Spontaneous ultraweak photon emission from rice (Oryza sativa L.) and paddy weeds treated with a sulfonylurea herbicide

All living organisms spontaneously generate ultraweak photon emissions, which originate from biochemical reactions in cells. Current research uses the ultraweak photon emissions from organisms as a novel tool to investigate the physiological states of plants. In this study, we found ultraweak photon emissions from leaf segments of rice and several paddy weed species treated with a sulfonylurea herbicide. There is a definite difference in photon emissions among plant species, and rice (Oryza sativa), barnyardgrass (Echinochloa crus-galli) and Cyperus serotinus showed extremely strong enhancement of photon emissions. Photon emissions from these three species treated with sulfonylurea herbicide were suppressed when the leaf segments were treated with the cytochrome P450 monooxygenase (P450) inhibitors, piperonyl butoxide and malathion. These results suggest that P450 inhibitors affect the ultraweak photon emissions from plants.     

New analogs of pochonicine,a potent β-N-acetylglucosaminidase inhibitor from fungus Pochonia suchlasporia var. suchlasporia TAMA 87

Three novel analogs of pochonicine (1) were isolated from a solid fermentation culture of the fungal strain Pochonia suchlasporia var. suchlasporia TAMA 87, and their structures were elucidated as 7-deoxypochonicine (2), 6-deoxypochonicine (3), and 6,7-dideoxypochonicine (4). These analogs were found to possess the same stereochemistry as pochonicine. Comparison of β-N-acetylglucosaminidase (GlcNAcase) inhibitory activity between these analogs and pochonicine suggested that the C-6 hydroxy group of pochonicine was essential to its potent GlcNAcase inhibitory activity and that the C-7 hydroxy group also contributed to the activity, but to a lesser extent than the C-6 hydroxy group.     

转cry1C,cry2A基因水稻对稻田浮游动物群落物种多样性的影响

转Bt基因水稻的非靶标效应一直以来都是人们关注的重点,为了解转Bt基因水稻对稻田浮游动物群落的安全性,本文以转cry1C基因水稻(T1C-19)、转cry2A基因水稻(T2A-1)及其非转基因亲本水稻明恢63(MH63)为材料,以浮游动物的种类和群落多样性为指标,研究并分析了转Bt基因水稻对稻田浮游动物群落物种多样性的影响。结果显示,水稻不同生长时期里稻田中浮游动物种类变化不明显,数量变化明显,Shannon-Wiener物种多样性指数和均匀性指数在拔节期达到最大值。另外由于孕穗期不得不施用农药防治蝗虫,导致孕穗期浮游动物的数量及物种多样性显著降低,并且在使用农药后,多样性指数和均匀性指数随着水稻的生长逐渐下降。但转Bt基因水稻对浮游动物种类和数量及群落物种多样性与对照非转基因水稻相比均无显著性差异。与施用农药对水体带来的负面影响相比,转Bt基因水稻对水体环境无负面影响。     

20%高渗乙蒜素乳油在水稻植株、稻米、稻壳、稻田水及土壤中的残留检测及消解动态

研究了20%高渗乙蒜素乳油在水稻植株、稻米、稻壳、稻田水和土壤中的残留分析方法及消解动态。样品采用二氯甲烷提取,液液分配净化,气相色谱(GC-ECD)测定。结果表明:乙蒜素在稻田水样中的平均添加回收率为85.3% ~89.1%,相对标准偏差(RSD)为4.3% ~6.3%;在植株中的平均添加回收率为87.0% ~90.3%,相对标准偏差为2.6% ~4.9%;在土壤中的平均添加回收率为90.3% ~95.4%,相对标准偏差为3.0% ~7.4%。乙蒜素的最小检出量为2.0 ×10-11 g,水样、土样、植株(以及稻米和稻壳) 中乙蒜素的最低检测浓度分别为0.002、0.01、0.02 mg/kg。湖南、天津、浙江、湖北 4地 的残留消解动态试验结果表明: 20%高渗乙蒜素乳油在稻田水样和水稻植株中的半衰期分别是 0.3 ~1.1 d和1.4~2.1 d。至水稻收获时,在稻田土壤、稻米和稻壳中均未检出乙蒜素残留,建议我国对乙蒜素在水稻中的最大允许残留限量(MRL)值可暂定为0.05 mg/kg。     

