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Diversidad Procariota En Ambientes Hipersalinos ESPECIAL TAXONOMÍA, FILOGENIA Y DIVERSIDAD SEM@FORO NUM. 65 | JUNIO 2018 cess of adaptation to an endophytic life style? PLoS Micromonospora lupini sp. nov. and Micromonospora Trujillo, ME, Riesco, R, Benito, P, and Carro, L. One 9(9): e108522. saelicesensis sp. nov., isolated from root nodules (2015). Endophytic actinobacteria and the interaction Trujillo ME, Kroppenstedt RM, Fernandez-Moline- of Lupinus angustifolius. Int J Syst Evol Microbiol of Micromonospora and nitrogen fixing plants. Front ro C, Schumann P y Martinez-Molina E (2007) 57:2799–2804. Microbiol 6, 1–15. Diversidad procariota en ambientes hipersalinos Cristina Sánchez-Porro, Rafael Ruiz de la Haba y Antonio Ventosa [email protected] [email protected] Departamento de Microbiología y Parasitología, Facultad de Farmacia, Universidad de Sevilla. [email protected] De izquierda a derecha: Blanca Vera Gargallo, Antonio Ventosa, María José León, Cristina Sánchez-Porro, Clara López Hermoso, Carmen Infante Domínguez, Ana Durán Viseras y Rafael Ruiz de la Haba. Nuestro grupo de investigación lleva activo campos tales como la genómica y la meta- tes hipersalinos. Estos estudios pusieron de desde hace más de 35 años; durante este genómica, los mecanismos de adaptación a manifiesto, entre otros muchos datos intere- tiempo hemos centrado nuestros estudios los ambientes extremos y las aplicaciones santes, la presencia de un nuevo grupo per- en aspectos relacionados con la diversidad biotecnológicas de estos microorganismos, teneciente a la clase Gammaproteobacteria, y taxonomía de bacterias y arqueas halófilas, especialmente con relación a la producción muy abundante en los estanques de salini- es decir, aquellas que habitan en ambien- de enzimas extracelulares por bacterias haló- dad intermedia, que no había sido ni aislado tes con elevadas concentraciones de sales. filas moderadas. ni caracterizado hasta la fecha. Tras un meti- Hasta la fecha nuestro grupo ha dado nom- culoso trabajo de aislamiento y caracteriza- bre a más de 130 especies de bacterias y ción describimos un nuevo género bacteria- arqueas, 34 géneros, varias nuevas familias EL NUEVO GÉNERO SPIRIBACTER no, al que denominamos Spiribacter, debido y una clase. Recientemente hemos partici- a su peculiar morfología celular en forma de pado en la actualización del Bergey’s Manual En los últimos años, hemos incorporado espiral. Hasta la fecha el género consta de of Systematics of Archaea and Bacteria, con las nuevas tecnologías de secuenciación cuatro especies: S. salinus, S. curvatus, S. 19 capítulos dedicados a las haloarqueas, masiva a nuestros estudios de diversidad roseus y S. aquaticus, todas ellas descritas que incluyen 10 géneros, 6 familias, 2 órde- procariota y hemos obtenido los metage- por nuestro equipo. En la actualidad esta- nes y una clase. Aunque nuestros estudios nomas de varios estanques de las salinas mos estudiando aspectos relacionados con se han centrado en el estudio taxonómico y de Santa Pola (Alicante) y de Isla Cristina la biología de este nuevo grupo bacteriano, de biodiversidad de procariotas de ambien- (Huelva). Su análisis nos ha permitido cono- como son su peculiar morfología, sus meca- tes acuáticos hipersalinos, también hemos cer en profundidad tanto la diversidad filoge- nismos de osmorregulación y su distribución realizado importantes aportaciones en otros nómica como metabólica de estos ambien- geográfica. 44 ESPECIAL TAXONOMÍA, FILOGENIA Y DIVERSIDAD SEM@FORO NUM. 65 | JUNIO 2018 FILOGENÓMICA DEL GÉNERO servación del orden de los genes entre repre- NUEVOS ESTUDIOS EN SUELOS SALINOS SALINIVIBRIO sentantes de cada filogrupo, observándose un elevado nivel de sintenia entre los miembros Aunque los ambientes hipersalinos acuá- Por otro lado hemos realizado una profun- del género Salinivibrio. ticos han sido nuestro principal objeto de da revisión del género Salinivibrio, habitante estudio, en los últimos años hemos ampliado habitual de las salinas solares. Tras numerosos nuestra línea de investigación incluyendo el muestreos y aislamientos, a partir de diferen- HALOARQUEAS: EL GÉNERO estudio de la diversidad procariota de suelos tes ambientes hipersalinos, obtuvimos una HALORUBRUM COMO MODELO salinos, concretamente de suelos hipersalinos colección de 70 cepas pertenecientes a este localizados en las Marismas del Odiel (Huelva). género. Además del estudio inicial basado en Con el objetivo de comprender mejor el Con este objetivo, inicialmente obtuvimos dos la comparación de secuencias del gen ARNr proceso de especiación en haloarqueas y metagenomas shotgun de estos suelos (con 16S de estas cepas, junto con las seis cepas cómo podría beneficiarse la taxonomía de una conductividad eléctrica de ~24 y 55 dS/m, tipo de las especies y subespecies del género, estudios de