Mycelial Growth Characteristics Research Articles

Biological Characteristics of the Mycelium and Optimization of the Culture Medium for Phallus dongsun.

This study aimed to elucidate the influence of various culture medium components, including carbon sources, nitrogen sources, inorganic salts, suspension agents, and temperature, on the mycelial growth characteristics of Phallus dongsun. Employing single-factor experiments and response surface methodology within glass Petri dishes, the research identified that carrot powder, soybean powder, and ZnSO4 notably enhanced the proliferation of aerial mycelium, significantly augmenting the growth rate of P. dongsun mycelium. The resultant mycelium was observed to be dense, robust, and fluffy in texture. In particular, ZnSO4 markedly accelerated the mycelium growth rate. Furthermore, xanthan gum was found to effectively modulate the medium's viscosity, ensuring a stable suspension and facilitating nutrient equilibrium. The optimal cultivation temperature was determined to be 25°C, with mycelial growth ceasing below 5°C and mycelium perishing at temperatures exceeding 35°C. The optimal medium composition was established as follows: wheat starch 5 g/l, carrot powder 5 g/l, soybean powder 7.50 g/l, glucose 10 g/l, ZnSO4 0.71 g/l, NH4Cl 0.68 g/l, xanthan gum 0.5 g/l, and agar 20 g/l. Under these optimized conditions, the mycelium of P. dongsun exhibited a rapid growth rate (1.04 ± 0.14 mm/day), characterized by a thick, dense, and well-developed structure. This investigation provides a theoretical foundation for the conservation, strain selection, and breeding of P. dongsun.

The Effects of Antofine on the Morphological and Physiological Characteristics of Phytophthora capsici.

Phytophthora capsici is an important plant pathogenic oomycete that causes great losses to vegetable production around the world. Antofine is an important alkaloid isolated from Cynanchum komarovii Al. Iljinski and exhibits significant antifungal activity. In this study, the effect of antofine on the mycelial growth, morphology, and physiological characteristics of P. capsici was investigated using colorimetry. Meanwhile, the activity of mitochondrial respiratory chain complexes of P. capsici was evaluated following treatment with a 30% effective concentration (EC30), as well as EC50 and EC70, of antofine for 0, 12, 24, and 48 h. The results showed that antofine had a significant inhibitory effect against P. capsici, with an EC50 of 5.0795 μg/mL. After treatment with antofine at EC50 and EC70, the mycelia were rough, less full, and had obvious depression; they had an irregular protrusion structure; and they had serious wrinkles. In P. capsici, oxalic acid and exopolysaccharide contents decreased significantly, while cell membrane permeability and glycerol content increased when treated with antofine. Reactive oxygen species (ROS) entered a burst state in P. capsici after incubation with antofine for 3 h, and fluorescence intensity was 2.43 times higher than that of the control. The activities of the mitochondrial respiratory chain complex II, III, I + III, II + III, V, and citrate synthase in P. capsici were significantly inhibited following treatment with antofine (EC50 and EC70) for 48 h compared to the control. This study revealed that antofine is likely to affect the pathways related to the energy metabolism of P. capsici and thus affect the activity of respiratory chain complexes. These results increase our understanding of the action mechanism of antofine against P. capsici.

Culture media and temperature effects on Neofusicoccum mediterraneum: assessing mycelial growth and colony characteristics

Culture media and temperature effects on Neofusicoccum mediterraneum: assessing mycelial growth and colony characteristics

Biology, morphology, and phylogeny of some strains of the Pleurotus eryngii species complex

Effective methods of preserving the gene pool of valuable edible and medicinal mushrooms are to maintain them in in vitro culture collections and to correctly identify and verify the preserved strains. It is necessary to consider not only the results of molecular genetic studies but also cultural, morphological and physiological characteristics as additional criteria. This article presents data on the colony and mycelial morphology, growth characteristics and temperature tolerance, and phylogenetical placement of four strains of edible and medicinal mushroom from the P. eryngii species complex received into the IBK Mushroom Collection as P. nebrodensis strains. All the studied strains are mesophiles with the fastest growth rate of 11.0 mm/day at 26?? and a lethal temperature of 40??. In addition to common anastomoses, mycelial strands and clamp connections, the vegetative mycelium of the studied strains formed single colorless round excretory cells on the lateral hyphal ramifications. All cultures were able to form primordia and fruit bodies on agar media. Phylogenetic analysis suggests that all four strains do not belong to P. nebrodensis, but two of them, IBK 1947, 2035 are P. eryngii var. ferulae, and two strains, IBK 1855 and 1927, belong to P. tuoliensis (P. eryngii var. tuoliensis).

