台湾虫草子实体人工培养条件的初步研究

目的研究人工培养条件对台湾虫草子实体形成的影响。方法通过黄山被毛孢的液体摇瓶培养,确定摇瓶种子的最佳培养时间,并研究了在人工诱导下台湾虫草子实体的形成条件,即子实体形成与营养液pH、营养液量、培养温度及光照时间之间的关系。结果黄山被毛孢液体摇瓶最佳培养时间为12 d,人工固体培养子实体所需营养液最佳pH为6.0,营养液量为40 mL,培养温度为25℃,菌丝培养阶段为完全黑暗培养,原基出现后给予光照,可获得较多的子实体。结论人工培养条件对台湾虫草菌丝体生长、原基形成、子实体产量具有显著影响。     

斜生褐孔菌子实体的诱导和发生条件

研究了斜生褐孔菌Phaeoporus obliquus在人工培养过程中适宜子实体形成、生长、发育的条件.主要包括子实体发生与培养基种类、培养方式、光照以及温度的关系.结果表明,甘露醇酵母膏培养基是适合斜生褐孔菌子实体发生的最佳培养基,在该培养基上斜生褐孔菌子实体的发生率可以达到100%.固体培养基适合子实体的发生和生长,第44天开始形成子实体,平均每瓶子实体的干重可达0.372g,子实体生物量对培养基中营养成分的转化率为5.81%.20℃是子实体发生的适宜温度;光照对子实体发生影响差异不明显.     

蛹虫草对锌的耐性与富集特征

在培养基内添加不同量的锌,研究其对蛹虫草子实体的形成、子实体和菌丝体生物量、子实体多糖含量和葡萄糖含量的影响,以及蛹虫草子实体和菌丝体对锌的富集能力。结果表明锌对上述各项都有影响。液体培养条件下,锌浓度在453906mg/L范围内可以促进菌丝体生长,锌浓度超过4077mg/L时,菌丝生长受到抑制。培养基锌的浓度在4077mg/L以下时,蛹虫草菌丝体锌的富集量随着液体培养基锌浓度的提高而提高。固体培养条件下,锌含量在226453mg/kg范围内可以促进蛹虫草子实体生长,并且在此含量范围内,蛹虫草子实体中葡萄糖含量较高。培养基锌含量在680906mg/kg时,子实体多糖含量较高。培养基锌含量在2038mg/kg以下时,蛹虫草子实体中锌的富集量随着培养基锌含量的提高而提高,在培养基锌含量为2038mg/kg时,子实体中锌的含量达到28570mg/kg(干重)。     

蜜环菌子实体的诱导和发生条件

研究蜜环菌子实体在人工诱导下的发生规律,主要包括子实体发生与培养基种类、培养基量、培养时间等的关系。麦芽汁培养基是适合蜜环菌子实体发生的培养基,在该培养基上蜜环菌子实体发生率可以达到100%,子实体最短可以在45天发生。子实体发生与培养基瓶装量有关,每瓶中培养基总固形物(琼脂除外)达到6.75g才足以发生子实体。子实体发生时间与25℃的培养时间有关系,培养时间不足使子实体发生时间延迟。菌体总生物量对培养基中总营养成分的转化率可以达到65.90%,子实体生物量对培养基中总营养成分的转化率为1.52%~9.11%。     

