Translocation and degradation of injected uniconazole in apple during a 4-month growing period

One-year-old ‘Golden Delicious’ apple trees grafted onto MM 106 rootstocks were injected in the rootstock stem (shank) with¹⁴C-uniconazole to determine the extent to which uniconazole is translocated and degraded over the length of an average growing season. In 4 months, 16% of recovered¹⁴C-activity was translocated to the new shoots. Most of the¹⁴C-activity remained in the rootstock. Chromatographic evaluation of shoot extracts demonstrated that the¹⁴C-activity associated with uniconazole decreased 49% in 4 months. However, shoot growth was still inhibited which suggests that the amount of uniconazole that was degraded did not interfere with the inhibition of gibberellin biosynthesis, probably due to the continuous translocation of uniconazole that occurred.     

Translocation of uniconazole after trunk injection of silver maple saplings

Four-year-old silver maple (Acer saccharinum L.) saplings were trimmed and trunk injected with¹⁴C-labeled uniconazole [(E)-1-(4-chlorophenyl)-4, 4-dimethyl-2-(1,2,4-triazol-1-yl)-1-penten3-ol] in May of 1987. Inhibition of shoot growth was observed in 1988 but not in 1987. The¹⁴C-activity detected in the foliage during 1987 and 1988 increased in a linear fashion, and the maximum concentration reached in 1988 was about one third that reached in 1987. Approximately 7.1 and 2.3% of the total¹⁴C-activity injected into the saplings was present in senescing foliage harvested in the fall of 1987 and 1988, respectively. Most of the¹⁴C-activity in the saplings 17, 134, or 500 days after injection remained around the injection site. Between 29 and 49% of the¹⁴C-activity found in the foliage collected in the fall was associated with metabolites rather than uniconazole, and there was no increase in the proportion in metabolites from 1987 to 1988, suggesting that metabolism occurred in the foliage and not the stem.     

Translocation of uniconazole after trunk injection of silver maple saplings

Four-year-old silver maple (Acer saccharinum L.) saplings were trimmed and trunk injected with¹⁴C-labeled uniconazole [(E)-1-(4-chlorophenyl)-4, 4-dimethyl-2-(1,2,4-triazol-1-yl)-1-penten3-ol] in May of 1987. Inhibition of shoot growth was observed in 1988 but not in 1987. The¹⁴C-activity detected in the foliage during 1987 and 1988 increased in a linear fashion, and the maximum concentration reached in 1988 was about one third that reached in 1987. Approximately 7.1 and 2.3% of the total¹⁴C-activity injected into the saplings was present in senescing foliage harvested in the fall of 1987 and 1988, respectively. Most of the¹⁴C-activity in the saplings 17, 134, or 500 days after injection remained around the injection site. Between 29 and 49% of the¹⁴C-activity found in the foliage collected in the fall was associated with metabolites rather than uniconazole, and there was no increase in the proportion in metabolites from 1987 to 1988, suggesting that metabolism occurred in the foliage and not the stem.     

Translocation and degradation of injected uniconazole in apple during a 4-month growing period

One-year-old Golden Delicious apple trees grafted onto MM 106 rootstocks were injected in the rootstock stem (shank) with¹⁴C-uniconazole to determine the extent to which uniconazole is translocated and degraded over the length of an average growing season. In 4 months, 16% of recovered¹⁴C-activity was translocated to the new shoots. Most of the¹⁴C-activity remained in the rootstock. Chromatographic evaluation of shoot extracts demonstrated that the¹⁴C-activity associated with uniconazole decreased 49% in 4 months. However, shoot growth was still inhibited which suggests that the amount of uniconazole that was degraded did not interfere with the inhibition of gibberellin biosynthesis, probably due to the continuous translocation of uniconazole that occurred.     

