Jatrorrhizine and columbamine alkaloids isolated from Argemone mexicana are inhibitory to spore germination of some plant pathogenic fungi

The isomeric alkaloids jatrorrhizine and columbamine isolated from the whole plant of Argemone mexicana were assessed against spore germination of some fungi, e.g. Alternaria cajani, Bipolaris sp., Helminthosporium sp., Fusarium udum and Curvularia sp. Both the alkaloids were effective against most of the fungi tested. Jatrorrhizine was highly effective against spore germination of Bipolaris sp., Fusarium udum and Curvularia sp. at 5000 ppm concentration, whereas columbamine was highly effective against Alternaria cajani, Helminthosporium sp. and Fusarium udum at 5000 ppm.     

Antifungal activity of the mixture of quaternary alkaloids isolated from Argemone mexicana against some phytopathogenic fungi

The anti-fungal activity of a mixture of quaternary alkaloids of Argemone mexicana was observed at different concentrations (1000, 2000, 3000, 4000 and 5000 μl/ml) against 10 fungi, viz., Ustilago cynodontis, Cercospora cajani, Sphaerotheca sp., Cercospora sp., Alternaria solani, Bipolaris sp., Helminthosporium sp., Curvularia sp., Fusarium udum and Alternaria cajani. Spore germination was inhibited at 2000, 3000, 4000, 5000 μg/ml. Analysis of phenolics by high performance liquid chromatography (HPLC) recorded 11 peaks in the alkaloids but only three could be identified, viz., tannic, caffeic and ferulic acids. The significant efficacy of the alkaloid under in vitro conditions may open the possibility of its use by farmers under field conditions for controlling some crop diseases.     

Effects of some carbon and nitrogen sources on spore germination,production of biomass and antifungal metabolites by species of Trichoderma and Gliocladium virens antagonistic to Sclerotium cepivorum

Conidia of Trichoderma pseudokoningii (IMI 322662) and T. viride (IMI 322659) were incubated in 1% bacteriological peptone at 25° C for 20 h and more than 95% of the spores germinated. In the same medium, only 35% of the conidia of Gliocladium virens (G20) and T. viride (IMI 322663) germinated but when 1% glucose was added, germination was increased to 70%. In the presence of glucose as a carbon source, maximal biomass production of G. virens (G20) after seven days at 25°C was obtained with either potassium nitrate or L‐alanine as the nitrogen source, whereas the Trichoderma isolates needed an organic nitrogen source. With L‐alanine as a nitrogen source, glucose, galactose and sucrose were readily utilized for biomass production by all fungal isolates. Maltose utilization by G. virens (G20) and T. pseudokoningii was incomplete after 21 days incubation, whereas glucose utilization was complete by this time. G. virens, T. pseudokoningii and T. viride (IMI 322663) produced antifungal metabolites which were effective at reducing radial growth of Rhizoctonia solani, Botrytis cinerea as well as S. cepivorum. The metabolites produced by G. virens were very active against all three pathogens but the metabolites produced by T. pseudokoningii and T. viride (IMI 322663) were less active. T. viride (IMI 322659) was a very poor antifungal metabolite producer.     

