RAPD技术在园艺植物育种资源中的应用

综述了随机扩增多态性DNA(RAPD)技术在多年生园艺植物(特别是果树)品种鉴定、种质资源及其育种研究中的应用现状.     

碱熔-离子色谱法测定土壤中全硫的研究

研究了用碱熔-离子色谱法测定土壤中全硫的方法,样品采用氧化镁—碳酸钠(2:1)混合熔剂,在恒温马弗炉中800℃加热1 h,使土壤中各种形态的硫转化为SO_4~(2-),冷却,用水超声浸提.离子色谱法测定.分析柱AS14A4-mm(4 mm×250mm),淋洗液0.008mol·L~(-1)Na_2CO_3、0.001 mol·L~(-1)INaHCO_3,再生液0.05mol·LH_2SO_4~(-1),流量1.0mL·min~(-1),进样体积20μL,外标峰面积定量.该方法测定土壤中硫检测限为0.8 mg·kg~(-1),在测量浓度范围内具有良好的线性关系.用该方法做精密度,相对标准偏差为3.91%.与不同地区的标准土做对比实验,所测结果跟标准值相比均在标准值范围内.实验证明该方法操作简便、快速.     

磷矿石的微生物风化作用——以一株黑曲霉(Aspergillus niger)为例

以直接和间接作用的方式研究培养基中黑曲霉Aspergillus niger对磷矿粉的风化作用。在装有100目磷矿石粉的液体培养基中接入该菌研究其对磷矿石粉的直接风化作用;同时将装有100目磷矿石粉的透析袋放入液体培养基中再接入该菌研究其对磷矿石粉的间接风化作用。按不同时间取培养液上清液,用电感耦合等离子体-发射光谱仪测定Ca2 、Mg2 、Al3 、Fe3 、K 和Mn2 浓度,用离子色谱法测定H2PO4-、SO42-和Cl-浓度。此外,黑曲霉风化作用后的矿物残渣用电子探针作表面微观形态分析和XRD矿物物相分析。结果表明:黑曲霉对磷矿石粉风化作用的直接作用强度大于间接作用;黑曲霉可风化磷矿石粉,但对矿粉中不同矿物的风化强度不同,风化过程中形成次生矿物水草酸钙。提出黑曲霉对磷矿石的风化作用源自黑曲霉生长导致的机械破坏作用、胞外分泌物的生化降解作用以及多种因素之间的协同作用。     

Protoplast fusion between Saccharomyces cerevisiae and Rhodobacter sphaeroides for the treatment of soybean wastewater

The kinetic parameters and the purification rates of the hybrid cell Foaz in soybean processing wastewater (SPW) were measured through a shaking reaction and in two automatic regulated control stable fermentation systems (ARCSFS). The maximum specific growth rate of Foaz was 0.576 h^‐1, higher than that of one of its parental strains Rhodobacter sphaeroides P9479 and lower than that of another parental strain Saccharomyces cerevisiae Y9407. The BOD₅ removal rate of Foaz in the No. 1 system was 61.3%, higher than those of both its parental strains when the influent BOD₅ concentration was 4600 mg/L. The results of this study suggest that the hybrid Foaz has a better capacity of the degradation of organic pollutants in SPW than its parental strains and it may be applicable to the treatment of high concentration organic wastewater.     

Identification of Tsuga Germplasm by Morphological Characters and RAPD Markers

Germplasm collection is important to preserve and maximize genetic diversity for germplasm conservation. Tsuga dumosa （ D. Don） Eichler in Engler ＆ Prantl. and T. chinensis var. forrestii （Downie） Silba germplasm was collected from three localities in China： Mt. Yulong, Wenfeng Temple and Mt. Dishiergu, Yunnan Province. Accessions were identified based on morphological characters and RAPD markers. The shapes of the apices and margins of needles were examined, and the length and width of needles, cones and seeds from accessions of mature plants were used to compare the morphological differences and to identify the germplasm. Molecular markers generated by randomly amplified polymorphic DNA （RAPD） were also used to characterize the taxa. Although the clustering based on RAPD markers was inconsistent with the morphological characters of the needles, based on the overall morphological characters and on RAPD markers, the accessions from Mt. Yulong and Wenfeng Temple were identified as T. chinertsis var. forrestii, and those from Mt. Dishiergu identified as T. dumosa. Taxonomic identification of the accessions was made based on morphology and by RAPD markers concurred. The results indicate that the shapes of the apices and margins of needles particularly from young plants could not be used as a possible key to identify T. dumosa and T. chinertsis var. forrestii. Fig 6, Tab 3, Ref24     

无花果曲霉对直接冻黄（G）的脱色特性研究

文章利用无花果曲霉对直接冻黄G进行脱色研究,在搅拌条件下,菌丝球能有效地使直接冻黄G脱色（＞90％在24h内）。在限氮培养基中,对影响脱色过程的因素进行了试验。这些因素包括：葡萄糖浓度、酒石酸 铵浓度、原初pH及温度。结果表明：该菌丝球对染料脱色的最佳温度为30℃,最佳pH5.0,培养基成分对脱色也有一定的影响。该菌对染料的脱色酶为组成酶,脱色酶在细胞内、外均有分布,染料对酶无诱导作用。菌丝球经二次脱色后能重新被利用。     