Growth Characteristics of Polyporales Mushrooms for the Mycelial Mat Formation

Mushroom strains of Polyporales from the genera Coriolus, Trametes, Pycnoporus, Ganoderma, and Formitella were explored in terms of mycelial growth characteristics for the application of mushroom mycelia as alternative sources of materials replacing fossil fuel-based materials. Among the 64 strains of Polyporales, G. lucidum LBS5496GL was selected as the best candidate because it showed fast mycelial growth with high mycelial strength in both the sawdust-based solid medium and the potato dextrose liquid plate medium. Some of the Polyporales in this study have shown good mycelial growth, however, they mostly formed mycelial mat of weak physical strength. The higher physical strength of mycelial mat by G. lucidum LBS5496GL was attributed to its thick hyphae with the diameter of 13 µm as revealed by scanning electron microscopic analysis whereas the hyphae of others exhibited less than 2 µm. Glycerol and skim milk supported the best mycelial growth of LBS5496GL as a carbon and a nitrogen source, respectively.

Comparison of Mycelial Growth Characteristics According to Culture Conditions of Ulleungdo Collection Strains

Comparison of Mycelial Growth Characteristics According to Culture Conditions of Ulleungdo Collection Strains

Influence of Carbon - Nitrogen Supplements and pH on Growth of Sugarcane Stem rot Pathogen Fusarium Solani NVS671

Fusarium solani NVS671 identified from infected sugarcane stem of Co 671 as a new pathogen was subjected to various cultural conditions to understand its physiological profile. In continuation with our previous work, cultural analysis was carried out under in vitro condition by supplementing various carbon and nitrogen sources in Czapek-Dox agar (CDA). Under different hydrogen ion concentrations, it was found that the growth of Fusarium solaniNVS671 was less at pH 4 and pH 10 and could reach up to 5 cm after 7 days of incubation on Potato Dextrose Agar (PDA). It is observed that the pH around 7 to 8 was optimum for the growth of F. solani. Ten different nitrogenous (N) sources and nine different carbon sources were tested on CDA medium to know their effect on the mycelial growth rate and characteristics of the fungus. Among the N sources evaluated, ammonium chloride (7.96±0.11 cm) was found to be most efficient for mycelial growth promotion followed by ammonium nitrate (7.7±0.1 cm) and ammonium sulphate (7.3±0.1 cm). The most preferred carbon source recorded to promote best radial mycelial growth was starch (7.96±0.05 cm) and sucrose (7.93±0.05 cm). Capabilities of using different carbon and nitrogen sources and ability to grow at different pH levels may allow species to adapt to specific soil conditions.This study is important to understand the physiology and metabolite preference of F. solani.

Pathogenicity of Phytophthora palmivora to fig fruits after its long-term storage in water

Long-term storage methods for Phytophthora palmivora were tested using five isolates stored in sterile distilled water (SDW) for up to 6 years and 8 months, and their growth and virulence were evaluated. All five cultures remained viable in SDW at room temperature and retained normal mycelial growth characteristics and morphology. All but one culture formed normal zoospores, and all five were still pathogenic to fig fruits. These findings suggest that this SDW storage is a simple, inexpensive method for long-term preservation of the pathogenic oomycete in the laboratory.