裸脚菇0612-9液体菌种优化及其对后续发酵产活性物质的影响

裸脚菇0612-9次级代谢产物具有强烈抗青绿霉活性,可作为微生物源防腐剂用于柑橘保藏,但是其发酵周期长,产出能耗大效率低。用摇瓶对裸脚菇0612-9的液体菌种培养基、培养条件进行优化并对优化后液体菌种接种种龄、接种量进行探索,最后用5L发酵罐进行放大发酵验证。取样计数测定菌丝球数量、过滤称重测定菌丝干重、HPLC监测活性物质Ⅱ的积累、牛津杯法评价抗青绿霉活性。经研究最佳碳源为玉米粉和麦芽糖,最佳氮源为蛋白胨,最佳液体菌种培养基组成为：玉米粉30g/L、麦芽糖10g/L、蛋白胨15g/L、KH₂PO₄ 2g/L、MgSO₄·7H₂O 1g/L;最佳培养条件：起始pH 5、接种3×Ф7mm菌块、装液量100mL/250mL三角瓶、温度28℃、转速160r/min;优化前菌丝球数46个/10mL,菌丝干重0.28g/100mL,优化后菌球数达985个/10mL,菌丝干重达0.69g/100mL,分别为优化前的21.4倍、2.43倍;后续发酵使用种龄9d的液体菌种、接种量7.5%。优化后液体菌种在发酵罐中后续发酵周期从10d缩短至5d,缩短50%,产量比优化前提高8.28%。     

低温振荡处理对香菇子实体的萌发,类脂化合物的构型和营养成分转移的影响

食用菌子实体的萌发常被许多环境因素所控制,诸如光、相对湿度、通风和温度等.就香菇而言,已报导了长满菌丝的培养料经进一步浸没在冷水中处理能够增加子实体的产量(Manashere,1977;Matsu-moto和Kitamoto,1987),并观察了它可能影响几丁质酶活力(Tok(?)mo(?),1988),但没有详细研究低温对子实体萌发的影响的生理学基础.本文报告了低温振荡处理生长在固体培养料和合成琼脂培养基中(体外)菌体对子实体的萌发以及振荡培养香菇对菌丝类脂化合物的构型和营养成分转移的影响.长满香菇菌丝的半固体培养基和固体培养基分别在5℃冷水中浸没24hr,结果发现子实体萌发得     

培养基及培养条件对冬虫夏草菌固体发酵产分生孢子的影响

为获得冬虫夏草菌固体发酵产分生孢子的最优工艺,以野生分离的冬虫夏草菌为材料,对其固体发酵产分生孢子的培养基及培养条件进行了研究。试验结果表明：泥炭土为最佳基础培养基,该培养基中冬虫夏草菌气生菌丝生长一般,但产分生孢子最多,可达4.2×10³个/g;泥炭土培养基中添加0.1‰ IAA（吲哚乙酸）、0.1‰ IBA（吲哚丁酸）和0.1‰ NAA（萘乙酸）能促进冬虫夏草菌气生菌丝的生长和分生孢子的产生,其分生孢子达8.1×10³个/g;该基础培养基中,冬虫夏草菌于18℃培养30d后,在10℃、相对湿度45%、蓝光照射进行诱导,分生孢子可达1.0×10⁴个/g。本研究建立了一种大量获取冬虫夏草菌分生孢子的方法,为冬虫夏草繁育奠定了基础。     

2,4-D对鱼腥藻增殖的效应

本文在实验室批量培养,室外小池大量培养和塑料大棚大面积培养的基础上,研究了植物激素2,4-D刺激鱼腥藻增殖的效应。浓度在0.01—2.00μg/mL范围内都具有刺激鱼腥藻增殖的效果,随着浓度增加,这种效果降低。品质分析结果表明,0.01μg/mL2.4-D可提高蛋白质和叶绿素a的含量;浓度为0.05μg/mL时,二者的含量与对照相差不大;浓度达到0.1μg/mL时,二者的含量降低。本文提出,2,4-D刺激鱼腥藻增殖的应用浓度应在0.05μg/mL以下,以0.01μg/mL效果最佳。     

冬虫夏草菌丝状菌源增殖培养的研究

冬虫夏草菌是珍稀濒危名贵中药材冬虫夏草的无性型菌种,是侵染蝠蛾幼虫的唯一菌种,其侵染后形成名贵中药材冬虫夏草。冬虫夏草菌在不同营养及条件下生长形态不同,主要有丝状体和菌球体两种生长形态。冬虫夏草菌丝状菌体能够侵染蝠蛾幼虫,有可能作为一种新的接种体被广泛应用;但丝状菌体对营养要求苛刻、生长缓慢、菌丝体得率低的特点,阻碍了冬虫夏草菌丝状菌体作为侵染蝠蛾幼虫的接种体的开发及应用,从而阻碍了冬虫夏草人工培殖产业化的进程。为了提高冬虫夏草菌丝状菌体的生物表达量,本文对营养及培养条件进行了优化研究,得出了最佳条件为:葡萄糖质量浓度40 g/L、酵母粉质量浓度65 g/L、培养温度17℃、MgSO_4质量浓度3 g/L、KH_2PO_4质量浓度1.5 g/L、培养时间12 d,按最优培养条件,冬虫夏草菌丝状菌体干重得率15.67 g/L,为下一步新的接种体的制备提供菌源保障。     

