Solid Culture of Lactarius deliciosus

The research explored the effects of carbon, nitrogen and inorganic salt nutritional factors on Lactarius deliciosus hyphae as per solid culture. The optimal recipe of culture medium was selected by observing hypha density and computing the growth index, which is the most suitable for hypha growth. The results indicated that glucose and peptone were optimal carbon and nitrogen for Lactarius deliciosus;MgSO4, CuSO4 and VB2 promoted hypha growth; FeSO4 and CoCl2 had inhibition effects. The orthogonal test concluded the optimal recipe contained glucose of 20 g/L, peptone of 1 g/L, KH2PO4 of 0.3 g/L, MgSO4 of 0.3 g/L, VB2 of 2 mg/L, and agar of 20 g/L, with original pH value adopted. After optimization, growth index of hyphae cultured at 28 ℃ was 4 055.9.

松乳菇Lactarius deliciosus与红汁乳菇L. hatsudake子实体内可培养细菌多样性的研究

分离松乳菇Lactarius deliciosus与红汁乳菇L.hatsudake子实体内可培养的细菌,进行分类鉴定,为后期细菌与乳菇的相互作用研究奠定基础。利用传统平板法分离培养乳菇子实体内的细菌,进行16S r DNA片段的扩增和测序及系统发育树的构建和物种多样性分析。从松乳菇中获得66株细菌,红汁乳菇中获得48株细菌。松乳菇子实体内细菌隶属于3个门6个属10个种,其中变形菌门的γ-变型菌纲为优势类群,占细菌总数的77.28%,β-变形菌纲的Pandoraea和拟杆菌门的金黄杆菌属Chryseobacterium分别占细菌总数的12.12%和10.6%。红汁乳菇子实体内共分离、鉴定出细菌3个门7个属7个种,其中变形菌门中的γ-变型菌纲为优势类群,占细菌总数的77.09%,厚壁菌门的芽胞杆菌属Bacillus和拟杆菌门的金黄杆菌属Chryseobacterium分别占细菌总数的14.58%和8.33%。松乳菇与红汁乳菇子实体内存在一定种类和数量的细菌,其中荧光假单胞菌Psedomonas fluorescens和美洲爱文氏菌Ewingella americana为优势细菌。

New Azulene-Type Sesquiterpenoids from the Fruiting Bodies of Lactarius deliciosus

In the^(1)H NMR-guided fractionation of extracts from the edible mushroom Lactarius deliciosus,two new azulene-type sesquiterpenoids,7-isopropenyl-4-methyl-azulene-1-carboxylic acid(1)and 15-hydroxy-3,6-dihydrolactarazulene(2),together with seven known compounds were characterized.Their structures were determined on basis of spectroscopic evidence,as well as by comparing with literature data.Amongst the known metabolites,the^(13)C NMR assignment of 15-hydroxy-6,7-dihydrolactarazulene(3)is reported here for the first time.Moreover,7-acetyl-4-methylazulene-1-carbaldehyde(5)displayed a moderate antibacterial activity against Staphylococcus aureus.

红汁乳菇(Lactarius hatsutake)的成功栽培—分子方法评估

自1998年始，我们已在湖南、江苏等地建立了65ha以上红汁乳菇马尾松种植园。这些林分一般三年后开始产菇，年产量可达675kg／ha。为了确认所产红汁乳菇子实体来源于接种的菌根苗而非自然侵入的孢子，我们进行了子实体、菌根苗菌根的分子研究。首次比较了试验地所产子实体与红汁乳菇-马尾松菌根苗培养钵内的菌根的ITS序列。同时，用RAPD法分析了取自于试验地内的10个红汁乳菇子实体。结果表明，10个子实体中的9个与起始菌种CX1相比具有75％～96％的相似性。其余一个及作为外群取自于340km外的广西融水的子实体与起始菌种分别具有61％和37％的相似性。我们认为，上述9个子实体与起始菌种的相关性比外群和第10个子实体更为紧密。推测认为起始菌株及其子代菌丝形成了上述9个子实体，而其余一个子实体可能来源于周边森林侵入的孢子。

乳香鱼LACTARIUS LACTARIUS(BLOCH et SCHNEIDERP)骨骼的研究

乳香鱼鼻骨为一小薄片.犁骨有钩状小齿.额骨背外侧有较高的弧形薄骨嵴,形成椭圆形的盘状凸起.缺眶上骨,眶下骨6块.上枕嵴特别发达.上、下颌前端各具2枚较大的犬齿.有辅上颌骨.角舌骨上有裂孔.具上后颞骨.

