利用DSC和EPR技术研究杜仲种子的贮藏条件

种子贮藏稳定性对于种质资源的长期保存具有重要意义，目前关于种子贮藏的最新理论为玻璃态理论，该理论认为种子的玻璃化有利于种子的长期贮藏。当种子处于玻璃态时，玻璃化物质的高度粘滞性降低了种子细胞内分子流动性，阻止了细胞质中分子的扩散，从而减少老化过程中细胞结构的损伤和化学组分的变化，延缓种子老化劣变反应速率，延长贮藏寿命。评价玻璃态的一个重要指标是玻璃化转变温度，当种子贮藏于玻璃化温度或以下10℃～30℃范围内时，种子具有最佳的贮藏稳定性。因此，检测种子的玻璃化转变温度对于种子的长期有效贮藏具有重要指导意义。 本研究将差示量热扫描技术（DSC）与电子顺磁共振波谱仪技术（EPR）应用于杜仲种子玻璃化转变温度方面的研究。在DSC方法中，选用4.4%～31.6%含水量范围的杜仲种胚分别进行了DSC图谱扫描。EPR方法选用3-羧基-2,2,5,5-四甲基吡咯烷-1-氧（3-carboxy-2,2,5,5-tetramethylpyrrolidine-1-oxyl，CP）和2,2,6,6-四甲基哌啶（4-hydroxy-2,2,6,6-tetramethyl-1-piperidinyloxy，TEMPO）作为探针标记杜仲种胚， 利用EPR技术测定不同含水量杜仲种胚的分子运动，通过对EPR图谱参数的分析计算，最终确定不同含水量杜仲种胚的玻璃化转变温度。 DSC实验结果显示，含水量为22.3%、28.0%、31.6%的杜仲种胚在0℃ 左右出现了一个水的熔融峰。该熔融峰的面积代表了自由水含量的多少，随着种胚含水量的降低该熔融峰面积减小。4.4%～31.6%含水量范围的杜仲种胚在-28℃左右还出现了一个熔融峰，推测此峰为杜仲种胚中某类物质熔融所形成的熔融峰。然而在此曲线上我们未观察到标志玻璃化转变的“台阶”出现。 CP-EPR实验的结果表明，利用EPR测定得到含水量为4.4%～11.6%的杜仲种胚在-110℃～20℃温度范围内，同一含水量的杜仲种胚随着温度的升高，分子运动速率加快;在同一温度条件下，高含水量的种胚比低含水量种胚的分子运动速率快。通过CP-EPR波谱两外缘峰最大距离（2Azz）的测定和数据统计分析，得到含水量为4.4%、5.7%、8.6%、10.3%、11.6%杜仲种胚的玻璃化转变温度分别约为44℃、25℃、4℃、-31℃、-43℃。可以把测定的杜仲种胚的这几个含水量的玻璃化转变温度与杜仲种子贮藏相结合，用于指导杜仲种子的贮藏。 TEMPO-EPR实验测定分析得到含水量为2.1%、3.4%、4.8%、8.3%、11.2% 的杜仲种胚的玻璃化转变温度分别为-21℃、-18℃、-24℃、-20℃、-27℃，玻璃化转变温度随含水量升高其变化的规律不明显，这与CP-EPR实验测得的结果有着较明显的差别。通过分析，认为对于脂质含量较高的杜仲种胚，随着含水量的降低，作为标记化合物的TEMPO随着脱水进入脂相，从而不能真实反映出不同含水量种胚的分子运动情况。与TEMPO标记相比，CP标记可能能够更真实地反映不同含水量杜仲种胚细胞质分子运动的情况，根据其分子运动情况得到的玻璃化转变温度更准确。

