In vitro and in vivo antifungal activities of decursin and decursinol angelate isolated from Angelica gigas against Magnaporthe oryzae, the causal agent of rice blast

Blast is considered the most important fungal disease of rice due to its wide distribution and destructiveness under favorable conditions. Development of new effective and environmentally benign agents against the causal pathogen, Magnaporthe oryzae, is of great interest. In the course of a search for natural antifungal compounds in medicinal plants, we found that the methanol extract of Angelica gigas roots showed a potent control efficacy against rice blast caused by M. oryzae. We isolated antifungal coumarins from the extract, and they were identified as decursin and decursinol angelate. Antifungal activities of these compounds, along with kasugamycin, were tested on M. oryzae in vivo and in vitro. In an in vivo assay, the three compounds effectively suppressed the development of rice blast at concentrations more than 100 μg/mL. Coumarins showed relatively weak inhibitory effect on fungal mycelial growth when compared to kasugamycin. However, they strongly inhibited M. oryzae spore germination, which was not observed in kasugamycin treatments. This is the first report demonstrating that decursinol angelate can provide control against rice blast and that the two coumarins inhibit M. oryzae spore germination. In addition, the wettable powder formulation of the crude extract of A. gigas prohibited the development of blast symptoms on rice plants more effectively than liquid concentrate formulation of kasugamin, a commercial fungicide. Based on our study, we propose that coumarin compounds as well as the A. gigas root crude extract can be used as natural, benign fungicides for controlling rice blast.     

福建省稻瘟病菌对主要抗瘟基因及主栽品种的致病性分析

为明确福建省各稻区的主要抗瘟基因及主栽品种的利用价值,2012—2014年采用喷雾法测定了丽江新团黑谷上的193个菌株对30个水稻抗瘟基因及93个主栽水稻品种的致病性。结果表明,供试稻瘟病菌对30个抗瘟基因表现为强致病力和较强致病力的出现频率分别为13.47%和52.85%,对93个主栽品种表现为强致病力和较强致病力的出现频率分别为1.55%和11.40%;供试稻瘟病菌对抗瘟基因Pi-k~m、Pi-7(t)、Pi-k~p和Pi-9(t)的毒力频率均低于20%;供试稻瘟病菌对谷优2329、谷优5138、昌优964等37个品种的毒力频率均低于20%,对谷优系列、全优系列、深优系列、泰丰优系列及天优系列水稻品种的毒力频率均低于20%。研究表明,在福建地区抗瘟基因Pik~m、Pi-7(t)、Pi-k~p和Pi-9(t)可作为抗源使用,且谷丰A、全丰A、深97A、泰丰A和天丰A仍是抗瘟性较好的育种亲本材料。     

云南省水稻稻瘟病菌无毒基因AVR-Pia变异及水稻抗性基因Pia的有效性

为进一步了解田间稻瘟病菌Magnaporthe oryzae群体中AVR-Pia基因的分布及变异,利用水稻单基因系IRBLa-C水稻品种对自云南省13个市(州)采集分离得到的471株稻瘟病菌菌株进行抗性基因Pia有效性测定;利用无毒基因AVR-Pia特异性标记对471株稻瘟病菌菌株进行PCR检测和测序,并分析稻瘟病菌群体中无毒基因AVR-Pia的分布及DNA结构变异;利用有效性结果和PCR检测结果对471株菌株进行反应型划分,筛选鉴定菌株;利用鉴定菌株对云南省112份地方稻种进行Pia基因鉴定。结果表明,在471株稻瘟病菌菌株中,对含有Pia基因的水稻单基因系IRBLa-C表现为抗病和感病的菌株数分别为139株和332株,所占比例分别为29.5%和70.5%;在471株稻瘟病菌菌株中,分别有244株和227株菌株含有无毒基因AVR-Pia和不含有无毒基因AVR-Pia,所占比例分别为51.8%和48.2%,无毒基因AVR-Pia主要为完全缺失变异;在471株稻瘟病菌菌株中,A-和V+反应型菌株数分别为56株和161株,共217株,占总菌株数的46.1%,在13个市(州)稻瘟病菌群体中,A-和V+反应型菌株所占比例差异较大,其中在普洱市、红河哈尼族彝族自治州、昭通市、玉溪市4个市(州)的比例较大,分别为77.8%、57.1%、52.1%和50.0%;在112份云南省地方稻种质资源中,有20份地方稻品种含有抗性基因Pia,主要分布在9个市(州)中。表明云南省13个市(州)绝大部分水稻产区水稻Pia基因已丧抗性,含Pia基因的水稻种质在云南省分布较广。     

