Cordyceps sensu lato contains more than 400 entomopathogenic and mycoparasitic species. It was traditionally classified in Clavicipitaceae of Hypocreales and recently phylogenetically separated into several genera such as Cordyceps Fr. 1818 emend. G. H. Sung et al.(Cordycipitaceae Kreisel ex Sung et al.), Elaphocordyceps G. H. Sung & Spatafora and Ophiocordyceps Petch emend. G. H. Sung et al.(Ophiocordycipitaceae G. H. Sung et al.), and Metacordyceps G. H. Sung et al . and Tyrannicordyceps Kepler & Spatafora (Clavicipitaceae (Lindau) Earle ex Rogerson). Some species of Cordyceps sensu lato (e.g., C. militaris and C. pruinosa) are highly regarded as medicinal mushrooms in East Asia. Beauveria species, on the other hand, are widely used as potential biocontrol agents for insects in agriculture and forest due to their wide host range and global distribution. A recently described Beauveria species, Beauveria sungii S.A. Rehner & Humber 2011, is here shown as the anamorph of Cordyceps scarabaeicola Y. Kobayasi 1976 based on the teleomorphic material and phylogenetic analysis of the B.sungii isolate KACC 47482. In Cordyceps sensu stricto clade, Cordyceps Fr. 1818 is an older generic name than Beauveria Vuill. 1912 .Hence ,C. scarabaeicola is suggested to be adopted for B. sungii on the priority basis of both generic name and species epithet in view of the recent revision of Article 59 of the Melbourne Code (recently changed to the ‘International Code of Nomenclature for algae, fungi, and plant’orICN). Based on one fungus = one name principle, all legitimate Beauveria species are also proposed to be transferred to Cordyceps. The consequences of abandonment of dual nomenclature for pleomorphic fungi are also briefly discussed with reference to Cordyceps and Beauveria.
Maintaining and preserving mushroom mycelial cultures for ensuring the genetic, morphological, physiological traits and viability over time are essential elements in mushroom production. A number of different mushroom species can be preserved by many means, such as cryopreservation, freeze-drying, and placement in preservation solutions like mineral oil, distilled water, or glycerol. This study tested simple and low-cost preservation methods of the mushroom strains from various species, namely Pleurotus ostreatus, Agaricus bisporus, Cordyceps militaris. Each mycelium was cultured and preserved in different storage temperatures and methods. Temperature were differed at 4, 14, 24, 34°C at agar slants. Different storage methods were cryobial tube with sterile water, cryobial tube + 10% glycerol, agar slant of test tube, and agar slant + mineral oil at 4°C for 2 years. The mushroom cultures were better in the viability and recovery at 4°C than those of preservation at higher temperatures. Result showed that all the mushroom strains tested remained viable without any contamination under all different storage methods over two year duration of the experiment. However, all the strains studied could be best stored in sterile water with 10% glycerol for the viability.
Cordyceps bassiana (asexual stage Beauveriabassiana) is a widely distributed entomopathogenic fungus that infects hundreds of various insects. It produces a number of secondary metabolites, including insect toxins beauvericin and bassianolide and pigment tenellin. In the present study, members of NRPS and PKS-NRPS hybrid genes were predicted from whole genome sequences of eighteen different fungal species belonging to five families of Hypocreales (Cordycipitaceae, Clavicipitaceae, Ophiocordycipitaceae, Hypocreaceae and Nectriaceae) using antiSMASH ver. 1.2.2. Phylogeny using all predicted adenylation (A) domains was drawn in order to identify potential orthologous genes of beauvericin synthetase (BbBEAS) and bassianolide synthetase (BbBSLS) in Hypocreales. The orthologs were examined to understand the evolution of beauvericin and bassianolide synthetase genes. Phylogenetic tree shows grouping of A1 and A2 domains of BbBEAS, BbBSLS and enniatin synthetases in two separate clades indicating the origin of genes from fusion of two distantly related modules. Species phylogeny of C. bassiana and its allies is congruent with gene trees of BbBEAS and BbBSLS, suggesting that the gene fusion event predates the species divergence. In the inferred phylogeny, A1 domain of BbBEAS showed highest similarity with that of BbBSLS, followed by those of Fusarium equiseti enniatin synthetase and Xylariasp. bassianolide synthetase. Syntenic analysis of beauvericin gene clusters of C. bassiana and closely related species represents that C. militaris lost the BbBEAS gene in evolution. Synteny of bassianolide gene clusters shows the loss of NRPS genes encoding BbBSLS in B. pseudobassiana, Isaria tenuipes, I. farinosa and C. pruinosa. BbBEAS and 2-ketoisovalerate reductase (kivr) genes are found conserved in beauvericin and enniatin synthesizing gene clusters.
