五助百里香植物愈伤组织培养及挥发油成份的生物合成研究

高等植物具有合成各种化合物的特殊功能。早在本世纪二十年代初期，许多有机化学家对这些复杂的天然次生代谢物提出了各种生源假说，阐述这些化合物的结构关系及其可能的生源途径。 目前，对于次生代谢产物生物合成研究主要有二个途径，整体方法和离体方法，其中离体方法包括非细胞系以及植物组织和细胞培养。 经过近一个世纪的努力，植物组织和细胞培养成了今天研究次生代谢产物生物合成的一个重要工具。目前这方面研究主要包括三个方面;﹤一﹥通过植物组织和细胞培养获得有价值的天然次生代谢产物;﹤二﹥研究次生代谢产物的生物合成途径;﹤三﹥生物合成新的有特殊价值的天然产物。 近二十年来，这一领域发展迅速，尤其是在寻找用植物细胞，像微生物那样，工业生产有价值的天然产物方面，为许多科学家所瞩目。 为了从植物组织和细胞培养中获得原植物特有的次生代谢产物，目前，主要包括四个方面的研究。﹤一﹥营养调节;﹤二﹥激素调节;﹤三﹥一级代谢对二级代谢的调节;﹤四﹥一些物理因子的调节。这些工作均在不同程度上取得一定的进展。 目前，在本领域中存在的主要问题是：﹤1﹥在植物组织培养中，许多植物特有的次生产物只能在在具有一定程度分化的细胞中才能产生;﹤2﹥在植物组织或细胞培养中，合成的次生产物含量较低;﹤3﹥许多植物组织或细胞培养不能合成原植物特有的次生代谢产物。 在我国，应用植物组织或细胞培养研究次生代谢产物合成的工作开展不多，尤其是有关精油的生物合成研究，目前才刚刚开始。 本实验应用两种化学型（芳樟醇型和香芹酚型）五肋百里香（Thymus guinguecostatus celak)植物的叶、茎、子房、花粉等诱导愈伤组织，以期探讨萜类及有关成份在愈伤组织中合成的规律;比较不同愈伤组织合成萜类及有关成份的差异;探讨不同化学型的遗传信息在愈伤组织中合成的作用。 在本实验中，我们的结果如下： 1.愈伤组织诱导 不同培养基对愈伤组织诱导不同，其中以添加6ppm 2,4-D和0.5ppm激动素的staba培养基的诱导率最高，接种17天后，诱导率达70%。老茎和老叶均不易产生愈伤组织。光照下培养也不易产生愈伤组织，诱导愈伤组织是在暗处进行的。 2.愈伤组织培养 愈伤组织在含有0.2ppm NAA和1ppm激动素的 staba 和 Ms 培养基中生长较好，五周后可长成约1克重。在光照下培养，茎的愈伤组织是绿色的，叶的愈伤组织为白色，无叶绿素，子房和花药愈伤组织含有少量叶绿素。光照下培养比暗处生长较快。在这两种培养基中添加200毫克/升亮氨酸时，茎的愈伤组织出现分化的不定芽，叶的愈伤组织不发生分化，但生长加快。 3.原植物精油成份 移栽于本所北京植物园的五肋百里香精油成份，经气-质联用鉴定,与原产地的精油成份相似。从山东烟台地区移栽的芳樟醇型，经气-质联用鉴定有13个化合物，除前已坚定的五个原产地精油成份，如芳樟醇，龙脑，莰烯，蒎烯和β-红波药烯外，本实验鉴定的成份。 从山东泰安地区移栽的为香芹酚型经气-质联用鉴定的精油成份有12个，除前已鉴定的香芹酚，百里香酚等9个成份外，本实验鉴定的成份有桧烯，α-侧柏烯和新异侧柏醇等。 4.愈伤组织中萜类和有关成份及其生源 芳樟醇型叶愈伤组织，经提取分离，气-质联合检出4个化合物：异戌酰胺，苯甲酸乙酯，橙花叔醇和香兰素。主要成份是异戌酰胺和苯甲酸乙酯。 在香芹酚型愈伤组织中，鉴定了一个化合物，异戌酰胺，它是该愈伤组织的主要成份。 在本实验中，没有检出单萜成份，但检出一个倍半萜，橙花叔醇。因此，我们认为单萜和倍半萜的生物合成部位可能是相互分离的。倍半萜可能是在细胞质中合成的。 在本实验中，我们认为主要成份异戌酰胺可能是由亮氨酸转化的;香兰素和苯甲酸乙酯可能是从肉桂酸途径衍生的。

