目的:全面了解菲牛蛭抗凝活性提取物蛋白质生物活性成分和功能.方法:比较了 30％～80％饱和度硫酸铵沉淀法、20％～60％乙醇沉淀法和60％丙酮联合等电点沉淀法分离纯化菲牛蛭抗凝活性物质的方法;采用Nano LC-MS/MS结合生物信息学分析鉴定提取物的蛋白质成分;对提取物进行了体外抗凝溶栓和细胞毒性的功能研究.结果:3种方法抗凝回收率分别为83％、66％和64％.三种方法提取物冻干粉颜色均较浅,无强烈腥臭味.硫酸铵沉淀法得到的冻干粉完全水溶,乙醇和丙酮法有少量不溶.从提取物冻干粉中共鉴定出397种蛋白质,其中生物活性物质除了含有水蛭素,还含有多种生物活性蛋白;冻干粉能延长血浆复钙时间PRT、凝血酶时间TT和凝血酶原时间PT;有体外溶栓效果;不影响正常的星形胶质细胞增殖,可呈浓度依赖性抑制胶质瘤细胞的生长增殖.结论:该研究建立了充分提取活体菲牛蛭抗凝活性物质的方法,抗凝提取物中含有多种具有潜在药用价值的蛋白质成分.该研究对菲牛蛭的应用奠定了基础.

通过校园绿化树种的调查统计,编写了校园绿化树种分种检索表,检索表中所用分类性状主要是容易观察到的茎叶特征,便于师生检索使用.

An identification key to woody plant species was developed based on the survey of plants in Dushuhu campus at Soochow University. The easily observed leaf and shoot traits were used as taxonomic characteristics in the identification key,which enable it to be followed by all kinds of people.

The in vitro culture of Echinodorus subalatus was established by using the unelongated stem as explants. The method of surface sterilization of explants,the effects of different plant growth regulators on the multiplication and rooting were investigated. The results showed that 20% sterile explants could be obtained by twice surface treatment with 0.1% HgCl2 for 8 min and 6-8 min with an interval of 24 h. When cultured on a 1/2 MS multiplication medium supplemented with 6.0 mg·L-1 of 6-BA for 42 d,the test-tube seedling exhibited a multiplication coefficient of 9.3. For optimal rooting,induction on a 1/2 MS rooting medium supplemented with 0.2 mg·L-1 of NAA for 28 d could produce rooted plantlets with 100% rooting rate,and each plantlet generated about 18 roots on average. Rooted plantlets were acclimatized and transferred into outdoor aquatic environment with 100% survival rate.

To investigate the relationship between salt stress and redox status in the hypocotyls of Vigna radiata,the levels of length and fresh weight along with the ratios of reduced and oxidized glutathione(GSH/ GSSG),ascorbic acid(AsA/DHA),nicotinamide adenine dinucleotide(NADH/NAD+) and nicotinamide adenine dinucleotide phosphate(NADPH/NADP+) were determined in the three-old-day seedlings treated with 150 mmol·L-1NaCl for 48 h.Above parameters of control and treatment at 48 h were equal to 493.1% vs 206.9%,661.0% vs 308.2%,118.1% vs 72.2%,444.4% vs 44.4%,96.6% vs 194.3% and 97.1% vs 154.4% of those at 0 h,respectively.The results suggested that the marked inhibition of growth in the hypocotyls of V.radiata by salt stress might be related to the significant variations of redox status.

A rolling leaf rice mutant was obtained by Ac/Ds transposon mutagenesis methods.In this research,we examined the phenotypes of this mutant,and visualized the anatomical structures in leaf tissues.The Ds insertion was also determined by PCR amplification,and the specific sequence flanking the Ds element was isolated by TAIL-PCR technique and then further sequenced.The site of Ds insertion was located by online BLAST sequence alignment algorithms.The results showed that the mutant exhibited a stable rolling leaf phenotype at both seedling and maturing stages,and the bulliform cells appeared abnormal in both number and shape,making them quite different from the fan-shaped structures found in wild-type leaves.Our results also indicated that the mutant genome harbored two components of Ac/Ds system,and the Ds element was identified to locate on chromosome 10.Therefore,it is suggested that this mutant is a kind of Ds-tagged rice rolling leaf mutant abnormal in bulliform cells.These results will provide a fundamental clue to define the roles of Ds-tagged gene in bulliform cell development and differentiation at the molecular level.

With an attempt to offer intensive skill training,microteaching technique is applied to develop mounting skill in the course of slide specimen preparation.The microteaching evaluation forms suitable for mounting skill training due to its superior operability and practicality are worked out by experimental design.Taking three turns within group is proved to be a successful microteaching training mode.