碎米莎草穗部总生物碱化感活性和抑菌效果的研究

以种子萌发率、幼苗根长、幼苗株高和单株鲜质量以及菌落直径为指标,对碎米莎草(Cyperus iria L. )穗部总生物碱的化感活性及抑菌效果进行了初步研究.结果表明,不同质量浓度的总生物碱溶液对6种供试植物的种子萌发和幼苗生长均有不同程度的影响;质量浓度为500～4 000 mg·L~(-1)的总生物碱溶液对水稻(Oryza sativa L. )种子萌发率和幼苗的株高和单株鲜质量无显著影响(P>0.05),但4 000 mg·L~(-1)总生物碱溶液对水稻幼苗的根长有显著的抑制作用(P<0.05);总生物碱溶液对烟草(Nicotiana tabacum L. )、鳢肠[Eclipta prostrata (L. ) L. ]、千金子[Leptochloa chinensis (L. ) Nees]、三叶鬼针草(Bidens pilosa L. )及丁香蓼(Ludwigia prostrata Roxb. )的种子萌发和幼苗生长的抑制作用随质量浓度的提高而增强,且当质量浓度达到4 000 mg·L~(-1)时, 种子萌发率和幼苗的根长、株高和单株鲜质量均最低, 与对照差异显著(P<0.05). 碎米莎草穗部总生物碱溶液对6种供试植物病原真菌的抑菌效果有一定的差异, 但总体上随质量浓度(25～400 mg·L~(-1))的提高抑菌效果逐渐增强;当质量浓度达到400 mg·L~(-1)时,总生物碱溶液对苹果轮纹病病菌(Physalospora piricola Nose)、油菜菌核病病菌[Sclerotinia sclerotiorum (Lib. ) de Bary]、水稻稻瘟病病菌[Magnaporthe grisea (Hebert) Barr. ]、小麦赤霉病病菌(Fusarium graminearum Schw. )和番茄早疫病病菌(Alternaria solani Jones et Grout. )的抑菌率最高,分别达到74.05%、64.73%、43.54%、35.09%和32.56%,与对照差异显著(P<0.05);但25～400 mg·L~(-1)总生物碱溶液对杨树溃疡病病菌(Dothiorella gregaria Sacc. )无显著的抑菌作用(P>0.05).研究结果显示,碎米莎草穗部总生物碱具有被开发为稻田用生物源农药的潜力.     

Effect of chitosan on hyphal growth and spore germination of plant pathogenic and biocontrol fungi

Aims: To investigate the toxic effect of chitosan on important root pathogenic and biocontrol fungi (nematophagous, entomopathogenic and mycoparasitic). Methods and Results: We have used standard bioassays to investigate the effect of chitosan on colony growth and developed bioassays to test spore germination. The results showed that the root pathogenic and mycoparasitic fungi tested were more sensitive to chitosan than nematophagous and entomopathogenic fungi. Chitosanases (and perhaps related enzymes) are involved in the resistance to chitosan. Two fungi, one sensitive to chitosan, Fusarium oxysporum f. sp. radicis‐lycopersici, and one less sensitive, Pochonia chlamydosporia, were selected for ultrastructural investigations. Transmission electron microscopy revealed differences in the ultrastructural alterations caused by chitosan in the spores of the plant pathogenic fungus and in those of the nematophagous fungus. Confocal laser microscopy showed that Rhodamine‐labelled chitosan enters rapidly into conidia of both fungi, in an energy‐dependent process. Conclusions: Nematophagous and entomopathogenic fungi are rather resistant to the toxic effect of chitosan. Resistance of nematophagous and entomopathogenic fungi to chitosan could be associated with their high extracellular chitosanolytic activity. Furthermore, ultrastructural damage is much more severe in the chitosan sensitive fungus. Significance and impact of the study: The results of this paper suggest that biocontrol fungi tested could be combined with chitosan for biological control of plant pathogens and pests.     

Effect of bovicin HC5 on growth and spore germination of Bacillus cereus and Bacillus thuringiensis isolated from spoiled mango pulp

AIMS: To use bovicin HC5 to inhibit predominant bacteria isolated from spoiled mango pulp. METHODS AND RESULTS: Bovicin HC5 and nisin were added to brain heart infusion (BHI) medium (40-160 AU ml(-1)) or mango pulp (100 AU ml(-1)) and the growth of Bacillus cereus and Bacillus thuringiensis was monitored. Cultures treated with bovicin HC5 or nisin showed longer lag phases and grew slower in BHI medium. Bovicin HC5 and nisin were bactericidal and showed higher activity in mango pulp at acidic pH values. To determine the effect on spore germination and D values, mango pulp containing bovicin HC5 was inoculated with 10(6) and 10(9) spores per ml(-1), respectively, from each strain tested. Bovicin HC5 reduced the outgrowth of spores from B. cereus and B. thuringiensis, but thermal sensitivity was not affected. CONCLUSIONS: Bovicin HC5 was bactericidal against B. cereus and B. thuringiensis isolated from spoiled mango pulp. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacillus cereus and B. thuringiensis had not been previously isolated from spoiled mango pulp and bovicin HC5 has the potential to inhibit such bacteria in fruit pulps.     