HACCP based management system for improving quality of freshwater giant prawn, Macrobrachium rosenbergii

The study was conducted on the present status of HACCP based quality management system of golda, Macrobrachium rosenbergii farms in Fulpur region of Mymensingh. Information was collected on general condition of farms, culture systems and post-harvest quality management. In almost all farms, there is no or inadequate infrastructure facilities such as, road access, electric supply, telecommunications, ice, feed storage facility, vehicle for golda transportation, washing and toilet facilities. The problems associated with sanitation and hygiene was: widespread use of cow dung, poultry manure and construction of open toilet within the vicinity of prawn culture pond. Different grades of commercially available and locally prepared feeds were used for golda culture in the pond. Golda post-larvae (PL) of 40-50 days old were stocked with carp species. The price of golda PL ranged from Tk. 1.00 to Tk. 1.25/piece. The pond size varied from 50 decimal (0.2 ha) to 2.5 acre (1.0 ha) with an average depth of 2-2.5 m. The culture period of golda varied from April-May to November-December and survival rate ranged between 75 and 80%. Production of golda varied from 250-500 kg/acre (625-1,250 kg/ha). Harvested golda were transported to city market within 4 h. Two size grading were generally followed during pricing, e.g. Tk. 500 to 550/kg for >100 g size and Tk. 300/kg for <100 g size. The cost-benefit ratio was found to remain around 1:1.25 depending on availability of PL. Water quality parameters such as, water temperature, pH, dissolved oxygen, total alkalinity and chlorophyll a in five golda farms in Fulpur region were monitored. Water temperature ranged from 29°C to 33°C, dissolved oxygen from 2.28 to 4.13 mg/l, pH between 6.65 and 7.94, alkalinity from 44 to 70 mg/l and chlorophyll a concentration from 61.88 to 102.34 µg/l in the five investigated ponds. The Aerobic Plate Count (APC) of the water sample was within the range of 2.0x10^6 - 2.96x10^7 CFU/ml and of soil samples within the range of 6.9x10^6 - 7.73x10^6 CFU/g. Streptococcus sp., Bacillus sp., Escherichia coli, Staphylococcus sp., Pseudomonas sp. and Salmonella sp. were isolated from pond water and sediment. Different feed samples used for golda was analyzed for proximate composition. Moisture content ranged around 14.14-21.22%, crude protein 20.55-44.1%, lipid 4.67-12.54% and ash 9.7-27.69%. The TVB-N values and peroxide values of feeds used as starter, grower and fish meal were found within the acceptable ranges and samples were free from pathogenic organisms. A training was organized for the golda farmers on HACCP, water quality and post-harvest quality management of prawn.

12种鳃形目鱼类mt DNA NI4 L- NI4基因的序列变异及其分子系统学

鳃形目鱼类是一类在鱼类乃至脊椎动物进化史上占有很重要地位的古老濒 危鱼类，长期悬而未决的系统演化关系为世人瞩目.采用DNA测序技术首次测定了 包括中国特有鳃形目鱼类在内的12种鳃形目鱼类的mtDNA ND4L和ND4基因(703 by)的序列，并进行了分子系统学分析.从分子系统学的角度得出了如下结论:(1) 鲤属可以归并到鳃属;(2)达氏鳃与中华鳃的亲缘关系最近，很有可能为中华鳃的 一陆封类型;(3)环太平洋地区的鳃科鱼类可能有共同的起源;(4) ND4L和ND4基 因是进行鳃形目鱼类系统演化研究很好的遗传标记.

