Anti-hyperlipidemic and anti-diabetic evaluation of ethanolic leaf extract of Catharanthus roseus alone and in combination therapy

The use of plants in disease treatment is cost effective and relatively safe. This study was designed to investigate anti-hyperlipidemic and anti-diabetic activity of ethanolic leaf extract of Catharanthus roseus alone and in combination therapy in hyperlipidemic & diabetic mice. Eight groups comprising five mice each were used. Group A was hyperlipidemic control, group B, C, D received atorvastatin (20 mg/kg), leaf extract (200 mg/kg) and leaf extract in combination with atorvastatin (200 mg/kg and 20 mg/kg) orally for 15 days. Group E was diabetic control. Group F, G, H received sitagliptin (40 mg/kg), leaf extract (200 mg/kg) and extract in combination with sitagliptin (200 mg/kg and 40 mg/kg) orally for 7 days. Blood cholesterol levels were measured at 1st, 5th, 10th and 15th day and fasting blood sugar levels were measured at 2, 12, 24, 72 and 168 hours during treatment. One-way ANOVA with tukey- kramer multiple comparison test was used. The chemical characterization of ethanolic extract of Catharanthus roseus leaves showed presence of alkaloids, saponins, tannins and flavonoids. Ethanolic extract of Catharanthus roseus has significant anti-hyperlipidemic & anti-diabetic effects (p<0.05, p<0.01) when compared with control but had not cause significantly increase in anti-hyperlipidemic effects of atorvastatin. While significantly increased the antidiabetic effect of sitagliptin (p<0.05)

Simultaneous quantitative determination of five alkaloids in Catharanthus roseus by HPLC-ESI-MS/MS / 中国天然药物

AIM@#To establish a method to simultaneously determine the main five alkaloids of Catharanthus roseus for trace samples, a high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS) analysis method was developed.@*METHOD@#The five Catharanthus alkaloids, vinblastine, vincristine, vinleurosine, vindoline, and catharanthine were chromatographically separated on a C18 HPLC column. The mobile phase was methanol-15 nmol·L(-1) ammonium acetate containing 0.02% formic acid (65 : 35, V/V). The quantification of these alkaloids was based on the Multiple Reaction Monitoring (MRM) mode.@*RESULTS@#This method was validated, and the results achieved the aims of the study. The intra- and inter-day precision and accuracy of the five alkaloids were within 1.2%-11.5% (RSD%) and -10.9%-10.5% (RE%). The recovery rates of the five alkaloids of samples were from 79.9% to 91.5%. The five analytes were stable at room temperature for 2 h, at 4 °C for 12 h, and at -20 °C for two weeks. The developed method was applied successfully to determine the content of the five alkaloids in three plant parts of three batches of C. roseus with a minute amount collected from three regions of China.@*CONCLUSION@#The HPLC-ESI-MS/MS method can be used for the simultaneous determination of five important alkaloids in trace C. roseus samples.

Identification and expression analysis of WRKY transcription factors in medicinal plant Catharanthus roseus / 生物工程学报

WRKY transcription factors, one of the largest families of transcriptional regulators in plants, involve in multiple life activities including plant growth and development as well as stress responses. However, little is known about the types and functions of WRKY transcription factors in Catharanthus roseus, an important medicinal plant. In this study, we identified 47 CrWRKY transcriptional factors from 26 009 proteins in Catharanthus roseus, and classified them into three distinct groups (G1, G2 and G3) according to the structure of WRKY domain and evolution of the protein family. The expression profiling showed that these CrWRKY genes expressed in a tissue/organ specific manner. The 47 CrWRKY genes were clustered into three types of expression patterns. The first type includes the CrWRKYs highly expressed in flowers and the protoplast treated with methy jasmonate (MeJA) or yeast extraction (YE). The second type contains the CrWRKYs highly expressed in stem and hairy root. The third type represents the CrWRKYs highly expressed in root, stem, leaf, seedling and the hairy root treated by MeJA. Real time quantitative PCR was employed to further identify the expression patterns of the 16 selected CrWRKY genes in various organs, the MeJA-treated protoplasts and hairy roots of Catharanthus roseus, and similar results were obtained. Notably, the expresion of more than 1/3 CrWRKY genes were regulated by MeJA or YE, indicating that these CrWRKYs are likely involed in the signalling webs which modulate the biosynthesis of terpenoid indole alkaloid and plant responses to various stresses. The present results provide a framework for functional identification of the CrWRKYs and understanding of the regulation network of terpenoid indole alkaloid biosynthesis in Catharanthus roseus.

