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.     

Concentration and release level of momilactone B in the seedlings of eight rice cultivars

The release levels of a growth inhibitor, momilactone B, from rice (Oryza sativa L.) seedlings of eight cultivars were compared with the endogenous concentrations of momilactone B in their seedlings. All rice cultivars contained momilactone B in the seedlings, and their concentrations differed between the cultivars. Momilactone B was also found in all culture solutions in which these rice seedlings were grown, and the concentrations differed between the cultivars. The momilactone B concentrations in the culture solutions were reflected in the momilactone B concentrations in the seedlings. These results suggest that all rice cultivars may produce momilactone B and release momilactone B into the culture solutions. In addition, the release level of momilactone B may depend on the production level of momilactone B in the seedlings, which may affect allelopathic potential of these rice cultivars because as a growth inhibitor, momilactone B is able to act as an allelochemical.     

The relation between growth inhibition and secretion level of momilactone B from rice root

Abstract

The growth inhibitory activity of seven rice (Oryza sativa L.) cultivars and the secretion level of momilactone B from these rice cultivars were determined to understand chemical basis of the interaction of rice with other plant species. All rice cultivars inhibited the growth of hypocotyls and roots of lettuce (Lactuca sativa L.) seedlings when the lettuce was grown together with the rice, and showed different range of the inhibitory activity. These results suggest that all rice cultivars may possess allelopathic activity and the activity may be cultivar dependent. Momilactone B, which is a potent growth inhibitor, was found in root exudates of all rice cultivars, and the momilactone B concentration was also cultivar-dependent. The allelopathic activity of each rice cultivar was closely correlated with momilactone B concentration in the root exudates. The present results suggest that rice cultivars possess various allelopathic activities and their allelopathic activity may depend on the secretion level of momilactone B. Therefore, momilactone B may play an important role in rice allelopathy and in the chemical interactions of rice with other plant species.     

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.     

Secretion of momilactone A from rice roots to the rhizosphere

The secretion levels of momilactone A from rice (Oryza sativa L.) seedlings of eight cultivars into the rhizosphere were compared with the endogenous momilactone A concentrations in their shoots and roots. All rice cultivars contained momilactone A in the shoots and roots, and concentrations differed among the cultivars. Momilactone A was also found in all culture solutions in which the rice seedlings were grown, and the concentrations differed among the cultivars. The momilactone A concentrations in the culture solutions were reflected in the momilactone A concentrations in the shoots. These results suggest that all rice cultivars may produce momilactome A and secrete momilactone A into the culture solutions. The secretion levels of momilactone A may be more dependent on their capacities for momilactone A production in the shoots than on their capacities for momilactone A transportation from the shoots into the environment via the roots. As momilactone A acts as an antimicrobial and allelopathic agent, the secretion of momilactone A into the rice rhizosphere may provide a competitive advantage for root establishment through local suppression of soil microorganisms and inhibition of the growth of competing plant species.     

Assessment of Allelopathic Potential of Root Exudate of Rice Seedlings

To determine the allelopathic potential of root exudate from early developmental stage of rice (Oryza sativa L), 6-d-old seedlings of eight cultivars were grown with 3-d-old alfalfa (Medicago sativa L.), cress (Lepidium sativum L.) or lettuce (Lactuca sativa L.) seedlings in Petri dishes under controlled condition. All rice cultivars (cv. Norin 8, Kamenoo, Nipponbare, Kinuhikari, Koshihikari, Sasanishiki, Yukihikari and Hinohikari) inhibited growth of roots, shoots and fresh mass of alfalfa, cress and lettuce seedlings. Effectiveness of cv. Koshihikari was the greatest and more than 60% inhibition was recorded in all bioassays, followed by that of cv. Norin 8 of which effectiveness was more than 40%.     

Identification of two phytotoxins, blumenol A and grasshopper ketone, in the allelopathic Japanese rice variety Awaakamai

Aqueous methanol extracts of the traditional rice (Oryza sativa) variety Awaakamai, which is known to have the greatest allelopathic activity among Japanese traditional rice varieties, inhibited the growth of roots and shoots of cress (Lepidium sativum), lettuce (Lactuca sativa), timothy (Phleum pratense), Digitaria sanguinalis, Lolium multiflorum and Echinochloa crus-galli. Increasing the extract concentration increased the inhibition, suggesting that the extract of Awaakamai contains growth inhibitory substances. The extract of Awaakamai was purified and two main growth inhibitory substances were isolated and determined by spectral data as blumenol A and grasshopper ketone. Blumenol A and grasshopper ketone, respectively, inhibited the growth of cress shoots and roots at concentrations greater than 10 and 30 μmol/L. The concentrations required for 50% growth inhibition on cress roots and shoots were 84 and 27 μmol/L, respectively, for blumenol A, and 185 and 76 μmol/L, respectively, for grasshopper ketone. These results suggest that blumenol A and grasshopper ketone may contribute to the growth inhibitory effect of Awaakamai and may play an important role in the allelopathy of Awaakamai.     