小麦秸秆对杂草种子萌发和土壤微生物代谢的影响

通过室内模拟小麦秸秆还田, 研究小麦秸秆对农田杂草种子萌发和土壤微生物代谢功能的影响。结果表明, 小麦秸秆对杂草种子的萌发抑制活性随秸秆还田的时间呈动态变化, 0~10 d抑草活性逐渐增强, 10 d后抑草活性逐渐降低。对狗尾草种子的萌发抑制效果最好, 在10 d时对狗尾草种子萌发抑制率高达83%。采用Biolog方法研究土壤微生物代谢功能, 10 d处理土壤微生物结构多样性及代谢活性显著高于其他处理。研究明确了小麦秸秆还田能够抑制杂草种子萌发, 并改变土壤微生物环境。     

稻田节肢动物群落多样性指数与植食类、捕食类亚群落发生量的相关性分析

为明确群落多样性指数在增强稻田生态系统抗性中的作用,运用线性相关性分析法,采用扫网、吸虫器和目测3种调查方法,对有机稻田和化学防治稻田中节肢动物群落多样性指数与植食类、捕食类亚群落的发生量进行了分析比较。结果表明：3种调查方法中有机稻田节肢动物群落多样性指数与植食类害虫发生量均呈负相关,2010年扫网法和2011年目测法的结果呈显著负相关,相关系数r分别为-0.77和-0.79;化学防治稻田中二者无显著相关性。有机稻田和化学防治稻田中节肢动物群落多样性指数与捕食类亚群落发生量均没有显著相关性。表明农业措施对节肢动物群落多样性与植食类亚群落发生量之间的相关性会产生直接效应制约,而对其与捕食类亚群落之间的相关性无直接效应制约。     

转Cry1Ab/Ac基因水稻对稻田底栖动物群落的生态安全性

为探究转Bt基因抗虫水稻对稻田水生生物的安全性,以转Cry1Ab/Ac基因抗虫水稻华恢1号(HH1)为材料,其非转基因水稻亲本明恢63(MH63)为对照,以底栖动物群落为指示生物,运用群落学方法研究了湖南湘潭转Bt水稻对稻田底栖动物群落的影响。结果显示,2012年在HH1与MH63稻田中,分别采集到底栖动物37种和38种,二者共有物种31种,群落物种相似度为0.8267,优势种相似度为0.8235;2013年在HH1与MH63稻田中分别采集到底栖动物40种和41种,二者共有物种33种,群落物种相似度为0.8148,优势种相似度为0.9231;2012与2013年在HH1与MH63稻田生境中,物种丰富度、个体数量、群落多样性指数、均匀性指数与优势集中性指数的时间动态趋势一致,且均无显著差异。研究表明,连续2年稻田种植转Cry1Ab/Ac基因抗虫水稻HH1,在群落水平上对湖南湘潭地区稻田底栖动物无显著影响。     

氯氟氰虫酰胺在稻田环境中的残留及消解特性

建立了一种快速、灵敏、可靠的用于分析稻田环境中氯氟氰虫酰胺残留的方法,同时研究了氯氟氰虫酰胺在稻田环境中的消解特性。田间样品经液-液分配及优化的Qu ECh ERS方法提取及净化,采用超高效液相色谱-串联质谱法(UPLC-MS/MS)进行定性、定量分析。结果表明:添加水平分别在0.002~0.5 mg/kg下,氯氟氰虫酰胺在稻田水、土壤以及水稻植株空白样品中的平均添加回收率为70%~101%,相对标准偏差(RSD,n=5)为0.7%~9.1%,其在田水、土壤和水稻植株中的最低检测浓度分别为0.002、0.005和0.01 mg/kg。该方法可满足水稻及其环境中氯氟氰虫酰胺残留量的检测要求。消解动态试验结果表明,氯氟氰虫酰胺在稻田水、土壤及水稻植株中的消解过程符合一级动力学方程,消解半衰期分别为4.8~7.7 d、5.2~8.3 d和1.5~15.4 d,属于易消解农药。     

