中国春孢锈菌属增补

本文对我国春孢锈菌属(形式属)增补了以下9个种：1．腺梗菜春孢锈菌Aecidium adenocauli Syd． 寄主：腺梗菜Adenocaulon himalaicum Edgew (菊科Compositae) 2．臭椿春孢锈菌(新种) Aecidium ailanthi J．Y．Zhuang sp．nov．寄主：臭椿 Ailanthus altissima Swingle (苦木科Simaroubaceae)3.筋骨草春孢锈菌 Aecidium alugae Syd．寄主：紫背金盘 Ajuga nioponensis Makino (唇形科 Labiatae) 4．八角枫春孢锈菌 Aecidium alangii Hirats．＆ Yosh 寄主：长毛八角枫 Alangium kurzii Craib,瓜木A platanifoHum Harms (八角枫科 Alangiaeeae) 5．紫珠生春孢锈菌(新种) Aecidium callicarpivola J Y．Zhuang sp．nov.寄主：杜虹花 Callicarpa oedunculata R,Br．(马鞭草科Verbenaeeae) 6．吴茱萸春孢锈菌(新种) Aecidium evodiae J．Y．Zhuang sp．nov． 寄主：吴茱萸 Evodia rutaecarva (Juss．)Benth．(芸香科Rutaceae) 7．杨叶木姜子春孢锈菌(新种) Aecidium litseae-populifoliae J．Y．Zhuang sp．Nov．寄主：杨叶木姜子 Litsea povulifolia Gamble (樟科Lauraceae) 8．青海春孢锈菌(新种) Aecidium qinghaiense J Y．Zhuang sp．nov． 寄主：莴苣属一种Lactuca sp．(菊科Compo sitae) 9．合掌消春孢锈菌 Aecidium vincetoxici P．Henn．＆ Shirai 寄主：牛皮消 Cynanchum auriculatum Royle (萝藦科 Asclepiadaceae)     

云南伞菌的十个新种

本文记述了云南伞菌中的十个新种：云南鸡油菌(Cantharelltcs yunnanensis sp. Nov．)、扁柄伞菌(Agaricus compressipes sp．nov．)、云南伞菌(Agarlcos yunnanensis sp．nov．)、黑点小脆柄菇(Psathyrella nigripunctipes sp．nov.)、粉柄粪伞(Bolbitius roseipes sp．nov．)、云南粪伞(Bolbitius yunnanensis sp．nov．)、赭枯滑菇(Hebeloma ochraceum sp．nov)、多鳞粘滑菇(Hebeloma squamulosum sp．nov．)、小鳞伞(Pholiota parvula sp．nov)及云南球盖菇(Stropharia yunanensis sp. Nov.)。     

斯氏假单胞菌(Pseudomonas stutzeri)A1501反硝化相关基因结构及功能分析

在A1501株的基因组上鉴定了4个反硝化相关的基因簇: nar, nir, nor和nos, 共40个基因, 基因的转录产物与其他种群中物质运输、基因调控以及还原酶类蛋白高度同源. nir, nor和nos基因簇在染色体上位置靠近, 与nar基因簇相隔较远. 与其他反硝化细菌比对的结果表明, A1501株中的40个反硝化基因组成了一套完整的反硝化催化系统. 在A1501株中, 该系统有以下特点: (ⅰ) nar基因簇中, narK基因只发现一个拷贝. (ⅱ) 在narK与narG之间有一个narM基因. (ⅲ) 在narX, narL基因的下游鉴定了两个基因dnrE和orf1, 其中dnrE基因是一个属于FNR家族的转录因子. (ⅳ) A1501株中nir基因有16个, 是所有已知反硝化细菌nir基因数量最多的. (ⅴ) 在A1501株中, 同时也是在假单胞杆菌属中首次报道norR基因. (ⅵ) nos基因簇相对保守, 无论在基因组成还是排列方式与参照的菌株都完全一致.     

