Molecular species of ceramides from the ascomycete truffle Tuber indicum

The ceramide fractions were isolated from the chloroform/methanolic extractable of the fruiting bodies of Tuber indicum and separated into three kinds of molecular species TI-1, TI-2, and TI-3 by normal and reverse phase silica gel-column chromatography. By means of (1)H NMR and (13)C NMR spectroscopy, fast atom bombardment mass spectrometry (FAB-MS), and chemical degradation experiment, their component sphingoid base for TI-1 and TI-2 was uniformly (2S,3S,4R)-2-amino-1,3,4-octadecantriol, while the sphingoid of TI-3 was d-erythro-sphingosine, and their structures have been determined unequivocally to be (2S,2'R,3S,4R)-2-(2'-d-hydroxyalkanoylamino) octadecane-1,3,4-triol, the fatty acid composition of which consists of 2-hydroxydocosanoic, 2-hydroxytetracosanoic, and 2-hydroxytricosanoic acids (from major to minor); (2S,3S,4R)-2-(alkanoylamino)octadecane-1,3,4-triol, the fatty acid composition of which is unusual and consists of docosanoic, hexadecanoic, tricosanoic, octadecanoic and nonadecanoic acids (from major to minor); and (2S,3R,4E)-2-(alkanoylamino)-4-octadecene-1,3-diol, the component fatty acids of which were hexadecanoic (predominant) and octadecanoic acids, respectively.     

适宜印度块菌生长的地形和土壤因子

在云南、四川和西藏印度块菌产区的9个地点进行调查和采样,选择地形因子和土壤因子为评价指标,采用主成分分析法对印度块菌生长适宜性进行综合评价,以确定印度块菌的生长与地形因子和土壤因子的关系.结果表明:1)地形因子和土壤因子15个指标中提取出的5个主成分,累计贡献率达到87.5％.地形因子中坡位对块菌生长的影响最大,坡位越高越不适宜块菌生长,以中坡和中下坡最佳.2)土壤因子中容重、粉粒含量、pH值、全氮含量、交换性钙镁含量是块菌生长的限制因子.容重为0.65～0.82 g· cm-3适宜块菌生长,而容重过高(＞1 g·cm-3)不利于块菌的生长;粉粒含量为30.0％、砂粒含量为55.0％左右适宜块菌的生长,而粘粒含量过高不利于块菌的生长.在土壤化学因子中,pH值在6.40左右、全氮为2.29 ～3.70 g·kg-1、交换性钙为22.91～37.17 cmol·kg-1、交换性镁含量为1.85～2.59 cmol·kg-1的环境条件适宜块菌生长.3)综合评价表明,在9个采集地点,云南省昆明市哨上村和西藏自治区林芝地区江色岗得分较高,其地形和土壤条件最适合块菌生长;而四川省攀枝花市二十九梁子和云南省楚雄州五顶山得分较低,其地形和土壤条件不太适宜印度块菌生长.     

Influence of pH on in vitro mycelial growth in three Japanese truffle species: Tuber japonicum,T. himalayense,and T. longispinosum

Truffle species indigenous to Japan include a white-colored truffle Tuber japonicum and two black-colored truffles, T. himalayense and T. longispinosum. As the fruiting bodies of these Tuber species are promising edible, studies on the artificial cultivation for these truffles are currently underway in Japan. In the present study, we investigated the influence of pH on in vitro mycelial growth in these Tuber species to determine the optimal pH conditions for the cultivation of these truffles. Mycelia of five strains from each species were cultured in modified Melin–Norkrans liquid medium at different pH values. Tuber japonicum grew well at pH 5.0 and 6.0, whereas T. himalayense and T. longispinosum grew well at pH 7.0. This results suggest that the optimal pH for mycelial growth varies among Tuber species. The growth data collected in this study can be used to design optimal pH conditions for artificial cultivation of these Japanese truffles.     

Phylogenetic study of two truffles, Tuber formosanum and Tuber furfuraceum, identified from Taiwan

Truffles are one of the most valuable edible fungi and have drawn extensive research interests worldwide. In Taiwan, two species of truffle, Tuber formosanum and Tuber furfuraceum , have been identified and reported. Although the morphological features of these two truffles have been described, lack of molecular identification has led to difficulties with firmly establishing their relatedness to other truffles. In this study, we utilized the internal transcribed spacer (ITS) and β-tubulin gene sequences to generate the phylogenetic relationship of T. formosanum and T. furfuraceum with other taxonomic relatives. Our analysis revealed five/three major phylogenetic clades according to the 5.8S-ITS2/β-tubulin gene sequences and corroborated with their morphological characterization. Tuber formosanum highly resembles the Tuber indicum B complex, while T. furfuraceum is most similar to Tuber huidongense . Based on a molecular clock, we estimated that T. furfuraceum and T. formosanum would have diverged from their close relatives in mainland China between 10.2 and 4.1 Ma, respectively. Based on the results, we propose that these two Tuber species found in Taiwan might originate from the common ancestors with some truffle species in China. However, due to a long divergence time and geographical separation, they have evolved into indigenous species of Taiwan.     

