Abstract
The inhibitory allelopathic activity of Pueraria montana (Kudzu), and activities of two putative allelochemical isoflavones, puerarin and daidzein, were evaluated using the protoplast co-culture method with digital image analysis using lettuce as a recipient (DIA-PP method). Cotyledon protoplasts of Kudzu were isolated using Cellulase R10 and Driselase 20 in 0.6 M mannitol solution. Optimal hormonal condition and density for growth of Kudzu protoplasts were surveyed. Medium for co-culture of Kudzu or isoflavones with lettuce protoplasts was 50 μl liquid MS basal medium containing 1 μM 2,4-dichlorophenoxyacetic acid, 0.1 μM benzyladenine, 3% sucrose, and 0.4 M or 0.6 M mannitol. Protoplast division of lettuce was strongly inhibited by Kudzu at a low density (104/ml). Slightly less inhibition by Kudzu on cell wall formation and yellow pigment accumulation stages of lettuce growth was also observed. Puerarin did not inhibit the growth of lettuce protoplasts at three growth stages but slightly stimulated growth at high concentrations. By contrast, daidzein, aglycon of puerarin, inhibited growth at three stages of lettuce protoplast growth and strongly inhibited cell division at 100 μM. Daidzein might be one cause of the strong inhibitory allelopathic activity of Kudzu. Grade of inhibitory activities was compared with that of other allelopathic plants including an invader plant and their allelochemicals studied using the DIA-PP method.
Highlights
Pueraria montana (Lour.) Merr. var. lobata (Willd.) (Kudzu), known as P. lobata Ohwi, and P. thunbergiana, is a leguminous plant, naturally grown in Japan and East Asia and introduced to other countries for its rapid growth in the field to stop soil erosion
We examined the effects of isoflavones, puerarin and daidzein using the same lettuce protoplast method to evaluate the putative allelochemicals of Kudzu, in comparison with the protoplasts of different plant species and different allelochemicals reported
In the protoplast cultures of Kudzu, optimal hormonal condition for cell division and colony development was a combination of 10 μM 2,4-D and 0.1 - 10 μM BA in Murashige and Skoog (MS) basal medium containing 0.6 M mannitol. 2,4-D only medium was good for early cell wall formation, but both 2,4-D and BA were needed for further growth
Summary
Pueraria montana (Lour.) Merr. var. lobata (Willd.) (Kudzu), known as P. lobata Ohwi, and P. thunbergiana, is a leguminous plant, naturally grown in Japan and East Asia and introduced to other countries for its rapid growth in the field to stop soil erosion. Lobata (Willd.) (Kudzu), known as P. lobata Ohwi, and P. thunbergiana, is a leguminous plant, naturally grown in Japan and East Asia and introduced to other countries for its rapid growth in the field to stop soil erosion. High content of an isoflavone, puerarin, was reported in the root [2] [3] [4] and stalk [5] of Kudzu, which is a glycoside of an aglycon, daidzein, though the content of daidzein was low. Different types of aglycons and their glycosides of isoflavones are known in different organs at different growth stages of Kudzu plants and tissue cultured cells [2]-[8]. In the green cotyledons-derived suspension-cultured cells of Kudzu, content of puerarin was the highest in isoflavones [2]
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