Occurrence of grapevine trunk diseases affecting the native cultivar Pedro Ximénez in southern Spain

‘Pedro Ximenez’ is a native grapevine cultivar from Cordoba province (Andalusia region, southern Spain), which has been seriously affected by fungal trunk pathogens causing general decline along the last decade. However, the occurrence of grapevine trunk diseases (GTDs) in Andalusia region has not been studied yet. Therefore, the main goal of this study was to evaluate the occurrence of GTDs affecting ‘Pedro Ximenez’, and to characterize the causal agents by morphological and molecular methods. Between 2016 and 2017, 25 commercial vineyards of cv. Pedro Ximenez showing decline symptoms were surveyed. Fungal isolations were done and 23 representative isolates of the fungal trunk pathogens associated with the different GTDs (black-foot and Petri disease, Botryosphaeria and eutypa dieback, and esca) were selected according to colony colour and mycelial growth development. Colony colour, mycelial growth, and conidial characteristics were defined. The effect of temperature on mycelial growth was evaluated. Phylogenetic analyses of ribosomal genes (ITS) and functional protein regions (ACT, HIS, TEF, TUB) were preformed to confirm their identification. The especies Cadophora luteo-olivacea, Cryptovalsa ampelina, Dactylonectria alcacerensis, Da. novozelandica, Diplodia corticola, D. mutila, D. seriata, Eutypa lata, Fomitiporia mediterranea, Ilyonectria liriodendri, Neofusicoccum mediterraneum, N. parvum, Phaeoacremonium iranianum, Pm. minimum and Phaeomoniella chlamydospora were identified. Despite grapevine fungal trunk pathogens being well known in the main grape growing areas worldwide, this present study is relevant because it has been focused in the native cultivar Pedro Ximenez, improving our knowledge about aetiology of GTDs in southern Spain towards make decisions about management strategies preventing infections.

Phenotypic variation of cabbage white mold pathogen, Sclerotinia sclerotiorum in the upcountry commercial cabbage fields in Sri Lanka

Sclerotinia sclerotiorum is a cosmopolitan, necrotrophic and soil borne plant pathogen, capable of infecting many economically important crops. Although it is a wellstudied pathogen elsewhere, there is no research conducted in Sri Lanka. The objectives of the current research were to study the phenotypic variations among isolates within the pathogen population in the upcountry commercial cabbage fields in Sri Lanka and to determine their sensitivity to a commonly applied fungicide, Mancozeb. Forty-six isolates obtained from infected cabbage heads were identified as S. sclerotiorum based on its unique morphological characteristics. Colony diameter was measured on potato dextrose agar (PDA) plates at 23 oC and mycelial colour was observed after two-week incubation. All the isolates were grown in pH indicator media to determine their acid producing abilities. Sensitivity of the isolates to Mancozeb was assayed in vitro. There was a significant difference in colony growth among the isolates and sensitivity to Mancozeb. Two isolates showed abnormal mycelial growth characteristics; a slower growth rate on PDA and reduced pathogenicity on cabbage leaves. Two levels of mycelial pigmentations were observed on PDA. All the isolates were acid producers, inferring that all of them were pathogenic. In summary, the upcountry S. sclerotiorum pathogen population was found to be diverse for selected phenotypic traits, especially for mycelial growth, and fungicide sensitivities inferring that the pathogen population is capable of adapting to changing environmental conditions. Therefore, disease management will be challenging if an epidemic occurs, and it is recommended that a proper management system should be identified before further expansion of upcountry cabbage cultivation.

RAPD analysis of Sclerotium rolfsii isolates causing collar rot of eggplant and tomato

Eight isolates of Sclerotium rolfsii from four strategically geographical sites of Bangladesh were characterized and their cultural properties like average linear mycelial growth, colony colour, colony consistency, growth pattern and sclerotia formation were studied. Isolates varied in mycelial growth and other growth characteristics and were grouped into three. The highest linear growth was displayed by S8. DNA concentration of eight isolates varied from 1150-7200 ng/?l. DNA fingerprinting by RAPD prompted the grouping of isolates. Selected 3 primers generated 20 bands with size ranging from 100-1500 bp. Out of the 20 bands, 9 bands (45%) were polymorphic and 11 bands (55%) were monomorphic among the eight isolates of Sclerotium rolfsii. The co-efficient of gene differentiation (Gst) was 1.000 reflecting the existence of high level of genetic variations among the 8 isolates. The lowest genetic distance and highest inter isolate similarity was found in S1 and S2 which would be homogeneous. The highest genetic distance and lowest inter isolate similarity found in S3, S7 and S3, S8 pair which would be most divergent isolates. The cluster analysis also revealed that S3, S7 and S8 belong to different clusters. All five varieties of eggplant and tomatoes were graded as susceptible when inoculated with eight isolates. Plant mortality 93.33% was recorded in S4, S6 and in S8. Considering the isolate factor the most virulent isolate would be S8 whereas the less virulent isolate would be S2 and S7. Host plant of S8 was tomato collected from Thakurgaon. S2 and S7 were collected from BAU farm and Dinajpur and host plants were lentil and tomato respectively. It is evident that Sclerotium rolfsii from Thakurgaon on host tomato is more virulent.Int. J. Agril. Res. Innov. & Tech. 6 (1): 47-57, June, 2016