中国链格孢属的研究I．

本文报道链格孢属的2个新种：寄生在苦术科(Simaroubaceae)臭椿[Ailanthus altissima(Mill)Swingle]上的臭椿链格孢(Alternaria ailanthi sp nov),寄生在桦术科(Betulaceae)黑桦(Betuladahurica Pall)上的桦术链格孢(A. betulae sp nov.),2个新组合：豆链格孢[A．Azulaae (Hara)comb．Nov],蔷薇生链格孢[A. rosicola(Rao) comb. Nov]和1个新名称红花链格孢(A．Carthami-tinctoriinom nov．)。文中为新种提供了拉丁文简介、描述和图。模式标本保藏在山东农业大学植物病理标本室(HSAUP)．     

中国单轴霉分类的研究*II.寄生在伞形科上的新种和已知种

本文报道在我国伞形科植物上寄生的4种单轴霉(Plasmopara)。其中寄生于鸭儿芹(Cryptotaenia japonica Hassk)上的鸭儿芹单轴霉(Plasmopara cyptotaeniae sp nov．)和寄生于水芹[Oenanthe iavanica(BI．)DC．]及卵叶水芹(O．Rosthornii Diels)上的水芹单轴霉(Pl-asmopara oenanlheae sp．nov．)是2个新种。寄生于变豆菜(Sanicttla chinensis Bunge)上的变豆菜单轴霉(Plasmopara saniculae Traian et O．Savulescu)是亚洲的新记录种。     

大肠杆菌-链霉菌高效接合载体的构建及其应用

以链霉菌质粒SCP2 的衍生质粒pHJL400为基础 ,构建了能够在大肠杆菌到链霉菌之间进行高效接合转移的质粒pGH112。pGH112含有在大肠杆菌和链霉菌中复制起始位点 ,以及分别在大肠杆菌和链霉菌中进行筛选的抗性标记。用pGH112转化EscherichiacoliET12567(pUZ8002 )后 ,与天蓝链霉菌 (StreptomycescoelicolorA3(2 ) )、除虫链霉菌 (Streptomycesavermitilis)、变铅青链霉菌 (StreptomyceslividansTK54 )、毒三素链霉菌 (StreptomycestoxytriciniNRRL15443)、委内瑞拉链霉菌 (Streptomyces.venezuelaeISP5230 )和红色糖多孢菌 (Saccharopolyporaerythraea)进行接合 ,发现本文构建的pGH112与pKC1139相比 ,接合转移效率较高 ,稳定性好 ,而且宿主范围较广。把组成型启动子ermE 与绿色荧光蛋白基因 (gfp)克隆到本文构建的pGH112 ,通过接合转移到链霉菌中 ,gfp获得表达,证明其可以用作基因接合转移的有效工具载体,这为研究链霉菌的基因功能创造了有利条件上。     

贵州灵芝科III．

本文继续报道贵州灵芝科(Ganodermataceae)3种,其中灵芝属(Ganoderma Karst.)2种,即新种镇宁灵芝(Ganoderma zhenningense He)和热带灵芝(Ganoderma troplcum(Jungh．)Bres．);网孢芝属(Humphreya Steyaert)1种,即咖啡网孢芝[Humphreya coffe-atum (Berk.)Steyaert]。以上引证标本全部保藏在贵州科学院生物研究所真菌标本室。     

峨眉山自然保护区的虫草及其它昆虫病原真菌I．

采自四川省峨眉山自然保护区的虫草及其相关昆虫病原真菌,其中除有常见的蛹虫草Cordyceps militaris (Vuill)Fr,粉被虫草(Cordyceps pruinosa Perch),蝉拟青霉Paecilomyces cicadae(Miquel) Samson,球孢白僵菌Beauveria bassiana(Bals．)Vuill等昆虫病原真菌外,将主要描述峨眉虫革新种(Cordyceps emeiensis A Y．Liu et Liang sp．nov),龙洞虫革新种(Cordyceps longdongensis A．Y Liu et Liang sp．nov),暗绿虫草新记录种 (Cordyceps atrovirens Kob et Shim．)和蛹虫草球头变种新记录种 Cordyceps militaris var sphaerocephala(Schw) Fr．等几种虫草菌。     

脱乙酰氧基头孢菌素C合成酶/羟化酶基因cefEF的克隆及序列分析

利用氯化苄分别从真菌顶头孢(Cephalosporium acremonium)和产黄头孢(Acremonium chrysogenum)中提取总DNA,通过PCR方法扩增脱乙酰氧基头孢菌素C合成酶/羟化酶基因cefEF,结果只能从产黄头孢DNA中扩增出cefEF基因。测序结果表明,其与已报道的基因序列只有3个碱基的差异,推断的氨基酸序列只有2个氨基酸有差异,并未涉及活性中心。同时表明,国外所指的与该酶有关的顶头孢(Cephalosporium acremonium或Acremonium chrysogenum)对应的是国内的产黄头孢(Acremonium chrysogenum)。     