The in vitro culture of Coleus blumei was established by using the nodal stem segments as explants. The method of surface sterilization of explants ,the effects of different plant growth regulators on the muhiplication and rooting were investigated. The results showed that 70% sterile explants could be obtained by surface treatment with 0.1% HgC12 for 8 minutes.When cultured on a MS multiplication medium supplemented with 2.0 mg/L of 6-BA and 0.2 mg/L of NAA for 30 d, the test-tube seedling exhibited a multiplication coefficient of 8.8. For optimal rooting, induction on a 1/2 MS rooting medium supplemented with 0.2 mg/L of 6-BA and 0.5 mg/L of NAA for 21 days could produce rooted plantlets and each plantlet generated 5.6 roots on average. Rooted plantlets were acclimatized and transferred into outdoor environment with 95% survival rate.

In recent years,we have practised management and reform in plant tissue culture research laboratory in opening pattern.Therefore,three models to opening laboratory for undergraduate students have been introduced strategically.It is emphasized that opening laboratory plays an important role in fostering and enhancing the innovation ability of undergraduate students.The measures of improving the management of plant tissue culture research laboratory in opening pattern are subsequently proposed.

1 植物名称 心叶日中花(Mesembryanthemum cordifolium L.F.),实验用植株购自苏州市相城区花木城. 2 材料类别 当年生枝条的带节茎段. 3 培养条件 以MS为基本培养基.(1)初代培养基:MS+6-BA 0.5 mg·L~(-1)(单位下同)+NAA 0.1;(2)增殖培养基:MS+6-BA 1.0+NAA 0.4;(3)生根培养基:1/2 MS+NAA 0.2.

Normal lettuce seeds(Lactuca sativa L.cv Guasihong) were used to understand the relationship between polyamines and nitric oxide(NO) in the process of seed germination after treatments with exogenous polyamines and NO donor SNP(sodium nitroprusside) together with NO specific scavenger cPTIO.The results showed that exogenous polyamines,especially Spd,had a certain role in promoting early germination and the optimal concentration was 0.5 mmol·L-1.Their effect was more significant before 48 h after seed imbibition.Consistent with the result,1.0 mmol·L-1 cyclohexylamine(CHA,spermidine synthase inhibitor) had a strong inhibitory effect on the germination of lettuce seeds.Like Spd,exogenous NO donor SNP also promoted seed early germination of lettuce,but in a concentration-dependent way,its function disappeared after 48 h.When treated with SNP and Spd,application of NO scavenger cPTIO could markedly reduce the SNP-or Spd-promoted effect on lettuce seed germination.Fluorescent detection using the DAQ,a NO-sensitive fluorescent probe,showed that there was strong NO fluorescence near seed embryo treated with Spd or SNP.The fluorescence could be eliminated by cPTIO concomitantly with its inhibitory effect on the early germination promoted by Spd or SNP.In this paper,we preliminarily approve that polyamines-,especially spermidine-induced early germination of lettuce seeds may be relevant to the NO induction.

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洞庭碧螺春是中国十大名茶之一,苏州太湖之东西洞庭山为碧螺春原产地[1].名茶的基础是良种,因此加强原产群体种茶树种质资源的保护和开发十分重要[2].虽然有关茶树的组织培养曾有报道[3],但碧螺春茶树的组织培养研究尚未见报道.本试验以从碧螺春原产地选出的早芽茶的腋芽为外植体,对无菌外植体的建立、试管苗的增殖和生根等各个阶段的培养条件进行优化,以期为碧螺春茶树优质种苗生产奠定基础.

The culture conditions of callus induction,plantlet regeneration and subculture were studied using stems of Pedilanthus tithymaloides(L.)Poit.var.nanus Dressler as explants.The results show that the optimal sterilizationmethod of stemsegments of P.tithymaloides var.nanus is soaking in 1.0 g·L-1 HgCl2 solution for 4-10 min.The optimal medium of callus induction is MS medium containing 1.5 mg·L-1 6-BA,0.10-0.15 mg·L-1NAA,30 g·L-1sucroseand 6 g·L-1 agarpowder(pH 5.78-pH 5.80).The optimal rooting medium with rooting rate of 93.3% is the rooting medium(1/2 MS,containing 15 g·L-1sucrose and 6 g·L-1 agar powder,pH 5.78-pH 5.80)containing 0.2 mg·L-1 NAA,and the survival rate of P.tithymaloides var.nanus plantlets reach above 95.0% after hardened and transplanted.The optimal subculture medium of P.tithymaloides var.nanus plantlets is the rooting medium containing 0.1 mg·L-1 NAA,and the optimal temperature and light intensity in subculture process is 15 ℃ and 20 μmol·m-2·s-1,respectively.Moreover,the callus of P.tithymaloides var.nanus can directly differentiate and form a large amount of rosette buds,so the multiplication goal of P.tithymaloides var.nanus plantlets can be achieved.