In the present study, the genetic diversity of 69 Ganoderma species from various regions was determined by different molecular markers, including the internal transcribed spacer (ITS) rRNA, a partial β-tubulin gene, and mitochondrial small-subunit ribosomal (SSU) rDNA gene as well as randomly amplified polymorphic DNA (RAPD) analysis. The size of the ITS rDNA regions and mitochondrial SSU rDNA gene from different Ganoderma species varied from 625 to 673 bp and 656 to 2,040bp, respectively, and those of the partial β-tubulin gene sequence were 419 bp. Phylogenetic analysis based on the ITS region, the partial β-tubulin gene, and the mitochondrial SSU rDNA gene reveal that Ganoderma species are largely divided into two groups. Interestingly, most of the Ganoderma lucidum strains could be classified into 1 group, while other Ganoderma species divided into several groups (4 or 5 groups) in phylogenetic tree. One fragment unique to G. lucidum was selected from the RAPD profile and then sequenced. One primer pair (designated as KGS-F and KGS-R) based on this specific fragment was designed to amplify a 559 bp DNA fragment within the sequenced region. A single band with the expected size of 559 bp was observed from G. lucidum, except for G. lucidum strains from China, Canada, and Taiwan. This specific marker for G. lucidum from RAPD analysis, also supported by the phylogenetic analysis of the ITS, partial β-tubulin gene, and mitochondrial SSU rDNA sequences, will be useful for the PCR-based identification of G. lucidum in research applications as well as in the market.
Armillaria속에는 수목병을 일으키는 종도 있고, 천마와 공생관계를 이루는 종도 있는 것으로 알려 져 있다. Armillaria gallica 가 Gastrodia elata와 공생관계에 있다는 것이 발견되면서, A.gallica는 연구적 중요한 가치를 가지게 되었다. 또한 천마는 그 약용적 가치로 재배수요가 늘고 있지만, 점차 인공재배시 생산량 감소로 인해 재배농가에서 어려움을 겪고 있다. 천마 인공재배를 성공적으로 이 끌기 위한 가장 큰 방법은 자마(어린 천마)가 A.gallica균을 접종한 참나무를 생육에 필요한 영양분 으로 이용하는 것이다. 현재 품종으로 개발된 천마균 1호는 오래 전에 개발되어 현재의 기후조건에 안정하지 않아, 우리 는 새로운 A.gallica계통의 품종을 육성하고자 한다. 그리하여 2012년에 1차적으로 83균주의 계통분 류 실험을 하였으며, 이번 2013년에는 1차 실험에서 누락된 한국자생균주를 비롯하여 인천대학교 버섯균주은행에서 분양받은 7균주를 더 추가하여 균주 선발에 있어 빠짐이 없도록 하였다. 이번 실 험에서 주목할 점은 A.gallica균을 3년 넘게 수집하여 계속하여 배양을 하던 중 균주에서 특이한 사 항이 발견된 데에 있다. 보통 A.gallica균은 검은색의 균사속이 뿌리처럼 굵게 자라는 것이 일반적 인데 반해, TC-4, TC-11, TC-12에서 오염된 것이 아닌 흰 색의 균사가 발생했다는 점이었다. 계대 를 반복하여 실험한 결과 오염된 부분이 아니라는 것이 밝혀졌고, 각 균주에서 검은 부분과 흰 부 분을 분리하여 실험을 하였다. (이하 검은 부분(B), 흰 부분(W) 표시) 그 결과 홍릉천마균(10042)은 천마균 1호와 큰 차이점이 있는 것으로 밝혀졌다. 현재까지 진행된 실험에서는 TC-4(W), TC-12(W)만 진행되어 ITS 결과를 보면 TC-4과 TC-12번의 결과가 다른 균주와 다른 종으로 판명된 듯 보이지만 RAPD 실험에서 보면 다시 동일한 균주인 것으로 보인다. 따르서 이번 결과를 통해 역시 처음에 예상한대로 검은 부분과 흰 부분간의 차이가 있는 것으로 예 상되어지면, 이에 대한 분석은 추후에 더 진행할 계획이다. 결과를 토대로 결론을 예상해보면 홍릉 천마균은 Armillaria속의 다른 종일 것이고, 나머지 수집균주들은 A.gallica일 것이다. 또한 흰색으로 관찰되는 TC-4, TC-11,TD-12는 A.gallica에 속하지만 정확히 A.gallica로 분류되지는 않는 2차적인 균일 것이라는 예측을 해본다.