类核环形结构在耐辐射球菌电离辐射抗性中不起主要作用

近来基于透射电子显微镜研究的论点“耐辐射球菌(Deinococcus radiodurans) R1类核致密的环形结构是其极端辐射抗性的关键因素”已引起广泛争议. 本文在以前研究基础上系统地比较了野生型R1菌株、pprI (DR0167)基因突变株YR1及pprI基因功能补偿株在电离辐射前后的类核结构的形态差异. 荧光显微镜观察结果显示: (ⅰ) 具有超强电离辐射抗性的野生型菌株R1细胞类核主要呈现紧凑的环形结构, 而同样有环形类核结构的pprI突变株YR1细胞却对辐射非常敏感; (ⅱ) 2个pprI基因功能补偿株, YR1001(pprI全基因功能补偿株)类核绝大多数呈现絮乱松散的不规则形态, YR1002(pprI部分基因功能补偿株)呈现约60%左右的环形类核结构, 这两个菌株都具有与野生型R1同样的电离辐射抗性; (ⅲ) 另一pprI部分基因功能补偿株YR1004 (敲除pprI基因3′端333个碱基对)对电离辐射极其敏感, 其约60%左右的细胞含有环形的类核结构. 上述结果表明, 耐辐射球菌类核的环形结构在辐射极端抗性中不起主要作用.     

Isolation,Culture and Plant Regeneration of Protoplasts from an Embryogenic Callus Tissue of Gossypium hirsutum

Protoplasts were isolated and cultured from hypocotyl embryogenic callus tissue of Gossypium hirsutum L. cv. "Lumian 6". The highest yields of viable protoplasts were obtained from a vigorous embryogenic callus 7 to 9 d old subcultured on MS medium supplemented with 2 mg/L IAA and 1 mg/L KT using a solution of 1% cellulase Onozuka R-10, 1% pectinase, 0.7 mmol/L KH2PO4, 2.5 mmol/L Ca2+ , and 0.5 mol/L osmoticum (mannitol), at pH 5.8 and at a temperature of 30 ℃. After separation and purification (in 21% sucrose floatation medium), the protoplasts were laid up in a quiet liquid protoplast culture medium containing K3 salts, NT vitamins with 0.1 mg/L 2,4-D, 0.2 mg/L KT and 0.45 mol/L glucose for 10 to 15 min. The protoplasts were fractioned into an upper and a lower layer in the centrifugal tube. Most of the protoplasts in the lower layer were smaller, round and rich in cytoplasts in which contain many granular substances. When this kind of protoplasts were cultured in the thin liquid protoplast culture medium with a density of 1 x l0s to 5 x los protoplasts/mL, the division and the callus formation of the regenerated cells were easily observed. The first divisions occurred in 3 days and small cell clusters could be seen after 2 to 3 weeks in the culture. At this moment, the addition of the protoplast culture medium with decreased osmoticum once or twice is needed for the continuous protoplasts division to form calli. Regenerated calli, 3 to 5 mm in diameter, were transferred in succession on MS medium with 2 mg/L IAA and 1 mg/L KT for the initiation of embryogenesis. The embryoids germinated on the hormonefree MS medium and a number of plantlets were obtained. It seems that using vigorous embryogenic callus and decreasing osmoticum are the two critical factors for plant regeneration of cotton protoplasts.     

从石刁柏原生质体培养再生植株

石刁柏,又名芦笋(Asparagus officinalisL.)是百合科天门冬属植物。其栽培品种含有丰富的维生素类及蛋白质。同时,石刁柏对于某些疾病有一定的药效,因此它已成为人们所喜爱的一种高级营养蔬菜。国外已有不少关于石刁柏试管苗繁殖的报告,但至今只有Bui Dang Ha等从石刁柏枝状叶分离的原生质体得到愈伤组织,并由此愈伤组织诱导获得了再生植株。此后,未见在石刁柏的原生质体培养方面再有新的工作。在本文中,我们利     

Somatic Embryogenesis and Plant Regeneration from Protoplasts of Cowpea (Vigna sinensis)