Efficacy of larvicidal and pupicidal activity of Catharanthus roseus aqueous and solvent extracts against Anopheles stephensi Liston and Culex quinquefasciatus Say (Diptera: Culicidae)

OBJECTIVE@#To investigate the larvicidal and pupicidal activities of aqueous, ethyl acetate and methanol extracts of Catharanthus roseus (C. roseus) against malaria and filariasis vectors.@*METHODS@#The larvicidal and pupicidal activities of C. roseus leaf extracts were tested against the fourth instar larvae and pupae of Anopheles stephensi (An. stephensi) and Culex quinquefasciatus (Cx. quinquefasciatus). The mortality was observed after 24 and 48 h post the treatment. The data were subjected to probit analysis to determine the lethal concentrations (LC50 and LC90) at which 50% and 90% of the treated larvae or pupae of the tested species were killed.@*RESULTS@#The larval and pupal mortality were observed after 24 and 48 h of exposure of aqueous, ethyl acetate and methanol extracts of C. roseus; no mortality was observed in the control group. The LC50 values against the fourth-instar larvae of An. stephensi were 68.62 and 72.04 mg/mL for the aqueous extract, 82.47 mg/mL for the ethyl acetate extract, and 78.80 and 86.64 mg/mL for the methanol extract, while the aqueous, ethyl acetate and methanol extracts had LC50 values of 85.21, 76.84 and 94.20 mg/mL against the fourth-instar larvae of Cx. quinquefasciatus. The aqueous, ethyl acetate and methanol extracts had LC50 values of 118.08, 182.47 and 143.80 mg/mL against the pupae of An. stephensi and 146.20, 226.84 and 156.62 mg/mL against the pupae of Cx. quinquefasciatus, respectively.@*CONCLUSIONS@#The aqueous and methanol extracts of C. roseus leaves had an excellent potential to control the malarial vector An. stephensi and filariasis vector Cx. quinquefasciatus.

Larvicidal efficacy of Catharanthus roseus Linn. (Family: Apocynaceae) leaf extract and bacterial insecticide Bacillus thuringiensis against Anopheles stephensi Liston

OBJECTIVE@#To explore the larvicidal activity of Catharanthus roseus (C. roseus) leaf extract and Bacillus thuringiensis (B. thuringiensis) against the malarial vector Anopheles stephensi (An. stephensi), when being used alone or together.@*METHODS@#The larvicidal activity was assayed at various concentrations under the laboratory and field conditions. The LC50 and LC90 values of the C. roseus leaf extract were determined by probit analysis.@*RESULTS@#The plant extract showed larvicidal effects after 24 h of exposure; however, the highest larval mortality was found in the petroleum ether extract of C. roseus against the first to fourth instars larvae with LC50=3.34, 4.48, 5.90 and 8.17 g/L, respectively; B. thuringiensis against the first to fourth instars larvae with LC50=1.72, 1.93, 2.17 and 2.42 g/L, respectively; and the combined treatment with LC50=2.18, 2.41, 2.76 and 3.22 g/L, respectively. No mortality was observed in the control.@*CONCLUSIONS@#The petroleum ether extract of C. roseus extract and B. thuringiensis have potential to be used as ideal eco-friendly agents for the control of An. stephensi in vector control programs. The combined treatment with this plant crude extract and bacterial toxin has better larvicidal efficacy against An. stephensi.

Nutrición y crecimiento en fase de vivero de Catharanthus roseus (L.) G. Don, Momordica charantia L. y Azadirachta indica A. Juss, en el Municipio Centro, Tabasco-México / Nutrition and growth in nursery phase of Catharanthus roseus (L.) G. Don, Momordica charantia L. Y Azadirachta indica A. Juss, in Centro, Tabasco-México

With the main objective of evaluate the principals agronomic parameters of soil and organic substrate, as well as nutritional indicators and biomass of medicinal plants used to control for diabetes mellitus: Catharanthus roseus (L.) G. Don, Momordica charantia L. and Azadirachta indica A. Juss, this research was conducted during 2010 and 2011 at El Colegio de la Frontera Sur (ECOSUR). The growth of potted plants was evaluated in nursery phase, analysis of soil, plants and organic substrate were performed according to NOM 021-RECNAT-2000, which allowed to identify significant differences between crop plants and substrates used. The use of organic matter in the soil improved the nutrition indicators in more than 50 percent as well as the biomass increased.