蜜环菌对镁的耐性和富集特性

研究了镁对蜜环菌生长的影响, 蜜环菌对于镁的耐性和富集规律, 以及高浓度镁胁迫下蜜环菌的抗氧化酶的变化情况。3?15 g/L的Mg浓度对于蜜环菌菌体的生长有促进作用。Mg浓度19 g/L以上时, 蜜环菌菌体的生长受到抑制。蜜环菌的子实体形成和子实体生物量在Mg的浓度为11 g/L以下时不受影响, 超过11 g/L则子实体不能萌发。皮壳状菌丝和菌索中Mg的含量随培养基中Mg浓度的增大而增大, 培养基Mg浓度达到16 g/L后, 菌丝、菌索中Mg的含量都不再上升。子实体对Mg的富集量比菌丝体小的多, 在培养基Mg浓度在9、10 g/L时, 子实体中Mg的含量与对照组有显著差异。随着培养基中Mg浓度的提高, 菌丝和菌索POD、CAT、SOD活性都有增加, 而且菌丝与菌索之间抗氧化酶活力的差异随着培养基中Mg浓度的提高而增大。     

巴西蘑菇菌体深层发酵培养条件的优化及成分分析*

对巴西蘑菇深层发酵条件进行了详细的研究,确定了巴西蘑菇菌体发酵的最优方案是:7%葡萄糖,1.5%酵母膏,0.3%磷酸氢二钾,0.1%硫酸镁,VB1 10mg/100ml,自然pH,接种量为15%,装液量为100ml/250ml三角瓶,150r/min,25℃恒温培养9天,菌丝干重达到1.8g每100ml发酵液。通过对巴西蘑菇子实体与深层培养菌丝体中蛋白质营养成分及多糖含量进行了分析比较,发现它们均是极好的营养食品之源。     

Somatic embryogenesis and plantlet regeneration from cell suspension cultures of Fagus sylvatica L

Summary Embryogenic cell suspension cultures and somatic embryos of five genotypes of beech, were obtained from aged cultures derived from immature zygotic embryos cultured on solid medium containing both 2, 4-dichlorophenoxyacetic acid and N6-benzyladenine. The origin of somatic embryos was traced from single cells. Embryos remained arrested at the globular stage on liquid media, further development was achieved after plating embryogenic aggregates on Murashige and Skoog's medium with half strength major salts supplemented with glutamine and low levels of growth regulators. Cultures of different genotypes showed significant differences in maturation frequency which was not affected by the hormone treatments assayed. The frequency of conversion of embryos into plantlets was low. This frequency increased after cold storage of embryos for up to 7 months.Abbreviations BA N6-benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - EtOH ethanol - GA₃ giberrellic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - MS Murashige and Skoog (1962) - NAA naphthalene acetic acid - WPM woody plant medium (Lloyd and McCown, 1980) - Z zeatin     

红豆草耐盐愈伤组织的筛选及植株再生

将红豆草种子在含１．２％ＮａＣｌ的ＭＳ培养基上萌发以消除盐敏感的幼苗,把存活的幼苗下胚轴切段在含１ｍｇ／Ｌ２,４－Ｄ、０．５ｍｇ／Ｌ６－ＢＡ及１．２％ＮａＣｌ的ＭＳ培养基上诱导愈伤组织,通过连续筛选得到可耐受１．８％ＮａＣｌ的愈伤组织,在有０．２ｍｇ／Ｌ ＮＡＡ和１ｍｇ／Ｌ ＩＡＡ存在下该愈伤组织分化出芽,待幼,待幼苗长至３ｃｍ左右时转至含２ｍｇ／ＬＮＡＡ和或ＩＢＡ的１／２ＭＳ培养基上生根。对对照     