酵母菌0732-1产生的抑细菌物质特性研究

<正>哈密瓜细菌性果斑病是近年来暴发流行的一种瓜类病害,也是我国对内对外检疫性重大植物病害。该病由燕麦嗜酸菌西瓜亚种(Acidovorax avenae subsp.citrulli,Aac)引起,具有发病快、危害广、损失重、防治难的特点,一旦发生往往给瓜产区造成严重经济损失。据报道[1],2007年新疆石河子市温室栽培的西瓜叶片上曾获得一株对Aac有     

An Ulva armoricana extract protects plants against three powdery mildew pathogens

The protective activity of a crude extract prepared from the green macroalga, Ulva armoricana, previously shown to induce plant defence responses, was evaluated on three plant species, common bean, grapevine and cucumber, cultivated in the greenhouse and inoculated with three powdery mildew pathogens Erysiphe polygoni, E. necator and Sphareotheca fuliginea respectively. Chemical analyses showed that the extract was enriched in ulvans, which are green algae polysaccharides essentially composed of uronic acid and sulphated rhamnose. Weekly applications were performed by spraying of the green algal extract at various dilutions on bean, grapevine and cucumber leaves. A significant effect (50% protection) was observed using a dilution corresponding to about 3 g l⁻¹ dry matter and up to 90% reduction of symptom severity was obtained for the highest concentration (1/9 dilution, 6 g l⁻¹ dry matter) for the three plant species. To study the natural variability of the protective activity, five extracts prepared from algae batches harvested at different year periods were evaluated. Although polysaccharide composition varied among batches, all extracts elicit a reporter gene regulated by a defence-gene promoter in a transgenic tobacco line, and protect cucumber plants against powdery mildew infection. Together, these data demonstrate that U. armoricana is a reproducible source of active compounds which can be used to efficiently protect crop plants against powdery mildew diseases.     

Natural occurrence of citrinin in rice grains and its biocontrol byTrichoderma hamatum

Paddy rice was sampled from El-Sharkia, El-Gharbia, El-Dakahlia and Kafr El-Shekh governorates, Egypt. Of the 30 samples taken, ten were contaminated with the mycotoxin citrinin. An average of 6.79 × 10⁴ fungal spores per gram rice was found. The isolated fungi represented 47 species and 28 genera. The predominant genera wereAspergillus, Cladosporium andPenicillium. Aspergilli were represented by 22 species;Aspergillus niger andA. flavus had the highest occurrence.Penicillium viridicatum produced the highest amount of citrinin on glucose ammonium nitrate salts broth and rice grains, and hence this isolate was selected as a good producer of citrinin in this study. The presence ofTrichoderma hamatum reduced the amount of citrinin produced byP. viridicatum compared with its respective control. The excessive growth ofT. hamatum onP. viridicatum was increased with time. Viability ofP. viridicatum conidia decreased byT. hamatum with an increase in the incubation period. Chitinases and 1,3-β-glucanase enzyme activity ofT. hamatum increased with extending the incubation period onP. viridicatum mycelia up to maximum values at 72 and 84 h, respectively.T. hamatum led to a decrease in the production of citrinin byP. viridicatum on rice grains compared with the respective control values. http://www.phytoparasitica.org posting Dec. 19, 2004.     

