Hisashi Kato-Noguchi

Rice seedlings release momilactone B into the environment.

Since the growth inhibitor momilactone B was found recently in root exudates of rice (Oryza sativa L.), 3-day-old rice seedlings were transferred to hydroponic culture and the level of momilactone B released into the environment from the seedlings was measured. At day 15 after transfer, the level of momilactone B in the culture solution was 1.8 nmol per seedling compared with endogenous levels of 0.32 and 0.63 nmol per root and shoot, respectively, suggesting that rice seedlings actively releases momilactone B into the culture solution. This release must occur from the roots because only rice roots were immersed in the culture solution. Momilactone B inhibited the growth of ten cress (Lepidium sativum L.) seedlings at concentrations greater than 3 microM. Ten rice seedlings were incubated with ten cress seeds in a Petri dish containing 1 ml of medium, the medium contained 18 nmol of momilactone B, which came to 18 microM. This level of momilactone B was enough to reveal growth inhibition of the cress seedlings. Release level of momilactone B and its effectiveness as a growth inhibitor suggest that it may play an important role in rice allelopathy.

The Role of Momilactones in Rice Allelopathy

Large field screening programs and laboratory experiments in many countries have indicated that rice is allelopathic and releases allelochemical(s) into its environment. A number of compounds, such as phenolic acids, fatty acids, phenylalkanoic acids, hydroxamic acids, terpenes, and indoles, have been identified as potential rice allelochemicals. However, the studies reviewed here demonstrate that the labdane-related diterpenoid momilactones are the most important, with momilactone B playing a particularly critical role. Rice plants secrete momilactone B from their roots into the neighboring environments over their entire life cycle at phytotoxic levels, and momilactone B seems to account for the majority of the observed rice allelopathy. In addition, genetic studies have shown that selective removal of the momilactones only from the complex mixture found in rice root exudates significantly reduces allelopathy, demonstrating that these serve as allelochemicals, the importance of which is reflected in the presence of a dedicated momilactone biosynthetic gene cluster in the rice genome.

Allelopathic substance in rice root exudates: Rediscovery of momilactone B as an allelochemical

Much research on rice allelopathy has been directed toward the selection of allelopathic rice strains and the identification of allelochemicals in rice. This paper briefly summarizes recent progress in the rice allelopathy and focuses on rediscovery of momilactone B as an allelochemical. A large number of rice varieties were found to inhibit the growth of several plant species when grown together under field and/or laboratory conditions. These findings suggest that rice probably produces and releases allelochemical(s) into the environment. The putative compound causing the inhibitory effect of rice was recently isolated from rice root exudates, and the chemical structure of the inhibitor was determined by spectral data as momilactone B. In addition, it has been found that momilactone B is released from rice roots into the neighboring environment, and the release level of momilactone B from rice may be sufficient to cause growth inhibition of neighboring plants. These findings suggest that momilactone B may play an important role in rice allelopathy.

Allelopathy of oats. I. Assessment of allelopathic potential of extract of oat shoots and identification of an allelochemical.

The allelopathic potential of oat (Avena sativa L.) extracts was investigated under laboratory conditions. The ethyl ether-, acetone-, and water-soluble fractions obtained from the extract of oat shoots inhibited the germination and growth of roots and hypocotyls of lettuce (Lactuca sativa L.). The inhibitory activity of the water-soluble fraction was maximum, followed by that of ethyl ether-soluble and acetone-soluble fraction. An active principle of the water-soluble fraction was isolated and its structure was determined by spectral data asl-tryptophan.l-Tryptophan inhibited the growth of hypocotyls and roots of lettuce seedlings at concentrations greater than 0.03 and 0.1 mM, respectively. These results suggested thatl-tryptophan may be an allelochemical which affects the growth or germination of different plant species.

Barnyard grass-induced rice allelopathy and momilactone B.

Here, we investigated chemical-mediated interaction between crop and weeds. Allelopathic activity of rice seedlings exhibited 5.3-6.3-fold increases when rice and barnyard grass seedlings were grown together, where there may be the competitive interference between rice and barnyard grass for nutrients. Barnyard grass is one of the most noxious weeds in rice cultivation. The momilactone B concentration in rice seedlings incubated with barnyard grass seedlings was 6.9-fold greater than that in rice seedlings incubated independently. Low nutrient growth conditions also increased allelopathic activity and momilactone B concentrations in rice seedlings. However, the increases in the low nutrient-induced allelopathic activity and momilactone B concentration were much lower than those in barnyard grass-induced allelopathic activity and momilactone B concentration. Root exudates of barnyard grass seedlings increased allelopathic activity and momilactone B concentration in rice seedlings at concentrations greater than 30 mg/L of the root exudates, and increasing the exudate concentration increased the activity and momilactone B concentration. Therefore, barnyard grass-induced allelopathic activity of rice seedlings may be caused not only by nutrient competition between two species, but also by components in barnyard grass root exudates. As momilactone B shows strong allelopathic activities, barnyard grass-induced allelopathic activity of rice may be due to the increased concentration of momilactone B in rice seedlings. The present research suggests that rice may respond to the presence of neighboring barnyard grass by sensing the components in barnyard grass root exudates and increasing allelopathic activity by production of elevated concentration of momilactone B. Thus, rice allelopathy may be one of the inducible defense mechanisms by chemical-mediated plant interaction between rice and barnyard grass, and the induced-allelopathy may provide a competitive advantage for rice through suppression of the growth of barnyard grass.

Contribution of momilactone A and B to rice allelopathy.