神农架巴山冷杉种子雨及种实特性研究

巴山冷杉主要分布于秦巴山地，是湖北神农架地区暗针叶林的建群种，构成了当地的优势群落，对神农架地区的植被景观具有决定意义。研究巴山冷杉的结实特性，种子雨时空格局，种子萌发能力和活力劣变过程，有利于进一步认识和分析这一物种的生活史特征，群落组成结构和更新动态。对这一重要物种的合理保护和有效利用具有促进作用。 在湖北神农架自然保护区选择巴山冷杉－箭竹、巴山冷杉－茵芋和巴山冷杉－陕甘花楸群落设置样地，通过布设种子收集器结合室内实验分析，对巴山冷杉种子雨的时空格局进行研究。待巴山冷杉球果成熟时，人工采集球果和种子，测量和比较巴山冷杉的结实特征，利用发芽实验研究巴山冷杉的萌发特性，并通过人工加速老化实验探讨巴山冷杉种子的劣变过程。 巴山冷杉种子雨始于10月初，持续超过2个月，不同类型的巴山冷杉种群表现出不同的种子雨时空格局。巴山冷杉－箭竹群落中巴山冷杉种子雨平均强度167.93±111.14粒/m2，有活力种子比例占22.31%，落种高峰期集中于10月27日到11月2日间，种子雨在种群中呈现聚集分布。巴山冷杉－茵芋群落种子雨强度在三个样地中最小，只有16.41±14.41粒/m2，有活力种子仅占3.05%。巴山冷杉－陕甘花楸群落，种子雨落种高峰为10月15日至10月21日，林冠内种子雨数量占到了收集总数的87.95%。扩散的种子数量与离开中心母树的距离间的关系近似正态分布。 对巴山冷杉的结实特性的分析结果表明，巴山冷杉球果近圆柱形，球果平均长5.37±0.75cm;平均宽3.01±0.32cm;重量介于5.9g～41.3g。巴山冷杉种子长1.08～8.68mm，种子宽1.16～6.42mm，种子厚0.66～3.48mm，种子千粒重7.30g。球果单果出种量在不同种群的频数分布格局不同，巴山冷杉球果和种子的各项指标在种群间和种群内存在显著差异。球果大小、出种量及单粒种子的质量间存在着极显著的正相关关系。此外，种子生活力水平在母树间差异较大，不同母树的种子发芽率和发芽势不等且随时间的萌发进程也不相同，相同种源的巴山冷杉种子在不同的培养条件下萌发能力不同，室温条件下具有最高的发芽率但在25℃，8h光照，恒温培养时的萌发最为整齐。 高温高湿条件能够加速巴山冷杉种子的老化进程，使巴山冷杉种子的含水量发生变化，种子的活力水平随着老化时间的延长而降低，40℃，100% 相对湿度处理6天后种子基本丧失活力。

RNAi沉默光合膜磷脂合成酶基因的研究

磷脂是动物和植物非光合组织细胞膜系统的主要组成成分，在细胞生命过程中扮演着重要角色。尽管绿色植物光合膜的的甘油脂主要是糖脂，但是它仍然含有大约10％的磷脂，说明磷脂在光合膜的结构和功能中起重要作用。构成生物膜的磷脂有多种，但是，光合膜只含有磷脂酰甘油（PG）一种磷脂。光合膜中的PG有其特殊性，即：在PG的sn-2位上总连着一个棕榈酸（16:0）或者反式十六碳烯酸（16:1trans），说明了这种具有特殊结构的甘油脂在维持类囊体膜的结构和功能方面具有重要的作用。 叶绿体中有两个重要酶参与了PG的生物合成，它们分别是胞嘧啶二脂酰甘油合成酶（CDS）和磷脂酰甘油合成酶（PGS）。本实验以烟草和马铃薯为材料，利用RNAi技术，对CDS和PGS基因的表达进行抑制，通过PG缺失突变体，研究其功能。 对转含有PGS片段的沉默结构的转基因烟草叶片膜脂进行了分析，结果表明，与野生型烟草相比较，其PG含量下降了约20%，同时，SQDG和PC的含量增加。PG含量的降低没有引起MGDG和DGDG含量的变化。另外，我们还对转基因植株目的基因片段的RNA表达水平进行了RT-PCR分析，发现其表达量大幅度降低。这些结果表明，在转基因株系中，PGS基因的表达受到了抑制，说明我们获得了PG部分缺失的烟草PGS突变体。 对烟草PG缺失体的PG脂肪酸组成进行分析，表明其特征性脂肪酸反式十六碳烯酸含量明显下降，比野生型降低了44%，C18:0、C18:1和C18:2的相对含量增加，整个变化与总脂脂肪酸变化基本一致。 为了研究PG缺失对光合作用的影响，我们分析了多种光合指标。对叶绿素含量的分析表明，PG含量的降低影响了光合色素的组成。PG部分缺失的转基因烟草中的叶绿素总的含量下降，其中叶绿素b含量下降更为明显，结果，叶绿素a与叶绿素b的比值较野生型高。转基因植株净光合速率下降，二氧化碳利用率降低;PSII的最大光化学效率（Fv/Fm）和实际光化学效率（фPSII）降低，光化学猝灭下降，非光化学猝灭增加，尤其老叶的变化更为明显。这些结果说明了PG的部分缺失影响了植株的光合能力，捕光色素蛋白复合体的结构受到了影响，PSII功能遭受损伤。 同时，我们根据已经报道的马铃薯CDS基因，克隆了一个片段，构建沉默结构，并对沉默结构进行了转化。通过抗性基因的筛选以及RT-PCR检测，证明了沉默结构转化成功，目的基因的表达受到抑制，获得了马铃薯CDS转基因植株。 对马铃薯野生型和CDS转基因植株进行膜脂和脂肪酸分析表明，转基因植株叶片的PE、PG和PC等磷脂含量降低，SQDG和DGDG含量增加;C16:1（3t）、C16:2、C16:3、C18:1和C18:2含量下降，C16:0和C18:3含量增加，而C16:1和C18:0变化不明显。马铃薯CDS转基因植株的叶绿素荧光分析表明，PSII最大光化学效率降低，从野生型的0.82下降到0.77。