Extraction and Antioxidant Activity Analysis of Crude Polysaccharides from Wild Lactarius volemus Fr. in Yunnan Province

[ Objective ] This study aimed to investigate the optimal extraction process of crude polysaccharides from wild Lactarius volemus Fr. in Yunnan Province and preliminarily analyzed its antioxidant activity in vitro. [ Method ] With water extraction and alcohol precipitation method, the optimal conditions for extracting crude polysaccharides from wild L. volemus Ft. were screened by single-factor and orthogonal experiments. The antioxidant activity of the extracted crude polysac- charides was determined with DPPH assay. [ Result ] The optimal conditions for pigment removal with activated carbon were： activated carbon amount of 20 g/L, water bath time of 40 min, water bath temperature of40 ℃ ; the optimal conditions for extracting crude polysaccharides from wild L. vo/emus Fr. with hot water ex- traction method were： hot water extraction time of 3 h, solid-liquid ratio of 1： 45, extraction frequency of twice. Under the optimized extraction conditions, the yield of crude polysaccharides was 21.33 mg/g. In addition, the antioxidant activity of 0. 665 mg/ml crude polysaccharides was 52.46% ; the amount of crude polysac- eharides was proportional to the antioxidant activity. [ Conclusion] Hot water extraction method can be used as a high-efficiency extraction technology of crude pol- ysaecharides from wild L. volemus Fr. with simple operation and low costs. Crude polysaccharides extracted from L. volemus Ft. exhibited certain antioxidant activi- ty in vitro.

Lactariolide,a New 14-Membered-Ring Compound from Lactarius Subvellereus

Lactariolide (I), a new 14-membered-ring lactone was isolated from the fruit bodies of Lactarius subvellereus (Russulaceae), and its structure was elucidated as (R)-13-hydroxy-trans-9, 11-octadecadienoic acid lactone by means of physical and spectral data.

Lactarius imbricatus名称的合格化(英文)

In our recent paper (Zhou & Wen 2007), a new species, Lactarius imbricatus M.X. Zhou & H.A. Wen was published with Latin and English descriptions, illustrations, and type designation. The name was not, however, validly published because the specimen designated as holotype was stated to be kept in two herbaria. This

Biodegradation and detoxification of the triphenylmethane dye coomassie brilliant blue by the extracellular enzymes from mycelia of Lactarius deliciosus

Fungi play an important role in dying wastewater treatment.In this work,the mycelia of Lactarius deliciosus exhibited an excellent capacity in decolorizing coomassie brilliant blue(CBB).The results demonstrated that the mycelia could treat CBB with high concentrations over a broad range of pH and temperature.The decolorization rate of 99.19%and the removal rate of 16.31 mg·L^(‒1)·h were realized.The mycelia could be recycled from decolorizing process for 19 times,indicating a good re-usability.It verified that the lignin peroxidase(121.65 U·L^(‒1))and manganese peroxidase(36.77 U·L^(‒1))were involved in the degradation and decolorization process of CBB.Toxicity assessments indicated the seed germination rate was up to 82.22%while inhibition to Escherichia coli decreased dramatically and no significant effect on Caenorhabditis elegans growth was found.The removal of CBB was a synergistic process accomplished by adsorption and biodegradation.The mycelia could be used for eco-friendly CBB treatment.

Immobilization and characterization of the Lactarius deliciosus mycelia-embedded polylysine-alginate beads and their decolorization performance

Liquid fermented fungal mycelia with decolorization capability have potential applications in scale-up. In this work, the Lactarius deliciosus mycelia were immobilized on ε-polylysine-alginate beads, and the decolorization effects of ε-polylysine-alginate beads were demonstrated along with Coomassie brilliant blue G-250 as a model dye. Morphology observation confirmed the beads had an exterior film and interior capsule with honeycomb microstructures suitable for mycelia growth. It was manifested that the maximum decolorization efficiency for mycelia was 98.5% at a removal rate of 0.68 mg·L^(-1)·h after 3 days. In comparison, the decolorization efficiency of the immobilized mycelia reached the maximum value of 97.3% at a removal rate of 6.1 mg·L^(-1)·h after 8 h. The enzyme activities of lignin peroxidase and laccase tested in the immobilized mycelia were significantly higher than in that of the free ones, such as the lignin peroxidase had the highest enzyme activity of 77.6 ± 7.4 U·L^(-1) in the former, while of 27.4 ± 8.7 U·L^(-1) in the latter. The immobilization of L. deliciosus mycelia could improve the decolorization of Coomassie brilliant blue G-250 efficiently. The prepared ε-polylysine-alginate beads embedded with L. deliciosus mycelia have very good reusability and a great potential in decolorizing analog dyes.