Phylogenetic relationships between Tuber pseudoexcavatum, a Chinese truffle, and other Tuber species based on parsimony and distance analysis of four different gene sequences

Phylogenetic relationships between Tuber pseudoexcavatum and other Tuber species were investigated by studying the sequences of four genes: 5.8S-ITS2, beta-tubulin, protein kinase C and elongation factor 1alpha. The four phylogenetic trees allowed to differentiate the black truffle clade, composed of two subclades, one comprising the Asian black truffles (T. indicum, T. sinense, T. himalayense) and the Perigord black truffle (T. melanosporum), the second comprising T. pseudoexcavatum and T. brumale. These two subclades diverged relatively early. We propose a common ancestor, located between Europe and China, to all the black truffles. The T. brumale/pseudoexcavatum subclade would have started to diverge and migrate first, T. brumale towards Europe through a northern route and T. pseudoexcavatum towards China. Later the T. melanosporum subclade would have started to migrate through the same route, T. melanosporum towards Europe and T. indicum towards China, leading to vicariant species.     

Quantitative response of cell growth and Tuber polysaccharides biosynthesis by medicinal mushroom Chinese truffle Tuber sinense to metal ion in culture medium

During the submerged fermentation of medicinal mushroom Chinese truffle Tuber sinense, there was no significant effect of metal ion on the cell growth and the production of intracellular polysaccharides, while metal ion and its concentration significantly affected the production of extracellular polysaccharides (EPS). By using the approach of "one-variable-at-a-time", 50 mM Mg2+ was identified to be the most favorable for EPS production, and the next was 10 mM K+. A mathematical model, constructed by response surface methodology combination with full factorial design, was applied to study the synergic effect of Mg2+ and K+. EPS production reached its peak value of 5.86 g/L under their optimal combination of 30 mM Mg2+ and 5 mM K+ predicted by the model, which was higher by 130.7% compared with the basal fermentation medium without metal ion. The validation experiment showed the experimental values agreed with the predicted values well. EPS production obtained in this work was the highest reported in the culture of T. sinense.     

中国块菌属多样性

在中国特别是西南地区块菌属Tuber资源调查的基础上,本文综合形态特征、生态特征以及分子系统学分析,对比研究了欧洲和北美的部分块菌属标本,对中国块菌属已知种类进行了分类订正和系统的总结.研究确认我国分布有17种块菌,其中3/4的种类分布于西南地区.尚有6种因未观察到可靠的标本而存疑,3种排除在我国分布.文中对该属在我国...     

成熟度对印度块菌香气成分的影响

研究成熟度对印度块菌Tuber indicum香气成分的影响,并测定成熟印度块菌的关键香气成分。将3种不同成熟度的印度块菌,以固相微萃取(SPME)技术为香气富集方法,利用气相色谱-质谱联用(GC-MS)分析其香气成分。结果表明未成熟印度块菌中仅检测出4种香气成分,中度成熟阶段检测出8种,成熟阶段检测出13种,而且成熟阶段检测出的香气成分大多都是前人报道过对块菌香气有贡献的成分;通过计算成熟块菌各香气组分的香气活度值(OAV),可知二甲基硫醚、2,3-丁二酮、3-甲基正丁醛、2-甲基正丁醛、己醛、1-辛烯-3-醇这6种物质是印度块菌的关键香气成分(OAV>1)。     

Analysis of gene expression in the vegetative and fructification phases of the white truffle, Tuber borchii Vittad., by mRNA differential display

The mRNA differential display technique was used to compare mRNA populations from fruit body and mycelium of a white truffle species in the attempt to identify and clone differentially expressed genes. The differential expression of five out of 30 amplicons was confirmed. One fragment (Tbm 56) corresponded to a part of the ribosomal genes. Three cDNA fragments (Tbf 12, Tbf 20, Tbf 21) were expressed only in the fructification phase, while the other cDNA (Tbf 55) was expressed strongly in fruit body and also detectable in the mycelium. These clones correspond to part of the single-copy genes in the Tuber borchii Vittad. genome.     

Experiments on the life cycle and factors affecting reproduction of Sphaerosporella brunnea provide evidence for rapid asexual propagation by conidiospores and for homothallism in an ectomycorrhizal competitor of cultivated truffle species

Sphaerosporella brunnea is a pioneer and opportunist ectomycorrhizal species, and the most common fungal competitor in nurseries producing plants mycorrhized with Tuber species. Our objective was to learn more about its life cycle as the first step to manage its presence in greenhouses. Conidiation and formation of resting spore-like structures were found to be triggered by aeration and to be highest on CMA medium. In pot experiments S. brunnea was able to form ectomycorrhizas and ascocarps rapidly, in 2 and 3 months respectively, if substratum moisture was high. Both mycelia and conidiospores were effective sources of inoculum for mycorrhization. This species seems to be homothallic as apothecia have been obtained after inoculations with single monospore isolates. Propagation by mitospores and homothallism are poorly documented in ECM fungi, therefore these results may be of fundamental interest beyond the question of greenhouse management.     