Effects of Capsaicin on Plant Growth

Rice (Oryza sativa L.) seedlings inhibited the growth of hypocotyls and roots of cress (Lepidium sativum L.) seedlings when both seedlings were grown together. Two growth inhibiting substances were found in the culture solution in which rice seedlings were hydroponically grown for 14 d. One growth inhibitor was further purified. This suggests that the rice seedlings may produce growth inhibiting substances, acting as allelochemicals to other plants, and release them from their roots into the environment.     

Diterpene phytoalexins are biosynthesized in and exuded from the roots of rice seedlings

Rice (Oryza sativa L.) produces a variety of diterpene phytoalexins, such as momilactones, phytocassanes, and oryzalexins. Momilactone B was previously identified as an allelopathic substance exuded from the roots of rice. We identified in this present study momilactone A and phytocassanes A-E in extracts of, and exudates from, the roots of rice seedlings. The concentration of each compound was of the same order of magnitude as that of momilactone B. Expression analyses of the diterpene cyclase genes responsible for the biosynthesis of momilactones and phytocassanes suggest that these phytoalexins found in roots are primarily biosynthesized in those roots. None of phytocassanes B-E exhibited allelopathic activity against dicot seedling growth, whereas momilactone A showed much weaker allelopathic activity than momilactone B. The exudation of diterpene phytoalexins from the roots might be part of a system for defense against root-infecting pathogens.     

Absorption of momilactone A and B by Arabidopsis thaliana L. and the growth inhibitory effects

Although most allelochemicals can potentially cause growth inhibition in receiver plants, there is little information available about the absorption of these allelochemicals by the receiver plants. The present research describes the absorption of momilactone A and B by Arabidopsis thaliana (L.) and effects of the absorption on Arabidopsis growth. Exogenously applied momilactone A and B inhibited the growth of Arabidopsis hypocotyls and roots at concentrations greater than 10 and 1μmol/L, respectively. The levels of momilactone A and B in Arabidopsis hypocotyls were approximately 3.2 and 2.4% of momilactone A and B, respectively, in the medium and those in Arabidopsis roots were about 3.9-3.4%, respectively. The absorption rates of momilactone A and B by Arabidopsis were not significantly different. The present research suggests that momilactone A and B may be absorbed in proportion to their applied levels, and the growth inhibitory effects of momilactone A and B may also correlated with their endogenous levels. However, the effectiveness of momilactone B on growth inhibition was much greater than that of momilactone A, and the sensitivities of hypocotyls to momilactone A and B were greater than those of roots. This is the first report describing the absorption of potent rice allelochemicals, momilactone A and B by receiver plants.     

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.     

UV-induced momilactone B accumulation in rice rhizosphere

UV-irradiation increased the concentration of momilactone B in shoots and roots of rice seedlings, and increasing the irradiation increased the concentration. The concentration in 90-min UV-irradiated shoots and roots, respectively, was 31.8- and 3.6-fold higher than that in non-irradiated shoots and roots. After UV-irradiation the concentration of momilactone B in rice shoots decreased. There was, however, an accumulation of momilactone B in the medium in which UV-irradiated seedlings were grown. Five days after UV-irradiation, momilactone B in the medium was at a level 2.5 times greater than on day 0, which was 47% of momilactone B in the seedlings, suggesting that rice may actively secrete momilactone B into medium. Therefore, UV-irradiation increased not only production of momilactone B in rice seedlings but also secretion of momilactone B into rice rhizosphere. As momilactone B acts as an antimicrobial and allelopathic agent, secretion of momilactone B into the rhizosphere may provide a competitive advantage for root establishment through local suppression of soil microorganism and inhibition of the growth of competing plant species.     