Con el objetivo de evaluar los principales parámetros agronómicos de suelo y sustrato orgánico, así como indicadores nutricionales y biomasa de las plantas medicinales utilizadas para el control de la diabetes mellitus: Catharanthus roseus (L.) G. Don, Momordica charantia L. y Azadirachta indica A. Juss, se realizó esta investigación durante 2010 y 2011 en El Colegio de la Frontera Sur (ECOSUR). El crecimiento de las plantas fue en macetas evaluándose en fase vivero, los análisis de suelo, sustrato orgánico y plantas se realizaron de acuerdo a NOM 021-RECNAT-2000; lo que permitió identificar diferencias significativas entre cultivos de plantas y sustratos utilizados. El uso de materia orgánica en el sustrato mejoró los indicadores nutricionales de los cultivos en más de un 50 por ciento al igual que la biomasa se incrementó.

Effects of water stress and nitrogen nutrition on regulation of Catharanthus roseus alkaloids metabolism / 中国中药杂志

<p><b>OBJECTIVE</b>Under various drought conditions and nitrogen application, the content of vindoline, catharanthine, vincristine and vinblastine in the leaf of Catharanthus roseus were illustrated to improve the content of alkaloid theoretically.</p><p><b>METHOD</b>Six groups were set in the experiment, which included: CK (natural control), CN (natural control + nitrogen), LK (low drought), LN (low drought + nitrogen), HK (high drought), HN (high drought + nitrogen) to discuss the change characteristics of total nitrogen, the activity of alkaline POD and TDC, the content of four alkaloids under the different conditions were measured.</p><p><b>RESULT</b>Under LK condition, the activity of POD, TDC were enhanced. In the early stage of stress (0-21 d), vindoline, catharanthine, vincristine and vinblastine accumulated, and reduced in the later stage (28-35 d). For all groups, adding exogenous nitrogen could improve the total content of nitrogen, vindoline and vinblastine, meanwhile the activity of POD and TDC were enhanced as well. The LN, HN treatments were beneficial to accumulating catharanthine and vinblastine.</p><p><b>CONCLUSION</b>Drought stress or additional nitrogen have an influence on both of the activities of POD and TDC, and the four alkaloids were affected as well. Thereinto, the LN condition was the most effective treatment for accumulating the four alkaloids (vindoline, catharanthine, vincristine and vinblastine), which were regulated by improve nitrogen content and enzymatic activity.</p>

Distribution and accumulation of vindoline, catharanthine and vinblastine in Catharanthus roseus cultivated in China / 中国中药杂志

<p><b>OBJECTIVE</b>The content of vindoline, catharanthine and vinblastine in the root, stem, leaf, flower and fruit of Catharanthus roseus at various developmental stages were determined, and the biomass allocation was also determined to find the best harvest time.</p><p><b>METHOD</b>The content of vindoline, catharanthine and vinblastine in the root, stem, leaf, flower and fruit of C. roseus were determined by HPLC.</p><p><b>RESULT</b>The content of these alkaloids were influenced by season and it varied in the different tissues of the plant. The content of vindoline and catharanthine in the leaves were the highest, and there was no vindoline detected in the root, but the content of vinblastine in the flower was the highest; the content of vindoline and catharanthine reached the maximum between the August and September, and the content of vinblastine reached the highest after the September. The biomass was the highest in the initial stage of September.</p><p><b>CONCLUSION</b>The best harvest time was in the initial stage of September.</p>

Monomeric indole alkaloids from the aerial parts of Catharanthus roseus / 药学学报

Catharanthus roseus (L.) G. Don is a plant of the Catharanthus genus of Apocynaceae which has been reported to have therapeutic effects of detoxication and anticancer. In order to further study the alkaloid constituents of C. roseus, the aerial parts of the plant were extracted with 95% EtOH, and then treated with 2% H2SO4 and NH3H2O to obtain total alkaloids. The total alkaloids were separated and purified by column chromatography over silica gel and prepared by high performance liquid chromatography (HPLC). Their structures were elucidated on the basis of physicochemical properties and spectral data. A new alkaloid together with five known compounds were isolated and identified as vindolinine B (1), lochnericine (2), horhammericine (3), vindorosine (4), vindoline (5), and coronaridine (6). Compound 1 is a new compound and named as vindolinine B.