Eight cultivars of rice (Oryza sativa L.) inhibited shoot and root growth of Echinochloa crus-galli when co-cultured with rice seedlings in a bioassay medium. Momilactone A and B were found in the bioassay medium of all rice cultivars, and concentrations of momilactone A and B in the medium were 0.21-1.5 and 0.66-3.8 micromol/L, respectively, indicating that all rice cultivars may secrete momilactone A and B into the medium. Exogenously applied momilactone A and B inhibited the growth of shoots and roots of E. crus-galli at concentrations greater than 30 and 1 micromol/L, respectively. The concentrations required for 50% growth inhibition of E. crus-galli shoots and roots, respectively, were 146 and 91 micromol/L for momilactone A and 6.5 and 6.9 micromol/L for momilactone B. Considering the growth inhibitory activity and concentrations found in the bioassay medium, momilactone A may have caused only 0.8-2.2% of the observed growth inhibition of E. crus-galli roots and shoots by rice. However, momilactone B in the medium was estimated to be able to cause 59-82% of the observed growth inhibition of E. crus-galli roots and shoots by the rice seedlings. In addition, the concentrations of momilactone B in the medium reflected the observed differences in the growth inhibition of E. crus-galli by the eight rice cultivars investigated. This suggests that the allelopathic activity of rice may depend primarily on the secretion level of momilactone B. Therefore, momilactone B may play a very important role in rice allelopathy.

Isolation and identification of an allelopathic substance from peel of Citrus junos.

The inhibitory effect of Citrus junos peel on plant growth using lettuce (Lactuca sativa L.) as a bioassay material was investigated, since the powder of the peel had been found to inhibit growth of weeds. Basic, neutral and acidic fractions were separated from the aqueous fraction obtained from the methanol extract of C. junos peel. All fractions inhibited the growth of lettuce seedlings, but by far the greatest inhibition was observed with the neutral fraction. Thus, the latter was further purified and an allelopathically active substance was isolated. The structure of the substance was determined from high-resolution MS and 1H and 13C NMR spectral data as abscisic acid-beta-D-glucopyranosyl ester (ABA-GE). ABA-GE inhibited hypocotyl and root growth of lettuce seedlings at concentrations greater than 0.3 microM, and the concentrations for 50% inhibition of hypocotyl and root growth were 2.3 and 1.4 microM, respectively. The effectiveness of ABA-GE on inhibition of growth and the occurrence of ABA-GE in the peel itself suggested that ABA-GE may play an important role in the allelopathic potential of C. junos peel. The peel may be potentially useful for weed management in a field setting.

Assessment of allelopathic potential of shoot powder of lemon balm

The allelopathic potential of lemon balm (Melissa officinalis) was investigated under laboratory conditions. The shoot powder of 30-day-old plants inhibited the germination and the growth of roots and shoots of Amaranthus caudatus, Digitaria sanguinalis and lettuce (Lactuca sativa). Significant reductions in the germination and growth of roots and shoots were observed as the powder concentration increased. The concentration-dependent responses of the test plants to the powder of lemon balm suggest that the powder might contain allelochemicals, which should be investigated further in the laboratory and the field for practical application of the powder as weed inhibiting agent.

Isolation and identification of an allelopathic substance in Pisum sativum

The residue of peas (Pisum sativum L.) has allelopathic activity and the putative compound causing this inhibitory effect was isolated from a methanol extract of pea shoots. Chemical structure of this compound was determined by high-resolution MS, IR and 1H NMR spectral data as pisatin. Pisatin inhibited growth of cress (Lepidium sativum L.) and lettuce (Lactuca sativa L.) seedlings at concentrations greater than 10 and 30 microM, respectively. The doses required for 50% growth inhibition of roots and hypocotyls of cress were 61 and 91 microM, respectively, and those of lettuce were 78 and 115 microM, respectively. The concentration of pisatin in the pea shoots was 32.7 nmol x g(-1) fresh weight. The effectiveness of pisatin on growth inhibition in cress and lettuce, and its occurrence in pea shoots suggest that it may contribute to the growth inhibitory effect of pea residue, and may play an important role in pea allelopathy.

Allelopathy of oats. II. Allelochemical effect ofL-Tryptophan and its concentration in oat root exudates.

Abstractl-Tryptophan caused growth inhibition of roots and hypocotyls (or coleoptiles) of cockscomb (Amaranthus caudatus L.), lettuce (Lactuca sativa L.), cress (Lepidium sativum L.), timothy (Phleum pratense L.), rice (Oryza sativa L.), wheat (Triticum aestivum L.), and oat (Avena sativa L.), increasing the dose ofl-tryptophan increased the inhibition. The concentrations for 50% inhibition of the root growth were 0.14, 0.15, 0.21, 0.79, 0.95, 1.7, and 2.4 mM for cockscomb, cress, lettuce, timothy, rice, wheat, and oat, respectively; the concentrations for 40% inhibition of the hypocotyl (or coleoptile) growth were 0.28, 0.33, 0.43, 2.7, 4.5, 7.2, and 15 mM for cockscomb, cress, lettuce, timothy, rice, wheat and oat, respectively. The levels ofl-tryptophan in oat seedlings and in its root exudates were 29.3 mg/kg fresh wt and 0.25 mM under light conditions, and 21.1 mg/kg fresh wt and 0.18 mM under dark conditions, respectively. The presence ofl-tryptophan in the root exudates coupled with its effect on growth suggested thatl-tryptophan may play an important role in the growth inhibition of other plants in nature.