The Monokaryotic isolates of two strains collected from two different strains 'SH' and 'MS of P. ostreatus were used to estimate the compatibility of matings. They were crossed in all combinations and dikaryon formation was assayed by the presence of clamp connections. All 134 dikaryotic F1 hybrids were obtained out of 144 monokaryotic crosses derived from two parent strains. The growth and density of the F1 hybrid mycelia differed considerably and the morphological characteristics of mycelial colony of F1 hybrids varied from cottony, feathery, puffy to streak. When the 12 monokaryons of 'SH' crossed with 12 monokaryons of 'MS', 122 strains were capable of normal fruiting and 12 were not fruited among 134 strains tested indicating 91% success rate. The reason for failure of fruiting body formation in 12 combinations may be not because of the incompatibility of mating types but unfavorable environmental condition for fruiting initiation and development. The pileus of F1 hybrids were varied with funnel 68.0%, hemispherical 25.4%, plane 6.6% in the shape and gray 59.8%, blue 35.3%, white 3.3%, brown 1.6% in the color.
틈새 버섯 소비시장을 겨냥하여 선호도가 높은 야생 식용버섯의 인공재배법을 확립하고자, 자주 방망이버섯속 균주를 수집하여 균사 생육특성을 조사하였고 또한 유연관계를 분석하였다. 시험균주는 모두 18균주(Lepista sordida 6, Lepista nuda 12)로 농촌진흥청 농업유전자원정보센 터에서 분양받아 사용하였다. 온도별 균사 생장은 PDA 배지를 사용하여 ∅8.5cm 샤레에서 15℃, 20℃, 25℃, 30℃에서 각각 12일간 배양 후 조사하였다. 배지 종류별 균사 생장은 배양 온도 25℃ 조건하에 배지 종류를 PDA, MCM, CDA, MEM, YM 5종을 사용하여 ∅8.5cm 샤레에서 각각 12일 간 배양 후 조사하였다. 유연관계는 UFPF-PCR법으로 UFPF4, UFPF7 2개의 프라이머를 사용하여 분석하였다. 시험균주 18종 중 Lepista sordida 3균주, Lepista nuda 8균주 등 총 11개 균주가 자주방망이버섯 속의 전형적인 균총색인 보라색을 띠었다. 온도별 균사 생육은 전반적으로 25℃에서 양호하였으나, LS-05와 LN-06은 15~30℃ 범위에서 균사 생장이 빠르고 활력도 좋았다. 배지 종류별 균사 생육은 전반적으로 PDA와 YM 배지에서 양호한 편이었고, CDA 배지에서는 균사 활력이 매우 떨어졌다. 균주별로는 LS-01, LS-05, LN-05, LN-06 등 4균주의 균사 생육이 양호한 편이었다. 이들 수집균 주의 PCR 분석 결과, 크게 5개의 그룹으로 나누어졌다.