Analysis of catharanthine content and agronomic traits in Catharanthus roseus / 中国中药杂志

Catharanthine content and agronomic traits in major Catharanthus roseus varieties were analyzed. It was found that there existed great difference in catharanthine content and agronomic traits among the varieties. Catharanthine content was the highest in variety Pacifica Polka Dot (PPD), reaching 3.79 mg g(-1) dry leaf weight, and the lowest in variety Cooler Pink (CP) with only 0.9 mg g(-1) dry leaf weight. Correlation existed in certain extent between catharanthine content and agronomic traits in C. roseus. Path analysis showed that among all the agronomic traits analyzed, internodal distance positively affected catharanthine content at significant level (P<0.05), with the path coefficient being 1.473. This study provides useful information for high-catharanthine content C. roseus introduction and breeding.

Oral glucose tolerance test [OGTT] in normal control and glucose induced hyperglycemic rats with Coccinia Cordifolia L. and Catharanthus roseus L

The aim of this study is to investigate the hypoglycemic effects of petroleum ether, chloroform and ethyl acetate fractions isolated from ethanolic extracts of Coccinia cordifolia and Catharanthus roseus on normal control and orally glucose-induced hyperglycemic rats. Single doses [150 mg/kg] of different fractions of C. cordifolia and C. roseus extracts were intraperitonelly administered. The serum blood glucose level was obtained by pricking the tail vein using glucometer at time 0, 30, 60, 90, 150 and 270 minutes.In the orally glucose induced hyperglycemic rats, chloroform-coccinia [CHCl[3]-CC] fraction showed maximum reduction of blood glucose level by 21.94% on 60 minute of the experiment. On the other hand maximum reduction [p<0.05] of 17.92% was observed for petroleum ether-catharanthus [PET-CR] on 30 minute of the experiment. Metformin HCl was used as standard drug.Our results indicate that the CHCl[3]-CC fraction is relatively more potent than other fractions of C. cordifolia. Similarly the PET-CR is found to be better than other fractions of catharanthus. Phytochemical screening test results showed that chloroform fraction of C. cordifolia contain saponins and flavonoids compounds, which are known to be hypoglycemic. On the other hand petroleum ether fraction of C. roseus contains tannins, flavonoids and alkaloid compounds produced varying degree of blood sugar reduction. On the pharmacological point of view C. cordifolia and C. roseus appears to be a valuable plant, which can be useful, at least as an adjunct, in the therapy of diabetes

Construction of genetic linkage map of the medicinal and ornamental plant Catharanthus roseus.

An integrated genetic linkage map of the medicinal and ornamental plant Catharanthus roseus, based on different types of molecular and morphological markers was constructed, using a F(2) population of 144 plants. The map defines 14 linkage groups (LGs) and consists of 131 marker loci, including 125 molecular DNA markers (76 RAPD, 3 RAPD combinations; 7 ISSR; 2 EST-SSR from Medicago truncatula and 37 other PCR based DNA markers), selected from a total of 472 primers or primer pairs, and six morphological markers (stem pigmentation, leaf lamina pigmentation and shape, leaf petiole and pod size, and petal colour). The total map length is 1131.9 cM (centiMorgans), giving an average map length and distance between two markers equal to 80.9 cM and 8.6 cM, respectively. The morphological markers/genes were found linked with nearest molecular or morphological markers at distances varying from 0.7 to 11.4 cM. Linkage was observed between the morphological markers concerned with lamina shape and petiole size of leaf on LG1 and leaf, stem and petiole pigmentation and pod size on LG8. This is the first genetic linkage map of C. roseus.

Impact of cadmium and lead on Catharanthus roseus--a phytoremediation study.