云南中部无量山黑长臂猿(Nomascus concolor jingdongensis)栖息地乔木层物种多样性和结构特征

无量山是黑长臂猿景东亚种的唯一分布区,现栖息着98群500余只黑长臂猿。长臂猿是典型的树栖性灵长类动物,通过对无量山西坡大寨子地区250个样方的调查,分析了随海拔梯度升高,黑长臂猿栖息地植被乔木层物种多样性和结构的变化情况。共记录乔木111种,隶属于33科;木质藤本23种,隶属于18科。其中优势科主要有樟科(Lauraceae)、木兰科(Magnoliaceae)、壳斗科(Fagaceae)、山茶科(Theaceae)和杜鹃花科(Ericaceae)。根据不同海拔乔木层物种组成的差异,该区可划分为半湿润常绿阔叶林(2100~2300m)和中山湿性常绿阔叶林(2400~2600m)两种植被类型。多样性指数(Shannon-Wiener指数)随海拔升高在两种植被类型中均呈下降趋势;均匀度指数(Pielou指数)在半湿润常绿阔叶林中呈下降趋势,在中山湿性常绿阔叶林中无明显变化。半湿润常绿阔叶林中,阳坡的乔木物种多样性低于阴坡;而阳坡物种多样性在两种植被类型间差异显著。随着海拔的升高,乔木层的平均高度和上层乔木(Ⅰ层)的比例均逐渐降低,但下层乔木却呈上升趋势。6种优势乔木树种的种群年龄结构分析表明,该区植被的乔木层呈...

赤水河上游云南光唇鱼年龄与生长研究

2007年10月至2008年5月间在赤水河国家级鱼类保护区上游的云南镇雄县坡头乡采集到云南光唇鱼(Acrossocheilus yunanensis)430尾(体长范围57mm～281mm),其中雄鱼77尾,雌鱼97尾,其余的性别不确定,样本为流刺网和电击捕获。通过鳞片对430号标本进行了年龄鉴定,首次对赤水河上游云南光唇鱼的年龄与生长特点进行了研究,分析了体长、体重、鳞径的关系及渔获物的年龄结构组成。所采集云南光唇鱼的年龄由0-5龄组成;体长与鳞径呈线性正相关关系,关系式为:L♀=0.0157R-0.0

浮游绿藻对沉水植物苦草生长的抑制作用

浮游植物大量生长引起的遮光通常被认为是导致富营养湖泊中沉水植物的衰退的主要原因,但是藻类对沉水植物的竞争抑制作用还包括化感作用和无机碳竞争等重要方面,它们可能产生很大影响．为了验证这一观点,本文测定了除遮光以外浮游植物对沉水植物的抑制作用．在相似遮光下(10%-33%),对比研究了不同遮光方式(黑塑料网的物理遮光/浮游绿藻的生物遮光)对苦草(Vallisneria natans)生长和光合作用的影响．结果表明在遮阳网作用下苦草的生长指标(叶数、叶长、叶片叶绿素含量和生物量)和净光合速率都显著高于浮游绿藻作