Somatic embryogenesis from leaf- and petiole-derived callus of Vitis rupestris

Somatic embryogenesis from leaf- and petiole-derived calli of Vitis rupestris was obtained with an efficiency of 3.2% and 4.2% of plated explants, respectively on two combinations of 6-benzyladenine and 2,4-dichlorophenoxyacetic acid (1/0.1 and 1/1 mgl^–1) added to MS medium. Embryogenic callus, embryo subcultures and somatic embryogenesis from somatic embryos were obtained either in the presence of 1 mgl^–1 indole-3-acetic acid or 0.1 mgl^–1 indole-3-butyric acid added to MS or NN media. Within a 4-month culture, embryo germination occurred at a frequency of 13% of explanted embryos when chilling at 4°C was provided for two weeks and a combination of 6-benzyladenine (1 mgl^–1) with indole-3-butyric acid (0.1 mgl^–1) was added to NN medium supplemented with casein hydrolysate (250 mgl^–1). A higher frequency (51%) was obtained in a longer culture time (9 months) when only indole-3-butyric acid was present in the medium and in absence of chilling.Abbreviations BA 6-benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - MS Murashige and Skoog (1962) - NN Nitsch and Nitsch (1969) - NOA 2-naphthoxyacetic acid     

Production of emetic alkaloid by in vitro culture of cephaelis ipecacuanha A. Richard

Callus and adventitious roots were induced on leaf segments from shoot culture of Cephaelis ipecacuanha A. Richard on Murashige-Skoog medium containing 2,4-dichlorophenoxyacetic acid, indole-3-acetic acid, 1-naphthaleneacetic acid and kinetin. The contents of emetic alkaloids in calli, roots and root suspension cultures were quantified by HPLC. Roots cultured in solid and liquid Murashige-Skoog media yielded emetine and cephaeline. The amount of the two alkaloids in the root suspension culture was very similar to that of roots from ipecac mother plant grown in a greenhouse. In contrast, calli subcultured on Murashige-Skoog media containing combinations of 2,4-dichlorophenoxyacetic acid and kinetin produced only trace amounts of emetic alkaloids.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - NAA l-naphthaleneacetic acid - Kin kinetin - MS Murashige-Skoog - EM emetine - CP cephaeline - DW dry weight.     

Somatic embryogenesis and bud formation from immature embryos of buckwheat (Fagopyrum esculentum Moench.)

Immature embryos of Fagopyrum esculentum cv. Pennquad were isolated from field-grown plants and cultured on media containing a high benzylaminopurine to indole-3-acetic acid ratio. Part of the embryos were grown in the presence of 2,4-dichlorophenoxyacetic acid and kinetin for the first 5 days, and then transferred to benzylaminopurine + indole-3-acetic acid medium. From callus tissues developed on hypocotyls and cotyledons, 3 types of tissue were selected in later subcultures: (a) callus tissue strains that produced buds, (b) embryogenic tissue, and (c) unorganized callus tissue, lacking any organogenic capacity. Pretreatment with 2,4-dichlorophenoxyacetic acid increased the number of explants which gave rise to bud forming and embryogenic tissue, but was not essential for morphogenesis. Somatic embryogenesis was confirmed by histological observation. Plantlets could be easily obtained by inducing adventitious roots on shoots, but spontaneous root development in somatic embryos was infrequent.Abbreviations BAP benzylaminopurine - IAA indole-3-acetic acid - 2,4-D dichlorophenoxyacetic acid - IBA indole-3-butyric acid     

Somatic embryogenesis and plant regeneration from immature embryos of western larch

