利用杀配子染色体2C诱导中国春-长穗偃麦草1E二体附加系染色体畸变的研究

偃麦草属具有许多优良的遗传性状,利用杀配子染色体诱导小麦-偃麦草的染色体易位,是创建小麦异源易位系、丰富小麦遗传资源的一个有效途径。杀配子染色体2C在单体附加时,可高效率地诱导染色体畸变,得到易位、缺失等畸变类型。本研究利用含有杀配子染色体的中国春-柱穗山羊草2C(Ae.Cylindrical2n=28CCDD)二体附加系与中国春-长穗偃麦草(E.elongate2n=14EE)1E二体附加系杂交。观察F1花粉母细胞减数分裂情况,发现存在到量的染色体异常行为。单价体数超过理论值,棒状二价体数明显增加,并出现一定频率的三价体和四价体,及大量的染色体断片、染色体桥和落后染色体、微核,并伴随发生多极分裂现象。经C-分带与荧光原位分子杂交鉴定83株杂交F2,共检测到5株易位、6株缺失。易位频率为6.02%,缺失率为7.22%,染色体畸变的总频率为13.24%。并对杀配子染色体的诱变机制及引起杂交后代结实率降低的原因进行了讨论。     

甘蓝型油菜与Brassica maurorum的异源六倍体后代及BC_2细胞学分析

将甘蓝型油菜品种中油821（2n=38,AACC）与芸薹属野生资源Brassica maurorum（2n=16,MM）的三倍体杂种进行染色体加倍,得到异源六倍体（2n=54,AACCMM）。该多倍体雄性高度不育,雌性育性极低。在其花粉母细胞的减数分裂终变期,染色体平均配对构型为1.17Ⅰ＋20.71Ⅱ＋0.56Ⅲ＋2.25Ⅳ＋0.08Ⅴ＋0.06Ⅵ。用甘蓝型油菜与该多倍体连续回交2次,均需借助幼胚培养才得到后代植株,BC1、BC2植株均雄性不育,雌性育性很低。BC2植株的形态特征接近甘蓝型油菜,但各植株形态有明显差异。基因组原位杂交分析表明,BC2植株含有2～5条M基因组的染色体,甘蓝型油菜染色体多配对形成二价体,附加的染色体多以单价体形式存在,或与甘蓝型油菜染色体发生联会配对。     

甘蓝型油菜与诸葛菜属间杂交F12群体的品质及遗传分析

以甘蓝型油菜（AACC,2n=38）品种Oro与诸葛菜（OO,2n=24）属间杂交F4世代中出现的五倍体单株（AACCO,2n=50）为起始材料连续自交选育、鉴定,直至获得F12群体。各F12株系间表现一定的形态差异,但株系内植株形态特征一致。细胞学观察表明85.3%的F12单株具有与甘蓝型油菜相同的染色体数目,花粉母细胞（PMCs）减数分裂中正常配对和分离;其余植株的染色体数目分别为37、39和40;红紫色植株在减数分裂中有落后染色体,四分体时期会出现微核。基因组原位杂交（Genomic in situ hybridization,GISH）分析未检测出整条的诸葛菜染色体;但AFLP分析表明存在渗入的诸葛菜DNA片段。F12植株的脂肪酸组成及硫苷含量表现较大的变异,有的植株油酸含量高于Oro而硫苷降低。     

2个甘蔗品种的遗传基础与核型分析

通过对2个甘蔗品种的遗传基础和染色体核型分析,研究其遗传组成源、染色体组成系统演化和血缘构成的基本特征。通过亲缘关系追溯分析,粤糖06-233和粤糖10-396这2个甘蔗品种的细胞资源均为班扎马新黑潭,其遗传组成源主要由热带种、割手密种、印度种和中国种组成。采用酶解去壁低渗法进行染色体数目与核型分析,粤糖06-233的染色体数2n=88,核型公式为K(2n)=88=82m+6sm,核型分类属于2C型;粤糖10-396的染色体数2n=106,核型公式为K(2n)=106=2M+98m+6sm,核型分类属于1B型。以上结果为甘蔗育种选配亲本提供遗传学和细胞学依据。     

秋水仙素诱导维多利亚葡萄试管苗多倍体研究

本实验采用0.05％、0.1％的秋水仙素溶液对维多利亚葡萄试管苗单芽茎段进行24 h、48 h浸泡处理来诱导发生多倍体.对诱变四倍体和二倍体进行染色体、保卫细胞叶绿体等进行比较.结果表明:0.1％的秋水仙素处理24 h对诱导葡萄发生多倍体效果最好.加倍率从M1代的40.9％提高到M9代的75.8％.根尖染色体压片检查表明,四倍体染色体数为2n=4x=76,二倍体对照为2n=2x=38.四倍体植株叶片表现气孔密度减小,气孔保卫细胞增大,保卫细胞内叶绿体数增加.     