黑曲霉吸附弱酸性艳蓝RAWL机理研究

考察了黑曲霉对染料吸附性能的pH响应模式,红外光谱表征了菌体表面的主要官能团组成,化学修饰定量研究了不同官能团的染料吸附贡献.结果显示,在低pH条件下,菌体表现出最优的染料吸附特性:随着pH值由2.0升至6.0,染料吸附量由39.6mg/g降至9.3mg/g.菌体表面含有氨基、羧基和磷酸根.高pH下,羧基和磷酸根电离导致菌体表面带负电并与染料发生静电斥力,使得染料吸附量下降;低pH下,氨基的质子化使得菌体带正电并通过静电吸引提高染料吸附.甲基化氨基在酸性条件下仍然可以质子化,故氨基甲基化修饰后染料吸附性能不变;乙酰化氨基在酸性体系中失去质子化能力,乙酰化修饰菌体染料吸附性能下降51.6%.氨基质子化引起的菌体正电性和染料负电性之间的静电引力是染料吸附的重要机制.图6参10     

Biosorption of low level Cd~（2＋） in water by Aspergillus sojae

以酱油曲霉（Aspergillus sojae）为吸附剂,对水溶液中微量Cd2＋进行了吸附研究.实验考察了预处理方法、菌体用量、pH、温度和吸附时间对吸附的影响,同时分析了其吸附动力学和吸附等温线特性,并对酱油曲霉菌体吸附Cd2＋的可能作用机理进行了探讨.结果表明,碱预处理的菌体吸附效果最好.在菌体用量为0.08 g时,对Cd2＋的吸附量达到最大,为0.095 mg.g-1.pH值在3.4—8之间,酱油曲霉菌体对Cd2＋的吸附效果比较稳定,其最大吸附率出现在pH 3.4.温度在15℃—30℃之间时,菌体对Cd2＋的吸附率差异很小,吸附率在88.3%—94.4%之间.二级动力学方程能很好地表征酱油曲霉吸附微量Cd2＋的过程,R2在0.99以上,其吸附平衡时间大约2.5 h,线性模型更适于描述该吸附平衡过程.SEM、FTIR分析得出,Cd2＋可能是与酱油曲霉菌体表面的羟基、酰胺基、羧基、磷酸基和胺基等基团相络合而被吸附,也可能是形成颗粒沉淀而附着在菌丝上.     

米曲霉氨基酰化酶酶法拆分制备D型芳香族氨基酸

以DL-苯丙氨酸为起始原料,乙酸酐乙酰化后得到N-乙酰DL-苯丙氨酸,利用米曲霉氨基酰化酶立体结构专一催化水解N-酰基-L-氨基酸的酰胺键的特点,分别得到L-苯丙氨酸和N-乙酰D-苯丙氨酸.通过结晶法分离两者后,N-乙酰D-苯丙氨酸经6 mol/L HCI水解得到D-苯丙氨酸.由此拆分DL-苯丙氨酸得到光学纯度分别为98%的L-苯丙氨酸(收率84.8%,以原料N-乙酰-DL-苯丙氨酸的0.5倍摩尔量为理论产率)和92.3%的D-苯丙氨酸(收率89.5%).参22     

Enhanced enzymatic removal of anthracene by the mangrove soil-derived fungus, Aspergillus sydowii BPOI

• A. sydowii strain bpo1 exhibited 99.8% anthracene degradation efficiency. • Four unique metabolic products were obtained after anthracene degradation. • Ligninolytic enzymes induction played vital roles in the removal of anthracene. • Laccase played a crucial role in comparison with other enzymes induced. The present study investigated the efficiency of Aspergillus sydowii strain bpo1 (GenBank Accession Number: MK373021) in the removal of anthracene (100 mg/L). Optimal degradation efficiency (98.7%) was observed at neutral pH, temperature (30℃), biomass weight (2 g) and salinity (0.2% w/v) within 72 h. The enzyme analyses revealed 131%, 107%, and 89% induction in laccase, lignin peroxidase, and manganese peroxidase respectively during anthracene degradation. Furthermore, the degradation efficiency (99.8%) and enzyme induction were significantly enhanced with the addition of 100 mg/L of citric acid and glucose to the culture. At varying anthracene concentrations (100–500 mg/L), the degradation rate constants (k₁) peaked with increasing concentration of anthracene while the half-life (t_1/2) decreases with increase in anthracene concentration. Goodness of fit (R² = 0.976 and 0.982) was observed when the experimental data were subjected to Langmuir and Temkin models respectively which affirmed the monolayer and heterogeneous nature exhibited by A. sydwoii cells during degradation. Four distinct metabolites; anthracene-1,8,9 (2H,8aH,9aH)-trione, 2,4a-dihydronaphthalene-1,5-dione, 1,3,3a,7a-tetrahydro-2-benzofuran-4,7-dione and 2-hydroxybenzoic acid was obtained through Gas Chromatography-Mass spectrometry (GC-MS). A. sydowii exhibited promising potentials in the removal of PAHs.