Somatic embryogenesis was initiated from immature embryos of western larch (Larix occidentalis Nutt.) on media containing 2,4-dichlorophenoxyacetic acid and N6- benzyladenine. The effects of explant type and ammonium nitrate and glutamine concentrations on initiation were tested. Although 21–93% of explants rendered cultures in various experiments, only 3% yielded sustainable embryogenic lines. Excised embryos at the early cotyledonary stage were optimal for initiation. Maturation of somatic embryos was promoted by abscisic acid. Response to abscisic acid concentrations and duration of exposure to abscisic acid varied with genotype. Maximal results were obtained with 0.025 M abscisic acid for 1 to 2 weeks followed by individual culture on medium without growth regulators. Mature somatic embryos developed into shoots with roots. Plantlets have been established in peat.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - BA N⁶-benzyladenine - IBA indole-3-butyric acid - ABA abscisic acid     

Stimulation of the formation of fruiting bodies of the fungusLentinus tigrinus (Bull.) Fr. by growth regulators

Growth regulators, indole-3-acetic acid (IAA), gibberellic acid (GA₃), and kinetin (KIN), were used in different concentrations to stimulate the initiation and further development of the fruiting bodies of the fungusLentinus tigrinus. Vegetative mycelium of the fungus was cultivated on cellulose cylinders soaked with a synthetic nutrient solution or with a 3% malt extract. When the mycelium covered the surface of the cylinders, further cultivation was carried out in graduated concentrations of the growth regulators mentioned above. The number of developed fruiting bodies showed that the optimum IAA and GA₃ concentrations were in both media 300 p.p.m. The optimum concentration of kinetin was 400 p.p.m. The individual growth regulators influenced characteristically also the shape of the fruiting bodies and changed in them the natural level of endogenous growth regulators. The addition of IAA into the medium raised the level of endogenous auxins in the cap. The presence of gibberellic acid and of kinetic in the medium resulted in an increase in the level of these regulators in the fruiting bodies.     

Shoot regeneration from petioles and leaves of Vitis X labruscana ‘Catawba’

Summary Shoot regeneration and normal plants were obtained from leaf and petiole explants derived from in vitro grown shoots of Vitis X labruscana Catawba. Regeneration was induced in the presence of both 6-benzylaminopurine and indole-3-butyric acid; combinations of 2,4-dichlorophenoxyacetic acid or 2-naphthoxyacetic acid with 6-benzylaminopurine did not permit regeneration from leaf explants. Up to 15% of leaf and 70% of petiole explants regenerated shoots on media with 5.0–10.0 M BA and 0.1–0.5 M IBA. Incubation in the dark was required to obtain regeneration. About 50% of shoots developed normally following transfer to light. An average of one shoot regenerated from leaf explants and 3.3 shoots regenerated per petiole explant. Regeneration from petioles and leaves was always from the basipetal end. The interaction of 6-benzylaminopurine with indole-3-butyric acid was also examined.Abbreviations BA 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - IBA indole-3-butyric acid - MS Murashige and Skoog (1962) medium - NN69 Nitsch and Nitsch (1969) medium - NOA 2-naphthoxyacetic acid     

Improvements in rooting regenerated safflower (Carthamus tinctorius L.) shoots

A continuing obstacle for regenerating safflower (Carthamus tinctorius L.) plants from cultured explants or callus has been a reliable method for rooting shoots. For shoots directly regenerated from primary explants, 76% of shoots rooted after a 7-d exposure to 10 mg/1 indole-3-butyric acid. Auxin source, concentration or exposure time did not greatly affect root formation or morphology, but strongly affected callus production. Shoots infected with Agrobacterium rhizogenes produced massive numbers of fibrous roots, but shoots did not elongate or survive transfer to soil. Shoot hyperhydricity symptoms were reduced by including 1 g/1 activated charcoal in rooting media. The optimal protocol for inducing root formation consisted of a 7-d exposure to 10 mg/l indole-3-butyric acid in root induction media, followed by incubation in media containing 15 g/l sucrose and 1 g/1 activated charcoal for 21 d.Abbreviations IBA indole-3-butyric acid - MS Murashige and Skoog (1962) medium - NAA anaphthalene acetic acid - POP 2,3,5-trichloro--phenoxypropionic acid