Characterization of fluorescent Pseudomonas spp. associated with roots and soil of two sorghum genotypes

Sorghum is used as bioenergy feedstock, animal feed, and food. Economical methods for disease prevention and control are valuable for producers. Fluorescent Pseudomonas spp. were isolated from sorghum roots and surrounding soil with the goal of finding isolates that significantly inhibited sorghum fungal pathogens. Fluorescent pseudomonads were collected from seedlings of sorghum cultivars RTx433 and Redlan and wheat cultivar Lewjain, grown in two soils. Lewjain is known to support growth of producers of the antibiotic, 2,4-diacetylphloroglucinol (2,4-DAPG). Isolates from all three plants were assessed for hydrogen cyanide (HCN) and extracellular protease production, and for a 2,4-DAPG gene, phlD. Both soil type and plant type affected HCN- and protease-production, but phlD was not affected. Subsets of phlD ⁺ isolates were chosen to determine phlD genotypes and to conduct in vitro inhibition assays against sorghum pathogens. Most isolates from sorghum and wheat were genotype D, previously associated with superior root colonization. phlD ⁺ sorghum isolates were co-cultured with five sorghum pathogens. One isolate from each sorghum line exhibited inhibition to all five pathogens but more Redlan isolates were inhibitory to the virulent pathogen, Fusarium thapsinum, than RTx433 isolates. Nearly all inhibitory isolates from either sorghum cultivar were from one soil type. This is consistent with what had been previously observed in field studies: that soil type played a significant role in determining characteristics of fluorescent Pseudomonas spp. isolated from roots or soil, but sorghum genotype also had a considerable effect.     

生防荧光假单胞菌CPF-10对西瓜根围土壤真菌群落的影响

探讨了生防菌荧光假单胞菌Pseudomonas fluorescens CPF-10对西瓜根围土壤真菌群落结构的影响。采用传统分离培养法,结合变性梯度凝胶电泳(DGGE)分析,研究了施用CPF-10后不同生育期西瓜根围土壤真菌群落结构的变化。结果表明:施入生防菌CPF-10后2周,其对土壤真菌有一定的促进作用;第3～7周时,CPF-10对土壤真菌尤其是部分病原菌有较强的抑制作用;CPF-10对丛枝菌根真菌(AMF)有一定的促进作用,可维持3周左右;收获期后则检测不到CPF-10对土壤真菌群落的影响。生防菌CPF-10对西瓜根围土壤真菌群落产生的短暂影响不会对土壤生态系统构成长期威胁。对比土著真菌及丛枝菌根真菌的DGGE图谱和切胶条带测序结果,发现DGGE技术更适用于分析小范围特定菌属如丛枝菌根真菌的变化。     

Inhibition of permethrin-hydrolyzing esterases from Wiseana cervinata larvae

Inhibition of permethrin-hydrolyzing enzymes from larvae of the porina moth Wiseana cervinata has been examined in vivo and in vitro. Significant inhibition was shown by carbaryl and pirimiphos-methyl. 1-Dodecylimidazole substantially inhibited permethrin hydrolysis only in liver insects. The triphenylmethane dye tetrabromophenolphthalein was a moderate inhibitor only in vitro. TMDM (bis(N-dimethyl-4-aminophenyl)methane) had little effect on hydrolysis. These observations extend the range of species and substrates for which the triphenylmethane dyes and 1-dodecylimidazole are useful inhibitors of insecticide metabolism.     

