Leila Garcês de Araújo

Increased enzymatic activity in rice leaf blast suppression by crude extract of Epicoccum sp

Epicoccum sp. showed in vitro antagonism to the rice pathogens Magnaporthe oryzae, Rhizoctonia solani, Sarocladium oryzae, Monographella albescens and Cochliobolus miyabeanus in dual culture. The colony growth of the rice blast fungus, M. oryzae, was reduced by 42.50%. The lethal doses (LD50) determined based on probit-log dosage response curves at 3 and 6 hours after germination were 393.0 and 326.6 ppm, respectively. The crude extract (600 ppm) reduced appressorial formation by 95.68%. A greenhouse experiment comparing the relative efficiency of conidial suspension and crude extract of Epicoccum sp. in the suppression of leaf blast showed no statistical difference between both application methods. However, the crude extract of Epicoccum sp. (4000 ppm) 48 hours before the application of M. oryzae induced resistance and suppressed leaf blast by 97.6%. Scanning electron microscopy of rice leaves inoculated with crude extract of Epicoccum sp. and challenged with M. oryzae showed appressorial deformation on penetration phase. Peroxidase and β-1,3-glucanase activities increased in plants sprayed with crude extract of Epicoccum sp., 24 hours after the application of the challenger. PAL as well as chitinase activities increased 72 hours after challenge inoculation. Epicoccum sp. was shown to be a potential antagonist and inducer of resistance against M. oryzae.

RAPD analysis of blast resistant somaclones from upland rice cultivar IAC 47 for genetic divergence

Seventeen somaclones of upland rice cultivar IAC 47 showing different plant types, and either resistance or susceptibility to leaf blast, were utilized for random amplified polymorphic DNA (RAPD) analysis. Somaclones exhibited differences in reaction to isolates of Pyricularia grisea. Two somaclones (SC02 and SC04) were resistant to all three field isolates of somaclones, while the cultivar IAC 47 was susceptible. The inheritance study of two distinct plant types, one with erect bright green leaves and the other with droopy yellow green leaves, showed that a single possibly different, dominant gene governs each plant type. Of 32 random decamer primers utilized, OPA02 and OPD02 detected polymorphisms between somaclones showing erect bright green leaves and droopy yellow green leaves. Reliable grouping exhibiting 80% similarity was achieved with 17 primers. Leaf blast resistance to race IC-2 of P. grisea was associated with the plant type of erect bright green leaves.

Expression of resistance in rice hybrids to Pyricularia grisea

Thirty-nine rice (Oryza sativa) hybrids and their restorers were assessed for vertical resistance to Pyricularia grisea in the rice blast nursery, and in artificial inoculation tests with two pathotypes, under controlled greenhouse conditions. The hybrids were developed from cytoplasmic genetic male sterile lines 046I and IR 58025A, derived from WA cytoplasm. In the rice blast nursery all hybrids showed susceptible reaction varying from 5 to 9. Compatible and incompatible leaf blast reactions of hybrids to two pathotypes, IC-1 and IB-45, were observed in inoculation tests. A majority of the hybrids were resistant when the restorer was resistant. However, seven of the 25 F1 hybrids exhibited susceptible reactions even when one of the parents was resistant to a pathotype. The partial resistance of 11 hybrids and their parents that showed compatible reactions to two pathotypes was analyzed. Differential interaction between isolates and genotypes was observed for partial resistance in relation to both disease severity and lesion number indicating the specific nature of partial resistance.

Tagging microsatellite marker to a blast resistance gene in the irrigated rice cultivar Cica-8

The rice cultivar Cica-8 exhibit differential reaction to several pathotypes of Magnaporthe oryzae. The objective of the present investigation was to determine the number of alleles involved in the expression of resistance to leaf blast and identify microsatellite markers linked to these alleles. A cross between cultivar Metica-1 and Cica-8 susceptible and resistant, respectively, to pathotype IB-1 (Py1049) was made to obtain F1, F2, BC1:1 and BC1:2 progenies. Greenhouse tests for leaf blast reaction showed that resistance is controlled by a monogenic dominant gene. For testing microsatellite markers, DNA of both resistant and susceptible parents and F1 and F2 populations was extracted. As expected for single dominant gene the F2 populations segregated at a ratio of 3:1. Of the 11 microsatellite markers tested, one marker RM 7102 was found to be closely linked to the resistant allele at a distance of 2.7 cM, in the cultivar Cica-8 to pathotype IB-1.

Use of local rice cultivars as additional differentials to identify pathotypes of Pyricularia oryzae

Eight additional rice cultivars as differentials were selected to identify pathotypes occurring in the Tocantins State. They were BRS Jaburu, BRS Taim, BRS Bigua, BR IRGA-417, Epagri 109, Javae, Metica-1 and Supremo. These differentials were used in addition to the international set to determine the pathogenic diversity of 193 Pyricularia oryzae isolates collected during 1994-2002 from irrigated rice cultivars. They were conserved in Pyricularia culture collection of Embrapa Rice and Bean Research center. From 193 P. oryzae isolates 38 pathotypes were identified based on leaf blast reactions of international set and 29 pathotypes based on these additional differentials. The predominant pathotypes (TI-1, TG-2, TD-15 and TF-2) were represented by 53% of the tested isolates. The major international pathotypes (IB-45, IB-41, II-1 and ID-13) were represented by 43% of the isolates tested. The virulence pattern of 28 isolates belonging to the pathotype IB-45 was further differentiated into nine local pathotypes using additional set of differentials. The irrigated rice cultivars used as additional differentials serve as complementary set to identify pathotypes of interest for breeding blast resistant cultivars for Tocantins State.

In vitro culture of Epidendrum nocturnum (Orchidaceae)occurring in the Cerrado in Central-West region

As orquideas podem ser propagadas in vitro em meios de cultivo assepticos ou em simbiose com fungos micorrizicos. Epidendrum nocturnum ocorre em areas de Cerrado e neste estudo objetivou-se a visualizacao de fungos micorrizicos em suas raizes bem como o cultivo assimbiotico de suas plântulas em diferentes meios e a micorrizacao in vitro. No desenvolvimento assimbiotico testou-se tres meios de cultura em um ensaio e, em outro ensaio, foram testadas formulacoes alternativas do meio Knudson C modificado, com a adicao de polpa de frutas. Fungos micorrizicos de orquideas foram usados na micorrizacao in vitro. Pelotons foram visualizados nas raizes de E. nocturnum. O meio Knudson C modificado diferiu estatisticamente dos demais, nas duas intensidades luminosas, no primeiro ensaio. No segundo ensaio, apos sete meses, os meios com formulacao alternativa nao diferiram estatisticamente entre si. Na micorrizacao in vitro as plântulas com o isolado fungico En07 (Waitea circinata) apresentaram maior vigor visual em relacao aos demais tratamentos, mas nao houve diferenca estatistica com o controle. Os meios com formulacao alternativa e o Knudson C modificado podem ser utilizados no desenvolvimento in vitro de E. nocturnum e o isolado En07 pode ser usado na micorrizacao visando estrategias de conservacao desta orquidea.