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Yolanda Rodrguez Pagaza, Lorenzo Guevara Olvera, Reyna Isabel Rojas Martnez, Emma Zavaleta Meja,
June K. Simpson, Mario Rocha Sosa
Condiciones ptimas para Inducir Germinacin Errumpente y No Errumpente de Sclerotium cepivorum y
Secuencias Asociadas a la Sntesis de Protenas Durante la Germinacin No Errumpente
Revista Mexicana de Fitopatologa, vol. 29, nm. 1, 2011, pp. 69-72,
Sociedad Mexicana de Fitopatologa, A.C.
Mxico
www.redalyc.org
Proyecto acadmico sin fines de lucro, desarrollado bajo la iniciativa de acceso abierto
REVISTA MEXICANA DE FITOPATOLOGA/69
de aminocidos y protenas. Los resultados sugieren que la processes in sclerotia of S. cepivorum germinated hyphally.
germinacin no errumpente es ms rpida que la germinacin
errumpente debido a un incremento en los procesos Additional keywords: Substractive hybridization, allyl
metablicos de los esclerocios de S. cepivorum germinados de disulfide, GenBank.
forma no errumpente.
It has been shown by studies performed in vitro that S.
Palabras clave adicionales: Hibridacin sustractiva, disulfuro cepivorum sclerotia reveal two germination types: eruptive
de alilo, GenBank. and hyphal. The determination of what genes are expressed
during either germination rate will help to elucidate the
Estudios realizados in vitro han mostrado que los esclerocios biochemical and physiological processes between the two
de S. cepivorum presentan dos tipos de germinacin: types of germination. Consequently, the present study is
errumpente y no errumpente. El determinar los posibles genes aimed to determine the optimal conditions to induce the S.
que se expresan durante uno y otro tipo de germinacin cepivorum in vitro eruptive and hyphal germination and to
ayudar a dilucidar los procesos bioqumicos y fisiolgicos identify possible genes differentially expressed during S.
entre los dos tipos de germinacin. Por esta razn, los cepivorum hyphal germination throughout the Suppression
objetivos del presente trabajo fueron determinar las Sustractive Hybridization (SSH) technique (Diatchenko and
condiciones ptimas para inducir la germinacin errumpente Siebert, 1998). A total of 5 treatments were tested in order to
y no errumpente de S. cepivorum in vitro y determinar los determine the optimal conditions for eruptive and hyphal
posibles genes que expresan de forma diferencial en la germination: sclerotia germinated in the presence of water
germinacin no errumpente de S. cepivorum mediante la (DAS 0%) at 17 and 22 C, sclerotia germinated in the
tcnica de Hibridacin Sustractiva bajo condiciones de presence of 1% allyl disulfide (DAS 1%) at 17 and 22 C; and
Supresin HSS (Diatchenko y Siebert, 1998). Se probaron 5 sclerotia germinated in 2% of allyl disulfide (DAS 2%) at 22
tratamientos para determinar las condiciones ptimas para la C. The sclerotia were placed individually in a drop of PDA
germinacin errumpente y no errumpente: esclerocios medium in Petri dishes. Approximately 100 sclerotia per
germinados en presencia de agua (DAS 0%) a 17 y 22 C, treatment were assessed individually every 24 h for 16 days to
esclerocios germinados en presencia de 1% de disulfuro de determine the form of germination.
alilo (DAS 1%) a 17 y 22 C; y esclerocios germinados en 2% The treatments selected as optimal were the sclerotia
de disulfuro de alilo (DAS 2%) a 22 C. Los esclerocios se germinated in DAS 0% at 22 C (96.7 % of hypal germination)
colocaron individualmente en una gota de medio PDA dentro and DAS 2% at 22 C (91.67% of eruptive germination) (Fig.
de cajas Petri. Se valoraron aproximadamente 100 esclerocios 1). The SSH was conducted using the PCR-SelectTM cDNA
por tratamiento de forma individual cada 24 h por 16 das para subtraction (Clontech) kit and it confirmed that the clones
determinar la forma de germinacin. were differential by means of Southern hybridization. A total
Los tratamientos que se seleccionaron como ptimos of 23 differential clones were obtained, which were purified,
fueron esclerocios germinados en DAS 0 % a 22 C (96.7 % de sequenced and compared with proteins at the GenBank; 2
germinacin no errumpente) y DAS 2% a 22 C (91.67% de redundant clones and 6 with no significant similarities were
germinacin errumpente) (Figura 1). La HSS se realiz
usando el Kit PCR-SelectTM cDNA subtraction (Clontech) y se
120
corrobor que las clonas fueran diferenciales mediante una
hibridacin tipo Southern. Se obtuvieron 23 clonas 100
Germinacin (%)