The in vitro culture of Limnophila sessiliflora was established by using the nodal stem segments from submerged branch as explants.The method of surface sterilization of explants,the effects of different plant growth regulators on the multiplication and rooting were investigated.The results showed that 15% sterile explants could be obtained by twice surface treatment with 0.1% HgCl2 for 4 min with an interval of 4 h.When cultured on a 1/2 MS multiplication medium supplemented with 2.0 mg·L-1 of 6-BA and 0.1-0.2 mg·L-1 of NAA for 35 d,the test-tube seedling exhibited a multiplication coefficient higher than 30.8.For optimal rooting,induction on a 1/2 MS rooting medium supplemented with 0.3 mg·L-1 of 6-BA and 0.5 mg·L-1 of NAA for 28 d could produce rooted plantlets with 3-5 lateral shoots,and each plantlet generated 4.8 roots on average.Rooted plantlets were acclimatized and transferred into outdoor aquatic environment with 100% survival ratio.

Insertion mutagenesis with T-DNA or transposable elements is currently one of the most powerful tools for screening various types of mutations in higher plants,and for analyzing functional information of genes related to those mutations.By using the Ac/Ds transposable elements system in rice,a great deal of Ds-inserted mutants may be efficiently generated,and used for functional identification of genes that influence important agronomic traits,by which the researches in rice functional genomics have been greatly promoted.As a key agronomic trait related to grain production in rice and other grass species and an important property parallel to branching in dicotyledonous plants,tillering in rice has attracted more research interests.In the present study,a rice tillering mutant was isolated from a collection of Ds-inserted mutant lines,and was designated dt1(double tillers mutant) according to its tillering phenotype of a major tiller and a minor tiller developed at the same tillering node.Both tillers were found to be productive with the ears normally emerged from each of them.The Ds-flanking sequence was cloned from this mutant by using TAIL-PCR amplification technique.The sequenced Ds-flanking sequence was used as a query to perform an online search for homologous sequence against nucleotide sequence database by NCBI-BLAST,the results indicated that the Dsflanking sequence showed 100% identity at the nucleotide level to the sequence of clone OJ1345H02(gi｜21281466) of rice chromosome 3.The structure of the Ds-inserted gene was predicted by using software of FGENESH and GeneMark,respectively,and gotten highly similar in the number,size and location of exons within the gene.The results also showed that the Ds element was inserted into the region between transcription start site and the first exon,at 173 bp upstream of the first exon.Moreover,the product encoded by the gene was predicted by NCBI Entrez server and Pfam.It was revealed that the deduced product was a rice formin-like protein containing a highly conserved FH2-domain.Given the fact that the Ds element as big as 5.9 kb in size was inserted immediately into upstream of the first exon,it is suggested that Ds insertion probably make the gene loss its function,and thereby generate the rice mutant dt1.In addition,the genotyping for Ds insertion in the progeny plants of dt1 mutant indicated that these progeny plants segregated into homozygous and heterozygous types for Ds insertion and Ds free plants,which suggested that the dt1 mutant was heterozygous for Ds insertion.

从正常生长的豌豆植株上采集具有各类不同侧生器官的茎段及节部,制备成透明、半透明标本及连续石蜡切片,通过观察,绘制出豌豆茎主要维管束走向、茎叶维管束联系、侧枝与主茎维管束联系的模式图,从解剖学角度阐明豌豆叶片同化产物分配规律的特殊性。

<正>1植物名称地中海绵枣儿(Scilla peruviana L.)。2材料类别鳞茎,植株购于苏州花鸟市场。3培养条件(1)初代培养基:MS+6-BA 0.5mg·L-1(单位下同)+NAA 0.1+GA3 0.1;(2)腋芽诱导培养

夏蜡梅(Sinocalycanthus chinensis Cheng et S. Y. Chang)为中国特有的三纪孑遗植物,特产于浙江临安县、昌化、天台,是蜡梅科唯一夏季开花的植物,但种源数量稀少,自然生境恶化,处于濒危状态.

通过对苏州市205个道路绿化植物群落的抽样调查,发现城市道路绿化树种达140余种,其中有123种绿化常用树种。在此基础上,统计道路绿化各常用树种频度、丰度以及各绿化植物群落的加权平均树种数等生态学指标,对苏州城市道路绿化树种资源的利用现状和存在的问题进行分析,提出城市道路绿化树种应用的若干发展对策。