利用代谢组学分析红汁乳菇-马尾松菌根苗根系分泌物

为探究红汁乳菇(Lactarius hatsudake)与宿主共生后菌根苗根系分泌物中促进红汁乳菇生长及菌根合成的潜在物质,利用非靶向代谢组(LC-MS/MS)分析红汁乳菇-马尾松菌根苗与马尾松非菌根苗的根系分泌物。结果表明:与马尾松非菌根苗相比,红汁乳菇-马尾松菌根苗根系分泌物在正、负离子模式下的差异代谢物共有352种,其中315种显著上调,37种显著下调;差异代谢物主要分布在代谢途径、抗生素生物合成、赖氨酸降解、次生代谢产物生物合成等28条通路中;差异代谢物2-异丙基苹果酸、磷酸、UDP-N-乙酰葡糖胺、茉莉酸、甲羟戊酸等参与多条代谢通路,可能在红汁乳菇-马尾松菌根形成中起重要作用。选取的9种差异显著代谢物中,1 mg·L^(-1)的D-天冬氨酸、DL-精氨酸、脱硫生物素、脱落酸对红汁乳菇JH5菌丝生长具有促进作用;1 mg·L^(-1)脱硫生物素、烟酸、脱落酸对红汁乳菇-马尾松菌根形成有促进作用。本研究可为红汁乳菇-马尾松菌根苗的高效培育提供参考。

基于菌株生长和菌根苗品质的红汁乳菇菌株筛选

【目的】筛选用于菌根苗接种的高质量红汁乳菇菌株,为提高红汁乳菇产量提供支持。【方法】以10个红汁乳菇菌株(JH1、JH2、JH4、JH5、JH7、JH8、JH10、JH11、JH12、JH13)为研究对象,将菌株分别在PDA和BAF培养基上培养后,测定菌株生长势指标(菌丝生长速度、菌丝生物量);将红汁乳菇液体菌种接种于马尾松幼苗根部合成菌根苗,以不接种为对照(CK),培养210 d后对菌根合成能力(菌根数量、菌根侵染率、菌根化率)和宿主植物生长量(苗高、主根长、地径、干质量)进行测定。对上述9个参数进行相关性分析,并用隶属函数法对菌株性状进行综合评价。【结果】①在PDA固体培养基上,以菌株JH1菌丝生长速度最快,达到3.17 mm/d;在BAF液体培养基上,以菌株JH5的菌丝生物量最大,为3.28 g/L。②所有菌株均能与马尾松根系形成外生菌根,但菌根形态并不相同。接种红汁乳菇菌株JH5的马尾松苗的菌根数量为45.65个/株,菌根化率为90.67%,菌根侵染率为79.60%,显著高于其他9个菌株,表现出较强的合成菌根苗能力。③接种不同红汁乳菇菌株的马尾松苗,在苗高、主根长、地径和干质量等指标上表现各异。接种菌株JH4的马尾松苗高、主根长和干质量表现最优,较对照分别提高24.45%,16.72%和53.33%;接种菌株JH5的马尾松苗地径生长量表现最优,较对照提高5.19%。④相关性分析表明,菌丝生物量与菌根化率和菌根苗干质量呈极显著正相关(P<0.01),菌根数量与菌根化率和菌根苗地径呈极显著正相关(P<0.01)。⑤基于隶属函数法的综合评价结果表明,红汁乳菇JH5为适合培育菌根苗的最佳菌株,JH4和JH7为次优菌株。【结论】筛选出1个优良红汁乳菇菌株JH5,该菌株的生长势和菌根苗品质整体表现较好。

