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An internal detoxification mechanism for AI was investigated in an AI-accumulating plant, hydrangea (Hydrangea macrophylla), focusing on AI forms present in the cells. l h e leaves of hydrangea contained as much as 15.7 mmol AI kg-' fresh weight, and more than two-thirds of the AI was found in the cell sap. Using "AInuclear magnetic resonance, the dominant peak of AI was observed at a chemical shift of 11 to 12 parts per million in both intact leaves and the extracted cell sap, which is in good accordance with the chemical shift for the 1 :1 AI-citrate complex. Purification of cell sap by molecular sieve chromatography (Sephadex C-1 O) combined with ion-exclusion chromatography indicated that AI in fractions with the same retention time as citric acid contributed to the observed "AI peak,in the intact leaves. l h e molar ratio of AI to citric acid in the crude and purified cell sap approximated 1. The structure of the ligand chelated with AI was identified to be citric acid. Bioassay experiments showed that the purified AI complex from the cell sap did not inhibit root elongation of corn (Zea mays L.) and the viability of cells on the root tip surface was also not affected. lhese observations indicate that AI is bound to citric acid in the cells of hydrangea leaves.A1 toxicity is primarily characterized by the inhibition of root elongation, with no appearance of clearly identifiable symptoms in plant tops. This is because A13+, a toxic ionic species, has a high binding ability with cellular components of roots, and usually shows little translocation to the upper parts of plants. Most plants contain not more than 0.2 mg AI 8-l dry weight. However, some plants, known as "A1 accumulators," may contain more than 10 times this leve1 of A1 without any AI injury. Tea plants are typical AI accumulators; the A1 content in these plants can reach as high as 30 mg 8-l dry weight in old leaves, although
A potential antagonist, Bacillus amyloliquefaciens strain RC-2, against Colletotrichum dematium, mulberry anthracnose fungus, was obtained from healthy mulberry leaves by in vitro and in vivo screening techniques. Application of culture filtrate of RC-2 inhibited disease on mulberry leaves, indicating that suppression was due to antifungal compounds in the filtrate. Development of mulberry anthracnose on mulberry leaves was inhibited only when the culture filtrate was applied before fungal inoculation, and it was not inhibited by application after inoculation. These results suggest that the antifungal compounds in the filtrate exhibit a preventive effect on the disease. Peptone significantly increased production of the antifungal compounds. The culture filtrate of RC-2 also inhibited the growth of several other phytopathogenic fungi and bacteria, such as Rosellinia necatrix, Pyricularia oryzae, Agrobacterium tumefaciens, and Xanthomonas campestris pv. campestris, in vitro. From the culture filtrate of RC-2, seven kinds of antifungal compounds were isolated by high performance liquid chromatography analysis, and one of the compounds was determined as iturin A2, a cyclic peptide, by nuclear magnetic resonance and fast atom bombardment mass analysis.
Pyrolysis of plant and animal wastes produces a complex mixture of phosphorus species in amorphous, semicrystalline, and crystalline inorganic phases, organic (char) components, and within organo-mineral complexes. To understand the solubility of different phosphorus species, plant (cottonseed hull) and manure (broiler litter) wastes were pyrolyzed at 350, 500, 650, and 800 °C and exposed to increasingly more rigorous extraction procedures: water (16 h), Mehlich 3 (1 mM EDTA at pH 2.5 for 5 min), oxalate (200 mM oxalate at pH 3.5 for 4 h), NaOH-EDTA (250 mM NaOH + 5 mM EDTA for 16 h), and total by microwave digestion (concentrated HNO3/HCl + 30% H2O2). Relative to the total (microwave digestible) P, the percentage of extractable P increased in the following order: M3 < oxalate ≈ water < NaOH-EDTA for plant biochars and water < M3 < NaOH-EDTA < oxalate for manure biochars. Solution phase (31)P NMR analysis of NaOH-EDTA extracts showed the conversion of phytate to inorganic P by pyrolysis of manure and plant wastes at 350 °C. Inorganic orthophosphate (PO4(3-)) became the sole species of ≥ 500 °C manure biochars, whereas pyrophosphate (P2O7(4-)) persisted in plant biochars up to 650 °C. These observations suggested the predominance of (i) amorphous (rather than crystalline) calcium phosphate in manure biochars, especially at ≥ 650 °C, and (ii) strongly complexed pyrophosphate in plant biochars (especially at 350-500 °C). Correlation (Pearson's) was observed (i) between electric conductivity and ash content of biochars with the amount of inorganic P species and (ii) between total organic carbon and volatile matter contents with the organic P species.
The forms of Al for uptake by the roots and translocation from the root to the shoot were investigated in a buckwheat (Fagopyrum esculentum Moench, cv. Jianxi) that accumulates Al in its leaves. The Al concentration in the xylem sap was 15-fold higher in the plants exposed to AlCl3 than in those exposed to an Al-oxalate (1:3) complex, suggesting that the roots take up Al in the ionic form. The Al concentration in the xylem sap was 4-fold higher than that in the external solution after a 1-h exposure to AICl3 solution and 10-fold higher after a 2-h exposure. The Al concentration in the xylem sap increased with increasing Al concentration in the external solution. The Al uptake was not affected by a respiratory inhibitor, hydroxylamine, but significantly inhibited by the addition of La. These results suggest that Al uptake by the root is a passive process, and La3+ competes for the binding sites for Al3+ on the plasma membrane. The form of Al in the xylem sap was identified by 27Al-nuclear magnetic resonance analysis. The chemical shift of 27Al in the xylem sap was around 10.9 ppm, which is consistent with that of the Al-citrate complex. Furthermore, the dominant organic acid in the xylem sap was citric acid, indicating that Al was translocated in the form of Al-citrate complex. Because Al is present as Al-oxalate (1:3) in the root, the present data show that ligand exchange from oxalate to citrate occurs before Al is released to xylem.
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