Spore germination of Frankia strains isolated from Colletia hystrix and Retanilla ephedra (Rhamnaceae)

Two Frankia strains, ChI1 from Colletia hystrix and ReI6 from Retanilla ephedra, were assayed to determine the germinability of their spores and the stages of the life cycle in different media. In BAP medium, both strains showed approximately 18% spore germination. However, in BAP medium lacking micronutrients and FeEDTA, strain ChI1 spores exhibited approximately 53% germination and strain ReI6 spores about 42%. The survey also showed that the spores were not heat-resistant, the ReI6 spores being more sensitive than ChI1 spores. Both strains exhibited a small increase in their percentage of germination with a 15-pulse ultrasonic treatment.     

Dynamics of spore germination and mycelial growth of streptomycetes under low humidity conditions

At extremely low values of moisture pressure (?96.4 MPa; a_w 0.50), the spores of xerotolerant streptomycetes (Streptomyces odorifer and S. rubiginosohelvolus) germinated, their germ lengths increased, and lateral branching of the mycelium was observed after 5 days of incubation in a thin layer of agarized nutrient medium. At ?22.6 MPa (a_w 0.86), the mycelium begins to branch after a 2-day incubation; over a 5-day incubation at ?2.8 MPa (a_w 0.98), it goes through a reproduction cycle, which culminates in spore formation. The mathematical model approach enabled us to elucidate the behavioral patterns of Streptomyces spores in a thin layer of agarized nutrient medium at low humidity levels. The dynamics of spore germination is governed by the exponential law, which allows calculation of the average duration of the period a before spore germination, as well as the time needed for 50% of viable spores to germinate.     

In vitro spore germination and infection of cultivars of Rubus and Rosa by downy mildews from both hosts

The cardinal temperatures for in vitro germination of conidia of imported and indigenous isolates of downy mildew from hosts in the genera Rubus and Rosa were similar. A high percentage of conidia germinated above 2°C and germination remained between 80% and 90% up to 15°C or 20°C, depending on the isolate. The highest incidence of disease on leaf disks of Tummelberry (blackberry × red raspberry) inoculated with an isolate of Peronospora rubi occurred at c. 15°C, with infection over a range from 2°C to 28°C. Tests on leaf disks in vitro, and leaflets of primocane and lateral shoots in plastic tunnels, with three hybrid berry (blackberry x red raspberry), six blackberry and nine red raspberry cultivars showed the hybrid berries to be most susceptible. In a plastic tunnel infected drupelets of red raspberry fruits developed more slowly and failed to ripen evenly compared with uninfected drupelets. Similar malformation of infected fruits occurred in a plantation of Tummelberry. An isolate of P. rubi attacked severely both Tummelberry and rose cv. Can Can. Fluorescence microscopy after staining with aniline blue showed that leaf disks of Tummelberry were extensively colonised by intercellular mycelium of P. sparsa isolated from rose, even though sporulation was sparse or absent. This supports the view that P. rubi and P. sparsa may be conspecific. Oospores of P. rubi were found routinely within leaf disks of Rubus cultivars inoculated in vitro and once in naturally infected leaflets of Tummelberry.     