贝莱斯芽胞杆菌ZF145抑菌物质提取方法比较及成分鉴定

为明确贝莱斯芽胞杆菌Bacillus velezensis菌株ZF145中可有效抑制黄瓜角斑病病原菌扁桃假单胞菌流泪致病变种Pseudomonas amygdali pv.lachrymans的主要物质,采用冻干法、酸沉法、絮凝法、萃取法和铵沉法提取菌株ZF145发酵液中的抑菌物质,通过室内盆栽试验验证粗提物对黄瓜角斑病的防治效果,综合比较不同方法的提取效果,并通过高效液相色谱-质谱（high performanceliquid chromatography-mass spectrometry,HPLC-MS）联用技术鉴定粗提物中的抑菌物质。结果表明,絮凝法最适用于贝莱斯芽胞杆菌菌株ZF145发酵上清液中抑菌物质的提取,通过絮凝法提取的粗提物对扁桃假单胞菌流泪致病变种的抑菌圈直径达到19.50 mm,对黄瓜角斑病的盆栽防治效果为42.85%,且粗提物中的主要抑菌物质为C₁₃~C₁₆的surfactin同系物,浓度和纯度均为最高,分别为2.77 g/L和69.28%。表明本研究优化的絮凝法可用于芽胞杆菌发酵液中抑菌物质的提取。     

A fungistatic chromene from<Emphasis Type="Italic">Ageratum conyzoides</Emphasis>

Ageratum conyzoides L. is an annual herb in the tropics and subtropics whose extracts are known to possess pharmacological and biocidal activity. We report on the bioactivity of a secondary metabolite (a chromene) isolated from the shoots ofA. conyzoides against some plant pathogenic fungi. Organic solvent extracts from the shoots were tested for antifungal activity against the plant pathogenic fungiRhizoctonia solani, Sclerotium rolfsii, Botryodiplodia theobromae, Phomopsis theae andFusarium species growingin vitro on potato dextrose agar medium. The cruden-hexane extract completely inhibited the growth ofR. solani andS. rolfsii. Then-hexane extract was chromatographed over a column of silica gel followed by activity-guided fractionation to give an antifungal principle. Structure elucidation by detailed analysis of¹H,¹³C NMR and mass spectroscopy identified the active compound as precocene II. The growth ofR. solani andS. rolfsii was completely inhibited by precocene II at a concentration of 80–100 ppm. The sclerotia ofR. solani andS. rolfsii were also completely suppressed by 150 ppm of precocene II. Sub-culture of these inhibited fungi onto precocene II-free medium restored growth of the fungus, indicating that precocene II is fungistatic. Crude or refined extracts fromA. conyzoides offer the possibility of biocontrol of plant pathogenic fungi. http://www.phytoparasitica.org posting Feb. 11, 2004.     

Transgenic cucumber expressing an endogenous class III chitinase gene has reduced symptoms from Botrytis cinerea

A class III chitinase gene (CHI2) is induced in cucumber plants (Cucumis sativa L.) in response to infection by pathogenic microorganisms. Infection of Botrytis cinerea, causal agent of gray mold disease on cucumber, also induces CHI2 expression. To investigate whether CHI2 is involved in resistance to gray mold disease, transgenic cucumber plants were produced to overexpress the CHI2 gene. One line was analyzed in detail in terms of disease resistance. The transgenic cucumber plant (CC2) constitutively expressed CHI2 and reduced the symptoms of B. cinerea for 4 days after inoculation compared with nontransgenic plants. However, this inhibitory effect was not absolute, and CC2 eventually developed serious disease symptoms. Chitinase activity of the crude extract from CC2 leaves was higher than that from nontransgenic plants. A high-molecular-weight fraction containing CHI2 from CC2 leaves had fungistatic activity against B. cinerea. Interestingly, the low-molecular-weight fraction from CC2 leaves with CHI2 removed also had fungistatic activity against B. cinerea. Not only the introduced chitinase activity but also the endogenous defense reactions activated by overexpression of CHI2 may be involved in the enhanced gray mold disease resistance in CC2.     

