小麦waxy蛋白亚基1D-SDS-PAGE分离方法改良

对小麦ｗａｘｙ蛋白的提取技术和电泳后胶片的银染方法进行了改良：将无胚的半粒种子研磨成粉,用洗液浸泡２ｈ代替蒸馏水浸泡过夜,并且去掉了用洗液反复洗涤和离心的程序;电泳后胶后的染色采用快速银染法代替常用的银染法,改良后的单向十二烷基酸钠聚丙烯酰胺胶电泳方法（１Ｄ－ＳＤＳ－ＰＡＧＥ）具有省时、方便、经济和有效的特点,可适用于大批小麦材料的ｗａｘｙ蛋白亚基的分析,图１参５。     

蔓茎堇菜细胞悬浮培养的研究

对蔓茎堇菜细胞悬浮培养的条件进行了选择,并测定悬浮培养的生长动态。结果表明：蔓茎堇菜细胞悬浮培养的最适接种量为２．０～３．５ｇ（５０ｍＬ）;最适培养基为ＭＳ＋２．０ｍｇ／Ｌ２,４－Ｄ＋１．０ｍｇ／Ｌ ＺＴ＋３００ｍｇ／Ｌ ＬＨ＋２００ｇ／Ｌ蔗糖（或葡萄糖）,ｐＨ５．８;细胞悬浮培养一个周期过程中细胞数目增长的动态基本上是一条Ｓ型曲线,对数生长期明显;细胞鲜重（ｍＦＷ）和细胞干重（ｍＤＷ）则与细胞     

用SPE法萃取,GC／NPD检测全血中氯氮平药物

本文报道了用固相萃取技术（ＳＰＥ）富集分离全血中的氯氮平药物，采用气相色谱氮磷检测器（ＧＣ／ＮＰＤ）分析方法凤ＳＫＦ５２５Ａ为内标物，在１ｍｌ全血中加入２．０μｇ药物，回收率为９０．０％，ＲＳＤ为７．９５％，血中最低检出浓度为５８ｎｇ．ｍｌ＾－１，该方法具有简单、快速、准确、１灵敏的优点。     

土壤中水溶性有机碳与铜的相互影响

通过向土壤溶液中加入Ｃｕ２＋和可溶性有机碳(ＤＯＣ),研究它们之间的相互作用．结果发现,加入铜可以显著改变土壤水溶性有机碳(ＷＳＯＣ)的吸着平衡,并降低了其水溶性．影响程度在０．８ｍｍｏｌ·ｌ－１Ｃｕ以下随铜浓度的增加而增加．加入０．５ｍｍｏｌ·ｌ－１铜的土壤中 ＷＳＯＳ含量从 ０．２７ｍｇ Ｃ·ｇ－１降至０．０４７ｍｇ Ｃ·ｇ－１,吸着系数从６．６×１０－３ｌ·ｇ－１降至３．３×１０－３ｌ·ｇ－１．另一方面,用Ｆｒｅｕｎｄｌｉｃｈ方程描述铜离子在土壤中的吸附行为(液相浓度单位取ｍｍｏｌ·ｌ－１,固相浓度取ｍｍｏｌ·ｇ－１土),ｋ＝９．ｌ×１０－３;ｎ＝３．４０． 加入ＤＯＣ后,ｋ＝９．２×１０－３;ｎ＝３．６６,没有明显改变土壤对铜的吸附性能．加入小同浓度的ＤＯＣ,结果仍说明该浓度范围内ＤＯＣ(＜８ｍｇ Ｃ·ｌ－１)的加入没有明显改变土壤对铜的吸附性能．     

