil, derived from several populations, were evaluated, in field conditions, as to tropical rust resistance caused by Physopella zeae. Twenty two experiments were carried out in randomized complete block designs, with two replications each, during 93/94 summer season, in Ribeirão Preto, State of Sao Paulo, Brazil. Resistant (Z 8568) and susceptible (P 3069) hybrids were used as controls. The evaluations were performed 30 days after flowering, according to a scale of notes from 1 to 9 (0%; 1%; 2.5%; 5%; 10%; 25%; 50%; 75% and >75% affected leaf area). All populations showed variability as to P. zeae reaction, suggesting that recurrent selection might be effective to achieve tropical rust resistance

Fungicidas no controle da ferrugem asiática (Phakopsora pachyrhizi) e produtividade da soja Fungicides on the control of soybean rust (Phakopsora pachyrhizi) and soybean yield

Neste trabalho, avaliou-se o efeito de fungicidas no desenvolvimento da ferrugem asiática, causada pelo fungo Phakopsora pachyrhizi, e na produtividade da soja. Os fungicidas foram pulverizados quando as plantas estavam no estádio de desenvolvimento R4. Utilizou-se a cultivar Fepagro-RS 10 e delineamento blocos ao acaso com quatro repetições e dez tratamentos. A avaliação da severidade da doença foi feita 30 dias após a pulverização, utilizando-se uma escala de notas de 0 a 5, considerando-se a porcentagem da área foliar infectada. Os tratamentos com os fungicidas azoxystrobin, difenoconazole + propiconazole, fluquinconazole, myclobutanil, pyraclostrobin + epoxiconazole, tebuconazole e trifloxistrobina + propiconazole, controlaram o patógeno em relação à testemunha, mostrando notas médias de severidade igual ou inferior a 2 e não diferiram entre si. O tratamento com difenoconazole (nota 3,05) controlou o patógeno, mas com nota inferior ao tratamento com carbendazin (nota 3,85) e à testemunha (nota 4,5). Todos os tratamentos mostraram produtividades superiores à testemunha, variando entre 14,5% e 27,3%. O peso de cem sementes foi superior ao da testemunha para todos os tratamentos (11,8% a 14,4%), com exceção do carbendazin (5,2%).<br>This work was done in order to evaluated the fungicides effect on the control of soybean rust caused by Phakopsora pachyrhizi, as well as their effect on yield. The fungicides were sprayed when the plants were at the growth stage R4. The cultivar used was Fepagro-RS 10. The experimental design used was a randomized blocks replicated four times and with ten treatments. The disease severity evaluation was done 30 days after spraying using a scale of notes (0 to 5), measuring the infected leaf area. The treatments with the fungicides azoxystrobin, difenoconazole + propiconazole, fluquinconazole, myclobutanil, pyraclostrobin + epoxiconazole, tebuconazole and trifloxystrobina + propiconazole controlled the pathogen when compared with the control, showing severity average notes equal or lower then 2, without difference among them. The treatment with difenoconazole (note 3.05) controlled the pathogen, with lower note than the carbendazin treatment (note 3.85) and non-treated plants (note 4.5). All fungicides treatments showed higher yield than non-treated plants varying from 14.5% to 27.3%. The weight of 100 seeds was higher than the control (11.8% to 14.4%), except for carbendazin (5.2%)

Intensidade da ferrugem asiática (Phakopsora pachyrhizi H. Sydow & P. Sydow) da soja [Glycine max (L.) Merr.] nas cultivares Conquista, Savana e Suprema sob diferentes temperaturas e períodos de molhamento foliar The effects of temperature and leaf wetness periods on the development of soybean rust in the cultivars Consquista, Savana and Suprema

A Ferrugem Asiática (Phakopsora pachyrhizi H. Sydow & P. Sydow), relatada em diversas regiões do globo terrestre de climas tropicais e subtropicais, causa redução significativa na produtividade da soja [Glycine max (L.) Merr.]. Fatores bióticos como interação patógeno-hospedeiro e abióticos influenciam o progresso da doença. Objetivou-se neste trabalho estudar os efeitos da temperatura e de períodos de molhamento foliar no progresso da Ferrugem Asiática nas cultivares Conquista, Savana e Suprema. O experimento foi conduzido no Departamento de Fitopatologia da Universidade Federal de Lavras, em junho de 2004, em câmaras de crescimento vegetal nas temperaturas de 15, 20, 25 e 30 &deg;C e períodos de molhamento foliar de 0, 6, 12, 18 e 24 horas. A inoculação foi realizada pulverizando-se as plantas com suspensão de 10(4) uredósporos de P. pachyrhizi.mL-1 de água. Dados da incidência e da severidade foram utilizados para avaliar o progresso da doença e integrados por meio da área abaixo da curva de progresso da incidência (AACPI) e da severidade (AACPS). Modelos de regressão não-linear foram ajustados para a AACPI e AACPS. Foi calculado o volume abaixo da superfície de resposta para incidência (VASRI) e severidade (VASRS) em relação à temperatura e molhamentos foliares com o objetivo de detectar diferenças entre cultivares. Molhamentos foliares acima de 15 horas e temperaturas próximas a 20 ºC, nas 3 cultivares avaliadas, determinaram maior intensidade da Ferrugem Asiática. Temperaturas próximas a 30 e 15 ºC ocasionaram menor intensidade da doença. Períodos de molhamento foliar abaixo de 6 horas reduziram a intensidade da doença. Todas as cultivares testadas foram suscetíveis à doença, entretanto, a cultivar Conquista apresentou maior VASRI e VASRS da Ferrugem Asiática comparada às cultivares Savana e Suprema, as quais não diferiram estatisticamente. Houve diferença entre as cultivares para AACPI em cada temperatura e molhamento foliar.<br>The asian rust (Phakopsora pachyrhizi Sydow) which has been reported in areas of tropical and subtropical climates around the world, causes significant soybean [Glycine max (L.) Merr.] yield reduction. The disease progress is influenced by biotic factors as interaction pathogen-host and abiotic factors of the environment. The objective of this work was to study the effects of temperature and leaf wetness period in the asian rust progress in the cultivars Conquista, Savana and Suprema. The experiment was conducted at the Department of Plant Pathology at Federal University of Lavras, in growth chamber at temperatures of 15, 20, 25 and 30 &deg;C and leaf wetness periods of 0,6, 12, 18 and 24 hours. The plants were inoculated by spraying a suspension of inoculum of P. pachyrhizi at concentration of 10(4) urediniospores.mL-1. Severity and incidence data were integrated by the area under disease progress curve for severity (AUDPCS) and incidence (AUDPCI). Non-linear regression models were adjusted for the disease severity (AUDPCS) and incidence (AUDPCI). Volume under the response surface of temperature and leaf wetness was calculated for incidence (VURSI) and severity (VURSS) to detect differences between cultivars. Higher soybean rust intensity occurred with leaf wetness above 15 hours and temperatures close to 20 ºC, for the three tested cultivars. Temperatures above 30 ºC and below 15 ºC reduced the disease progress. Disease intensity was reduced in leaf wetness below 6 hours. All cultivars were susceptible, but higher VURSI and VURSS occurred in Conquista cultivar, followed by Savana and Suprema cultivars. Disease intensity was not statistically different between Savana and Suprema cultivars. Temperature and leaf wetness were different among cultivars for AUDPCI