Association between elevated activity of aryl acylamidase and propanil resistance in Jungle-rice,Echinochloa colona

Aryl acylamidase (aryl-acylamine amidohydrolase, EC 3.5.1.13) activity has been measured in crude extracts from leaves of propanil-susceptible (S) and propanil-resistant (R) biotypes of the grass weed. Echinochloa colona (L.) Link from Columbia. Both specific and total amidase activity increased with plant age up to 15 days (four-leaf stage), then decreased beyond 20 days to about 50% of the maximum at 36 days in both R and S E. colona biotypes. Specific activity with propanil in the R biotype was about 80% of that obtained for rice (Oryza sativa L.), compared to 25% in the susceptible biotype. The specific activity of the propanil amidase was three-fold higher in the R biotype than in the S. Partially purified amidase extracts from rice and both S and R biotypes of E. colona were compared biochemically. Both rice and E. colona amidases had a pH optimum of 7.5 and native relative molecular masses, estimated by gel filtration, of 179 000 and 181 000, respectively. Out of six substrates tested, three produced appreciable activity (propanil, 4-chloroacetanilide and acetanilide) in both rice and E. colona. Michaelis constants showed that the rice amidase had a higher affinity for propanil (0.36 mM) than had the E. colona enzyme (1.1 mM). Carbamates and organo-phosphorus pesticides were shown to inhibit amidase activity in partially purified rice and E. colona extracts. Additional preliminary data have implicated peroxidase in the next step of propanil metabolism in vitro. These data demonstrate that increased aryl acylamidase activity contributes to resistance to the herbicide propanil in E. colona weeds. Also, a biochemical comparison of purified aryl acylamidases from S and R biotypes of E. colona is presented for the first time.     

实时荧光定量PCR法检测十字花科细菌性黑斑病菌

为有效防控我国的检疫性有害生物十字花科细菌性黑斑病菌Pseudomonas syringae pv.maculicola在国内的传播与蔓延,通过设计1对特异性引物3539,利用132株靶标和非靶标菌为模板进行PCR扩增,建立了实时荧光定量PCR法,并进行了模拟种子带菌试验。结果显示,引物3539为只针对十字花科细菌性黑斑病菌扩增出的特异性产物;在模拟种子带菌检测中,常规PCR对菌悬液的检测限为10~5CFU/m L,实时荧光定量PCR的检测限为10~3CFU/m L,其中10~8CFU/m L菌液的Ct值最低,为22.90,10~3CFU/m L菌液的Ct值最高,为35.73,且不同浓度菌液间的Ct值均有显著差异;不同带菌率模拟种子的检测结果表明,常规PCR和实时荧光定量PCR能检测到的带菌率分别为0.5%和0.1%。研究表明,实时荧光定量PCR法不仅可用于病种的检测,也可用于病害的早期诊断。     

Spore cell wall components ofAspergillus niger elicit downy mildew disease resistance in pearl millet

Elicitors derived from the cell wall of fungi are shown to be active in eliciting resistance in plants against a wide range of pathogens. In the present study carbohydrate components from the autoclaved spore cell wall ofAspergillus niger were prepared as aqueous suspensions and tested for defense response in pearl millet (Pennisetum glaucum (L.) R.Br.) against the oomycetous downy mildew pathogenSclerospora graminicola (Sacc.) Schroet. The aqueous suspension derived from the spore cell wall ofA. niger was used as a seed soak treatment at concentrations of 0.25, 0.5, 1.0, 1.5 and 2.0 mg ml⁻¹ for time intervals of 3, 6, 9 and 12 h. The concentration of 0.5 mg ml⁻¹ for a 6 h soaking period offered 94% seed germination and seedling vigor index increased to 1526. The seed germination and the seedling vigor were significantly higher than the untreated check. Spore cell wall suspension as seed treatment at a concentration of 0.5 mg ml⁻¹ required a 3-day time interval to provide 67% protection against downy mildew. Histological and biochemical studies were conducted to elucidate the mechanism of defense response in treated seedlings uponS. graminicola infection. Resistance host response was detected in the form of lignin and callose deposition in the epidermal cell wall of pearl millet seedlings, which is the site ofS. graminicola infection. A time course study showed rapid and localized deposition of lignin and callose in epidermal cell wall of carbohydrate components-treated pearl millet seedling coleoptiles. Increased levels of the defense-related enzyme peroxidase were detected in the treated seedlings. Peroxidase activity in elicitor-treated samples reached a peak at 8 h post-infection, which was 45% more than in their respective uninoculated control. Characterization of peroxidase isoforms by isoelectric focusing revealed 16 different isoforms, of which pI 6.8, 7.2 and 8.7 increased in elicitor-treated samples uponS. graminicola infection. http://www.phytoparasitica.org posting Nov. 14, 2005.     