烟草单倍体植株减数分裂的研究

本文阐明了烟草单培体花粉植株小孢子母细胞的减数分裂和小孢子发育的过程。发现由于前期Ⅰ时没有同源染色体配对,中期Ⅰ时染色体分散于细胞质中,绝大多数细胞的染色体不排列于赤道板上,仅少数细胞有赤道板的形成。后期Ⅰ时单价染色体随机分布两极,故末期Ⅰ形成大小不等的双核或多核。减数分裂Ⅱ为两条染色单体分离,表现基本正常,形成含有两两相等染色体数目的四核或多核。减数分裂还发现了异常的连续型分裂和四分孢子呈平面体型。小孢子较小,平均直径为16μ,与二倍体烟草的小孢子相比,经测定其差异达到极显著水平。小孢子的细胞质退化,未发现有丝分裂,基本上形成不育的小孢子。     

糖甜莱(Beta Vulgaris L)三体选育初报1.2.8.9号四种三体选育

1982年在黑龙江省拉哈,以三倍体品种双丰303(2n=27)和二倍体品种双丰5号(2n=18)为杂交亲本,然后按单株收获种子。同年将这批种子在江苏省徐洲地区种植。在一定时期,田间取心叶,采用苏木精分带和前期核型的分析方法,鉴别出附加1、2、8、9号染色体的四种甜莱三体,并得到这四种三体的清晰的细胞染色体图。采用鉴别体细胞同源染色体的途径,划分糖甜菜三体类型,从而将糖甜菜三体类型和具体的染色体联系在一起,这在世界上是首次报道。根据这一成果,本文讨论了Levan.Butterfass以及Bormotov等人以形态特征划分糖甜菜三体的结果,将染色体途径和形态特征途径划分三体的类型做了对比,找到了1、2、8、9号四种糖甜菜三体的标志性形态特征。     

具翼烟草(Nicotiana alata)核型、C带与rDNA染色体定位研究

采用去壁低渗法、BSG法和双色荧光原位杂交技术,分别开展了具翼烟草（Nicotiana alata）的核型分析、C显带与45S和5S rDNA染色体定位研究。4个具翼烟草收集系根尖细胞有丝分裂中期染色体平均核型为2n=2x=12m+6st,染色体臂比的变异幅度为1.14~4.22,平均臂比值为2.17,最长与最短染色体之比为2.16,属2B型。C带研究结果显示,具翼烟草中期染色体C带丰富,多态性良好,共显带25条,包括7条着丝粒带、15条端带和3条中间带。45S和5S rDNA染色体定位研究结果显示,具翼烟草根尖细胞有丝分裂中期染色体细胞相分别显示了3对45S rDNA和2对5S rDNA基因位点信号,45S rDNA基因位点分别位于I S、III S、V L上,2对5S rDNA基因位点分别位于I L、IV S上（字母L和S分别代表染色体长臂和短臂,罗马数字代表同源染色体序号）。在I号同源染色体上同时存在45S和5S rDNA基因位点。研究结果对烟草属系统起源、遗传进化以及遗传育种等领域的进一步研究具有重要的参考价值。      

以子房为材料制备烟草染色体标本的方法

为使烟草染色体标本制备过程中取材更方便,本研究以异源五倍体烟草（2n=5x=58）为材料,观察了不同大小子房、不同预处理方法对烟草染色体制片效果的影响。首先,子房大小按花冠与花萼长度比例分为6类;其次,预处理方法分为2类,即：（1）室温下于0.002 mol/L的8-羟基喹啉中预处理2、4和6 h;（2）7℃下于0.002 mol/L的8-羟基喹啉中预处理12、24和36 h。结果显示,当花冠长度小于或等于花萼长度的1/2时,子房中分裂期细胞的比例最大,为97.00±17.82个/1000个。该时期子房于7℃经24 h预处理后,染色体数目与形态清晰可辨。利用上述方法制作云烟87（2n=4x=48）和N.plumbaginifolia（2n=2x=20）的染色体标本,均获得较好的结果。此外,将上述方法制作的五倍体烟草染色体经5S rDNA-FISH,获得了清晰可辨的5个不同信号,与预期一致。可见,幼嫩子房可用于烟草染色体标本的制作,且用改良的预处理方法制备的染色体标本可用于染色体计数和核型分析,也可用于原位杂交分析。     

