Naturally and artificially aged seeds of rape, Brassica napus L., produced less ethylene than freshly harvested seed during the early stage of germination. With (9); it also breaks the dormancy of seed of various species (4,9,10,18,19). Stewart and Freebaim (17) showed that heat-treated lettuce seed which became insensitive to gibberellic acid germinated well subsequent to treatment with exogenous ethylene. Ruge (15) showed earlier that ethylene enhanced the percentage germination of aged oat seed. In the present paper, the pattern of ethylene production by rape seedlings in the early stage of germination and the stimulation of the germination of aged seeds by exogenous ethylene were studied.
MATERIALS AND METHODSMaterials used in the experiments were the seeds of rape, Brassica napus L., variety Chisaya-natane, harvested in 1965 and seeds were sealed in tin cans and were put with the other seed at 3 C until the experiment was started.A germination test of the seeds was conducted with three replications (100 seeds X 3) on December 10, 1969. The blotter method at 25 C was used. The first count was on the 3rd day, and the final count was on the 7th day. The number of normal seedlings, abnormal seedlings, and slowly germinating seeds was counted.For observing ethylene production by seedlings and effects of exogenous ethylene on aged seeds, all experiments were conducted in Erlenmeyer flasks equipped with rubber serum stoppers. To prevent mold growth, the flasks, stoppers, and seeds were sterilized at the start of the experiment. Flasks containing filter paper or agar media were autoclaved at 15 psi for 20 min. The rubber stoppers were immersed in 95% ethanol and dried under sterile conditions. The seeds were treated with 95% ethanol for 5 sec, rinsed with sterile water, sterilized with 0.1% sodium hypochlorite for 30 min, and washed three times with sterile water. Seedlings were germinated and grown under continuous fluorescent light in a room maintained at 25 C. Samples of ethylene given off from seedlings in a flask were withdrawn with a 1-ml syringe through the rubber stopper. The amount of ethylene was determined by gas chromatography, using an alumina column maintained at 50 C and a flame ionization detector. Each experiment was replicated three times.The first two experiments were conducted to follow the trend of ethylene production during germination. In the first trial, the seeds were maintained in a closed system in 25-ml Erlenmeyer flasks containing two layers of wet filter paper. A 1-ml gas sample was withdrawn daily for 9 days from each flask for ethylene determination. In the second trial, the seeds were transferred aseptically to 125-ml Erlenmeyer flasks containing 25 ml of 1% agar. In contrast to the first trial, a 1-ml gas sample was taken every 6 or 12 hr for ethylene analysis, after which the gaseous content of the flask was flushed with fresh ethylene-free air. This trial was conducted for 120 hr.In the third experiment, ethylene was introduced into the flask containing aged seeds t...