Effect of biopesticides applied separately or together on nutritional indices of the rice leaffolderCnaphalocrocis medinalis

Laboratory assays were done to evaluate the effect of neem seed kernel extract (Azadirachta indica A. Juss),Vitex negundo L. (Lamiales: Verbenaceae) leaf extract, andBacillus thuringiensis (Berliner), applied separately or together, on nutritional indices of the rice leaffolderCnaphalocrocis medinalis (Guenée) (Lepidoptera: Pyralidae). All three biopesticides suppressed feeding and larval growth and low concentrations affected the larval performance. The combined effect of the three biopesticides resulted in a considerable decrease in nutritional indices, indicating strong deterrence. Dose response relationships were established with respect to frass production and larval growth. The efficiency of conversion of ingested and digested food was considerably reduced.     

Selection and Screening of Endorhizosphere Bacteria from Solarized Soils as Biocontrol Agents Against Verticillium dahliae of Solanaceous Hosts

Verticillium dahliae antagonistic endorhizosphere bacteria were selected from root tips of tomato plants grown in solarized soils. Fifty-three out of the 435 selected bacterial isolates were found to be antagonistic against V. dahliae and several other soilborne pathogens in dual cultures. Significant biocontrol activity against V. dahliae in glasshouse trials was demonstrated in three of 18 evaluated antagonistic isolates, provisionally identified as Bacillus sp. Although fluorescent pseudomonads were also isolated from root tips of tomato plants, none of the tested isolates exercised any significant antagonistic activity against V. dahliae in dual cultures. So these isolates were not tested in glasshouse trials in this study. Finally, two of the most effective bacterial isolates, designated as K-165 and 5-127, were shown to be rhizosphere colonizers, very efficient in inhibiting mycelial growth of V. dahliae in dual cultures and successfully controlling Verticillium wilt of solanaceous hosts. In glasshouse experiments, root dipping or soil drenching of eggplants with bacterial suspension of 10⁷cfu ml^–1 resulted in reduced disease severity expressed as percentage of diseased leaves (40–70%) compared to the untreated controls under high V. dahliae inoculum level (40 microsclerotia g^–1 soil). In heavily Verticillium infested potato fields, experiments with potato seeds dusted with a bacterial talc formulation (10⁸cfu g^–1 formulation), showed a significant reduction in symptom development expressed as percentage of diseased potato plants and a 25% increase in yield over the untreated controls. As for their effectiveness in increasing plant height, both bacterial isolates K-165 and 5-127 produced indolebutyric, indolepyruvic and indole propionic acids. Both antagonists are considered as plant growth promoting rhizobacteria bacteria since significantly increased the height of treated plants compared with the untreated controls. Chitinolytic activity test showed that both isolates were able to produce chitinase. Testing rhizospheric and endophytic activity of the antagonists it was shown that although the bacteria are rhizosphere inhabitants they also preferentially colonize the endorhizosphere of tomatoes and eggplants. Fatty acid analysis showed that isolate K-165 could belong to Paenibacillus alvei while 5-127 to Bacillus amiloliquefaciens.     