Study on the quality of beche-de-mer in trade and shrinkage of specimens during processing

The beche-de-mer industry in India is a cent percent export oriented industry being confined to south east coast in Palk Bay and Gulf of Mannar in Tamil Nadu. Chemical quality of 180 trade samples of beche-de-mer of four sizes collected from the beche-de-mer curing centres of Ramanathapuram district was studied. Moisture ranged from 6.2 to 24.4% and sand content from 0.11 to 20.42% for all grades. Mean values of sand content are for grade 1=3.47%, grade 2=4.50%, grade 3=3.68%, grade 4=6.87%. Sodium chloride was almost constant for all grades at 5.7%. TVBN values ranged from 10 to 78.4 mg%. 44 laboratory samples of different grades were prepared following trade practice and examined for chemical quality. Mean moisture values are for grade 1=13.4%, grade 2=12.44%, grade 3=12.62%, grade 4=12.08% and mean values of sand are for grade 1=0.70%, grade 2=0.90%, grade 3=1.16%, grade 4=2.15%. The percentage of shrinkage of the animals ranged from 56% to 60% for dried beche-de-mer of 7.5 cm size and above.

抗HIV p24多肽单克隆抗体和抗血清的制备及初步应用

检测p2 4抗原的诊断试剂在AIDS研究和防治方面具有重要意义。为研制国产HIVp2 4抗原诊断试剂 ,根据国内外文献 ,合成了HIVp2 4抗原的六个肽段 :G -A 12 ,D -R 16 ,P -S 18,A -G 2 3(HIV - 2ROD) ,P -S 18,N -I 15以及D -C 2 2。六个肽段中 ,N -I 15肽和D -C 2 2肽用于制备McAb ,其余 4个用于制备山羊抗血清。除D -R 16肽的滴度较低 (1∶2 0 0 0 )外 ,其余三个肽的抗血清滴度都在 1∶10 0 0 0以上。McAb中 ,N -I 15肽的反应比D -C 2 2肽要强 ,但两者的腹水滴度相同 ,均为 1∶10 0 0。单抗铺板检测病毒 p2 4抗原的非特异性反应比较强。用山羊抗血清铺板 ,1∶80 0 0的稀释度检测效果最好。用我们研制的抗体检测不出裂解HIV - 1ⅢB中 p2 4 ,却能够测出Abbott公司p2 4抗原诊断试剂盒的阳性对照 (HIVAG - 1)。用北海道大学免疫科学研究所研制的单抗和人阳性血清做的交叉对照实验表明 ,可能是由于合成肽免疫产生的抗体与裂解病毒p2 4的亲和力较差引...

The zonation of soft-bottom sediments in Gorgan Bay (Caspian Sea) using macrobenthos as indicator of ecological health

In this study, in order to assess the ecological health status and zoning of soft bottom of Gorgan Bay, the spatial and temporal distribution of macrofauna and their relationship with environmental stress were investigated. Sediment samples were collected using a Van Veen grab at 22 sampling points, seasonally during 2012-2013. The averages (±SD) of the percentages of sand, silt, clay and TOM (Total Organic Matter) in the sediment samples were determined (44.4± 15, 53.4 ± 14, and 2.2 ±2.2 and 7.2% ± 1.6, respectively). Our results showed that mean (range) of Al, As, Cu, Fe, Ni, Pb and Zn in the sediment samples were 1.2 % (0.4-2.1), 4.8 (2.5- 10.3) ppm, 10.5 (4.4-16.9) ppm, 1 (0.4 – 1.6) % , 13.6 (6.2-21.5) ppm, 9.1 (4.7-12.9) ppm and 23.9 (3.1-39.4) ppm, respectively. In spring, both Al and Ni were higher than the guideline level. In the event that arsenic was exceeds the guidelines in summer. In this study, 14 species of macrofauna from 12 families were identified. Polychaeta with 3 species was the most dominant group in terms of abundance. The four most abundant taxa making up 85% of all specimens (Streblospio gynobranchiata, Tubificidae, Hediste versicolor and Abra segmentum). The western area were characterized by the higher species diversity (H', 1.94). So Gorgan Bay presents transitional macrobenthic assemblages that are spatially distributed along substrate gradients .The mean of Shannon index, BENTIX, BO2A, AMBI and M-AMBI in the bay was 1.3, 2.2, 0.4, 3.2 and 0.65 respectively. According to the results of these indices, ecological status of the western part of the bay assessed better than the other parts. According to the results of the nmMDS (non-metric Multidimensional Scaling), PCA (Principal Components Analysis), the map of distribution of heavy metals and the map of the ecological status , it seems Gorgan Bay is divided into two separate zones (the eastern and the western parts).M-AMBI finaly introduced reliable index for assessing the ecological status of the Bay.

