Identification of chitinase mRNA in abscission zones from bean (Phaseolus vulgaris Red Kidney) during ethylene-induced abscission

By:

GOMEZ L., M.A

Contributor(s):

Material type: Article

ArticleLanguage: English Description: 10(9):741-746Subject(s): LOC classification:

33182

In: Plant, Cell and EnvironmentSummary: En diferentes momentos se extrajo el ARN total de las zonas de abscision de hojas de frijol, despues de la induccion de la abscision por etileno. El ARN se traslado al sistema de germen de trigo y los productos se analizaron por SDS-EGPA. Los productos de p.mol. 42, 32 y 17 kilodaltones se acumularon sustancialmente durante la induccion. Se intento establecer que la especie ARNm que origino el producto de 32 kilodaltones estaba codificada por la enzima quitinasa regulada a su vez por el etileno. La quitinasa madura (30 kilodaltones) se purifico de zonas de abscision tratadas por etileno y se uso para crear anticuerpos monoespecificos en conejos. Estos anticuerpos reconocieron el producto de 32 kilodaltones y la quitinasa madura. La diferencia de 2 kilodaltones en el p.mol. se debio a la presencia de la secuencia de senal que las membranas microsomicas podian eliminar. Tambien se detecto quitinasa mediante la prueba enzimatica y la inmunodeteccion en soporte o matriz de homogenatos crudos de las zonas de abscision tratadas con etileno. La quitinasa parece ser ubicua en plantas de frijol y probablemente no tiene una funcion directa en la abscision. (RA-CIAT)Summary: Total RNA was extracted from bean leaf abscission zones at different times after the induction of abscission by ethylene. The RNA was translated in the wheat germ system and the products analyzed by SDS-PAGE. Products of mol.wt. 42, 32, and 17 kilodaltons were seen to accumulate substantially during induction. An attempt was made to establish that the mRNA species which produced the 32 kilodaltons product, was coded for the ethylene- regulated enzyme chitinase. Mature chitinase (30 kilodaltons) was purified from ethylene-treated abscission zones and used to raise monospecific antibodies in rabbits. These antibodies recognized the 32 kilodaltons product and mature chitinase. The 2 kilodaltons difference in mol.wt. was due to the presence of the signal sequence which could be removed by microsomal membranes. Chitinase was also detected by enzymatic assay and immunoblotting of crude homogenates from ethylene-treated abscission zones. Chitinase appears to be ubiquitous in bean plants and probably does not have a direct role in abscission. (AS)

Tags from this library: No tags from this library for this title.

Average rating: 0.0 (0 votes)

Holdings ( 1 )
Title notes ( 2 )

Holdings
Item type	Current library	Collection	Call number	Copy number	Status	Date due	Barcode	Item holds
Journal Article	CIAT Library Document collection CINFOS	Document Collection CINFOS	33182 (Browse shelf(Opens below))	c.1	Short Loan		100046327

Total holds: 0

En diferentes momentos se extrajo el ARN total de las zonas de abscision de hojas de frijol, despues de la induccion de la abscision por etileno. El ARN se traslado al sistema de germen de trigo y los productos se analizaron por SDS-EGPA. Los productos de p.mol. 42, 32 y 17 kilodaltones se acumularon sustancialmente durante la induccion. Se intento establecer que la especie ARNm que origino el producto de 32 kilodaltones estaba codificada por la enzima quitinasa regulada a su vez por el etileno. La quitinasa madura (30 kilodaltones) se purifico de zonas de abscision tratadas por etileno y se uso para crear anticuerpos monoespecificos en conejos. Estos anticuerpos reconocieron el producto de 32 kilodaltones y la quitinasa madura. La diferencia de 2 kilodaltones en el p.mol. se debio a la presencia de la secuencia de senal que las membranas microsomicas podian eliminar. Tambien se detecto quitinasa mediante la prueba enzimatica y la inmunodeteccion en soporte o matriz de homogenatos crudos de las zonas de abscision tratadas con etileno. La quitinasa parece ser ubicua en plantas de frijol y probablemente no tiene una funcion directa en la abscision. (RA-CIAT)

Total RNA was extracted from bean leaf abscission zones at different times after the induction of abscission by ethylene. The RNA was translated in the wheat germ system and the products analyzed by SDS-PAGE. Products of mol.wt. 42, 32, and 17 kilodaltons were seen to accumulate substantially during induction. An attempt was made to establish that the mRNA species which produced the 32 kilodaltons product, was coded for the ethylene- regulated enzyme chitinase. Mature chitinase (30 kilodaltons) was purified from ethylene-treated abscission zones and used to raise monospecific antibodies in rabbits. These antibodies recognized the 32 kilodaltons product and mature chitinase. The 2 kilodaltons difference in mol.wt. was due to the presence of the signal sequence which could be removed by microsomal membranes. Chitinase was also detected by enzymatic assay and immunoblotting of crude homogenates from ethylene-treated abscission zones. Chitinase appears to be ubiquitous in bean plants and probably does not have a direct role in abscission. (AS)