Environmental fate of the triazole fungicide propiconazole in a rice-paddy-soil lysimeter

Environmental fate of the triazole fungicide propiconazole, 1-[[2(2,4-dichlorophenyl)-4-propyl-1,3-diox olane-2-yl]methyl]1H-1,2,4-triazole, in soil was investigated using lysimeters simulating a rice-paddy-soil conditions. Two lysimeters composed of different soil types, a sandy loam (lysimeter A) and silty clay (lysimeter B), were used. Propiconazole (Tilt 250^R EC) plus [U-¹⁴C]-propiconazole was applied over a two-year period to the soil surface of the lysimeters. Propiconazole fate in the lysimeters was assessed by measuring total radioactivity in the leachate, evolved ¹⁴CO₂, and ¹⁴C-residues in the soil and rice plants. The amounts of applied ¹⁴C in the leachate from lysimeter A were 4.4 and 5.2% in the first and second year, respectively. A background level of (0.00005% of applied) ¹⁴C in the leachate from lysimeter B was detected, suggesting negligible movement of the fungicide to groundwater in the silty clay soil. The amount of ¹⁴CO₂ evolved from lysimeter A accounted for 7.8 and 12.2% of applied ¹⁴C in the first and second year, respectively, whereas those from lysimeter B were 5.7 and 7.1%. Total ¹⁴C detected in the rice plants grown in lysimeter A were 7.3 and 9.8% of applied ¹⁴C in the first and second year, respectively, which compared to 3.0 and 7.6% in lysimeter B. Most of the applied ¹⁴C was detected in the top 10 cm soil layer, suggesting that propiconazole remains close to the soil surface after application in soil. Degradation products of propiconazole identified in the lysimeter soils were 1-[[2(2,4-dichlorophenyl)-2-(1,2,4-triazole -1-yl) ketone (DP-1), 1-(2,4-dichlorophenyl)-2-(1,2,4-triazole-1- yl) ethanol (DP-2) and 1-[[2(2,4-dichlorophenyl)-4-hydroxypropyl-1,3-dioxolane-2-yl]methyl]1H-1,2,4-triazole (DP-3 and DP-4).     

Biological Control of Rice Blast byPseudomonas fluorescensStrainPf7–14: Evaluation of a Marker Gene and Formulations

Pseudomonas fluorescensstrainPf7–14 was evaluated for biological control of rice blast in field experiments. StrainPf7–14 was formulated in methylcellulose:talc (1:4) and applied to IR50 rice (Oryza sativa) seeds as a seed treatment and as foliar sprays in seedbed and field experiments. When applied as a seed treatment followed by three foliar applications, the strain provided a 68.5% suppression of rice blast in the seedbed experiment and a 59.6% suppression in the field experiment. The persistence and migration ofPf7–14 on the rice plant was studied with the aid oflacZYgenes inserted into the bacterium. In greenhouse experiments,Pf7–14gal was detected on rice roots at 10⁶to 10⁵cfu/g of root tissue for 110 days, the duration of the rice crop. Migration of the strain from the seeds to the leaves occurred only until the seedlings were 16 days old. WhenPf7–14 was applied to the rice plants by foliar sprays, 10⁴cfu of the bacterium per gram of leaf tissue was detected for the next 40 days. The limited migration of the bacterial biocontrol agent emphasizes the need for multiple foliar applications of the bacterium to sustain the bacterial population for effective suppression of rice blast.     

Untersuchungen über Stoffwechselbeziehungen zwischen Parasit und Wirt am Beispiel von Puccinia graminis var. tritici auf Weizen

Summary Rust infected leaves of wheat plants were incubated with glucose-¹⁴C. Uredospores which were formed during the application of the tracer were analyzed. All isolated compounds were labeled with ¹⁴C. When germinating uredospores were incubated directly with ¹⁴C-glucose, the isolated glutamic acid, arginine and lysine had practically no radioactivity. These compounds did, however, contain considerable ¹⁴C-activity when they were isolated from uredospores formed on leaves that had been treated with the tracer. We therefore conclude that these amino acids were synthesized in the host and were taken up by the haustoria of the mycelium.High ¹⁴C-radioactivity was also found in all carbohydrates (chitin, glucomannan, polyols etc.). Hexoses isolated from the spore constituents chitin and glucomannan showed the same distribution of radioactivity as the applied glucose-1-¹⁴C or glucose-6-¹⁴C. It follows that the rust mycelium takes up glucose or a similar monosaccharide from the wheat plant. The C-6-skeleton is not degraded to smaller metabolites before it is taken up.     