The genetic diversity of Acipenser stellatus populations in the Caspian Sea was studied using microsatellite technique

A total of 361 caudal fin samples were collected from adult A. stellatus specimens caught in the north Caspian Sea, including specimens from Kazakhstan (Ural River), Russia (Volga River), Azerbaijan (Kura River), specimens caught in the south Caspian Sea including specimens from Fishery Zone 1 (from Astara to Anzali), Fishery Zone 2 (from Anzali to Ramsar), Fishery Zone 3 (from Nowshahr to Babolsar), Fishery Zone 4 (from Miyankaleh to Gomishan) as well as from specimens caught in Turkmenistan (all specimens were collected during the sturgeon stock assessment survey). About 2 g of fin tissue was removed from each caudal fin sample, stored in 96% ethyl alcohol and transferred to the genetic laboratory of the International Sturgeon Research Institute. Genomic DNA was extracted using phenol-chloroform method. The quality and quantity of DNA was assessed using 1% Agarose gel electrophoresis and Polymerase Chain Reaction (PCR) was conducted on the target DNA using 15 paired microsatellite primer. PCR products were electrophoresed on polyacrylamide gels (6%) that were stained using silver nitrate. Electrophoretic patterns and DNA bands were analyzed with BioCapt software. Allele count and frequency, genetic diversity, expected heterozygosity and observed heterozygosity allele number, and the effective allele number, genetic similarity and genetic distance, FST and RST were calculated. The Hardy Wienberg Equilibrium based on X2 and Analysis of Molecular Variance (AMOVA) at 10% confidence level was calculated using the Gene Alex software. Dendrogram for genetic distances and identities were calculated using TFPGA program for any level of the hierarchy. It is evident from the results obtained that the 15 paired primers studied, polymorphism was observed in 10 pairs in 12 loci, while one locus did not produce DNA bands. Mean allele number was 13.6. Mean observed and expected heterozygosity was 0.86 and 0.642, respectively. It was also seen that specimens from all regions were not in Hardy Wienberg Equilibrium in most of the loci (P≤0.001). Highest Fst (0.063) was observed when comparing specimens from Fishery Zone 2 and Fishery Zone 4 (Nm=3.7) and lowest FST (0.028) was observed when comparing specimens from the Volga River and those from the Ural River (8.7). Significant differences (P<0.01) were observed between RST recorded in the specimens studied. Highest genetic distance (0.604) and lowest genetic resemblance (0.547) were observed between specimens from Fishery zones 2 and 4. Lowest genetic distance (0.311) and highest genetic resemblance (0.733) was observed between specimens from Turkmenistan and specimens from Fishery zone 1. Based on the genetic dendrogeram tree derived by applying UPGMA algorithm, A. stellatus specimens from Fishery zone 2 or in other words specimens from the Sepidrud River belong to one cluster which divides into two clusters, one of which includes specimens from Fishery zones 1, 3 and 4 and specimens from Turkmenistan while the other cluster includes specimens from Ural, Volga and Kura Rivers. It is thus evident that the main population of this species belongs to the Sepidrud River. Results obtained from the present study show that at least eight different populations of A. stellatus are found in the north and south Caspian Sea, four of which are known populations including the Ural River population, the Volga River population, the Kura River population and the Sepidrud River populations. The four other populations identified belonging to Fishery zones 1, 3, and 4 and to Turkmenistan are most probably late or early spawners of the spring run and autumn run of each of the major rivers mentioned. Specific markers were also identified for each of the populations identified. The Ural River population can be identified using primers Spl-68, 54b and Spl-104, 163 170, 173, the Volga River population can be identified using primers LS-54b and Spl-104, 170, 173 113a and similarly the population from the Kura River can be identified using primers LS-34, 54b and Spl-163, 173 and that from the Sepidrud River can be identified using primers LS-19, 34, 54b and Spl-105, 113b. This study gives evidence of the presence of different populations of this species and calls for serious measures to be taken to protect the genetic stocks of these populations. Considering that the population of A. stellatus in Fishery zone 2 is an independent population of the Sepidrud River in the Gilan Province, the catch of these fishes in the region needs to be controlled and regulated in order to restore the declining stocks of this species.

