Alfalfa (Medicago sativa L.) plants have been reported to be autotoxic as well as allelopathic. Laboratory and greenhouse experiments through petri-dish and pot test were conducted to determine autotoxic effects of alfalfa leaf and soil extracts on the germination or early seedling growth of alfalfa, and to evaluate allelopathic effects of alfalfa leaf residues on alfalfa, barnyard grass, com, eclipta and soybean. Alfalfa seed germination was delayed depending on aqueous extract concentration, with no difference in final germination after 48 hours. Alfalfa root length was more sensitive to the autotoxic chemicals from leaf extracts than was germination or shoot length. Root growth of alfalfa was significantly inhibited at extract concentration of more than 1 g dry tissue/L (g ~textrmL-1 ). Hypocotyl growth, however, was not affected by all the concentrations of leaf extracts. Soil extracts from 4-yr-old alfalfa stand significantly reduced alfalfa root length by 66%, while soil extracts from 0,1, and 3yr-old stand stimulated root length up to 14-32% over the control. Residue incorporation with dry matters of alfalfa leaf at 100 g ~textrmkg-1 reduced seedling length of several crop and weed species, ranging from 53 to 87% inhibition. Addition of nutrient solution into alfalfa leaf extracts alleviated alfalfa autotoxic effect. This result indicates alfalfa leaf and soil extracts or residues could exert autotoxic as well as allelopathic substances into soil environments during and after establishment.

Autotoxicity restricts reseeding of new alfalfa(Medicago sativa L.) after alfalfa until autotoxic chemical(s) breaks down or is dispersed into external environments, often requiring up to a year or more. One solution for reducing autotoxicity would be to select germplasms or cultivars with tolerance to the autotoxic chemical(5) and use genetically breeding program. Bioassay of seed germination and early seedling growth was conducted to evaluate autotoxic responses of 3 varieties of alfalfa to the water-soluble extracts(at 4 and 8g/L) from alfalfa ‘Cody’leaf by using agar and filter paper medium in a petri-dish assay. Root length at 5 days after seeding was more sensitive to the extract than was hypocotyl length or seed germination, and was a better parameter of autotoxic effects of alfalfa leaf extracts. Use of an agar medium gave better sensitivity of root length than did use of filter paper. Evaluating tolerance with percent of control was more important indicator than was mean of root length because of significant variation among varieties in root length of control treatment. Bioassay ranked varieties in the following order of tolerance on the basis of relative root length; “Cody” >“ Pioneer 5373” >“ Alfagraze”. Seedling growth from old “Cody” seed was more sensitive to the autotoxic chemical(5) than was that from newly produced seed.

본 실험은 알파파 꽃 추출물로부터 Allelopathy와 Autotoxicity에 관련되는 활성물질을 분리, 동정하기 위하여 실시하였다. 알팔파 꽃 200g을 80% MeOH로 추출하여 CHCl3 층을 silica gel thin layer chromatography (TLC), microcrystalline cellulose thin layer chromatography (MCTLC)와 droplet countercurrent chromatography(DCCC)의 분획을 이용하여 생물검정을 행하고 DCCC의 생물검정에서 가장 억제력을 보였던 분획 5와 6은 HPLC로 활성물질을 동정하였다. HPLC분석 결과 분획 5에서는 ferulic acid, caffeic acid, vanillic acid, rutin, narringin 이, 분획 6에서는 ferulic acid, caffeic acid, vanillic acid, rutin, coumarin이 동정되었다. 이들 동정된 물질을 이용한 알팔파와 돌피에 대한 발아실험에서도 모두 알팔파와 돌피의 발아와 생육에 억제적으로 작용하였으며, 이중 coumarin의 처리가 가장 억제적이었다. 따라서 이들 물질이 알팔파와 돌피의 autotoxicity와 allelopathy에 관련하는 것으로 생각되었다.