Kosumi Yamada

Gene activation by cytoplasmic acidification in suspension-cultured rice cells in response to the potent elicitor, N-acetylchitoheptaose

N-Acetylchitoheptaose strongly induces a set of defense reactions in suspension-cultured rice cells including cytoplasmic acidification (K. Kuchitsu, Y. Yazaki, K. Sakano, and N. Shibuya, Plant Cell Physiol. 38:1012-1018, 1997) and the accumulation of mRNAs for two rapidly activated genes, EL2 and EL3 (E. Minami, K. Kuchitsu, D.-Y. He, H. Kouchi, N. Midoh, Y. Ohtsuki, and N. Shibuya, Plant Cell Physiol. 37:563-567, 1996), as well as phenylalanine ammonia-lyase (PAL), chitinase and β-1,3-glucanase. Treatment of cells with propionic acid resulted in the accumulation of the mRNAs for EL2, EL3, and PAL in a manner similar to the accumulation induced by N-acetylchitoheptaose. Concomitantly, there was a rapid decrease in the cytoplasmic pH as detected with in vivo 31P-nuclear magnetic resonance (NMR) spectroscopy. Interestingly, K-252a, a potent inhibitor of Ser/Thr protein kinases, strongly inhibited gene induction by N-acetylchitoheptaose, but showed much less inhibition of gene induction caused by propionic ...

Phototropic stimulation induces the conversion of glucosinolate to phototropism-regulating substances of radish hypocotyls

The distribution of natural growth inhibitors, the raphanusanins (isomers of 3-(methylthio)methylene-2-pyrrolidinethione) and their precursors (4-methylthio-3-butenyl glucosinolate (MTBG) and 4-methylthio-3-butenyl isothiocyanate (MTBI), between illuminated and shaded halves of radish hypocotyls during phototropic curvature was analyzed using a physicochemical assay. Phototropic stimulation rapidly decreased MTBG content, and abruptly increased contents of MTBI and raphanusanins in the illuminated halves of radish hypocotyls within 30 min after the onset of unilateral illumination. Content in the shaded halves was similar to that in dark controls. When MTBG, MTBI, and raphanusanins at endogenous levels were applied unilaterally to etiolated hypocotyls, MTBI and raphanusanins caused hypocotyls to bend but MTBG showed no activity. Blue illumination promoted myrosinase (thioglucosidase) activity, which releases MTBI from MTBG, in hypocotyls after 10 min, although enzyme activity in dark controls did not change. These results suggest that phototropic stimulation promotes myrosinase activity in the illuminated side of radish hypocotyls, releasing bioactive MTBI from inactive MTBG and simultaneously producing bioactive raphanusanins.

Allelopathic substance exuded from a serious weed, germinating barnyard grass (Echinochloa crus-galli L.), roots

Abstract. The allelopathy of a serious weed, barnyard grass (Echinochloa crus-galli L.), was investigated. Root exudates of young barnyard grass showed allelopathic effects and plant-selective activity and inhibited root elongation of all plants tested. With respect to shoot growth, the exudates did not show inhibition of barnyard grass only. The allelopathic substance was isolated and identified as p-hydroxymandelic acid by NMR. p-Hydroxymandelic acid strongly inhibited shoot growth and root elongation of all plants tested. The effects of three congeners of p-hydroxymandelic acid were tested on rice shoot growth. In the biological activity exhibited in rice, shoot growth was related to the hydroxyl groups.

An allelopathic substance exuded from germinating watermelon seeds

When watermelon seeds were cultured in a Petri dish together with amaranth, barnyard grass, cockscomb, lettuce or tomato seeds, the shoot growth of amaranth and cockscomb was markedly promoted, whereas the shoot growth of lettuce and tomato was inhibited. The shoot growth of barnyard grass was not affected. These results suggest that plant-selective allelopathic substance(s) affecting the shoot growth of other plant seedlings were exuded from watermelon seeds. An allelopathic substance was isolated from the exudates of germinating watermelon seeds and identified as vanillic acid by its spectral analysis and Rf value on TLC. Vanillic acid promoted the shoot growth of cockscomb at the concentrations of 300 to 10 mg/l and that of amaranth at the concentrations of 30 to 3 mg/l, although the shoot growth of amaranth was inhibited by 300 mg/l of vanillic acid. The shoot growth of lettuce and tomato was inhibited at the concentrations higher than 30 mg/l by vanillic acid. However, the shoot growth of barnyard grass was not affected at the concentrations used. All these results suggest that vanillic acid may play as a major component of allelopathic substance(s), which shows plant-selective activity, in the exudates of germinating watermelon seeds.

Isolation and identification of lateral bud growth inhibitor, indole-3-aldehyde, involved in apical dominance of pea seedlings

A lateral bud growth inhibitor was isolated from etiolated pea seedlings and identified as indole-3-aldehyde. The indole-3-aldehyde content was significantly higher in the diffusates from explants with apical bud and indole-3-acetic acid treated decapitated explants, in which apical dominance is maintained, than in those from decapitated ones releasing apical dominance. When the indole-3-aldehyde was applied to the cut surface of etiolated decapitated plants or directly to the lateral buds, it inhibited outgrowth of the latter. These results suggest that indole-3-aldehyde plays an important role as a lateral bud growth inhibitor in apical dominance of pea seedlings.