An allelopathic substance in red pine needles (Pinus densiflora)

Aqueous methanol extracts of red pine (Pinus densiflora) needles inhibited the growth of roots and shoots of cress (Lepidium sativum), lettuce (Lactuca sativa), alfalfa (Medicago sativa), ryegrass (Lolium multiflorum), timothy (Pheleum pratense), Digitaria sanguinalis and Echinochloa crus-galli. Increasing the extract concentration increased inhibition, suggesting that the pine needles may have growth inhibitory substances and possess allelopathic potential. The aqueous methanol extract of the pine needles was purified, and a main inhibitory substance was isolated and determined by spectral data as 9alpha,13beta-epidioxyabeit-8(14)en-18-oic acid. This substance inhibited root and shoot growth of cress and Echinochloa crus-galli seedlings at concentrations greater than 0.1 mM. The endogenous concentration of the substance was 0.13 mmol/kg pine needle. These results suggest that 9alpha,13beta-epidioxyabeit-8(14)en-18-oic acid may contribute to the growth inhibitory effect of the pine needles and may play an important role in the allelopathy of red pine.     

Plant Growth Inhibitory Compounds from Aqueous Leachate of Wheat Straw

When seedlings of lettuce, cress, rice and wheat were incubated with the leachate of wheat straw, the roots growth of lettuce and garden cress were particularly inhibited. The leachate of wheat straw (100 g eq./l) showed 80.5 and 79.4% inhibition for lettuce and cress roots, respectively. The inhibitory activity was stronger as the concentration of wheat straw leachate was greater. This result indicates that allelochemical(s) inhibiting the roots growth of lettuce and cress are leached from the wheat straw into the water. Two potent compounds were isolated from the leachate of the wheat straw and identified as syringoylglycerol 9-O-β-d-glucopyranoside and l-tryptophan by spectral analyses. Syringoylglycerol 9-O-β-d-glucopyranoside inhibited the roots growth of lettuce and cress at concentrations greater than 0.1 and 10.0 μM, respectively. On the other hand, l-tryptophan inhibited the roots growth of lettuce and cress at concentrations greater than 0.1 and 1.0 μM, respectively. The content of syringoylglycerol 9-O-β-d-glucopyranoside and l-tryptophan in the leachate of wheat straw (100 g eq./l) was 18.4 ± 0.7 and 6.2 ± 0.6 μM, respectively. Syringoylglycerol 9-O-β-d-glucopyranoside (18.4 μM) showed 21.5 and 13.5% inhibition in the lettuce and cress roots assay, respectively. On the other hand, 6.2 μM of l-tryptophan showed 47.5 and 35.0% inhibition in the lettuce and cress roots assay, respectively. These results suggested that l-tryptophan may be a major contributor to the allelopathy in aqueous leachate of wheat straw and syringoylglycerol 9-O-β-d-glucopyranoside may be a minor contributor.     

Diterpene Phytoalexins Are Biosynthesized in and Exuded from the Roots of Rice Seedlings

Rice (Oryza sativa L.) produces a variety of diterpene phytoalexins, such as momilactones, phytocassanes, and oryzalexins. Momilactone B was previously identified as an allelopathic substance exuded from the roots of rice. We identified in this present study momilactone A and phytocassanes A–E in extracts of, and exudates from, the roots of rice seedlings. The concentration of each compound was of the same order of magnitude as that of momilactone B. Expression analyses of the diterpene cyclase genes responsible for the biosynthesis of momilactones and phytocassanes suggest that these phytoalexins found in roots are primarily biosynthesized in those roots. None of phytocassanes B–E exhibited allelopathic activity against dicot seedling growth, whereas momilactone A showed much weaker allelopathic activity than momilactone B. The exudation of diterpene phytoalexins from the roots might be part of a system for defense against root-infecting pathogens.     

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.     

Possible involvement of momilactone B in rice allelopathy

Since residues and extracts of rice plants were known to inhibit the germination and growth of several plant species, the possible involvement of a growth inhibitor, momilactone B, in rice allelopathy was discussed. Momilactone B was found in shoots and roots of rice plants over their entire life cycle. The level of momilactone B in shoots and roots increased with rice plant growing until flowering initiation, and then decreased. The highest level of momilactone B in the shoots and roots at the day of flowering initiation was 245 and 64.1 nmol g(-1) fresh weight, respectively. Thus, 1 kg of rice shoots and roots, respectively, may be able to release 245 and 64.1 micromol of momilactone B into the soil or neighboring environment by decomposition of their residues, which may be sufficient to cause growth inhibition of their neighboring or successional plants. The growth inhibitory activity of momilactone B and the occurrence of momilactone B in rice plants suggest that momilactone B may contribute the growth inhibitory effect of rice residues and extracts, indicating that momilactone B may have an important role in the rice allelopathy.     