Allelopathy and the active metabolites of the endophytic fungus,Alternaria J46, from Platycladus orientalis

The endophytic fungus, Alternaria J46, was isolated from the stem of the medicinal plant, Platycladus orientalis. A suspension of Alternaria J46 mycelial segments and the culture filtrates of the fungi exhibited marked seed germination inhibition against the monocot wheat, large crabgrass, bromegrass, rice and barnyardgrass and weak inhibition against the dicot redroot pigweed and morning glory, but it was safe for use on soybean, rape, cucumber, tomato, lettuce and radish crops. It is possible to use J46 culture filtrates in order to prevent monocot weeds in dicot cropland. Three active metabolites were isolated from an extract of the fungus cultures and elucidated as 3‐acetyl‐5‐sec‐butyltetramic acid (1, tenuazonic acid), 3‐acetyl‐5‐iso‐butyltetramic acid (2, vivotoxin II) and cyclo‐(L‐leucyl‐L‐proline) (3). Among these three compounds, compounds 1 and 2 showed significant phytotoxic effects on the seed germination of large crabgrass, while compound 3 exhibited weak activity, and all were safe for lettuce at 100 μg mL^?1. Accordingly, compounds 1 and 2 were the main active metabolites that were responsible for endophytic fungus Alternaria J46's strong seed germination inhibition against monocotyledons.     

Evaluation of rice allelopathy in hydroponics

The inhibitory activity of water extracts from the shoots and roots of three rice cultivars, Taichung native 1 (TN1) and IAC165 (both allelopathic rice) and AUS196 (non-allelopathic rice), grown in hydroponics was evaluated. The release of germination inhibitors by allelopathic rice plants into hydroponic solution was also determined with freshly collected solution and XAD-4 resin desorbate. The degree of the inhibition was quantified in terms of root growth in Echinochloa colona, Echinochloa crus-galli, Echinochloa crus-galli var. oryzicola, Triantema portulacastrum and Lactuca sativa. The allelopathic activity of rice was species specific, and depended on source and concentration. Root length of all test species was inhibited by the different concentrations of shoot extract of allelopathic and non-allelopathic rice. However, of the three cultivars, TN1 showed higher inhibition than IAC165 and AUS196 in all test species. Water extracts of shoots and roots significantly inhibited root growth in E. crus-galli but the shoot extract gave a greater inhibitory effect on E. crus-galli than the root extract. Root exudate of TN1 inhibited root elongation of E. crus-galli from 2 weeks after transplanting (WAT) and the inhibition continued for 4 WAT. The results confirmed the previous finding of a laboratory bioassay that the TN1 had allelopathic activity and produced allelochemicals that inhibit growth of some weed species.     

Translocation and degradation of pyrazosulfuron-ethyl in rice soil

BACKGROUND: Pyrazosulfuron‐ethyl {ethyl 5‐[(4,6‐dimethoxypyrimidin‐2‐ylcarbamoyl)‐sulfamoyl]‐1‐methylpyrazole‐4‐carboxylate} is a new rice herbicide belonging to the sulfonylurea group. This study reports the translocation of ¹⁴C‐pyrazosulfuron‐ethyl to rice plants and its degradation in rice‐planted and unplanted soil. RESULTS: Pyrazosulfuron‐ethyl did not show any appreciable translocation to rice shoots, as ¹⁴C‐activity translocated to the aerial portion never exceeded 1% of the initially applied ¹⁴C‐activity over a 25 day period. Results suggested that the dissipation of pyrazosulfuron‐ethyl from soils followed first‐order kinetics with a half‐life of 5.5 and 6.9 days in rice‐planted and unplanted soils respectively. HPLC analysis of the organic extract of soil samples showed the formation of three metabolites, namely ethyl 5‐(aminosulfonyl)‐1‐methyl‐1‐H‐pyrazole‐4‐carboxylate, 5‐[({[(4,6‐dimethoxy‐2 pyrimidinyl)‐amino]‐carbonyl} amino)‐sulfonyl]‐1‐methyl‐1H‐pyrazole‐4‐carboxylic acid and 2‐amino‐4,6‐dimethoxy pyrimidine, in both rice‐planted and unplanted soils. CONCLUSION: The study indicates that pyrazosulfuron‐ethyl was a short‐lived compound in the soil and was degraded relatively faster in rice‐planted soil than in unplanted soil. The herbicide did not show any appreciable translocation to rice plants. Copyright © 2011 Society of Chemical Industry     