Eco-friendly methods to control infection of Botrytis cinerea during propagation of grapevines

Grey mould (Botrytis cinerea Pers.) is a pathogenic fungus which causes damage to the production of grapevine nursery plant materials especially on the stored canes or on graftings during the callusing period. The conditions, increased temperature and humidity, are ideal for the pathogen during the pre-forcing stage and in the storage Botrytis cinerea can easily infect the outbursting buds as large amounts of grapevine canes are stored in a relatively small place. The fungicide-based management is general in the prevention of gray rot infection but the palette of authorised chemical compounds is narrowed year by year due to the regulations of the European Union. Our aim is to develop an eco-friendly method which combines the use of natural materials with techniques used in organic farming. Effect of the ultraviolet-c light on the Botrytis cinerea was studied. The development characteristics of the pathogen were examined under daylight and dark conditions and experiments were set up with cow’s milk and acetic acid. UV-C light destroyed the developed conidia, however, the radiation stimulates the development of immature propagules. Larger quantities of conidiophores and conidia were formed in daylight compared to culture in the dark, while different conditions did not signifi cantly change the mycelial growth characteristics. The developed bacteria prevented the spread of pathogen mycelia during the test with cow’s milk in Petri dishes, although the smooth development of propagules that occurred did not change the vitality of the fungal colony. Furthermore the growth of Botrytis cinerea fungus mycelia was strongly inhibited by acetic acid.

천마버섯(Armillaria gallica) 자실체 조직배양체의 특성

자연 상태에서 발생한 천마버섯 자실체의 다양한 부위로부터 분리한 조직을 버섯완전배지 및 감자배지에 배양하여 균사생장 속도, 균총 형태, 균사속 형성 정도에 따라 12종류로 구분할 수 있었다. 자실체배양체(fruiting culture isolate; FC)의 균사생장과 균사속은 감자배지보다 버섯완전배지에서 빠르고 잘 형성되었다. 이들을 다시 2차 계대 배양한 결과 버섯완전배지에서 11번 균주가 균총 분리 되면서 다시 분화되어 성장하였다. 균총 분리된 한 쪽의 균총이 12번과 유사하였고 이 12번 균주는 다른 종류와는 전혀 다른 특이한 양상을 나타내었다. 또한 RAPD에 의한 DNA 패턴 분석 결과 자실체 조직배양체는 천마균1호 품종과 동일한 양상을 나타냈으나 12번은 달랐다. 자실체 조직배양체 중에서 기존 천마균1호 품종보다 균사생장과 균사속 형성이 양호한 것을 선발할 수 있었다. 따라서 이들 조직배양체는 새로운 우량 계통으로 이용될 수 있을 것으로 사료된다. The fruiting body of honey mushroom, Armillaria gallica, was collected from Gastrodia elata cultivated fields. Pure cultures were isolated from fruiting body tissue of the mushrooms, and cultured on MCM (mushroom complete medium) or PDA (potato dextrose agar) medium. Then, 12 different types of mycelial growth characteristics such as growth rate, colony morphology and rhizomorph formation were obtained. The vitality of the mycelial growth and rhizomorph formation of the fruiting body culture isolates were better on MCM than PDA, suggesting that the optimal culture medium for A. gallica mycelia was MCM. To observe the feature of colony morphology, the subculture of isolates were incubated on MCM. Consequently, we could find the segregated or differentiated colony morphology from isolate type 11 that was similar morphology to isolate type 12. For phylogenetic analysis of the 12 isolates, RAPD (Random Amplified Polymorphic DNA) were performed. The isolate type 12 was not only shown different band patterns of RAPD variation in other 11 isolates, but also commercial strain known as Chunmagyun No. 1. Among the tissue culture isolates of fruiting, strains with better mycelial growth characteristics than Chunmagyun No. 1 were selected. We expect that the new strain can be substituted to commercial strain Chunmagyun No. 1.