Populations and activity of carbofuran-degrading microorganisms in soil

Kendaia clay loam contained more than 10⁵ microbial cells per g able to convert ¹⁴C-carbonyl-labelled carbofuran (2,3-dihydro-2,2-dimethylbenzofuran-7-yl methylcarbamate) to ¹⁴CO₂ but never more than 130 cells per g transforming ¹⁴C-ring-labelled carbofuran to CO₂. The sizes of the population rarely increased as a result of addition of the insecticide to soil. Mineralization of these compounds proceeded with little or no acclimation phase, and subsequent additions were usually etabolized more readily, except at 10 mg of carbofuran per kg or if subsequent additions of the pesticide were made long after the first. More than 60% of the ¹⁴C in the carbonyl but less of the ¹⁴C in the ring was microbiologically converted to ¹⁴CO₂ in this soil. Streptomycin and cycloheximide each inhibited conversion of the carbonyl or ring carbon to CO₂. Urea but not NH₄NO₃ markedly inhibited the conversion of the carbonyl-labelled insecticide to ¹⁴CO₂. The addition of glucose and succinate together with the insecticide did not enhance mineralization of ring- or carbonyl-labelled carbofuran. The data suggest that soils containing a large population of microorganisms able to convert the carbonyl carbon to CO₂ will not show a marked effect of prior treatment with the insecticide and that few organisms individually are able to mineralize the ring.     

Toxicity assessment of insecticides commonly used in rice pest management to the fry of common carp, Cyprinus carpio, a food fish culturable in rice fields

Toxicity of four insecticides commonly used in rice pest management, chlorpyrifos, dimethoate, carbaryl and carbosulfan, to the fry of common carp was assessed through median lethal concentrations (LC₅₀) and in vivo inhibition of the brain acetylcholinesterase (AChE) enzyme at sublethal concentrations. The 96‐h LC₅₀ values for these four insecticides were determined to be 0.008, 26.11, 7.85 and 0.60 mg L^?1 respectively. Exposure of fish to a series of sublethal concentrations (0.5–5% LC₅₀) of each insecticide for 14 days resulted in concentration‐dependent inhibition in AChE activity in comparison with the controls. AChE activity was greatly inhibited in the fish exposed to sublethal concentrations of chlorpyrifos. Upon transfer to insecticide‐free water, AChE activities in fry exposed to 0.5 and 1% LC₅₀ concentrations of carbaryl and carbosulfan were restored to the control level within 7–21 days whereas the fish exposed to chlorpyrifos or dimethoate did not fully recover from the insecticide‐induced anticholinesterase action. Of the four insecticides tested, chlorpyrifos was the most toxic for the fry of common carp. Although dimethoate was least toxic for the fish under acute exposure, the restoration level of normal AChE activity was slower under chronic exposure in comparison with carbaryl and carbosulfan. Hence, the use of carbamates, especially carbaryl, to control insect pests of rice in rice‐cum‐carp culture systems is recommended when considering survival, restoration of the normal AChE activity and stamina of the cultured fish.     

Untersuchunge über Stoffwechselbeziehungen zwischen Parasit und Wirt am Beispiel von Puccinia graminis var. tritici auf Weizen

Summary Rust infected wheat plants were incubated with different ^14C-labelled amino acids. Uredospores that were formed during the incubation contained ¹⁴C-activity. By analysis of these spores it was investigated whether the parasitic mycelium of Puccinia graminis takes amino acids from the host. It could be demonstrated that the applicated amino acids were taken up directly from the wheat leaf. The carbon sceletons of applicated lysine and arginine showed only little randomization of ¹⁴C-activity. Glutamic acid, alanine and glycine isolated from uredospore protein showed a very strong alteration of the original label. The pools of free amino acids in the host and the parasite are in isotope equilibrium. This demonstrates, that synthesis of amino acids in the mycelium is quantitatively not important. By following the kinetics of incorporation of an amino acid it could be demonstrated that the amino acids enter the parasite as free amino acids and not in the form of proteins.     

Mineralization of Parathion in the Rice Rhizosphere

We studied ¹⁴CO₂ evolution from ring-labeled [2,6-¹⁴C]parathion (O,O-diethyl-O-p-nitrophenyl phosphorothioate) in the rhizosphere of rice seedlings. The soil samples (nonflooded [60% water-holding capacity] and flooded) were treated first with technical parathion (20 μg/g) and then after 10 days with ring-labeled [¹⁴C]parathion. In unplanted soil, less than 5.5% of the ¹⁴C in the parathion was evolved as ¹⁴CO₂ in 15 days under both flooded and nonflooded conditions. In soil planted with rice, 9.2% of the radiocarbon was evolved as ¹⁴CO₂ under nonflooded conditions, and 22.6% was evolved under flooded conditions. These results suggest that soil planted with rice permits significant ring cleavage, especially under flooded conditions.     