菜籽粕对异育银鲫免疫应答能力的影响

试验鱼以投喂饲料的不同和是否注射抗原共分为10组,即免疫组:A、B、C、D、E组,免疫对照组:a、b、c、de、组,饲料对照组:A、a组。饲料试验组B、C、D、E、bc、、d、e组。其中,饲料对照组以豆粕和鱼粉为基础蛋白源,饲料试验组分别以双低菜籽粕和普通菜籽粕等氮替代对照组中50%(B、b;D、d)和100%(C、c;E、e)的豆粕蛋白,测定异育银鲫血液白细胞和头肾吞噬细胞的吞噬活性、血清溶菌酶活性、血清补体(C3,C4)含量、血清凝集抗体效价及免疫保护率。结果表明:从免疫后21 d开始,饲料试验组E、

Headspace solid-phase microextraction with novel sol-gel permethylated-beta-cyclodextrin/hydroxyl-termination silicone oil fiber for determination of polybrominated diphenyl ethers by gas chromatography-mass spectrometry in soil

A simple, sensitive, and accurate method for determination of polybrominated diphenyl ethers (PBDEs) in soil has been developed based on headspace solid-phase microextraction (HS-SPME) followed by gas chromatography-mass spectrometry (GC-MS). Permethylated-beta-cyclodextrin/hydroxyl-termination silicone oil (PM-beta-CD/OH-TSO) fiber was first prepared by sol-gel technology and employed in SPME procedure. By exploiting the superiorities of sot-gel coating technique and the advantages of the high hydrophobic doughnut-shaped cavity of PM-beta-CD, the novel fiber showed desirable operational stability and extraction ability. After optimization on extraction conditions like water addition, extraction temperature, extraction time, salts effect, and solvents addition, the method was validated in soil samples, achieving good linearity (r>0.999), precision (R.S.D. < 10%), accuracy (recovery>78%), and detection limits (S/N =3) raging from 13.0 to 78.3 pg/g. (c) 2007 Published by Elsevier B.V.

Allelopathic interactions between Potamogelon maackianus and Microcystis aeruginosa

The allelopathic interactions between Potamogeton maackianus and toxic cyanobacteria (Microcystis aeruginosa) were studied. P maackianus inhibited the growth of M. aeruginosa, both in a coexistence culture system and in exudates experimental culture system. M. aeruginosa also showed effects on the secondary metabolic biosynthesis and secreting behavior of P maackianus. The main lipophilic components of the hexane extracts and the exudates from the macrophyte were analyzed through GC-MS determination. The lipophilic components of the hexane extracts and the exudates from P. maackianus were influenced by M. aeruginosa or their released chemicals. Comparing the lipophilic constituents of the hexane extracts with those in the exudates, the results showed that weak polar compounds contained in the macrophytes can be secreted into the surrounding water. Many compounds, such as N-phenyl-2-naphthalenamine and isopropyl myristate, were detected both in the hexane extracts and the exudates. The changes of lipophilic components in the hexane extracts would be a response to the stress of toxic cyanobacteria or their released toxins. Those changes of exudates, especially the increased content of N-phenyl-2-naphthalenamine, might also be an induced defensive behavior mediated by the released toxins from M aeruginosa. The study results about reciprocal allelopathic responses between macrophytes and cyanobacteria can help in the management of eutrophic waters, and is also important information concerning strategies for recovering eutrophic waters.

Bibarba bibarba: A new genus and species of Cobitinae (Pisces : Cypriniformes : Cobitidae) from Guangxi Province (China)

A new genus of Cobitinae, Bibarba gen. n., and a new species, B. bibarba sp. n., were discovered and are described for the Chengjiang River, a tributary of the Hongshuihe River in Guangxi Province of southern China. This river region is characterized by a Karst landscape, and the river that is inhabited by the new genus is a slowly moving stream with arenaceous and cobblestone beds. The new genus resembles Cobitis Linnaeus, 1758 (subfamily Cobitinae) in the shape and pigmentation pattern of their body, the absence of scales on their head, and the presence of a suborbital spine, but differs from it by a single Lamina circularis on the third pectoral fin ray instead of on the base of the second pectoral fin ray; two pairs of barbels (one rostral pair and one maxillo-mandibular pair) instead of three pairs of barbels (one rostral pair, one maxillary pair, and one maxillo-mandibular pair); a relatively thick and short suborbital spine with a strong medio-lateral process instead of a suborbital spine without or with a weakly formed medio-lateral process as in Cobitis; and the lack of a black stripe extending from the occiput through the eye to the insertion of the rostral barbel. The first two characters have not been reported in any other genus of the subfamily Cobitinae. A morphometric character analysis based on PCA reveals differences between B. bibarba and C. sinensis in body size, barbel length, interorbital width, pectoral fin length in males, and the position of the dorsal and ventral fins. Type specimens of the new species are kept in the Freshwater Fishes Museum of the Institute of Hydrobiology at the Chinese Academy of Sciences in Wuhan, Hubei Province. (c) 2007 Elsevier GmbH. All rights reserved.