黄荆种子总生物碱对棉蚜无翅成蚜的毒力及其对几种酶活力的影响

采用酸水溶液提取法获得黄荆(Vitex negundo L.)种子总生物碱粗品,通过生物测定和生化分析方法研究了黄荆种子总生物碱对棉蚜(Aphis gossypii Glover)无翅成蚜的生物活性.结果显示:黄荆种子总生物碱对棉蚜无翅成蚜的触杀毒力较高,对单头棉蚜无翅成蚜的LD50为0.073 μg;用质量浓度0.500、1.000、2.000、4.000和8.000g·L-1总生物碱处理棉蚜无翅成蚜后24、48和72 h的忌避中浓度(AFC50)分别为1.256、1.720和3.923 g·L-1,具有一定的忌避和持续控制作用;用总生物碱LD30剂量(每头0.045 μg)处理棉蚜无翅成蚜,其存活时间较对照缩短7d,单头雌成蚜的繁殖力较对照降低了30.43％;单头棉蚜无翅成蚜每天的平均排蜜频率和排蜜量分别较对照减少了4.26滴和27.10 μg,差异达显著水平.在离体条件下,黄荆种子总生物碱对棉蚜无翅成蚜的羧酸酯酶、乙酰胆碱酯酶和谷胱甘肽-S-转移酶的活力有显著的抑制作用,IC50分别为0.778、1.901和4.216 g·L-1;但对多酚氧化酶的活力则有明显的激活作用,激活程度呈明显的量效关系.用总生物碱LD50剂量(每头0.073 μg)处理棉蚜无翅成蚜后3、6、12和24h,成蚜体内羧酸酯酶、乙酰胆碱酯酶和谷胱甘肽-S-转移酶的活力显著低于对照,而体内的多酚氧化酶活力则显著高于对照.研究结果表明:黄荆种子总生物碱对棉蚜无翅成蚜的杀虫作用机制与其对几种酶活力的影响效应相关.     

The effects of spermidine biosynthetic inhibitor methyl bis-(guanylhydrazone) on spore germination,growth and ethylene production in Alternaria consortiale

The spermidine synthesis inhibitor methyl bis-(guanylhydrazone) was found to reduce spore germination, hyphal and mycelial growth in Alternaria consortiale. The addition of spermidine to the culture medium resulted in a promotion of growth. Methyl bis-guanylhydrazone and spermidine did not change ethylene production.The data suggest that spermidine plays a role in the development of Alternaria consortiale independent of ethylene.Abbreviations MGBG methyl bis-(guanylhydrazone) - SPD spermidine - ACC 1-aminocyclopropane-1-carboxylic acid - PDA potato dextrose agar     

Anti-trichomonal, biochemical and toxicological activities of methanolic extract and some carbazole alkaloids isolated from the leaves of Murraya koenigii growing in Nigeria

The methanolic extract of Murraya koenigii leaf was screened for toxicological and biochemical effects on rats because of the folkloric uses as an anti-dysentery and anti-diabetes. The extract was moderately toxic (LD(50)=316.23 mg/kg body weight) to rats and had appreciable effect on the liver and kidney at higher doses leading to liver inflammation. It had little or no effect on haematology and relative organ weight of lungs, heart and spleen. Acute doses (500 mg/kg) reduced significantly serum globulin, albumin, urea, glucose, total protein, aspartate transaminase (AST), and increased cholesterol and alanine transaminase (ALT) indicating hepatic injury. However, chronic administration for 14 days gave a significant (p<0.05) reduction in the serum cholesterol, glucose, urea, bilirubin, ALT and AST showing that the plant has hypoglycaemic and hepatoprotective effects after prolonged use. The activity demonstrated by some of the isolated carbazole alkaloids and their derivatives against Trichomonas gallinae confirmed that the anti-trichomonal activity of the leaf may be due to its carbazole alkaloids. The order of activity was C(18)>C(23)>C(13). Girinimbine and girinimbilol with IC(50) values of 1.08 and 1.20 microg/ml were the most active. Acetylation of girinimbilol and mahanimbilol improved their activities to 0.60 and 1.08 microg/ml.     

The antitumoral, trypanocidal and antileishmanial activities of extract and alkaloids isolated from Duguetia furfuracea

The alkaloid extract and five alkaloids isolated from subterranean stem bark of Duguetia furfuracea (Annonaceae) were investigated for the following activities: antitumoral, trypanocidal and leishmanicidal. Dicentrinone showed weak cytotoxicity, but it had the strongest leishmanicidal activity (IC₅₀ 0.01 μM). Duguetine and duguetine β-N-oxide caused considerable antitumoral activity in every cell lines evaluated, although duguetine was more active against trypomastigote forms (IC₅₀ 9.32 μM) than other alkaloids tested.