蚜虫病原细菌的分离鉴定及其对褐飞虱的杀虫活性初探

为寻找有效防治褐飞虱的病原微生物,采用平板划线法和稀释涂布法,从感病死亡的蔷薇蚜虫体中分离筛选出一株对褐飞虱具有较高致病性的菌株JSA1;通过形态学特征、生理生化特性及16S r DNA序列分析,对菌株JSA1进行了初步鉴定;采用盆栽法和稻苗浸渍法,对其杀虫活性进行了测定。鉴定结果显示菌株JSA1为铜绿假单胞菌Pseudomonas aeruginosa。杀虫活性测定结果表明:菌株JSA1对褐飞虱具有杀虫作用,且致死率随菌液浓度的升高和作用时间的延长而提高,但致死率低于40%;菌株JSA1发酵液的上清液和沉淀破碎液对褐飞虱3龄若虫均具有致病力,72 h后的校正死亡率分别为30.72%和82.53%,表明铜绿假单胞菌JSA1在褐飞虱的生物防治方面具有一定的应用前景。     

Multidrug-resistance plasmid of <Emphasis Type="Italic">Enterobacter cloacae</Emphasis>: transfer to <Emphasis Type="Italic">Erwinia herbicola</Emphasis> on the phylloplane of mulberry and weeds

A mulberry epiphytic Enterobacter cloacae MUL1 harbors plasmid pMUL1 encoding five drug-resistance genes. This plasmid was examined upon its conjugal transfer into epiphytic Erwinia herbicola on the phylloplane of mulberry and 12 species of weeds. The plasmid was transferred into Er. herbicola at a frequency of 10^–5–10^–3/recipient in mulberry and Lolium multiflorum LAM. 1–8 days after wound inoculation with 10⁶–10⁸/ml suspensions. In Chenopodium album L. and C. album L. var. centrorubrum, however, it was transferred only after wound inoculation with a 10⁸/ml suspension, but not with 10⁷/ml or 10⁶/ml suspensions, owing to the weak epiphytic fitness of Ent. cloacae on these weeds. Transconjugants were also obtained for seven other species of weeds in the case of inoculation with a 10⁸/ml suspension. In contrast, when bacterial suspensions were sprayed on mulberry leaves with or without fresh wounds, transconjugants were obtained only in wounded leaves, which were considered suitable for bacterial conjugation. These findings suggest that epiphytic bacteria, including Ent. cloacae and Er. herbicola, may be carriers of drug-resistance genes distributed among plant pathogenic bacteria in nature.     

Isolation and purification of growth‐inhibitors from Vietnamese rice cultivars

Bioactive compounds that are produced by select rice cultivars have the potential to control barnyardgrass, a major rice yield‐limiting factor. In this study, the Vietnamese rice cultivars, OM 3536, OM 4498 and OM 5930, were selected and their inhibitory activity on cress, lettuce, canola and barnyardgrass was evaluated by using donor–receiver bioassays. These bioassays revealed that OM 3536 and OM 5930 exerted the greatest inhibitory activity, with an average growth inhibition of 57.2% on the roots and shoots of barnyardgrass. A bioassay‐guided approach was used to identify the allelopathic fractions from OM 5930 and was coupled with reverse‐phase chromatography in order to isolate several growth‐inhibitory fractions. The effective dose of the aqueous methanol extract of the rice plants that was required for 50% inhibition (ID₅₀) of the shoot and root growth of the four test plant species, as determined by a logistic regression analysis, averaged 0.091 and 0.062 g mL^?1 for OM 5930 and 0.112 and 0.072 g mL^?1 for OM 3536, respectively. The ID₅₀ of the barnyardgrass roots and shoots in the assays, as determined by a logistic regression analysis, was 0.044 and 0.149 g mL^?1 for OM 3536 and 0.052 and 0.114 g mL^?1 for OM 5930, respectively. A growth‐inhibitory fraction in the aqueous methanol extract of OM 5930 was isolated and the inhibitory activity on lettuce seedlings was determined. At a concentration of 100 p.p.m., this growth‐inhibitor inhibited lettuce root growth by up to 80.2%, compared to the control plants. These findings demonstrate that the dried plant tissues of Vietnamese rice cultivars could contain bioactive compounds that strongly inhibit plant growth.