Effect of Temperature,pH, and Media on the Mycelial Growth of Tuber koreanum

Members of the genus Tuber are ectomycorrhizal fungi; this genus includes more than 180 species worldwide. In the present study, the optimal pH, temperature, and medium suitable for the mycelial growth of the Korean truffle, Tuber koreanum, were determined. Mycelium of T. koreanum, isolated from fruiting bodies collected in Korea, was used to investigate the effects of these environmental factors. The results showed that malt extract agar and potato dextrose agar were the most suitable for the mycelial growth of T. koreanum when cultured at a pH of 6.0 at 25 °C for 30 days.     

块菌人工栽培现状及菌根苗培育方法的研究进展

松露(块菌)是名贵的地下真菌,因其独特的香味和口感享誉全球,具有重要的经济价值、食药用价值和生态价值。块菌必须与宿主植物共生形成外生菌根后才能形成子实体(子囊果)。由于块菌具有较高的经济价值而野生块菌资源被严重破坏,以块菌菌根苗制备技术为基础的人工栽培手段受到广泛关注。本文以块菌菌根苗培育为切入点,阐述了块菌资源面临的较大威胁、人工栽培手段的进步和人工种植园的快速发展,通过综述菌根苗培育前对共生组合的选择,菌根苗培育时使用的基质、接种剂、培养方式、与其他生物间的互作,以及菌根苗培育后进行检测的方法等研究进展,为外生菌根食用菌的菌根苗培育技术和人工栽培推广提供参考。     

盐碱地耐盐小麦覆膜栽培高产机理的研究

通过测定不同ＮａＣｌ浓度下不同小麦品种的发芽率（Ｇｒ）、发芽指数（Ｇｉ）和活力指数（Ｖｉ）及盐渍土壤上的产量筛选出耐盐小麦品种德抗９６１。研究了覆膜穴播对土壤温度、土壤含水量、土壤含盐量、产量及其构成因素、旗叶离子含量和旗叶光合作用特性的影响。结果表明,覆膜明显提高土壤温度和土壤含水量,抑制返盐。覆膜减少Ｎａ＾＋在旗叶中的积累,增加旗叶Ｋ＾＋含量。覆膜显著增加旗叶净光合速率、蒸腾速率,气孔导度和细     

Morphological,mycorrhizal and molecular characterization of Finnish truffles belonging to the Tuber anniae species-complex

The truffle species Tuber anniae was originally described from the U.S. Pacific Northwest and is purported to be uncommon. Here, we report for the first time on the fruiting of closely related taxa in Baltic Rim countries. These truffles were found in a forest dominated by Scots pine in eastern Finland. Mycorrhizal analyses confirmed its symbiosis with Pinus sylvestris. Morphological observations of ascomata and mycorrhizae, and phylogenetic analyses confirmed that these white truffles belong within the group of Tuber puberulum (i.e., Puberulum clade). Further, they group in Clades II and III of the T. anniae species-complex. With the inclusion of sequences from GenBank we are able to demonstrate that the previously unnamed environmental clade (Clade II) has been found as ectomycorrhiza in symbiosis with pine, birch, oak, aspen and even orchids in Europe. Thus, the T. anniae species-complex as a whole (and two of the three clades within) exhibit considerable geographic disjuncts: Northwestern North America and the Baltic Rim of Europe. Clade II, which was collected in agricultural soils in Finland and along roadsides in Alaska, may also be adapted for colonization into new habitats. This may help to explain its presence in New Zealand (where Tuber is not native), which most likely resulted from human-mediated dispersal of these fungi through forestry or the nursery trade. Based on our results, we hypothesize that management practices such as organic and lime amendments, along with aeration, are beneficial to the fruiting of T. anniae. Further research is needed to determine the edibility of these species and whether commercial markets can be developed.     

Interaction of leachable model compounds and their impact on Chinese hamster ovary cell cultivation

The presence of leachables in biopharmaceutical processes using single-use technologies (SUT) is well known. For the detection and quantification of the latter, extractable studies of SUT are very common nowadays. Although a mixture of compounds is regularly found in extractable studies, research has only been carried out regarding the effect of individual compounds on cell culture and the cumulative effect of a mix of leachables has not been investigated yet. In this study, a set of leachable model compounds (LMCs) was chosen and the effect of the LMCs on a Chinese hamster ovary DG44 cell line producing an IgG antibody was investigated concerning cell growth, cell cycle distribution and productivity. It was shown that even if worst-case concentrations were used, the LMCs solely impact cell growth. Additionally, interaction studies revealed that the inhibiting effect of the mix is lower than the expected cumulative effect. A strong antagonism between the antioxidant butylated hydroxytoluene and the plasticizer Tris(2-ethylhexyl)trimellitate was found using an isobologram analysis.     

羊肚菌的多样性、演化历史及栽培研究进展

羊肚菌属Morchella真菌是一类珍稀食用和药用真菌,具有重要的经济和科研价值。从羊肚菌属的分类研究、物种多样性、物种分布与生态多样性、演化历史、人工栽培等方面,对新近的研究成果进行了综述,总结了世界羊肚菌研究中取得的主要成绩,指出了仍然存在的主要问题及解决这些问题的相应思路和对策。