兔受精卵显微注射外源基因后体外培养的卵裂发育

从超数排卵的14只母兔获得438枚受精卵, 卵龄16-22h。显微操作在带微分干涉和相差的Nikon倒置显微镜下进行。注射针的尖端外径0.5#mu#m, 离尖端40和80#mu#m处的外径分别为4.2和6.5#mu#m。注射用外源基因是绵羊生长激素基因与MT-1启动基因藕连的线状DNA溶液(1ng/#mu#l)。140枚注射的受精卵和未注射的145枚受精卵(对照), 在Ham^sF-10培养液(补充生长因子)中培养(38℃, 5％的CO_(2))。结果培养48h后, 注射组卵裂发育率分别是: 未卵裂7.9％(11/140)、卵裂至2-4细胞期11.0％(16/140)、卵裂至8-16细胞期80.7％(113/140)。对照组相应的卵裂率分别是4.1％(6/145)、12.4％(18/145)和83.4％(121/145)。两组卵裂发育率相近。

广西华缨鱼属鱼类一新种记述

在整理华缨鱼属标本时发现，1993年9月在广西壮族自治区天峨县红水河水系地下河采集到的一批 标本为一个未经发表的新种，新种订名为大眼华缨鱼(＆加c瑚soc^eiz獬胧gnzDp^f^8h邯)。其下咽齿2行，可与 下咽齿3行的7种华缨鱼相区别，而与属内同样具2行下咽齿的贵州华缨鱼(s．肛妇b淝瑚括)、小口华缨鱼 (Js．m幻M幻，抛￡琳)和宽唇华缨鱼(5．z06缸t琳)亲缘关系较近。但：(1)新种胸鳍中点上方无黑斑，背鳍分枝 鳍条7，腹鳍分枝鳍条7，背鳍前鳞15—16，眼大，头长为眼径2．5—3．1倍，眼径为头宽44．7％一57．8％，吻 须后伸至前后鼻孔之间或眼前缘，口角须后伸至眼前缘至眼中之问或眼中至眼后缘之间，体长为尾柄高8．9一 lO．7倍，头长为吻长2．5—3．7倍，可与贵州华缨鱼(胸鳍中点上方有一明显黑斑，背鳍分枝鳍条8，腹鳍分枝 鳍条8，背鳍前鳞12—14，头长为眼径4．O一5．0倍，眼径为头宽16．6％一20．7％，吻须后伸不达后鼻孔后缘， 口角须后伸至眼前缘，体长为尾柄高7．2—8．2倍，头长为吻长1．9—2．2倍)相区别；(2)新种背鳍前鳞15— 16，侧线鳞39～40，侧线上鳞4．5—5．5，背鳍分枝鳍条7，鳃耙13，腹鳍末端伸达肛门，眼径为头宽44．7％一 57．8％，可与宽唇华缨鱼(背鳍前鳞22，侧线鳞42—45，侧线上鳞6．5，背鳍分枝鳍条8，鳃耙10，腹鳍末端 不达肛门，眼径为头宽23．3％一30．O％)相区别；(3)新种与同水系的小口华缨鱼在鳍条数、侧线鳞、体色、 斑纹等方面最为相似，但新种围尾柄鳞14／16，眼大，头长为眼径2．5—3．1倍，腹鳍末端伸达肛门，口角须后 伸至眼前缘至眼中之间或眼中至眼后缘之间，吻皮边缘深裂成小穗，背鳍起点距吻端较距尾鳍基为近，背鳍长 大于头长，体长为尾柄长4．8—5．9倍，头长为吻长2．5—3．7倍，尾柄长为尾柄高1．6—2．1倍，可与之(围尾 柄鳞12，眼小，头为眼径4．4—4．6倍，腹鳍末端不达肛门，口角须伸达眼下方，吻皮边缘不开裂或开裂不明 显，背鳍起点距吻端等于距尾鳍基，背鳍条约等于头长，体长为尾柄长6．1—6．4倍，头长为吻长2，1—2．4倍， 尾柄长为尾柄高1．4—1．5倍)相区别。