Characteristics of Metacordyceps yongmunensis, a New Species from Korea

Metacordyceps yongmunensis is a newly reported species from Korea, which is very similar to Cordyceps species in morphological characters. It grows on large lepidopteran pupa, and numerous white stromata grow on a single host. Mycelial growth characteristics of M. yongmunensis isolates were studied in different media and at different temperatures. Also, different carbon sources, nitrogen sources, and mineral salts were tested for mycelial growth of M. yongmunensis. Schizophyllum (mushroom) genetics complete medium plus yeast extract, Schizophyllum (mushroom) genetics minimal medium, and Martin’s peptone dextrose agar produced longer colony diameters and more compact mycelial density than other media. The optimum temperature for mycelial growth was 25°C. Carbon sources such as sucrose, soluble starch, dextrose, glucose, dextrin, maltose, and fructose showed better mycelial growth, whereas peptone, yeast extract and tryptone resulted in the best mycelial growth of all of the nitrogen sources tested. All of the mineral salts tested showed similar growth as the control, except K2HPO4 which showed longer colony diameter and more compact mycelial density. The compact colonies were white and cottony with a greenish margin. The results showed that M. yongmunensis is an easy fungus to growas it grew from 30 to more than 50 mm in 2 wk.

풀버섯의 균사배양 적합조건 및 우량균주 선발

This study was carried out to obtain basic data on mycelial growth characteristics for an artificial cultivation of Volvariella volvacea. Twelve URP primers were used to assess genetic relationship of V. volvacea and its result was divided into two groups. But there was nothing different in morphological characteristics in V. volvacea. Among five kinds of mushroom media, MCM medium was selected as the favorable culture medium. The optimal range of temperature and pH for mycelial growth on V. volvacea were 35oC and pH 6~8, respectively. Carbon sources had not an effect on 10 strains of V. volvacea, and nitrogen source for the optimum mycelial growth was yeast extract. Also, we selected GMVV 79004 as a superior strain of 10 strains V. volvacea based on the mycelial growth and yield.

Optimal Conditions for the Mycelial Growth of Coprinus comatus Strains

The principal objective of this study was to acquire basic data regarding the mycelial growth characteristics for the artificial cultivation of Coprinus comatus. 12 URP primers were employed to evaluate the genetic relationships of C. comatus, and the results were divided into three groups. Among six kinds of mushroom media, MYP medium was selected as the most favorable culture medium for C. comatus. The optimal temperature and pH ranges for the mycelial growth of C. comatus were 23~26°C and pH 6~8, respectively. The carbon and nitrogen sources for optimal mycelial growth were sucrose and tryptone, respectively.

Profile of the extracellular lignocellulolytic enzymes activities as a tool to select the promising strains of Volvariella volvacea (Bull. ex Fr.) sing

Out of 26 strains of Volvariella volvacea used, 18 were of 'typical' type and possessed all the characteristics of a normal V. volvacea mycelium, while the rest 4 'atypical' type strains showed completely distinct mycelial growth characteristics. The remaining 4 strains grew very slowly and exhibited growth characteristics of single spore isolates of V. volvacea. Strains varied in their extracellular lignocellulolytic activities and strains; OE-274, OE-272 and OE-210 with high ligninase enzymes (laccase and polyphenol oxidase) activities, gave highest mushroom yield on pasteurized paddy straw substrate. On the composted paddy straw substrate, additional two strains, OE-213 and OE-215 with lower activities of ligninases also gave higher mushroom yield. Mushrooms were harvested 3 to 4 d early from the composted substrate than on the pasteurized substrate. Activities of endoglucanase, laccase and polyphenol oxidase were found to be more crucial for mushroom yield on pasteurized substrate, while xylanase and β-glucosidase were more important for composted substrate. Strains also varied in their fruiting bodies quality and the substrate used for mushroom cultivation also affected the fruiting body quality. The superior yielding strains varied in shape, size, weight, colour and contents of sodium and potassium in their fruiting bodies; while contents of carbon, calcium and protein did not vary much with the strains.