松乳菇菌丝生长促生细菌的筛选与鉴定

[目的]从乳菇(Lactarius sp.)菌柄基部土壤分离并筛选出对松乳菇菌丝生长有促进作用的细菌(MHB),以提高松乳菇菌根苗人工合成效率。[方法]从乳菇菌柄基部土壤分离得到细菌,将纯化后的细菌液体培养离心后获得胞外代谢产物,以添加不同菌株和不同浓度胞外代谢产物的PDA培养基接种松乳菇菌丝块,并以未添加胞外代谢产物PDA平板接种松乳菇菌丝为对照,比较25℃培养30 d后松乳菇菌丝生长的直径大小和菌丝密度;另一方面,基于形态特征和16S rDNA序列鉴定对松乳菇菌丝生长有促进作用的细菌。[结果]共筛选出3株可促进松乳菇菌丝生长的芽胞杆菌属细菌,即菌株L1、L2和L3。当3株菌株的胞外代谢产物添加量为2.5%时,均能显著促进松乳菇菌丝生长,其菌落直径分别增加33.6%、55.6%和75.4%。当3株菌株胞外代谢产物以不同组合等比例混合后,在混合物添加量为5.0%时,L1-L3,L1-L2-L3两个组合显著促进松乳菇菌丝生长,松乳菇菌丝直径分别增加36.9%和42.5%。L3菌株的胞外代谢产物在添加量为2.5%、5.0%和7.5%时均能增加松乳菇菌丝密度,且添加量为7.5%时增加松乳菇菌丝密度效果最佳;L1-L3和L1-L2-L3两个组合也能增加松乳菇菌丝密度。通过对3株菌株形态特征鉴定和16S rDNA序列测定,L1菌株为高山芽孢杆菌(Bacillus altitudinis),L2菌株为蜡样芽胞杆菌(B.cereus),L3菌株为环状芽孢杆菌(B.zanthoxyli)。[结论]筛选出3株可促进松乳菇菌丝生长的芽孢杆菌,其中B.zanthoxyli促进松乳菇菌丝生长效果最好。

雁鹅菌菌丝培养条件优化研究

为优化雁鹅菌菌丝的培养条件,以菌丝生长速度为评价指标,通过单因素试验分别确定雁鹅菌菌丝生长培养基的最佳碳源、氮源以及无机盐,并通过比较不同的营养元素浓度对雁鹅菌菌丝生长速度的影响,确定各营养元素的最佳添加区间,后续采用正交试验法优化雁鹅菌菌丝生长培养基中各营养元素的组合。结果表明,最佳碳源为可溶性淀粉,最佳氮源为胰蛋白胨,最佳无机盐为氯化钙。雁鹅菌菌丝生长培养基的最优组合为可溶性淀粉浓度5 g·L^(-1),胰蛋白胨浓度3 g·L^(-1),培养基中氯化钙浓度1.00 g·L^(-1)。

橙黄乳菇-马尾松菌根广靶代谢组学比较与分析

【目的】分析比较橙黄乳菇-马尾松菌根合成前后根系代谢物差异,为探究调控橙黄乳菇-马尾松共生的潜在根系代谢物提供参考。【方法】在纯培养条件下进行橙黄乳菇-马尾松菌根合成,基于液质联用技术测定马尾松及橙黄乳菇-马尾松菌根根系代谢物成分,利用KEGG和HMDB数据库进行代谢物的定量与注释,采用主成分分析、正交偏最小二乘-判别分析等多元统计方法进行代谢组数据的比较,并筛选差异代谢物。【结果】共检测出代谢物627种,其中差异代谢物95种,上调39种,下调56种,主要集中在黄酮类化合物、萜类化合物、氨基酸及其衍生物、酚类化合物、生物碱类化合物、有机酸及其衍生物、核苷酸及其衍生物、香豆素和木脂素、植物激素等。KEGG代谢通路分析显示次生代谢产物的生物合成、苯丙烷类化合物的生物合成、类黄酮生物合成、神经活性配体-受体相互作用、铁载体基非核糖体肽的生物合成、苯甲酸盐降解途径、植物激素的生物合成、莽草酸途径生物碱的生物合成和氨基苯甲酸降解等9条差异代谢途径在橙黄乳菇-马尾松菌根形成后差异代谢物显著富集,可能对橙黄乳菇-马尾松菌根形成具有重要调节作用。【结论】利用广泛靶向代谢组学技术可在整体水平上揭示橙黄乳菇-马尾松菌根形成前后的代谢组学差异,研究结果可为明晰橙黄乳菇-马尾松共生机制及橙黄乳菇-马尾松菌根合成提供科学依据。