转化血型用蕃茄α－D－半乳糖苷酶的分离、纯化及理化性质

成熟蕃茄匀浆后，经硫酸铵盐析，ＤＥＡＥ－ＳｅｐｈａｄｅｘＡ－５０离子交换层析，ＳｅｐａｄｅｘＧ－１００凝胶过滤和Ｍｅｌｉｂｉｏｓｅ－Ａｇａｒｏｓｅ亲和层析，获得了α－Ｄ－半乳糖苷酶（Ｃ．Ｅ．３．２．１．１１）。酶制剂经ＰＡＧＥ检测为一条带；ＳＤＳ－Ｇ－ＰＡＧＥ测得酶Ｍｒ为３４０００；比活力５２．９Ｕ／ｍｇ·；提纯倍数为５２９０１产率为４５％．酶专－催化以α－Ｄ－半乳糖为末端ａ－（１，３）连接的糖苷键，以ＰＮＰＧ（对硝基苯－α－Ｄ－半乳糖昔）为废物，酶催化反应的Ｋｍ＝０．１１ｍｍｏｌ／Ｌ，Ｖｍａｘ为６７μｍｏｌ·ｍｇ１－·ｍｉｎ－１．ｔ稳定范是０～３５℃；ＰＨ稳定范围是４．０～７．０．最适ｐＨ为５．１．半乳糖是酶的竞争性抑制剂；Ｃｕ２＋、Ｚｎ２＋、Ｍｎ２＋、Ｆｅ３＋、Ａｇ＋和ＥＤＴＡ对酶活性无影响．纯酶制剂可作为Ｂ型血向Ｏ型血转化的工具酶液．     

麻黄愈伤组织细胞的悬浮培养

用中麻黄幼苗的３种外植体进行了离体培养。不同外植体诱导愈伤组织的结果表明：最佳培养基为ＭＳ＋２ｍｇ／Ｌ２,４－Ｄ＋１ｍｇ／Ｌ６－ＢＡ;最佳外植体为下胚轴;用源于下胚轴的愈伤组织进行悬浮培养,从第３代开始建立起稳定的悬浮系。愈伤组织、悬浮培养细胞及培养液中都含有麻黄碱,悬浮培养液中的麻黄碱含量高于愈伤组织和悬浮培养细胞。表２参１２     

六六六热解废渣中2,3,7,8—取代PCDDs和PCDFs的测定

采用ＨＰＬＣ和ＧＣ／ＭＳ法对六六六热解废渣中的ＰＣＤＤｓ和ＰＣＤＦｓ进行了测定，使用此种方法，于废渣中检出了所有２，３，７，８－取代的同系物。此外，还根据各ＣＤＤｓ，ＣＤＦｓ的毒性当量因子ＴＥＦｓ，计算了废渣的毒性当量ＴＥＱｓ，约为６１２－９２４μｇ／ｇ。分析表明，ＯＣＤＤ是含量最丰富的导构体，而１，２，３，４，６，７，８－ＨｐＣＤＤ，１，２，３，４，６，７，８－ＨｐＣＤＦ和１，２，３，４，７，８     

草浆漂白过程中二恶英类生成机理探讨

本文以氯苯,氯酚,二苯并二恶英和二苯并呋喃为ＰＣＤＤ／ＦＳ前生体,模拟工业草浆漂白条件,探讨了次氯酸盐和氯气漂白苇浆过程中二恶英类的生成机制,研究表明,在苇浆中加入二恶英类前生体加ＤＢＤ／Ｆ,其氯化产物与在木浆中有所区别。在实验条件下,二恶英类的生成量分别为;１５μｇＰＣＤＤＤｓ．ｍｇ＾－１ＤＢＤ,１．５μｇＰＣＤＦｓ．ｍｇ＾－１ＤＢＦ。     

单甲脒农药对土壤微生物种群和土壤酶活性的影响

研究了单甲脒（Ｎ＇－２，４－二甲苯基－Ｎ－甲基甲脒，简称ＤＭＡ）对土壤微生物种群和土壤酶活性的影响．田间试验表明，喷洒单甲脒盐酸盐（ＤＭＡＨ）于棉花植株对土壤微生物种群和土壤酶活性无明显影响．实验室试验表明，投加ＤＭＡＨ于土壤对土壤微生物种群有不同的抑制作用；当ＤＭＡＨ在土壤中的浓度为２０ｍｇ／ｇ时，好氧异养菌总量开始降低，硝化细菌和放线菌、真菌的土壤ＤＭＡＨ抑制浓度分别为０．５ｍｇ／ｇ和０．１ｍｇ／ｇ．ＤＭＡＨ对土壤酶活性的影响亦不相同，脱氢酶对土壤中ＤＭＡＨ含量为１０ｍｇ／ｇ时其活性即有明显的减弱，而过氧化氨酶对ＤＭＡ的耐受性相当高，其活性的抑制浓度达５００ｍｇ／ｇ．试验还证明了土壤中ＤＭＡ的微生物降解作用．当ＤＭＡ在土壤中的浓度为５０～５００ｇ／ｇ时，其消解速率可从８．７５ｍｍ·ｋｇ－１·ｄ－１提高到４８ｍｇ·ｋｇ－１·ｄ－１．     