Immature cotyledons of cowpea (Vigna sinensis Endlo) were used for protoplast isolation. Enzyme solution for protoplast isolation contained 40% cellulase Onozuka R-10,0.30% Macerozyme R-10 and 2% hemicellulase. The purified protoplasts were cultured in Bs,MS or KM8p liquid medium in dark (25℃) at a density of 1 × 105–5 × 105/ml. The protoplasts started cell division in 3–5 days . Sustained cell divisions resulted ill formation of cell clusters and small calli,with cell division frequency reaching 23%–28% in MS medium . Calli of 2 mm in size were transferred onto MSB (MS salts+B5 vitamins) medium with 2 mg/L 2,4-D, 0. 5mg /L BA forfurther growth. Embryogenic calli appeared on this medium. After passage to fresh medium with the same composition, the embryogenic calli were transferred into MSB liquid medium to establish suspension culture. When the suspended calli were transferred back onto MSB agar medium with 0. 1 mg /L IAA, 0.5mg/L KT, 5% mannitol (cultured in light,2000 lx,12h/d), a lot of adventitious roots formed in 7–10 days, and then somatic embryos formed from the protoplast derived calli. But only a few embryoids developed further into the cotyledonary stage ,and the others died at globular, heart-shaped, or torpeto stage . Finally, some cotyledonary embryoids germinated and developed into plantlets or shoots with leaves.     

Study on Tissue and Protoplast Culture of Wild Cotton (Gossypium davidsonii)

This paper deals with the study on the condition of callus formation, embryogenesis, organogenesis, plant regeneration and protoplast culture of wild cotton (G. davidsonii) Callus cultures derived from several organs such as root, stem, leaf, cotyledon and hypocotyl. The results obtained in these cultures showed that the modified MS medium containing 2,4-D 1.0+KT 0.1; 2,4-D 0.1+KT 0.01; NAA (IAA) 2.0+KT 0.1 and NAA (IAA) 1.0+KT 0.1 mg/L were favorable to callus formation. Modified MS medium containing 2,4-D was suitable for initiated callus of G. davidsonii Besides, suspension cultures from callus of G. davidsonii were saccessfully initiated. Optimum concentration of 6BA (or ZT, or 2ip) and NAA (IAA) was for shooting, somatic embryo or leaf formation. Plantlets regenerated from somatic embryo at lower concentration of 6BA, or ZT, or 2ip. As to protoplast culture of this species, the age and physiological condition of callus or suspension cells and concentration of enzymes used for protoplast isolation affected the yield and survival of protoplasts. Protoplast of this species cultured in modified MS medium containing 2,4-D 0.5+NAA 0.5+ZT 0.1–0.2 mg/L. and divied after 3–4 days. The rate of division was 3--4% and cell cluster formed after 14 days, then these cells died.     

陆地棉体细胞胚胎发生与植株再生

利用陆地棉品种下胚轴为外植体进行体外培养研究。激素和品种是影响愈伤诱导和胚胎发生的主要因素。去除激素后胚性愈伤在固体培养基上只能形成少量的成熟胚。悬浮培养是获得大量成熟胚的中间步骤。悬培两周后,悬培物转到固体培养基上促进胚状体成熟,30—60目之间的悬培物比大于30目的悬培物易形成成熟胚。KT 0.1ppm、Zea 0.1ppm分别有效地促进了胚状体成熟。活性碳250mg/L、NAA 0.1ppm、IBA 0.1ppm和IAA 0.1ppm能使胚状体萌发并健壮生长。目前已得到100多株幼苗,大苗已达八片真叶。     

Plants Regenerated from Mesophyll Protoplasts of Cottonwood New Clone

Poplar NL-80106 (Populus deltoides×P, simonii) mesophyll protoplasts were isolated from leaves of 30 days-old sterile shoot, with 4 × 107/g fr. wt of protoplast yield after purification. The protoplasts were cultured in KM8p and MS liquid media containing 2 mg/L 2, 4-D, 0. 5 mg/L NAA and 0.5 mg/L KT. Higher plating density and lower osmatic pressure (0.45 mol/L) were proved to be favourable to division of protoplast-derived cells. The first division initiated 5 days after culture, and the division frequency reached 4.5 % on the 10th day. A number'of cell colonies and microcalli was formed in 12 weeks. Using organic nitrates and glucose in protoplast culture medium was beneficial to increase division frequency and plating efficiency. The calli were allowed to grow to 4--6 mm in height with red colour and compact structure on the gelrite-sohdified NLZ1 proliferation medium in 3 weeks and were transferred onto NLF differentiation medium where the frequency of shoot formation could reach 100%. The 3 cm high shoots were then cut off from the callus and rooted on 1/2 MS medium.     