Plant growth-promoting oligosaccharides produced from tomato waste

Tomato juice waste was hydrolyzed with acid. Tomato juice waste (500 g; wet weight) was heated with 0.5 N HCl (2.5 l) at 70 degrees C for 4 h. After neutralization, the growth-promoting extracts (300 g; dry weight) in the plants were produced from the tomato waste. The acid extract significantly promoted the growth of cockscomb (Celosia argentea L.) and tomato (Lycopersicon esculentum L.) seedlings. We have recognized potent plant growth-promoting substances in the acid extract from tomato waste. The most effective components in the active fraction were almost all oligogalacturonic acids (DP 6-12). This paper is the first report that plant growth-promoting oligosaccharides can be directly produced from tomato juice waste. It is possible that the substances from the tomato waste can become useful plant growth regulators in the agriculture field in the future.

ppGpp Controlled by the Gac/Rsm Regulatory Pathway Sustains Biocontrol Activity in Pseudomonas fluorescens CHA0

In Pseudomonas fluorescens CHA0 and other fluorescent pseudomonads, the Gac/Rsm signal transduction pathway is instrumental for secondary metabolism and biocontrol of root pathogens via the expression of regulatory small RNAs (sRNAs). Furthermore, in strain CHA0, an imbalance in the Krebs cycle can affect the strains ability to produce extracellular secondary metabolites, including biocontrol factors. Here, we report the metabolome of wild-type CHA0, a gacA-negative mutant, which has lost Gac/Rsm activities, and a retS-negative mutant, which shows strongly enhanced Gac/Rsm-dependent activities. Capillary electrophoresis-based metabolomic profiling revealed that the gacA and retS mutations had opposite effects on the intracellular levels of a number of central metabolites, suggesting that the Gac/Rsm pathway regulates not only secondary metabolism but also primary metabolism in strain CHA0. Among the regulated metabolites identified, the alarmone guanosine tetraphosphate (ppGpp) was characterized in detail by the construction of relA (for ppGpp synthase) and spoT (for ppGpp synthase/hydrolase) deletion mutants. In a relA spoT double mutant, ppGpp synthesis was completely abolished, the expression of Rsm sRNAs was attenuated, and physiological functions such as antibiotic production, root colonization, and plant protection were markedly diminished. Thus, ppGpp appears to be essential for sustaining epiphytic fitness and biocontrol activity of strain CHA0.

Leaching of the allelopathic substance, -tryptophan from the foliage of mesquite (Prosopis juliflora (Sw.) DC.) plants by water spraying

The allelopathic potential of -tryptophan in the leachate from the foliage of mesquite (Prosopis juliflora (Sw.) DC.) plants was investigated. Distilled water (500 ml) was sprayed on mesquite plants from above for 50 min and water passing through the foliage was collected. The content of -tryptophan in the sample (30 ml eq.) was analyzed using a physicochemical method and the concentration was shown to be 17.9 µM. The sample (30 ml eq.) caused 69.8% inhibition of the root growth of barnyard grass (Echinochloa crus-galli L.). The concentration of authentic -tryptophan required for 69.8% inhibition of the root growth of barnyard grass was estimated at 20.0 µM from the dose-response curve. Moreover, to establish the origin of -tryptophan in the leachate, the amount of -tryptophan in both the leaves and the leachates was determined. The amount of -tryptophan containing in the foliage significantly decreased by soaking with time, whereas in the leachates the amount increased. After 60 min, its content in the leachates was nearly equivalent to that of the leaves. These results suggest that -tryptophan leached from the foliage may play an important role in the allelopathy imposed by mesquite.

Isolation and identification of plant growth inhibitors as candidate(s) for allelopathic substance(s), from aqueous leachate from mesquite (Prosopis juliflora (Sw.) DC.) leaves

As candidate(s) for allelopathic substance(s), two plant growthinhibitors were isolated from aqueous leachate of leaves of mesquite, whichshowa strong allelopathy, and which were identified as syringin and(–)-lariciresinol by their spectral analyses. Syringin inhibited root andshoot growth of lettuce seedlings at concentrations greater than 0.8μM, and root and shoot growth of barnyard grass seedlings atconcentrations greater than 2.7 and 26.9 μM, respectively. Onthe other hand, (–)-lariciresinol inhibited root and shoot growth oflettuce seedlings at concentrations greater than 2.8 and 0.8μM,and root and shoot growth of barnyard grass seedling at concentrations greaterthan 0.8 and 2.8 μM, respectively. The contents of syringin and(–)-lariciresinol in the rhizosphere soil of mesquite were 0.34 and 0.38μg/g soil, respectively. These results indicate that syringinand (–)-lariciresinol are allelopathic substance(s), and may play rolesinthe allelopathy of mesquite.

A growth-inhibitory substance exuded from freeze-dried mesquite (Prosopis juliflora (Sw.) DC.) leaves

When lettuce seeds were incubated in a concentric circle aroundfreeze-dried mesquite leaves in a dish containing agar culture medium, thegrowth, especially radicle growth, of the seedlings was inhibited, the morestrongly the closer they were to the leaves. This result indicates thatallelopathic substance(s) inhibiting the lettuce growth are exuded from themesquite leaves. A potent substance was isolated from the exudates of thefreeze-dried mesquite leaves and identified as L-tryptophan by spectralanalyses. L-Tryptophan inhibited the radicle growth of lettuce and barnyardgrass at concentrations greater than1.5 × 10−3 . The content ofL-tryptophan in the exudates of freeze-dried mesquite leaves (1 eq.) was 4.8 × 10−3 . Theseresults suggest that L-tryptophan may play an important role in the allelopathyof mesquite leaves.