烟草花粉母细胞减数分裂的观察方法

用烟草花蕾观察研究烟草花粉母细胞减数分裂,对于鉴定和分析烟草核型以及染色体在减数分裂过程中的行为,是一种有价值的实验方法。现将观察方法和我们工作中的点滴体会介绍于后。一、取材取材包括所采花蕾的大小、取样的时间和取样时田间温度。而花蕾的大小则是观察减数分裂的关键。据我们6月4日以晾晒烟(黄花烟草,2n=48)为材料,从7时半到18时半(夏时制),每隔1小时取样同时测定田间温度,对12个样品进行观察的结果表明,     

Reliable identification of grapevine rootstock varieties using RAPD PCR on woody samples

Since grapevine ( Vitis spp .) rootstock material is being traded increasingly as disbudded woody material a lack of distinctive morphological features on such material necessitates an alternative and reliable means of identification. Methods described here were developed for rapid and efficient extraction of DNA from woody samples rich in phenolic compounds and polysaccharides, and for subsequent identification of varieties by RAPD PCR. Using these methods, and with the application of only one selected RAPD primer, we were able to differentiate sixteen rootstock varieties, including the seven varieties most commonly used in Germany. Problems commonly encountered with reproducibility of RAPD patterns were avoided by choosing primers with a dinucleotide sequence and a high G/C content that allowed a rather high annealing temperature of 45°C. Methods described here should also be useful for other horticultural crops, especially those with woody tissues rich in phenolic compounds and polysaccharides.     

An internet compendium of analytical methods and spectroscopic information for monomers and additives used in food packaging plastics

An internet website (http://cpf.jrc.it/smt/) has been produced as a means of dissemination of methods of analysis and supporting spectroscopic information on monomers and additives used for food contact materials (principally packaging). The site which is aimed primarily at assisting food control laboratories in the European Union contains analytical information on monomers, starting substances and additives used in the manufacture of plastics materials. A searchable index is provided giving PM and CAS numbers for each of 255 substances. For each substance a data sheet gives regulatory information, chemical structures, physico-chemical information and background information on the use of the substance in particular plastics, and the food packaging applications. For monomers and starting substances (155 compounds) the infra-red and mass spectra are provided, and for additives (100 compounds); additionally proton NMR are available for about 50% of the entries. Where analytical methods have been developed for determining these substances as residual amounts in plastics or as trace amounts in food simulants these methods are also on the website. All information is provided in portable document file (PDF) format which means that high quality copies can be readily printed, using freely available Adobe Acrobat Reader software. The website will in future be maintained and up-dated by the European Commission's Joint Research Centre (JRC) as new substances are authorized for use by the European Commission (DG-ENTR formerly DGIII). Where analytical laboratories (food control or other) require reference substances these can be obtained free-ofcharge from a reference collection housed at the JRC and maintained in conjunction with this website compendium.     

Chlorohydrins of bisphenol A diglycidyl ether (BADGE) and of bisphenol F diglycidyl ether (BFDGE) in canned foods and ready-to-drink coffees from the Japanese market

BADGE.2HCl and BFDGE.2HCl were determined in 28 samples of ready-to-drink canned coffee and 18 samples of canned vegetables (10 corn, 5 tomatoes and 3 others), all from the Japanese market. HPLC was used as the principal analytical method and GCMS for confirmation of relevant LC fractions. BADGE.2HCl was found to be present in one canned coffee and five samples of corn, BFDGE.2HCl in four samples of canned tomatoes and in one canned corn. No sample was found which exceeded the 1mg/kg limit of the EU for the BADGE chlorohydrins. However the highest concentration was found for the sum of BFDGE.2HCl and BFDGE.HCl.H₂O at a level of 1.5mg/kg. A Beilstein test confirmed that all cans containing foods contaminated with BADGE.2HCl or BFDGE.2HCl had at lest one part coated with a PVC organosol.     

Aroma characterization of various apricot varieties using headspace–solid phase microextraction combined with gas chromatography–mass spectrometry and gas chromatography–olfactometry

The characterization of the aromatic profile of several apricot cultivars with molecular tracers in order to obtain objective data concerning the aromatic quality of this fruit was undertaken using headspace–solid phase microextraction (HS–SPME). Six apricot cultivars were selected according to their organoleptic characteristics: Iranien, Orangered, Goldrich, Hargrand, Rouge du Roussillon and A4025. The aromatic intensity of these varieties measured by HS–SPME–Olfactometry were defined and classified according to the presence and the intensity of grassy, fruity and apricot like notes. In the six varieties, 23 common volatile compounds were identified by HS–SPME–GC–MS. Finally, 10 compounds, ethyl acetate, hexyl acetate, limonene, β-cyclocitral, γ-decalactone, 6-methyl-5-hepten-2-one, linalool, β-ionone, menthone and (E)-hexen-2-al were recognized by HS–SPME–GC–O as responsible of the aromatic notes involved in apricot aroma and considered as molecular tracers of apricot aromatic quality which could be utilized to discriminate apricot varieties.     