The Madagascar Periwinkle, Catharanthus roseus (L.) G. Don (a valued medicinal plant) was exposed to different concentrations ofheavymetals like, CdCl, and PbCl, with a view to observe their bioaccumulation efficiency. Germination was inhibited by both the heavy metals in the seeds previously imbibed in GA, and KNO, for 24 hr. EC50 (the effective concentration which inhibits root length by 50%) was recorded as 180 microM for CdCl2, and 50 microM for PbCl2. Both alpha-amylase and protease activity were reduced substantially on treatment of seeds with increasing concentrations of CdCl2, and PbCl2. Malondialdehyde (MDA) a product of lipoxigenase (LOX) activity also increased due to the treatment of both CdCl, and PbCl2. When two-months-old plants grown in normal soil were transferred to soils containing increasing amounts of these two heavy metals, senescence of lower leaves and extensive chlorosis were noticed after four days of transfer However, plants gradually acclimatized and after 20 days the chlorophyll content was almost comparable to normal. Plants receiving CdCl2 treatment (250 microg g(-1) and less) became acclimatized after two weeks and started normal growth. But PbCl2 of 432 microg g(-1) and less could not affect the plant growth throughout, after a preliminary shock was erased. In case of CdCl2 treatment, a stunted growth with reduced leaf area, reduced biomass and sterility were recorded after six months, while plants show normal growth and flowering in case of PbCl2 treatment. Total alkaloid was also found to be decreased in the roots of CdCl2 treated plants. No change was observed in case of PbCl2. GA3 treatments to the CdCl2 treated plants show internode elongation and increase in leaf area with relatively elongated leaves and thinning of stem diameter AAS analyses of leaves of treated plants exhibited 5-10% accumulation of cadmium, but there was no accumulation of lead at all.

Hypoglycemic effect of Catharanthus roseus in normal and streptozotocin-induced diabetic rats.

The effects of crude juice (at 0.5 and 1 ml/kg b.w.) and aqueous extract (at 0.30 and 0.45 gm/kg b.w.) of leaves of Catharanthus roseus on serum glucose level in streptozotocin induced diabetic rats were examined at 8 hours, 12 hours and 24 hours following single oral administration. The administration of crude juice at 1 ml/kg b.w. continued for another 9 doses (total 10 single morning doses given) and its effect was examined on the 4th and 11th day. The rats were made diabetic by single intraperitoneal injection of streptozotocin at 45 mg/kg b.w. Glibenclamide was used in the study for comparison. The crude leaf juice at 0.5 and 1 ml/kg b.w. reduced the serum glucose level in streptozotocin induced diabetic rats throughout the 24-hour period significantly (P varies between 0.05 and 0.001 at different times). The aqueous extract at 0.30 and 0.45 gm/kg reduced the serum glucose level in streptozotocin diabetic rats at 8 and 12 hour significantly (P varies between 0.05 to 0.01 at different times) but not at the 24 hour. Glibenclamide, at 500 mug/kg, also reduced the serum glucose level in streptozotocin induced diabetic rats throughout the 24-hour period (P<0.001).The crude leaf juice at 1 ml/kg also significantly reduced the serum glucose level in the streptozotocin induced diabetic rats on the 4th and 11th day (P<0.001 on both occasions). The effect of crude leaf juice at 1 ml/kg b.w administered daily orally over a 10 day period was also examined on a group of normal rats at different times. The study showed significant reduction at 8 hr (P<0.05), 12 hr, 24 hr and on the 4th day (P<0.01 on these 3 occasions) and also on the 11th day (P<0.001).

Isolation of poly (A)+ mRNA for downstream reactions from some medicinal and aromatic plants.

In the present protocol for extraction of RNA, hexadecyltrimethylammoniumbromide (CTAB) and insoluble polyvinylpyrrolidone were used followed by LiCl precipitation, CsCl ultracentrifugation and finally poly (A)+ mRNA was isolated with the help of oligo(dT)-cellulose columns. The isolated poly (A)+ mRNA was found to be suitable for cDNA-AFLP and suppression subtractive hybridization applications. It is a modified and consolidated protocol based on previously described methods for isolated steps and works better for medicinal and aromatic plants. High yield of poly (A)+ mRNA coupled with its amenability for downstream reactions like RT-PCR, northern blotting and cDNA synthesis for library construction is a key feature of the present protocol.

Protective effects of Piper nigrum and Vinca rosea in alloxan induced diabetic rats.

In the present study aqueous extract of Piper nigrum seeds and Vinca rosea flowers were administered orally to alloxan induced diabetic rats once a day for 4 weeks. These treatments lead to significant lowering of blood sugar level and reduction in serum lipids. The levels of antioxidant enzymes, catalase and glutathione peroxidase decreased in alloxan induced diabetic rats however these levels returned to normal in insulin, P. nigrum and V. rosea treated rats. There was no significant difference in superoxide dismutase activity in all groups compared to controls. Lipid peroxidation levels were significantly higher in diabetic rats and it was slightly increased in insulin, P. nigrum and V. rosea treated rats as compared to control rat. These results suggest that oxidative stress plays a key role in diabetes, and treatment with P. nigrum and V. rosea are useful in controlling not only the glucose and lipid levels but these components may also be helpful in strengthening the antioxidants potential.