青叶胆愈伤组织的诱导与液体继代培养

以青叶胆(Swertia mileensis)幼叶为外植体,在添加不同生长调节剂组合的MS培养基上进行愈伤组织诱导。结果表明,最有效的生长调节剂组合为IAA 1.0 mg/L+NAA 1.0 mg/L+KT 1.0 mg/L和NAA 4.0 mg/L+KT 2.0 mg/L+GA₃ 1.0 mg/L,其诱导率分别为96.6%和96.0%。使用以上两种生长调节剂组合进行愈伤组织的液体继代培养,10 d可使愈伤组织增殖率分别达257.3%和310.2%。     

羊草与灰色赖草F1代细胞悬浮系的建立及植株再生

本研究以羊草(L eym us ch inensis)与灰色赖草(L eym us cinereus)杂种F1代幼穗为外植体诱导愈伤组织,在3.0 m g/L 2,4-D M S培养基上继代1次后,转入不同浓度激素(2,4-D、IAA、KT)配比和不同浓度蔗糖的M S液体培养基进行振荡培养,建立杂种F1代细胞悬浮系和植株再生体系.结果表明,细胞悬浮培养时,M S 1.0 m g/L2,4-D 0.1 m g/L KT 4%蔗糖的液体培养基最佳;悬浮细胞分化时,1.0 m g/L 2,4-D 0.1 m g/L KT 4%蔗糖 M S和1.0 m g/L 2,4-D 4%蔗糖 M S培养的悬浮细胞在1.0 m g/L NAA 0.5 m g/L KT M S分化培养基上的绿苗分化率分别达到83%和80%.细胞悬浮系及再生体系的建立为杂种F1代育性恢复的研究奠定了基础.     

紫云英叶片及叶肉原生质体的培养

本工作研究了豆科植物紫云英的叶片及叶肉原生质体的培养。叶片培养实验表明,诱导愈伤组织的最适培养基为MS加1.0-2.0毫克/升2,4-D和0.25毫克/升KT;诱导根分化需加1.0—5.0毫克/升NAA和0.5毫克/升BA;而苗分化则以0—0.5毫克/升IAA和0.5毫克/升BA为好。高浓度的NAA有利于根分化而抑制茎芽形成;高浓度的IAA对根和芽分化都有抑制作用。叶肉原生质体分离和培养试验表明,紫云英叶肉原生质体的释放及其培养活力受叶龄、植株生理状态和酶浓度的影响。叶肉原生质体在改良的KM8P培养基中能分裂。用改良KM8细胞培养基定期稀释,可使分裂持续进行而得到细胞团。BA和2,4-D为诱导紫云英叶肉原生质体分裂所必需。其最佳组合激素为BA 0.21毫克/升和2,4-D 1.13毫克/升。葡萄糖作为渗透压稳定剂时,其浓度明显影响原生质体的存活率。弱光条件下培养比黑暗培养有利于叶肉原生质体分裂。由叶肉原生质体形成的愈伤组织能形成瘤状结构和根。     

Stem Apex Culture and Quality Improvement of Tubercles of Pinellia ternata

Formation of Plantlets was achieved when stem apex of Pinellia ternata Brier. Cultured in vitro on MS medium with KT 0. 5 mg/L + NAA 0.2 mg/L (MSI). With petioles of the plantlet as explants callus could be induced after cultured for a week on MS medium with 2, 4-D 2.0 mg/L + KT 0.5 mg/L (MSII). Calli were subcultured once in every month. After 3--4 months a kind of friable calli could be selected, from which the tubercles could be differentiate and the plantlets formed when transfered onto MSI. But before callus differentiation, a lot of roots were formed on callus. The plantlets could be produced directly from the petiole segment. It was found that the stem growing tip was always covered by the leaf primordium and the former leaf primordium was covered by the latter leaf primordium during the differentiation of the apical bud of tubercle. The frenquency of plantlet differentiation from callus and petioles was over 70%. The rate of regeneration of plantlet on liquid static culture was twice as much as that on solid culture. All plantlets grew well after being transfered into the plot. The fresh weight of tuber-plant was 103 % higher than that of control (cultivated plant come from tubers). The alkaloid content of tubers come from tuberplant was 0. 344%, that of control was 0. 203% and 0. 264% for the wild tuber.