五氯酚及其钠盐中氯代二恶英类分析

采用ＨＰＬＣ和ＧＣ／ＭＳ的方法，对国产ＰＣＰ及其钠盐中的ＰＣＤＤｓ，ＰＣＤＦｓ进行了分析。测定了所有２，３，７，８－取代同系物。此外，还根据各ＣＤＤｓ，ＣＤＦｓ的毒性当量因于ＴＥＦｓ，计算了其毒性当量Ｉ－ＴＥＱｓ，分别为１４２，９２ｎｇ．ｇ＾－１。在前述基础上，仨算了由于ＰＣＰ及其钠盐的应用，引起的ＰＣＤＤｓ，ＰＣＤＦｓ环境输入。约为２４０ｋｇ／ａ。     

CMS高粱保持系叶绿体基因ps1A1和ps1A2片段的克隆与序列比对

以15种包括同质异核和同核异质高粱材料的总：DNA为模板进行RAPD分析，196个随机引物中，引物Y-19扩增得到一个很有规律的差异片段SAY-192300。该片段只出现在4个保持系(B)和3个恢复系(R)中，而不出现在6个不育系(A)和2个杂种F1中，进一步对cms—Al、cms—A2两套8个材料(A／B／R／F1)的总DNA、mtDNA和cpDNA用引物Y-19进行扩增，发现片段SAY-192300仅在cpDNA得到，Southern杂交结果证实了片段SAY-192300的叶绿体来源，同时也表明在cms胞质中该片段所在区未发生插入、缺失等大的结构变异,DNA序列测定结果表明，该片段为叶绿体ps1A1和ps1A2基因的部分序列。图6表2参19。     

可吸入尘中苯并（a）芘超声提取的高效液相色谱测定法

本文介绍一种简便、快速的可吸入尘中苯并(a)茈高效液相色谱测定方法.本法仅需5mL乙腈,超声处理10min即可完成提取过程.提取液离心后,上清液可直接进行HPLC分析.色谱条件:反相C18柱,用乙腈/水梯度洗脱,紫外检测器,λ=254nm.方法的最小检出量10~(-10)g.回收率95%,变异系数4、3%.此方法可用于大气可吸入尘中Bap的监测.     

Mitochondrial plasticity in brachiopod (Liothyrella spp.) smooth adductor muscle as a result of season and latitude

Habitat temperature and mitochondrial volume density (Vv_(mt,mf)) are negatively correlated in fishes, while seasonal acclimatization may increase Vv_(mt,mf) or the surface density of the mitochondrial cristae (Sv_(im,mt)). The effect of temperature on invertebrate mitochondria is essentially unknown. A comparison of two articulate brachiopod species, Liothyrella uva collected from Rothera Station, Antarctica in summer 2007, and Liothyrella neozelanica collected from Fiordland, New Zealand in winter 2007 and summer 2008, revealed a higher Vv_(mt,mf) in the Antarctic brachiopod. The Sv_(im,mt) was, however, significantly lower, indicating the Antarctic brachiopods have more, less reactive mitochondria. L. uva, from the colder environment, had larger adductor muscles in both absolute and relative terms than the temperate L. neozelanica. Furthermore, a seasonal comparison (winter vs. summer) in L. neozelanica showed that the absolute and relative size of the adductor increased in winter, Vv_(mt,mf) was unchanged, and Sv_(im,mt) was significantly increased. Thus, seasonal acclimatization to the cold resulted in the same number of more reactive mitochondria. L. neozelanica was clearly able to adapt to seasonal changes using a different mechanism, i.e. primarily through regulation of cristae surface area as opposed to mitochondrial volume density. Furthermore, given the evolutionary age of these living fossils (i.e. approximately 550 million years), this suggests that mitochondrial plasticity has roots extending far back into evolutionary history.     