Analogy in the mode of action of fluotrimazole and clotrimazole in Ustilago avenae

Fluotrimazole [BUE 0620; 1-(3-trifluoromethyltriphenyl) 1,2,4-triazole] (20 μg/ml of nutrient solution) and clotrimazole [Bay b 5097; bisphenyl(2-chlorophenyl)-1-imidazolyl methane] (5 μg/ml) did not inhibit dry weight increase and only slightly reduced multiplication of sporidia of Ustilago avenae during the first doubling period (about 4 hr). After 8 hr, both fluotrimazole and clotrimazole more strongly inhibited sporidia multiplication than dry weight increase. As a consequence of treatment with both fungicides the usually single-celled sporidia appear swollen, multicellular, and branched. Both chemicals at a concentration range of 5–100 μg/ml did not affect oxidation of glucose. The effect of fluotrimazole and clotrimazole on protein, DNA, and RNA synthesis was similar to that on dry weight. Following a 6-hr incubation period total lipid synthesis was quantitatively unaffected by both chemicals. As the analysis of major fatty acids of total lipids revealed fluotrimazole substantially induced the synthesis of 20:4 carbon fatty acids, while in clotrimazole-treated sporidia the pattern of fatty acids did not differ from that of control sporidia. Fluotrimazole and clotrimazole produced a higher quantity of free fatty acids in sporidia of U. avenae. Gas-liquid chromatographic analysis of sterol fractions in treated and control sporidia (6 hr) indicated that both fluotrimazole and clotrimazole seriously inhibited ergosterol biosynthesis and concomitantly caused an accumulation of immediate ergosterol precursors which represent C-4-methyl and 4,4-dimethyl sterols. Incorporation of [¹⁴C]acetate for 2 hr into various lipid fractions of sporidia of U. avenae also revealed that radioactivity in C-4-desmethyl sterols in both fluotrimazole- and clotrimazole-treated sporidia was drastically reduced, while the radioactivity of C-4-methyl and 4,4-dimethyl sterols distinctly increased. The data suggest that fluotrimazole and clotrimazole are specific inhibitors of the oxidative demethylation of the C-14-methyl group during ergosterol biosynthesis in U. avenae.     

稻瘟酰胺在水稻植株及稻田水、土壤中的 消解动态研究

为评价稻瘟酰胺在水稻上使用的安全性,建立其使用规范,于2006-2007年研究了35%酰胺·稻瘟灵乳油(稻瘟酰胺∶ 稻瘟灵=30∶ 5,质量比)中稻瘟酰胺在浙江和福建两地水稻植株及稻田水、土壤中的消解动态。结果表明:稻瘟酰胺标准溶液的线性方程为y=246 192x+925.88(r=0.999 7),线性范围为 0.005~1 mg/L。残留分析中样品采用乙腈和乙酸乙酯提取,经中性氧化铝和弗罗里硅土柱层析净化,气相色谱检测。当样品中添加水平为0.001~1 mg/kg时,采用该方法测得的平均回收率为78.6% ~108.4%,相对标准偏差(RSD)在3.5% ~23.4%之间;其在水稻植株、土壤和田水中的最低检测浓度(LOQ)均为0.001 mg/kg(L)。田间试验结果表明,稻瘟酰胺在稻田样品中的消解动态曲线符合一级动力学方程,其在水稻植株、土壤以及田水中消解迅速,半衰期分别为5.2~9.5、3.8~7.3、0.8~3.2 d。     

Antifungal mode of action of imazalil

Imazalil had no effect on the initial growth of mycelia of Penicillium italicum (for 10 hr) or Aspergillus nidulans (for 2 hr). In P. italicum during this period neither respiration nor cell permeability was affected, but uptake of [³²P]phosphate, [¹⁴C]leucine, or [¹⁴C]uridine was partially inhibited. The initial (5 hr) inhibition of substrate uptake coincided with a 50% reduction in ergosterol content. Within 0.5 hr, incorporation of [¹⁴C]acetate into C-4-desmethyl sterols was strongly inhibited in mycelia of A. nidulans treated with 0.5 μg/ml of imazalil. However, radioactivity in C-4-methyl and dimethyl sterols exceeded that of control cultures. Concentrations of imazalil as low as 0.005 μg/ml caused short-term (1 hr) declines of incorporation into desmethyl sterols and increases into the C-4-methyl and dimethyl sterols. Incorporation into phospholipids, triglycerides, and free fatty acids was not affected. These data suggest that the primary antifungal action of imazalil is inhibition of demethylation in the biosynthesis of ergosterol.     