橙黄乳菇对马尾松和华山松幼苗生长及根系代谢物的调控

【目的】探究接种橙黄乳菇对马尾松和华山松幼苗生长、根系发育及代谢物的影响,为橙黄乳菇与2种松树菌根化苗木培育及共生栽培提供参考。【方法】以马尾松和华山松幼苗为研究对象,基于大棚盆栽试验,测定分析接种和未接种橙黄乳菇的2种松树幼苗菌根侵染情况、地上和地下生长指标、根系构型性状(根长、表面积、根体积、根尖数、分叉数)、形态性状(比根长、组织密度、比表面积)及根系代谢物差异。【结果】橙黄乳菇与2种松树幼苗均形成明显的菌根形态,并有较高的菌根侵染率和菌根依赖性。接种橙黄乳菇对2种松树幼苗生长具有一定促进作用,但存在差异,接种后马尾松幼苗株高和地上部分干质量显著增加,华山松幼苗株高和地下部分干质量显著增加,2种松树幼苗总根长、根表面积、根体积、根尖数和分叉数均明显提高,根系活力和阳离子交换量降低。接种橙黄乳菇明显影响2种松树幼苗根系代谢物含量,接种后马尾松幼苗根系总酚和总黄酮质量分数降低,总三萜、总生物碱和总甾醇质量分数升高,华山松幼苗根系代谢物变化趋势与马尾松相反。【结论】接种橙黄乳菇可改善2种松树幼苗根系形态、提高生物量、影响根系主要代谢物含量,在松树菌根化苗木培育即外生菌根真菌共生栽培方面具有可行性。

4种乳菇属真菌的形态学与rDNA-ITS鉴定

为明确内蒙古赤峰地区常见有毒乳菇种类,准确、快速地识别有毒乳菇,将采集的4个乳菇属Lactarius真菌标本采用传统形态学分类方法进行鉴定,同时提取子实体DNA,使用真菌通用引物ITS1/ITS4进行PCR扩增,用Blast对扩增产物纯化测序所得的序列进行比对,采用MEGA5.1软件构建系统发育树确定属和种。结果表明,2种方法鉴定结果一致,标本CFSZ10167为毛头乳菇[Lactarius torminosus(Schaeff.)Pers.],标本CFSZ10216为绒边乳菇[Lactarius pubescens(Fr.ex Kronbh.)Fr.],标本CFSZ13021为灰褐乳菇[Lactarius pyrogalus(Bull.)Fr.],3种均为胃肠炎型毒菌,CFSZ10044为皱乳菇(Lactarius ruginosus Romagn),为中国新记录种,主要特点是菌盖棕褐色、浅褐色,有绒质感,盖缘呈齿状,孢子球形具小刺,食毒不明。对4个种的形态学特征进行了详细描述,并附有生境图片和显微图像,研究结果为乳菇属物种鉴定和资源分布提供了重要补充,并为常见毒菌识别提供了理论依据。

鲜艳乳菇母种培养基的改良与优化

以恩施土家族苗族自治州农业科学院分离保存的鲜艳乳菇(Lactarius hysginus)菌株LWL-1为试材,从维生素、生物因子、氮源种类及其添加量3个方面对报道的培养基配方进行改良与优化,以期使菌丝快速健壮生长。试验结果表明,氮源相同时,4种维生素和生物因子的改变不会使菌丝生长速率产生显著差异。改变氮源添加量,菌丝生长速率随氮源添加量的增加而逐渐降低;氮源添加量相同时,以大豆蛋白胨为氮源的菌丝生长速率快于蛋白胨,且菌丝也较为健壮。因此,选择以大豆蛋白胨添加量为2 g/L的培养基较为适合。

松乳菇营养成分分析及松乳菇多糖的提取分离

以野生鲜松乳菇子实体、发酵所得菌丝体为材料,对其粗蛋白、氨基酸、脂类总量、多糖、灰分及微量元素的含量进行测定,并以正交法确定了松乳菇多糖提取的最佳工艺条件,以层析法定性分析了多糖的组成。

松乳菇组织分离菌株的rDNA ITS序列分子鉴定

对松乳菇子实体和组织分离菌株rDNA ITS区域进行测序分析,通过和GenBank中已知的乳菇属的种进行序列比对,构建系统发育树,通过树图分析,鉴定出分离菌株就是松乳菇的纯菌种。