金针菇子实体富硒栽培特性及HPLC-ICP-MS法对硒的分布研究

以金针菇子实体为富硒载体,比较研究其富硒及生长特性,同时采用高灵敏度分离和超痕量元素分析技术（HPLC-ICP-MS）分离检测其含硒生物大分子化合物,确定其分子量及硒含量。结果表明：金针菇子实体对硒的富集能力与培养基中硒浓度有关。样品中的硒浓度与培养基中硒浓度呈正相关性,同一培养基子实体中硒的分布为：菌冠＞菌柄＞菌根;培养基中硒浓度小于30mg/kg时对金针菇的生长有促进作用,富硒系数在2.8－9.9之间;培养基中硒浓度在40－200mg/kg范围时,对金针菇的生长产生抑制作用;高于200mg/kg时,不适于富硒金针菇的培养。对富硒金针菇子实体以Tris-HCl浸提,采用体积排阻色谱法（SEC-HPLC）根据蛋白分子量标样检测浸提液中可溶性生物大分子化合物的分子量分布在25kDa以下;同时与电感耦合等离子体质谱联机（SEC-HPLC-ICP-MS）将不同分子量的含硒化合物中的硒进行高温电离检测各形态硒化合物中硒的含量在68.74－2,986.00μg/kg之间,占浸提液中硒含量的71.87%;其中硒蛋白含硒量占4.88%。因此,以金针菇为载体进行硒的生物有机化不仅成本低,含硒量高,而且硒的主要存在形态安全无毒、人体利用率高。     

钙对金针菇生长发育的影响

研究了CaCO3粉末对金针菇(Flammulina velutipes)生长发育的影响及子实体的富钙作用。结果表明,钙能显著刺激金针菇菌丝体和子实体的生长,缩短出菇时间,提高产量和子实体含钙量。这些生理生化变化与钙浓度密切相关,低质量分数(0~5%)和高质量分数(5.5~10%)的CaCO3粉末造成了相似的变化。初步分析认为,这可能与金针菇细胞内存在两种不同的钙信号转导途径有关。     

金针菇栽培基质替代原料初步筛选研究

针对近年金针菇栽培主要原料棉籽壳价格大幅度上涨,导致栽培成本不断增加、栽培经济效益不断下滑的现实问题,在以棉籽壳为主要栽培料配方（CK1）的基础上,用新型基质花生壳、木屑、玉米芯、油菜杆、黄豆杆、猕猴桃枝、桑枝屑、高粱壳等资源部分替代棉籽壳,设计了8个金针菇栽培料配方,并以常用配方CK2为辅助对照,进行金针菇（川金3号）栽培试验。结果表明,配方6（花生壳）的产量虽未超过对照CK2,但其产量显著高于对照CK1（高33.11%）,转化率达到对照CK2的水平,其栽培效益超过对照CK1（高629.03%）和CK2（高15.90%）,且子实体品质好,所以在成都平原冬末早春季出菇时,用30%的花生壳替代棉籽壳栽培金针菇,可以显著降低材料成本,增加栽培效益。     

中国五种重要食用菌学名新注

基于最新的分子系统发育分析研究、《国际藻类、菌物和植物命名法规》和《汉语学名法规》,对毛木耳、玉木耳、金针菇、阿魏侧耳和白灵侧耳等5种重要食用菌的学名进行了解析和介绍,建议使用下述规范名称：毛木耳Auricularia cornea Ehrenb.,玉木耳Auricularia cornea Ehrenb.,金针菇Flammulina filiformis (Z.W. Ge et al.) P.M. Wang, Y.C. Dai, E. Horak & Zhu L. Yang,阿魏侧耳（阿魏菇）Pleurotus eryngii var. ferulae (Lanzi) Sacc.,白灵侧耳（白灵菇）Pleurotus tuoliensis (C.J. Mou) M.R. Zhao & Jin X. Zhang。虽然拉丁名称发生了很大变化,但为了保持汉语学名的稳定性,上述5种食用菌的汉名不变。     

Expression and purification of a recombinant Fip-fve protein from Flammulina velutipes in baculovirus-infected insect cells