poblaciones, hemos examinado respectivamente). La comparación de estos se llevó a cabo un estudio MLSA (MultiLocus diferentes comunidades naturales perte- resultados con los obtenidos de los ambien- Sequence Analysis), basado en las secuencias necientes al género Halorubrum. Para ello, tes acuáticos ha puesto de manifiesto que la individuales y concatenadas de los genes gyrB, partimos de cepas aisladas y cultivadas a diversidad procariota presente en los suelos recA, rpoA y rpoD, que nos permitió clasificar- partir de diversas salinas solares de España, salinos es mucho mayor, pudiendo identificar- las en cuatro filogrupos diferentes, a excepción Namibia e Irán, comparándolas con las cepas se hasta 29 phyla diferentes, en contraste con de la cepa tipo de la especie Salinivibrio sha- tipo de 30 especies descritas previamente de los 8-9 phyla generalmente encontrados en los rmensis, que no pudo ser incluida en ningún dicho género mediante cinco aproximaciones ambientes acuáticos estudiados. Por otra parte, filogrupo, constituyendo, por tanto, un filotipo diferentes: i) análisis filogenético en base al a partir de estos metagenomas hemos logra- independiente del resto. Con la finalidad de gen ARNr 16S, que mostró una escasa capa- do reconstruir cuatro genomas de las especies poder emplear este esquema MLSA con fines cidad de resolución para diferenciar adecua- más abundantes en estos suelos, las cuales taxonómicos en la descripción de nuevas damente los nuevos aislados; ii) MLSA en están relacionadas con los phyla Balneolaeota especies de Salinivibrio, realizamos una com- base a la concatenación de los genes atpB, y Bacteroidetes y con la clase Halobacteria, y paración y posterior validación de este estudio EF-2, glnA, ppsA y rpoB’, que sí nos permitió cuyo metabolismo y estrategias de osmoadap- MLSA frente a los datos de hibridación ADN- agrupar las cepas en distintos clústeres; iii) tación están muy relacionados con las propie- ADN (DDH). En todos los casos, los valores perfil de lípidos polares (obtenidos mediante dades intrínsecas de este ecosistema. Por otro de DDH confirmaron los filogrupos obtenidos cromatografía en capa fina de alta resolu- lado, hemos determinado la influencia de la en base al MLSA, constituyendo cada uno de ción, HPTLC), que permitió observar peque- salinidad y de otros parámetros fisicoquímicos ellos una especie diferente. Con estos resul- ñas diferencias entre cepas pertenecientes en la estructura de la comunidad procariota tados hemos propuesto una correlación entre al mismo filogrupo, posiblemente debidas a de dichos suelos salinos. Actualmente estamos los valores de MLSA y DDH, considerando un la adaptación al nicho ecológico; iv) identidad trabajando en el aislamiento y caracterización 97 % de semejanza entre la secuencia conca- media de nucleótidos (ANI), que confirmó las de nuevos grupos de arqueas y bacterias a tenada de los cuatro genes empleados como clústeres obtenidos en base al estudio MLSA; partir de estos ambientes salinos, que cons- valor de corte para la delineación de especies v) estudios de hibridación ADN-ADN, median- tituyen representantes de nuevos taxones no del género Salinivibrio. Para completar el estu- te los que también se observó la heteroge- descritos hasta la fecha. dio hemos secuenciado los genomas de 33 de neidad y divergencia de cepas aisladas en estas cepas pertenecientes a distintos filogru- diferentes localizaciones geográficas a pesar pos y, según los diferentes índices de relación de pertenecer al mismo filogrupo. Estos estu- PUBLICACIONES RECIENTES genómica (ANIb, ANIm, OrthoANI y DDH in dios nos han permitido delinear y describir SELECCIONADAS silico) y tras un análisis filogenómico, se con- nuevas especies del género Halorubrum y firmaron nuevamente los filogrupos definidos establecer diversas sinonimias entre especies Amoozegar MA, Khansha J, Mehrshad M, Shahzadeh Fazeli SA, Ramezani M, de la Haba RR, Sánchez-Po- anteriormente en base al MLSA. Estos estudios previamente descritas. Actualmente estamos rro C y Ventosa A. (2017). Soortia roseihalophila gen. han llevado a la reclasificación de la especie realizando el aislamiento de nuevas cepas nov., sp. nov., a new taxon in the order Balneolales isolated Salinivibrio costicola subsp. vallismortis, como pertenecientes al género Halorubrum para from a travertine spring, and description of Soortiaceae una sinonimia de la especie S. proteolyticus, ampliar estos estudios mediante una apro- fam. nov. Int J Syst Evol Microbiol 67: 113-120. y a la descripción de una nueva especie, ximación genómica. Hemos secuenciado el Amoozegar MA, Siroosi M, Atashgahi S, Smidt H y Ventosa A. (2017). Systematics of haloarchaea and Salinivibrio kushneri, constituida por 10 cepas genoma completo de un elevado número de biotechnological potential of their hydrolytic enzymes.
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