Influence of Temperature and Seed Ripening on the in-vivo Incorporation of CO(2) into the Lipids of Oat Grains (Avena sativa L.)

To elucidate the influence of growth temperature and of stage of maturity on lipid synthesis in seeds, oat plants (Avena sativa nuda L., variety NOS) were fed with ¹⁴CO₂ at different stages after flowering, and the ¹⁴C-incorporation into the grain lipids was determined at 2, 24, and 48 hours after the end of ¹⁴CO₂-application. By changing growth temperature from 12 C to 28 C after the application of ¹⁴CO₂ to intact plants, a higher ¹⁴C-labeling of saturated fatty acids was found at the higher temperature. At 28 C, palmitic and stearic acids contained 23% and 9% respectively of total fatty acid-¹⁴C shortly after the ¹⁴CO₂-application, whereas at 12 C the corresponding values were 19% and 4%, respectively. Within 2 days ¹⁴C-activity of saturated fatty acids decreased at both temperatures, but to a lesser degree at 28 C. The higher ¹⁴C-labeling of saturated fatty acids and its lower decrease within 2 days at 28 C clearly show a direct influence of temperature on fatty acid biosynthesis in oat grains.     

The significance of denitrification of applied nitrogen in fallow and cropped rice soils under different flooding regimes I. Greenhouse experimènts

The role of nitrification-denitrification in the loss of nitrogen from urea applied to puddled soils planted to rice and subjected to continuous and intermittent flooding was evaluated in three greenhouse pot studies. The loss of N via denitrification was estimated indirectly using the¹⁵N balance, after either first accounting for NH₃ volatilization or by analyzing the¹⁵N balance immediately before and after the soil was dried and reflooded. When urea was broadcast and incorporated the loss of¹⁵N from the soil-plant systems depended on the soil, being about 20%–25% for the silt loams and only 10%–12% for the clay. Ammonia volatilization accounted for an average 20% of the N applied in the silt loam. Denitrification losses could not account for more than 10% of the applied N in any of the continuously flooded soil-plant systems under study and were most likely less than 5%. Intermittent flooding of soil planted to rice did not increase the loss of N. Denitrification appeared to be an important loss mechanism in continuously flooded fallow soils, accounting for the loss of approximately 40% of the applied¹⁵N. Loss of¹⁵N was not appreciably enhanced in fallow soils undergoing intermittent flooding. Apparently, nitrate formed in oxidized zones in the soil was readily denitrified in the absence of plant roots. Extensive loss (66%) of¹⁵N-labeled nitrate was obtained when 100 mg/pot of nitrate-N was applied to the surface of nonflooded soil prior to reflooding. This result suggests that rice plants may not compete effectively with denitrifiers if large quantities of nitrate were to accumulate during intermittent dry periods.     

Uptake, translocation and metabolism of fluazifop-butyl in Setaria viridis

The uptake and translocation of fluazifop-butyl was investigated in Setaria viridis. Young plants (three to four leaves) with a portion of the second, third or fourth leaf covered, were sprayed with a dose equivalent to 0.25 kg a.i. ha-¹. ¹⁴C-fluazifop-butyl was subsequently applied to the unsprayed area and the treatment resulted in plant death within 2 wk. Uptake by leaf 3 was rapid, with less than 5% of the applied dose remaining on the leaf surface after 24 h. The highest proportion of ^14C-activity was retained in the treated portion of the leaf. Only 2% of the applied dose was translocated from leaf 3 and 0. 76% accumulated in the apical meristem. Uptake by the younger leaf 4 was more rapid and the pattern of translocation differed in that more ¹⁴-^-activity accumulated in apical meristematic tissue. 6–30% of the applied dose was undetected and this was greatest when foliar uptake was slow. Artificial leaf surface experiments indicated that this undetected activity may have been due to volatility of fluazifop-butyl or degradation to volatile products. ^14C-activity extracted from treated leaves was identified as fluazifop-butyl, fluazifop acid and polar conjugates. The major ¹⁴C-activity extracted from the apical meristem was fluazifop acid and no fluazifop-butyl was detected in this extract.     