Characterization of sulfonylurea-resistant Schoenoplectus juncoides having a target-site Asp376Glu mutation in the acetolactate synthase

Schoenoplectus juncoides, a noxious weed for paddy rice, is known to become resistant to sulfonylurea (SU) herbicides by a target-site mutation in either of the two acetolactate synthase (ALS) genes (ALS1 and ALS2). SU-resistant S. juncoides plants having an Asp₃₇₆Glu mutation in ALS2 were found from a paddy rice field in Japan, but their resistance profile has not been quantitatively investigated. In this study, dose–response of the SU-resistant accession was compared with that of a SU-susceptible accession at in vivo whole-plant level as well as at in vitro enzymatic level.     

Chemical structure-activity relationships of the inhibition of sterol biosynthesis by N-substituted morpholines in higher plants

Tridemorph and fenpropimorph as well as several related N-alkyl morpholines have been tested in vitro on the cycloeucalenol-obtusifoliol isomerase, a microsomal enzyme involved in higher plant sterol biosynthesis. The results showed that N-substituted morpholines inhibit powerfully the enzyme (I₅₀ = 0.4 μM for fenpropimorph). The following important molecular parameters of the inhibition could be determined: (i) the inhibitory capacity was probably related to the presence of a positive charge on the nitrogen atom, (ii) the length of the alkyl group was critical, with a maximum activity for n = 13 carbons in the case of a linear hydrocarbon chain, (iii) the presence of bulky substituents at the proximity of the nitrogen atom led to a strong decrease of the inhibitory power, (iv) in the fenpropimorph series where a chiral center is present at C-2 of the alkyl chain, a remarkable enantiomeric selectivity of the inhibition was observed, (v) the N-oxide derivative of fenpropimorph was shown to be as active as the parent compound. The N-alkyl morpholines have been also assayed on suspension cultures of bramble cells and led to a strong accumulation of 9β, 19-cyclopropyl- and Δ⁸-sterols. This result confirmed that the cycloeucalenol-obtusifoliol isomerase was a major target of the N-substituted morpholines and suggested that the Δ⁸ → Δ⁷-sterol isomerase was also a target for these chemicals. The molecular parameters implied in the in vivo accumulation of 9β, 19-cyclopropyl sterols were very similar to those resulting from the in vitro study. The chemical structure-inhibitory activity relationship of N-alkyl morpholines was discussed with respect to their fungicidal activity which has been described in a previous study [E. H. Pommer, Pestic. Sci. 15, 285 (1984)]. The comparison revealed that the better the inhibitory capacity on the cycloeucalenol-obtusifoliol isomerase was, the higher was the fungicidal activity in vivo.     

秸秆还田对水稻根际微生物多样性和水稻纹枯病发生的影响

为探究稻麦轮作模式下秸秆还田对水稻根际微生物多样性与水稻纹枯病发生的影响,本研究采用Illumina测序技术结合生物信息学比较分析了秸秆还田与不还田处理的水稻根际微生物群落多样性差异,并采用实时荧光定量PCR （real-time fluorescent quantitative PCR,RT-qPCR）对这2种稻田土壤中水稻纹枯病菌Rhizoctonia solani含量进行测定,并采用随机五点取样法调查稻田中水稻纹枯病发病率和病情指数。结果表明,秸秆还田后水稻根际细菌群落Chao1丰富度指数和Shannon多样性指数均有所增加,分别增加14.42%和4.39%,其中前者增幅显著;根际真菌群落Chao1丰富度指数降低29.60%,而Shannon多样性指数略有增加,增加5.96%;在门水平上,秸秆还田和不还田处理的水稻根际土壤优势细菌门均为变形菌门、放线菌门、绿弯菌门和酸杆菌门,优势真菌门均为子囊菌门、接合菌门和担子菌门;在属水平上,秸秆还田处理的硝化螺旋菌属和芽胞杆菌属相对丰度分别比不还田处理的显著增加96.11%和106.19%;在水稻孕穗期秸秆还田土壤中水稻纹枯病菌含量比不还田处理的显著增加120.20%;秸秆还田后水稻纹枯病发生加重,病情指数比不还田处理的增加18.10%~45.41%。     