Development of Molecular and Biochemical Markers for Selecting a Potential High Yielding Strain of Paddy Straw Mushroom (Volvariella volvacea)

Twelve ‘normal type’ strains of paddy straw mushroom (Volvariella volvacea) varied in their mycelial growth characteristics, and the fast growing strains, OE-215, OE-209 and OE-212 exhibited higher activity of cellulases, but not of laccase and polyphenol oxidase. In protein profiles, out of twelve, two strains, OE-210 and OE-273 varied only at one protein band with respect to its intensity. The four strains, OE-272, OE-140, OE-209 and OE-55 with higher activity of endoglucanase, β-glucosidase, xylanase and laccase, were also recorded to have higher protein content in their mycelial cultures. The RAPID profiles obtained with 5 OPB series primers varied in different strains and the combined phylogenetic analysis of the RAPID profiles revealed 5 distinct phylogenetic clusters, where in the strains with common origin showed 85% to 95% similarity. The strain, OE-210, with a low intensity protein band in its protein profile also varied in mycelial growth rate on two different media, possessed highest activity of endoglucanase, β-glucosidase, laccase and polyphenol oxidase and also showed highest dissimilarity of 28% with other strains in phylogenetic analysis. The present study thus, reveals that strains with faster mycelial growth rate, forming aerial mycelia and chlamydospores, also exhibited higher activities of xylanase, exoglucanase, endoglucanase or β-glucosidase, while simply higher activity of an individual enzyme was not found sufficient to support the mycelial growth and mushroom yield. A relationship between origin of the strains and their phylogenetic belongingness was also recorded, as the strains from different origins formed different clusters in combined phylogenetic analysis.

잎새버섯 균주의 균사체 생육 최적조건

잎새버섯의 우량계통을 선발하여 농가에 보급하기 위하여 수집균주들의 균사체 생육 최적조건을 검토한 결과는 다음과 같다. 잎새버섯 수집균주의 생장에 적합한 배지는 PDA배지가 선발되었으며, 균사배양 적온은 <TEX>$25^{\circ}C$</TEX>로 나타났다. 잎새버섯의 균사생장에 적합한 수소이온 농도는 pH <TEX>$4{\sim}5$</TEX>로 약산성에서 균사생장이 우수하였다. 균사생장에 적합한 탄소원으로는 glucose와 fructose, 질소원으로는 peptone이 우수하였으며 C/N율은 <TEX>$10{\sim}20$</TEX>으로 질소 요구도가 다소 높은 것으로 나타났다. This study was carried out to obtain basic data on mycelial growth characteristics for an artificial cultivation of Grifola frondosa. Ten strains of G. frondosa were collected from Korea, China and Japan and investigated its optimal culture condition. Among four kinds of mushroom culture media, PDA medium was selected as the suitable culture medium. The optimal conditions for the mycelial growth of G. frondosa in PDA medium were <TEX>$25^{\circ}C$</TEX> and <TEX>$4{\sim}5$</TEX> of pH, respectively. The carbon and nitrogen sources for the optimum mycelial growth were fructose and peptone, respectively, and the highest mycelial growth was observed when the C/N ratio was <TEX>$10{\sim}20$</TEX>.

Peroxidase Activity in Poplar Inoculated with Compatible and Incompetent Isolates of Paxillus involutus

Peroxidase activity of the hybrid poplar Populus×canescens (Ait.) Sm. (= P. tremula L. × P. alba L.) inoculated with compatible and incompetent isolates of Paxillus involutus (Batsch) Fr. was investigated. Screening of the ectomycorrhizal fungal isolates was initiated with exploration of mycelial growth characteristics and mycorrhizal ability in vitro with poplar. Both traits varied within the fungus although they did not seem to be genetically correlated. While isolates SCO1, NAU, and 031 grew faster than others, only isolates MAJ, SCO1, and 031 were able to form ectomycorrhiza with poplar. Isolates MAJ (compatible) and NAU (incompetent) were subsequently selected for further experiments. Activity of peroxidase, one of the defense-related enzymes, was examined in pure culture and short root components of compatible and incompetent interactions between poplar and P. involutus. Peroxidase activities increased significantly in poplar inoculated with incompetent isolate of the fungus compared to control, while induction of the same enzyme was not detected in compatible associations.