A simultaneous extraction method for metabolome and lipidome and its application in cry1Ac and sck-transgenic rice leaf treated with insecticide based on LC–MS analysis

Abiotic stress caused by insecticide treatment is an interesting and challenging topic in plant research. Here a simultaneous extraction method with methyl tert-butyl ether for metabolomics and lipidomics analysis by using rapid resolution liquid chromatography quadrupole time-of-flight mass spectrometry was developed and validated. The extraction efficiency of lipidome and metabolome based on the current developed method showed an obvious improvement compared with literatures. About 300 metabolites were identified in rice leaf. Method validation was performed and analytical properties including the linearity (R², 0.991–1.000), repeatability (over 87 % peaks with CV < 20 % accounting for over 92 % of total response) and recovery (74.4–119.6 %) were satisfactory. The method was then applied to investigate time-course changes caused by insecticide treatment in transgenic rice with cry1Ac and sck genes and its wild counterpart. Antioxidants including ferulic acid and sinapic acid were down-regulated at 24 h after insecticide treatment. Signaling metabolites including salicylic acid and nicotinamide adenine dinucleotide were significantly up-regulated at 12–24 h after treatment in transgenic rice. More flavonoids were significantly changed in transgenic plants. Results indicated that gene transformation affected the metabolic response of rice to insecticide stress.     

Comparison of ethylenediaminetetraacetate-enhanced exudation from detached and translocation from attached bean leaves

A technique for collection of phloem exudate from detached leaves using 20 millimolar EDTA (pH 7.0) has previously been developed (King, Zeevaart 1974 Plant Physiol 53: 96-103). It was the aim of the present study to determine the efficiency of this technique in relation to undisturbed export from attached leaves. Paired primary leaves of bean seedlings (Phaseolus vulgaris L. cv Montcalm) were used to minimize variations in plant material. Attached leaves, exposed to ¹⁴CO₂ for 10 minutes with subsequent excision of one of the leaves and collection of the exudate over a 12-hour period, showed a 25% export of total assimilated ¹⁴C from the attached versus 15% of total assimilated ¹⁴C in the form of exudation from the detached ones. Leaf excision changed the labeling pattern within the leaf, increasing% total leaf ¹⁴C-activity in the ethanolic fraction, while decreasing activity in the starch fraction, as compared to attached leaves. This was presumably caused by a lack of translocation from the detached leaves. Excision did not affect dark respiration. However, measurements of total nonstructural carbohydrates in leaf starch and neutral fractions indicated no significant differences between attached and leaves detached in EDTA. Thus, in terms of actual carbon export, and accompanying distribution of nonexported carbohydrate within the leaf, EDTA-enhanced exudation compares favorably with translocation from attached leaves.     

Residue dynamics and risk assessment of pyraclostrobin in rice,plants, hulls,field soil,and paddy water

A modified quick, easy, cheap, effective, rugged and safe–ultra-performance liquid chromatography–tandem mass spectrometry method was developed and validated for the analysis of pyraclostrobin residue in rice plants, hulls, soil, and paddy water. Pyraclostrobin residue dynamics and final residues were studied in supervised field trials under Good Agricultural Practice conditions in rice, plants, hulls, soil, and paddy water. The levels of detection of all chemicals in five types of matrices were in the ranges of 2 × 10^?7 to 1 × 10^?5 mg/kg with recoveries in the ranges of 74.1–107.4%. The dissipation experiments showed that the half-lives (T_1/2) of pyraclostrobin in the plants, soil, and paddy water were 6.3–13.9, 5.0–34.7, and 6.9–11.6 days, respectively. At pre-harvest intervals (PHIs) of 14, 21, and 28 days, pyraclostrobin residue levels were <0.01–0.19 mg/kg in rice. The residue levels at 35-d PHI were all below 0.1 mg/kg (maximum residue limit of the USA), and all the health risk quotient values calculated were below 0.027. The environment risk of pyraclostrobin in soil and paddy water was also monitored throughout the rice-growing period, and the results showed that the risk level was between low and medium in the soil and high in the paddy water.     