Fast-growing transgenic common carp mounfing compensatory growth

Compensatory growth is a phase of accelerated growth apparent when favourable conditions are restored after a period of growth depression. To investigate if F-2 common 'all-fish' growth hormone gene transgenic common carp (Cyprinus carpio) could mount compensatory growth, a 9 week study at 29 degrees C was performed. The control group was fed to satiation twice a day throughout the experiment. The other two groups were deprived of feed for 1 or 2 weeks, respectively, and then fed to satiation during the re-feeding period. At the end of the experiment, the live masses of fish in the deprived groups were still significantly lower than those of the controls. During the re-feeding period, size-adjusted mean specific growth rates and mean feed intakes were significantly higher in the deprived fish than in the controls, indicating a partial compensatory growth response in these fish. No significant differences were found in food conversion efficiency between the deprived and control fish during re-feeding, suggesting that hyperphagia was the mechanism responsible for increased growth rates. The proximate composition of the deprived fish at the end of the experiment was similar to that of the control fish. This study is, to our knowledge, the first to report that fast-growing transgenic fish can achieve partial compensation of growth following starvation. (c) 2007 The Authors Journal compilation (c) 2007 The Fisheries Society of the British Isles.

Progress in the evaluation of transgenic fish for possible ecological risk and its containment strategies

Genetically improved transgenic fish possess many beneficial economic traits; however, the commercial aquaculture of transgenic fish has not been performed till date. One of the major reasons for this is the possible ecological risk associated with the escape or release of the transgenic fish. Using a growth hormone transgenic fish with rapid growth characteristics as a subject, this paper analyzes the following: the essence of the potential ecological risks posed by transgenic fish; ecological risk in the current situation due to transgenic fish via one-factor phenotypic and fitness analysis, and mathematical model deduction. Then, it expounds new ideas and the latest findings using an artificially simulated ecosystem for the evaluation of the ecological risks posed by transgenic fish. Further, the study comments on the strategies and principles of controlling these ecological risks by using a triplold approach. Based on these results, we propose that ecological risk evaluation and prevention strategies are indispensable important components and should be accompanied with breeding research in order to provide enlightments for transgenic fish breeding, evaluation of the ecological risks posed by transgenic fish, and development of containment strategies against the risks.

Pressure-assisted field-amplified sample injection with reverse migrating micelles for analyzing trace steroids in MEKC

This paper describes a novel method that applies pressure-assisted field-amplified sample injection with reverse migrating micelles (PA-FASI-RMM) for the online concentration of neutral analytes in MEKC with a low-pH BGE. After injection of a plug of water into the separation capillary, negative voltage and positive pressure were simultaneously applied to initialize PA-FASI-RMM injection. The hydrodynamic flow generated by the positive pressure compensated the reverse EOF in the water plug and allowed the water plug to remain in the capillary during FASI with reverse migrating micelles (FASI-RMM) to obtain a much longer injection time than usual, which improved stacking efficiency greatly. Equations describing this injection mode were introduced and were supported by experimental results. For a 450-s online PA-FASI-RMM injection, three orders of magnitude sample enhancement in terms of peak area could be observed for the steroids and an achievement of detection limits was between 1 and 10 ng/mL.

Redescription of Nais inflata Liang (Annelida : Oligochaeta) from China

Nais inflata Liang 1963 is redescribed on the basis of fully mature specimens collected from several localities of the Yangtze River and Yellow River. The observations and redescription resulting from the study of mature specimens supports N. inflata as a valid species. This species is characterized by a thickened body, the ventral crotchets all of the same type with distal tooth usually longer than proximal one, a greater number of dorsal chaetae per bundle, stout dorsal needles with equal minute bifid teeth (<1 mu m), presence of penial chaetae, and prostate glands on vasa deferentia instead of atria. The species is most closely related to N. communis Piguet, 1906 and N. variabilis Piguet, 1906. A table comparing allied species is provided.