Growth promotion of rice seedlings by allelopathic polysaccharide from Welsh onion

The underground portion of the Welsh onion (Allium fistulosum) was extracted and separated into seven fractions to purify allelopathic compounds that were found to promote the growth of rice seedlings (Oryza sativa). The 80% (v/v) ethanol-insoluble fraction of the hot water extract showed the highest growth-promoting effect. Purification by DEAE-cellulose chromatography gave four fractions, AD-1, 2, 3 and 4. AD-3 at 1000 ppm stimulated root and shoot growth by about 2.4 and 1.5 times over the respective water and sucrose-treated controls. The molecular weight of AD-3 was roughly estimated as 630 kDa by gel permeation chromatography. Seventy two percent (by wt.) of AD-3 was a carbohydrate and no proteins were detected. A GC analysis of the neutral sugar composition revealed the presence of Gal (50.1%), Man (17.9%), Ara (10.4%), Rha (8.8%), Glc (7.2%) and Xyl (5.6%). About 3% (by wt.) of the total carbohydrate was uronic acid, which was identified as GalU by a GC analysis. The remainder of AD-3 was extracted in ethyl acetate after its hydrolysis with 2 M HCl. The major component of the ethyl acetate-soluble fraction was identified as unsaturated linear primary alcohols, 1-tetradecanol and 1-octadecanol (16% and 84% by peak areas from GC, respectively) by means of NMR and GC-MS. Sole or combined treatment of these alcohols did not affect the growth of rice seedlings. After partial hydrolysis with 0.2 M trifluoroacetic acid or by a sodium periodate treatment, the activity of AD-3 had completely disappeared, suggesting that the sugar moiety and/or molecular size were important for the activity.