In vivo stimulation of ovarian development in the red swamp crayfish,Procambarus clarkii (Girard), by 5-hydroxytryptamine

Summary

The possibility that biogenic amines affect ovarian development in the red swamp crayfish, Procambarus clarkii, was investigated. Females were administered 15 μg/g body weight (bw) of norepinephrine, dopamine, 5-hydroxytryptamine (5-HT) or octopamine on days 1, 5 and 10 and were sacrificed on day 15. Crayfish given 5-HT showed significant increases in ovarian index (30.5%) and oocyte size (34.0%) over the concurrent controls, while norepinephrine, dopamine and octopamine did not significantly affect either the ovarian index or oocyte size. Significantly more labeling by ¹⁴C-leucine of ovarian proteins was found in ovaries of crayfish that were injected with 5-HT in vivo, but when ovarian lobes from crayfish that had not been injected with 5-HT were incubated in vitro with 5-HT added to the incubation medium, no significant change in the level of incorporation of ¹⁴C-leucine into ovarian proteins occurred.

The 5-HT receptor blocker LY53857 (25 μg/g bw) retarded ovarian development. The 5-HT releaser fenfluramine and the 5-HT potentiator fluoxetine (both 15 μg/g bw) were also used. Crayfish given fenfluramine, fluoxetine, fenfluramine plus 5-HT or fluoxetine plus 5-HT showed significant increases of ovarian index (24.0–102.8%), oocyte size (20.0–87.4%) and in vitro ¹⁴C-leucine labeling of ovarian proteins (30.6–123.6%) over the concurrent controls. The ovaries of crayfish that received the 5-hydroxytryptaminergic neurotoxin 5,6-dihydroxytryptamme (10 μg/g bw) did not show any significant change as compared with the initial control. These findings are consistent with the hypothesis that 5-HT, which is present in the central nervous system of Procambarus clarkii, exerts its stimulatory effect on the ovary of this crayfish indirectly by triggering release of the ovary-stimulating hormone.     

灭虫精的杀虫活性及田间防治褐飞虱的应用研究

灭虫精(Imidacloprid通用名称吡虫啉)系一种内吸性杀虫剂,属硝基亚甲基化合物,对稻飞虱具有极强的生物活性。室内测定褐飞虱付Nilapavarta lugens Stal 3龄与5龄若虫LD₅₀值分别为2.55x10^-5μ/头与6.78x10^-5μg/头,其毒力回归方程分别为Y=r5.6231+1,0486X和Y=5.2091+1.2389X。实验表明,该药具有极好的触杀和胃毒作用,稻田应用的有效剂量为15～30g/hm²,是扑虱灵常规用量的1/5～1/10。田间药效特征突出地表现为速效(1-3d)、持效(4-10d)和残效(11d以上)的三段效应,90%以上的防治效果可维持在40d以上.试验还表明,灭虫精田间应用表现出在飞虱和天敌之间的良好选择性,对稻田益蛛基本无害,是一种很有前途的稻田应用杀虫剂。     

"双季稻-鸭"共生生态系统C循环

"稻鸭共生"是对我国传统农业稻田养鸭的继承与发展。在双季稻主产区湖南布置了稻田养鸭田间对比试验,以常规稻作为对照,采用投入产出法,分析"稻鸭共生"生态系统C的输入输出及循环情况。结果表明:"早稻-鸭"生态系统碳输出中,水稻籽粒C占42.21%;水稻秸秆C占38.42%;气态C(CH4和CO2)占18.50%;鸭产品C仅占0.87%。"晚稻-鸭"生态系统碳输出中,水稻籽粒C占53.80%;水稻秸秆C占35.12%;气态C占8.67%;鸭产品C仅占1.07%。两季稻作里,"稻鸭共生"土壤截存C量是2103.2 kg/hm2,水稻植株地上部分固定的C量是15109.96 kg/hm2,水稻根固定的C量是1261.34 kg/hm2,归还给土壤的鸭粪C量是229.87 kg/hm2。鸭子系统C输入主要来自系统外投入的饲料C,早稻季鸭所食的杂草C和害虫C分别为60.53 kg/hm2和2.75 kg/hm2,晚稻季鸭所食的杂草C和害虫C分别为3.64 kg/hm2和6.73 kg/hm2。对"双季稻-鸭"共生生态系统的碳收支与平衡的分析表明,"稻鸭共生"生态系统是碳汇,且固碳潜力大于常规稻作。