Potential allelopathic effects of Suregada multiflorum and the influence of soil type on its residue's efficacy

This experiment was conducted to determine the potential allelopathic effects of the aqueous extracts and dried leaf powder of Suregada multiflorum and to find the herbicidal effects of its granule on the weed species, barnyardgrass (Echinochloa crus‐galli) and slender amaranth (Amaranthus viridis), in various growth media (germination paper, sand, and soil). Observations in this study demonstrated that the potential and magnitude of their inhibitory effects varied among the plant parts. The leaves and branches possessed the strongest inhibition levels, followed by the bark. The aqueous extract from the leaves of S. multiflorum had a remarkably stronger inhibitory effect on the germination and growth of slender amaranth, compared with barnyardgrass; when applied at the concentration of 100 g L^–1, it completely inhibited the germination and initial seedling growth of slender amaranth. In addition, all the treatments of the dried leaf powder had stronger inhibition effects than the aqueous extract. Interestingly, the inhibitory effects of the dried leaf granule of S. multiflorum were stronger than the effects of the dried leaf powder at equal rates. The degree of inhibitory effect of the S. multiflorum granule, applied in various growing media, can be classified in order of decreasing inhibition as germination paper = sand > clay soil. The present study indicates that the S. multiflorum granule product that was developed in this research has the potential to provide a powerful organic herbicide for controlling slender amaranth.     

Herbicidal and biological characteristics of a new benzenesulfonylurea compound K-11451 under submerged paddy conditions

The new benzenesulfonylurea K-11451, an α-hydroxy-β-fluoropropyl- compound, applied at 3–9 g ha ⁻¹, controlled annual and perennial weeds grown in submerged paddy soil under greenhouse conditions. It effectively controlled barnyardgrass at growth stages varying from pre-emergence to the five-leaf stage. The compound inhibited acetolactate synthase, I₅₀ values for the enzyme isolated from barnyardgrass and rice being 56 and 67 nM , respectively. K-11451 inhibited the growth of rice when it was transplanted at a shallow depth (0–1 cm) and water leaching from the paddy soil was high (3–5 cm per day). With a water depth of 3 cm, the compound appeared to move readily down into the paddy soil and had the relatively short half-life of 15.2 days under submerged paddy conditions. A mixture of K-11451 + mefenecet + daimuron (9 + 250 + 250 g ha⁻¹) controlled almost all weeds in the paddy field without injury to rice, so that the combination could be used as a ‘one-shot’ herbicide in rice culture.     

特丁硫磷在环境中的迁移性研究

采用室内模拟试验方法,研究了有机磷类农药特丁硫磷在不同环境介质中的挥发性以及在3种不同类型土壤中的吸附性、移动性及其影响因素。结果表明:特丁硫磷具有较强的挥发性,其在空气、土壤表面和水中的挥发率分别为87.1%、38.6%和2.51%;在江西红壤、太湖水稻土与东北黑土中的吸附性较好地符合Freundlich方程,3种土壤中的吸附系数Kd值分别为6.56、11.34和15.80;薄层层析试验显示,当溶剂展开至12.0 cm时,特丁硫磷在江西红壤、太湖水稻土与东北黑土中最远可分别移至6~8、4~6和6~8 cm处,土柱试验结果显示,江西红壤、太湖水稻土与东北黑土淋出液中特丁硫磷质量分数分别为5.77%、1.13%和1.22%,表明特丁硫磷具有一定的移动性。影响特丁硫磷在土壤中吸附性和移动性的主要因素为土壤有机质含量,其次为土壤质地。特丁硫磷具有高挥发性及一定的移动性,其对人畜的安全以及地下水的危害应引起高度重视。