香溪河库湾轮虫现状及水质评价初探

香溪河是长江三峡水库湖北省最大的支流。通过对香溪河库湾(下游河段)一周年的调查,发现轮虫80种,其中占优势的有13种。轮虫密度在不同样点差异显著,样点Ⅸ最低(仅为68.96 ind./L),样点Ⅵ最高(为1505.83ind./L);对9个样点进行聚类分析,可分为4组,第1组为样点Ⅰ至Ⅴ,第2组为样点Ⅵ,第3组为样点Ⅶ、Ⅷ,第4组为样点Ⅸ;除样点Ⅸ外,沿着水流方向,轮虫密度逐渐降低;在时间上差异也显著(P<0.01),9月份最高(2664.17ind./L),12月份最低(85.00ind./L)。水质评

密度和温度对萼花臂尾轮虫产卵量和混交雌体的影响

用小球藻(Chlorella sp.)为食培养萼花臂尾轮虫(Brachionus calyciflorus),食物浓度约1×10~6cells/mL,温度25℃,光照强度约40001x,昼长比LD=18:6。在1.0mL、0.5mL和0.25mL培养液中同时进行单个体和群体培养。结果发现:1)单个体培养时每个母体平均产卵量分别为12.0,13.8和7.8个;后代个体中混交雌体的百分比分别是46.38％,53.49％和55.83％。2)群体培养时,每个母体平均产卵量分别为8.7、3.1和2.65个,随密度增

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Changes in plasma thyroid hormones and cortisol levels in crucian carp (Carassius auratus) exposed to the extracted microcystins

The endocrine response of crucian carp injected intraperitoneally with extracted microcystins (MC) was investigated in this study. Fish were injected intraperitoneally either with 0.75% NaCl (control) and Microcystis extract corresponding to 150 and 600 mu g microcystins per kg body weight. The plasma levels of triiodothyronine (T-3), thyroxine (T-4), free triiodothyronine (FT3), free thyroxine (FT4), and cortisol were determined at 0, 1, 3, 12, 24. and 48 h post-administration of MC-containing extract. Treated fish displayed abnormal behaviors, Such as a startle response and disoriented swimming, as well as changes in ventilation rates. Plasma cortisol concentrations of fish in both dose groups significantly increased after administration of extracted MC and remained high throughout the experiment, which suggested that MC elicited a stress response in treated fish. The profiles of cortisol changes in treated fish appeared to be dose dependent, indicating that fish in the high dose group experienced greater MC-incluced disturbance. Mortality occurred after 12 h in the high dose group. Plasma levels of T-4, T-3, FT4, and FT3 did not vary significantly between the control fish. In contrast to this, fish exposed to MC-containing extract showed significant declines in T-3, FT4, and FT3 levels in a dose-depenclent manner throughout the experiment. Plasma T4 levels, however, did not vary significantly in the low dose group, whereas they decreased significantly it 48 h post injection in the high dose group. This study demonstrates that administration of microcystins-containing extract causes a stress response and reduces the plasma levels of thyroid hormones in crucian carp. These results illustrate that microcystins exerted potent effects on the endocrine system of crucian carp, through activating their hypothalamus-pituitary- interrenal axis and disturbing thyroid function. (c) 2008 Elsevier Ltd. All rights reserved.