Isolation and identification of a potent allelopathic substance in Bangladesh rice

Aqueous methanol extracts of Bangladesh rice (Oryza sativa L. cv. BR17) inhibited the growth of roots and shoots of cress (Lepidium sativum), lettuce (Lactuca sativa), alfalfa (Medicago sativa), timothy (Phleum pratense), Digitaria sanguinalis, Echinochloa crus-galli and Echinochloa colonum. Increasing the extract concentration increased the inhibition, suggesting that the BR17 may have growth inhibitory substances and possess allelopathic potential. The aqueous methanol extract of the BR17 was purified and a main inhibitory substance was isolated and determined by spectral data as 2,9-dihydroxy-4-megastigmen-3-one. This substance inhibited root and shoot growth of cress and E. crus-galli seedlings at concentrations greater than 0.03 and 3 μM, respectively. The concentrations required for 50% growth inhibition on cress roots and shoots were 0.22 and 0.47 μM, respectively, and on E. crus-galli roots and shoots were 36 and 133 μM, respectively. These results suggest that 2,9-dihydroxy-4-megastigmen-3-one may contribute to the growth inhibitory effect of BR17 and may play an important role in the allelopathy of BR17. Thus, Bangladesh rice BR17 may be potentially useful for weed management in a field setting. An erratum to this article can be found at     

Allelopathy and the allelothathic activity of a phenylpropanol from cucumber plants

The growth inhibitory effect of cucumber (Cucumis sativus L.) plants after crop harvested was investigated. Aqueous methanol extracts of the cucumber plants inhibited the growth of roots and shoots of cress (Lepidium sativum L.), lettuce (Lactuca sativa L.), alfalfa (Medicago sativa L.), ryegrass (Lolium multiflorum L.), timothy (Pheleum pratense L.), crabgrass (Digitaria sanguinalis L.), Echinochloa crus-galli (L.) Beauv and Echinochloa colonum (L.) Link, and increasing the extract concentration increased the inhibition. These results suggest that cucumber plants may possess allelopathic activity. The aqueous methanol extract of cucumber plants was divided into ethyl acetate and aqueous fractions, and the growth inhibitory activity of ethyl acetate fraction was greater than that of aqueous fraction. Thus, ethyl acetate fraction was further purified and a main allopathically active substance in the fraction was isolated and determined as (S)-2-benzoyloxy-3-phenyl-1-propanol by spectral data. This substance inhibited root and shoot growth of cress seedlings at concentrations greater than 10 μM, and the concentration required for 50% inhibition of root and shoot growth was 21 and 23 μM, respectively. These results suggest that (S)-2-benzoyloxy-3-phenyl-1-propanol may contribute to the growth inhibitory effect of cucumber plants and may play an important role in cucumber allelopathy. Thus, cucumber plants may be potentially useful for weed management in a field setting. An erratum to this article can be found at     

The chemical-mediated allelopathic interaction between rice and barnyard grass

Aims

The possible involvement of the chemical-mediated interaction in allelopathy between rice and barnyard grass was investigated.

Methods

Effcts of rice seedlings and rice root exudate on the alleloapthic activity of barnyard grass were determined and a key compound invovled in the allelopathic interaction between rice and barnyard grass was isolated.

Results

Allelopathic activity of barnyard grass was increased by the presence of rice seedlings. Rice root exudates also elevated the allelopahtic activity of barnyard grass. A key compound, which increased the allelopathic activity of barnyard grass, in the rice root exudates was isolated and determined as momilactone B. Momilactone B increased the allelopathic activity of barnyard grass at concentrations greater than 3 μM, and increasing the momilactone B concentration increased the activity.

Conclusions

Momilactone B is known to act as a potent rice allelochemical and to possess strong growth inhibitory activity against barnyard grass. The present research suggests that barnyard grass may response to the presence of neighboring rice by sensing momilactone B in rice root exudates and increase allelopathic activity. Thus, momilactone B may not only act as a rice allelochemical but also play an important role in rice-induced allelopathy of barnyard grass. The induced-allelopathy may provide a competitive advantage for barnyard grass through the growth inhibition of competing plant species including rice. Barnyard grass allelopathy may be one of the inducible defense mechanisms by chemical-mediated plant interaction between rice and barnyard grass. Rice allelopathy was also reported to be increased by the presence of barnyard grass through increasing production and secretion of momilactone B into surrounding environments. During the evolutional process, rice and barnyard grass may have developed the chemical cross talk to activate the defense mechanisms against some biotic stress conditions by detection of certain key compounds.