湖流域土壤微生物基因组总DNA分离纯化及其质粒文库的初步构建

采用研磨／冻融和SDV蛋白酶K热处理以及CTAB处理等理化方法，直接从太湖流域土壤样品中抽提得到微生物总基因组DNA，所得粗DNA用透析袋法以及Nucleaotrap suspension DNA纯化试剂盒进行了纯化，纯化过的DNA能够满是常用限制性内切酶酶切、细菌16S rDNA通用引物和随机引物进行PCR扩增的要求，将所得纯品DNA经过EcoRI部分酶切之后与酶切并脱磷酸的pSK^ 空载体连接，再以大肠杆菌JM109为宿主细胞初步构建了土壤微生物基因组文库，图6参7     

Atmospheric sulfur deposition onto different ecosystems over China

The regional acid deposition model system (RegADMS) was applied to simulate the air sulfur deposition onto different landuse types over China, in which the dry deposition velocities of SO2 and sulfate aerosol (SO4(2-) were estimated by use of a big leaf resistance analogy model and the wet scavenging coefficients were parameterized in terms of precipitation rate. Investigations show that the annual total sulfur deposition over mainland China is 7.24mt (1 mt = 10(6) ton) , in which dry deposition and wet deposition accounts for 56 and 44%, respectively. The sulfur deposition onto agriculture land, grass land, and forest land is 1.09, 3.6 and 1.41 mt, respectively, which sums 6.1 mt and accounts for 84% of the total sulfur deposition. The modeled sulfur deposition was in agreement with the measurement conducted at farmland in Yingtan, a typical read soil region in Jiangxi province of China, during the period of November 1998--October 1999. The total sulfur deposition at the Yingtan site is about 10.3 gm(-2) year(-1) of which 83% is dry deposition. The modeling sulfur deposition at the same site is 8.4 g m(-2) year(-1) Furthermore, the comparison between RegADMS and RAINS-ASIA on modeling regional sulfur deposition shows the consistence of the two models. The correlation coefficient between the simulated sulfur deposition at the medium-large cities reaches 0.72.     

Modified pretreatment method for total microbial DNA extraction from contaminated river sediment

Extraction of high-quality microbial DNA from contaminated environmental samples is an essential step in microbial ecological study. Based on previously published methods for soil and sediment samples, a modified pretreatment method was developed for extracting microbial DNA from heavily contaminated river sediment samples via selection of optimal pretreatment parameters (i.e., reagent solution, reaction duration, and temperature). The pretreatment procedure involves washing the river sediment sample for three times with a solution containing 0.1 mol·L^-1 ethylene diamine tetraacetic acid (EDTA), 0.1 mol·L^-1 Tris (pH 8.0), 1.5 mol·L^-1 NaCl, 0.1 mol·L^-1 NaH₂PO₄, and Na₂HPO₄ at 65°C with 180 r·min^-1 for 15 min to remove humic materials and heavy metals prior to the employment of standard DNA extraction procedures. We compared the results of standard procedure DNA extraction following pretreatment, without pretreatment, and with using a commercial PowerSoil^TM DNA Isolation Kit. The results indicated that the pretreatment significantly improved the DNA quality based on DNA yield, DNA fragment length, and determination of prokaryotic diversity. Prokaryotic diversity exhibited in the DNA with the pretreatment was also considerably higher than that extracted with the PowerSoil^TM DNA Isolation Kit only. The pretreatment method worked well even with a small amount of sediment sample (0.25 g or even lower). The method provides a novel, simple, cost-effective tool for DNA extraction for microbial community analysis in environmental monitoring and remediation processes.     

ABA对米槠和格氏栲种子发育的影响

采用酶联免疫分析法（ＥＬＩＳＡ） 提取和测定米槠和格氏栲发育过程中果实内ＡＢＡ含量,并用不同浓度ＡＢＡ喂养,测定果实不同发育期对外源ＡＢＡ的敏感性．用放射性同位素法测定ＡＢＡ对贮藏蛋白质的积累作用,结果表明：在米槠和格氏栲种子发育过程中内源ＡＢＡ 含量逐渐上升,两者均在１０ 月５ 日达到峰值,米槠为１６４８ ｎｇｇ,格氏栲为１ ４６８ ｎｇｇ,随后ＡＢＡ含量均逐渐下降．果实成熟采收时,ｃ（ＡＢＡ） 仅为高峰时的１９．随着果实的发育对外加ＡＢＡ的敏感性逐渐变小,ｃ（ＡＢＡ）＝１０－４ ｍｏｌＬ－１ 可以完全抑制１０ 月２５ 日（ 米槠） 和１０ 月１５ 日（ 格氏栲） 前种子发芽,但对完全成熟的种子ｃ（ＡＢＡ）＝１０－２ ｍｏｌＬ－１ 亦不能抑制发芽．ＡＢＡ 对成熟前期胚的贮藏蛋白质合成无影响,但能促进成熟中后期胚的贮藏蛋白质的积累作用,ＡＢＡ维持米槠和格氏栲贮藏蛋白质合成和积累作用表现在转录水平上．     