A study on the hairy root culture and antitumor alkaloids production of Catharanthus roseus / 中国中药杂志

<p><b>OBJECTIVE</b>To establish transformation system and obtain alkaloids from the hairy root of Catharanthus roseus.</p><p><b>METHOD</b>Hairy roots were obtained by infecting the different explants of C. roseus. Culture conditions of hairy root were optimized.</p><p><b>RESULT</b>The best transformation condition was leaf infected by two-day's pre-culture and two-day's co-culture and additional A(S) (hydroxyacetosyringone) 100 mg x L(-1). The inducing rate of hairy root was up to 86.25%. The best condition of hairy root culture was MS medium with sucrose as carbon material and lactalbumin as nitron material. The analysis result showed that the contents of total alkaloids in hairy roots were higher than explants and calli.</p><p><b>CONCLUSION</b>Hairy root of C. roseus will be useful for the production of active components in C. roseus.</p>

Advances in the study of vincristine: an anticancer ingredient from Catharanthus roseus / 中国中药杂志

Vincristine is a dimer-indo-alkaloid which is extracted from the leaves of Catharanthus roseus. It is effective to treat acute lymphocytic cell leukemia, Hodgkin disease and non-Hodgkin disease clinically. But the severe side effects, such as neurotoxic and tissue damage, limit its application. In this paper, we summarize physical, chemical, pharmacological and pharmacokinetical properties of VCR and advances in decreasing its side effects. In clinic, association with other medication is adopted. In pharmaceutics, people adopt some new methods and technology such as conjugation with the antibody, encapsulation in liposomes or controlled release films.

Industrialization of medicinal plant tissue culture / 中国中药杂志

Compared with other countries, the industrialization of medicinal plant tissue culture is more important for our country because China is consuming and exporting the most amounts of herb materials in the world. Each year, many papers and patents are published on cell cultures of popular medicinal plants, such as Taxus sp., Catharanthus roseus, and Panax ginseng, and, meanwhile, the research on organ cultures of medicinal plants is increasing very quickly, which is deepening the study of medicinal plant tissue culture. During the past 30 years, Chinese scientists have cultured many medicinal plant cells, organs and hairy roots. In addition, the large-scale cultures have been tested on medicinal plants, such as Catharanthus roseus, Panax notoginseng, Anisodus acutangulus, Lithospermum erythrorhizon, and Taxus chinensis. However, the bioreactor size is not big enough for the commercial cultivation and we have not mastered the culture technique on a large scale. We should clearly understand the importance and great potential benefit of medicinal plant tissue culture and develop the tissue culture techniques for the modernization of TCM. To develop the technique that we have the property right, the pioneering spirit is needed in our research, and, meanwhile, it should be pointed out emphatically the collaboration is indispensable among scientists from different research fields.

Effects of Plant Extracts on Conidial Germination, Mycelial Growth and Sporulation of Fungi Isolated From Poultry Feed

Effect of ethanolic extracts of Lawsonia inermis, Azadirachta indica, Vinca rosea, Tagetes patula, Ocimum sanctum, Colocasia antiquorum, Adhatoda vasica, Moringa oleifera, Datura metel and Curcuma longa leaf on conidial germination, mycelial growth and sporulation of Aspergillus flavus, A. niger and A. fumigatus were examined. The conidial germination of A. flavus and A. fumigatus were most inhibited by the extract of L. inermis, while that of A. niger was inhibited by A. indica. Other tested plant extracts have a good effect on conidial germination on the selected fungi. The highest mycelial growth of A. flavus (37 mm) was found in V. rosea, but in case of A. niger and A. fumigatus it (38 and 39 mm) was found in D. metel. The lowest (4, 9 and 6 mm) respectively mycelial growth of these fungi found in L. inermis. The highest sporulation (75 x 10(4)/ml) of A. flavus was counted in V. rosea, but in case of A. niger and A. fumigatus those (45 x 10(4) and 55 x 10(4)/ml) were in D. metel and the lowest (5 x 10(4), 12 x 10(4) and 9 x 10(4)/ml) respectively sporulation of these fungi counted in L. inermis plant extract medium.