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Molecular identification of Echinochloa oryzicola Vasing. and E. crus‐galli (L.) Beauv. using a polymerase chain reaction–restriction fragment length polymorphism technique
Author(s) -
YASUDA KENTARO,
YANO AZUSA,
NAKAYAMA YUICHIRO,
YAMAGUCHI HIROFUMI
Publication year - 2002
Publication title -
weed biology and management
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.351
H-Index - 36
eISSN - 1445-6664
pISSN - 1444-6162
DOI - 10.1046/j.1445-6664.2002.00041.x
Subject(s) - biology , primer (cosmetics) , amplified fragment length polymorphism , restriction fragment length polymorphism , polymerase chain reaction , microbiology and biotechnology , restriction site , restriction enzyme , polymorphism (computer science) , genetics , restriction fragment , dna , genotype , gene , population , chemistry , demography , organic chemistry , sociology , genetic diversity
Polymerase chain reaction (PCR) and polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) techniques were applied for establishing the reliable practice in identification of Echinochloa oryzicola Vasing. and E. crus‐galli (L.) Beauv. (barnyardgrass). Total DNA was extracted from 18 accessions and 86 individuals of E. oryzicola , 33 accessions and 140 individuals of E. crus‐galli var. crus‐galli , 23 individuals of E. crus‐galli var. praticola , and six individuals of E. crus‐galli var. formosensis that were collected from Japan. A partial region of intergenetic spacer between trn T and trn L, and an intron of trn L were amplified separately using a trn‐a and trn‐b1 primer set, and a trn‐c and trn‐d primer set, respectively. All individuals of E. oryzicola showed the same fragment amplified by the trn‐a and trn‐b1 primer set. The fragment was 481 bp in length, and was undigested by Eco R I, whereas all individuals of E. crus‐galli , including three botanical varieties, showed the same fragment with a 449‐bp length. The fragment was digested by Eco R I into two fragments (178 and 271 bp). The fragment amplified by the trn‐c and trn‐d primer set in all individuals of E. oryzicola was digested by Alu I into two fragments (174 and 452 bp), but undigested by Dra I. In contrast, the fragment amplified by the trn‐c and trn‐d primer set in all individuals of E. crus‐galli was digested by Dra I into two fragments (134 and 487 bp), but undigested by Alu I. There was no intraspecific variation in these regions; thus, these two species are easily identifiable by using our method.