荔枝胚性愈伤组织体胚发生系统的优化及转化抗性愈伤组织培养再生植株

首先优化了荔枝(Litchi chinensis Sonn.)幼胚胚性愈伤组织(embryogenic calli,EC)体胚发生的条件，在附加5mg L^-1 ZT的高糖(60g L^-1)高琼脂(10g L^-1)的MS培养基上获得了荔枝高频率(100％)体胚发生；同时，对荔枝体胚成熟和成苗的关键问题作了分析，建立了一套完整的荔枝体胚发生与再生系统，在附加10％椰子汁的高糖(6％)MS培养基上诱导体胚正常成熟，并在无激素MS培养基中诱导体胚萌发，为荔枝转基因研究提供了良好的受体系统．利用此再生系统对荔枝转基因抗性愈伤组织进行体胚发生与再生植株研究，获得了大量再生植株．图1表7参11     

大肠杆菌JM105中硝酸盐还原酶的制备及其性质

近年来,硝酸盐和亚硝酸盐的污染已引起普遍关注。目前,蔬菜和粮食的NO3-含量过高主要由于农药和化肥的使用、工业废水或生活污水灌溉等,食品类主要来自腌制食品和食品添加剂或防腐剂等。另外,向乳制品或食品中参碱、食盐、化肥、脏水、碱性水等也是硝酸盐污染的来源之一。因此,迫切需要快速、简便、可用于现场的硝酸盐检测方法。这就需仰赖生物酶方法,而生物酶方法的关键是酶制剂的制备和检测方法的建立。本研究通过筛选、厌氧和硝酸盐诱导培养、超声波细胞破碎和差速离心提取等方法,从大肠杆菌(Escherichia coli)JM105细胞膜中制备了硝酸盐还原酶并对其性质进行了研究。结果表明:从大肠杆菌JM105中制得的酶制剂活力很高,且在酶过量的情况下可将NO3-完全转化为NO2-,在用磷酸缓冲液清洗并冷冻保藏过夜后不含有亚硝酸盐还原酶。在加入黄素单核苷酸辅酶(FMN)后,该酶的活力可提高64%,比活力达0.42U·mg-1蛋白。该酶十分稳定,在40℃下24h活力无影响,在浓度为1mmol·L-1的Cu2 、Fe3 、Ca2 、Zn2 、Mg2 和Mo6 存在下,其活力亦不改变。因此,该酶可用于测定食品、蔬菜和环境中硝酸盐的含量。     

Sperm motility and longevity in the giant cuttlefish, Sepia apama (Mollusca: Cephalopoda)

The sperm kinetics and fertilisation literature in marine invertebrates is heavily biased toward free-spawning species. Nonetheless, many species (e.g. cephalopods) transfer and/or fertilise gametes in confined external spaces or internally, creating very different selective pressures on sperm storage, sperm longevity and hence sperm competition. Here we report the results of an investigation into the effects of sperm age, water temperature and sperm concentration on sperm motility in the giant cuttlefish (Sepia apama). Significant positive correlations were found between percent motility and sperm concentration, and between sperm motile speed and sperm concentration. Mean percent motility of cuttlefish sperm suspension was still 9% eight hours after being released from the spermatophore and diluted into filtered seawater at 12°C (ambient field temperature during the spawning season). Sperm resuspended from spermatangia taken from (mated) females in the field were motile for up to 100 hours. When spermatophores were stored at 4°C motility was still observed in resuspended sperm after two months. Our results show that spermatangia and spermatophores can retain and release live sperm for long periods. The observed longevity of sperm in S. apama greatly increases the potential for sperm competition in this species.