Aims: To develop an efficient and facile expression system supply of high purity and stable activity of rFip-fve for oral administration, medicinal study and applications. Methods and Results: A recombinant virus that contained the chimera gene, encoding a bombyxin signal peptide sequence fused to a Fip-fve-6His sequence, was constructed. The rFip-fve was purified from the supernatant of the infected Sf21 cells using a nickel-chelated affinity column, and was verified by Western blot and MALDI-MS (matrix-assisted laser desorption ionization mass spectrometry) analyses. Results showed that a glycosylated mature rFip-fve was produced and secreted into the infected cell supernatant. The immunomodulatory activity of rFip-fve was evaluated by measuring the amount of interleukin-2 released from murine splenocytes. Conclusions: A reliable scheme to express and purify active rFip-fve in a baculovirus/insect cell system for medicinal applications and genetic study is a feasible means of solving potential problems related to the production and activity of rFip-fve protein. Significance and Impact of the Study: The rFip-fve expressed in insect cells was processed and modified in a manner more similar to that of its native counterpart than that in bacterial cells. Therefore, the potential applications of rFip-fve that is generated in Sf21 cells can be more effectively evaluated that produced in Escherichia coli.     

Optimization of submerged culture conditions for the production of angiotensin converting enzyme inhibitor from <Emphasis Type="Italic">Flammulina velutipes</Emphasis>

The angiotensin-converting enzyme (ACE) inhibitory effect was tested in the culture broth from submerged mycelial cultures of 20 basidiomycetes. The ACE inhibitory effect of culture broth from Flammulina velutipes strain 414 was the highest (52.8%), followed by Lentinus edodes strains 2 (44.4%) and 16 (41.3%). Nutritional requirements for the production of ACE inhibitory substance from F. velutipes were studied. Sucrose, ammonium acetate, and glutamic acid were chosen for the maximum production of ACE inhibitory substance. The optimal medium composition was (g/l): sucrose 20, ammonium acetate 5, glutamic acid 2, KH₂PO₄ 3, MgSO₄·7H₂O 0.8, and yeast extract 0.5. Under optimal culture conditions, the ACE inhibitory effect was more than 80%. Received 04 May 2002/ Accepted in revised form 11 June 2002     

金针菇保鲜贮藏研究

金针菇经过PE袋包装、减压、充气、低温等处理进行保鲜贮藏,经测定结果表明,各处理组的水份丧失、干物质消耗、呼吸强度和粗纤维的含量均比对照低,可溶性固形物、可溶性糖、蛋白质和维生素C的含量虽比贮前略低,但均比对照高。从外观观察,各处理均有较好的保鲜作用。     

荧光标记金针菇原生质体融合

以金针菇（Flmmulina velutipes）为材料,经异硫氨酸荧光素（FIC）标记的金针菇单核W19菌株的原生质体与未经标记的单核Y7菌株的原生质体在聚乙二醇（PEG）的诱导下进行融合,利用显微操纵仪直接挑取一个带有荧光而另一个不具荧光的原生质体对,培养后,根据融合形成的汉核菌丝具有“锁状连合”这一形态学特征选择、鉴定融合菌株,酯酶同工酶电泳分析结果表明,融合菌株具双亲互补酶带。经测定,多数融合菌株菌丝生长速度和生物量较亲株高,且出菇正常。     

聚类分析在金针菇杂交育种中的应用

对12个金针菇菌株两种生化标记（酯酶和可溶性蛋白）进行了筛选研究．采用多元统计方法,进行聚类分析。结果表明,供试菌株可分为四个类群。按照亲本的遗传相似系数,对各杂交组合的杂种优势进行了检测,实际结果表明,不同杂交组合的产量与预测结果基本一致。群内组合为低产组合,群间组合杂种优势较强。     

大型真菌冬菇103工业发酵产物及生物活性的研究

本研究表明大型真菌冬菇103 Flammulina velutipes(Curt.: Fr.)Sing.-103工业发酵性能良好,具备生产周期短、产量高、产物种类多等特点,发酵时间72—96小时,在深层发酵培养基中,每升可形成9—15g干菌丝体;菌丝体的稠密度为每升含有40—42g干物质。并能产生多种功能的生理活性物质。从菌丝体和发酵液中分析出16种氨基酸、14种微量元素、4种维生素、其中包括脂溶性维生素E醋酸酯、αE、K_1和水溶性维生素B_2此外还提取出具有抗菌活性和免疫功能的粗多糖和菌丝干粉(包括菌丝的提取液),它们在抗菌活性方面对大肠杆菌(Eschericha coli);八叠球菌(Sarcina later);枯草杆菌(Bacillus subtilis);油桐炭疽菌(Colletotrichum Sp.c-837-1)显示出不同程度的抗菌活性。在促进免疫功能方面,可促进小鼠吞噬鸡红血球功能和促进体液免疫小鼠溶血空斑的形成,菌丝干粉经小鼠急性毒性LD_(50)测定表明,小鼠以5g/kg剂量为最高档剂,给药后未见死亡,故103菌丝干粉对小鼠的最大安全量为5g/kg以上。