European priorities for research to support legislation in the area of food contact materials and articles

A strong science base is required to underpin the planning and decision-making process involved in determining future European community legislation on materials and articles in contact with food. Significant progress has been made in the past 5 years in European funded work in this area, with many developments contributing to a much better understanding of the migration process, and better and simpler approaches to food control. In this paper this progress is reviewed against previously identified work-areas (identified in 1994) and conclusions are reached about future requirements for R&D to support legislation on food contact materials and articles over the next 5 or so years.     

Tests of potential functional barriers for laminated multilayer food packages. Part I: Low molecular weight permeants

The advent of the functional barrier concept in food packaging has brought with it a requirement for fast tests of permeation through potential barrier materials. In such tests it would be convenient for both foodstuffs and materials below the functional barrier (sub-barrier materials) to be represented by standard simulants. By means of inverse gas chromatography, liquid paraffin spiked with appropriate permeants was considered as a potential simulant of sub-barrier materials based on polypropylene (PP) or similar polyolefins. Experiments were performed to characterize the kinetics of the permeation of low molecular weight model permeants (octene, toluene and isopropanol) from liquid paraffin, through a surrogate potential functional barrier (25 μm-thick oriented PP) into the food simulants olive oil and 3% (w/v) acetic acid. These permeation results were interpreted in terms of three permeation kinetic models regarding the solubility of a particular model permeant in the post-barrier medium (i.e. the food simulant). The results obtained justify the development and evaluation of liquid sub-barrier simulants that would allow flexible yet rigorous testing of new laminated multilayer packaging materials.     

Textural Properties of Cold-set Gels Induced from Heat-denatured Whey Protein Isolates

A 9% whey protein (WP) isolate solution at pH 7.0 was heat-denatured at 80°C for 30 min. Size-exclusion HPLC showed that native WP formed soluble aggregates after heat-treatment. Additions of CaCl2 (10–40 mM), NaCl (50–400 mM) or glucono-delta-lactone (GDL, 0.4–2.0%, w/v) or hydrolysis by a protease from Bacillus licheniformis caused gelation of the denatured solution at 45°C. Textural parameters, hardness, adhesiveness, and cohesiveness of the gels so formed changed markedly with concentration of added salts or pH by added GDL. Maximum gel hardness occurred at 200 mM NaCl or pH 4.7. Increasing CaCl2 concentration continuously increased gel hardness. Generally, GDL-induced gels were harder than salt-induced gels, and much harder than the protease-induced gel.     

Influence des ions sur le pouvoir hydratant de l'ure e: e tude sur peau de porc ex vivo

This study deals with the influence of ions (NaCl and MgSO4) in a W/O emulsion containing 10% urea. Moisturization kinetics are assessed by corneometry on pig skin ex vivo. The formula's influence on urea penetration is measured by infrared spectrometry with an ATR device and the stripping method. Corneometry and spectroscopy were chosen to record simultaneously the hydratation levels and urea localization into superficial cell layers. Urea crystallization after evaporation of emulsions and aqueous solutions is described. Results show that urea does not hydrate nor penetrate when applied to the skin through an aqueous gel. In a W/O emulsion, sodium chloride increases the ability of urea to moisturize without improving penetration. In vitro urea crystallization is disturbed by sodium chloride or magnesium sulphate for solutions and emulsions. This stabilization by ions is correlated with good moisturization values. The stabilization of urea in the solute state provided by ions increases its water epidermal binding capacity without enhancing penetration.     

Occurrence of bisphenol-F-diglycidyl ether (BFDGE) in fish canned in oil

The levels of bisphenol-F-diglycidyl ether (BFDGE) were quantified as part of a European survey on the migration of residues of epoxy resins into oil from canned fish. The contents of BFDGE in cans, lids and fish collected from all 15 Member States of the European Union and Switzerland were analysed in 382 samples. Cans and lids were separately extracted with acetonitrile. The extraction from fish was carried out with hexane followed by re-extraction with acetonitrile. The analysis was performed by reverse phase HPL C with fluorescence detection. BFDGE could be detected in 12% of the fish, 24% of the cans and 18% of the lids. Only 3% of the fish contained BFDGE in concentrations considerably above 1mg/kg. In addition to the presented data, a comparison was made with the levels of BADGE (bisphenol-A-diglycidyl ether)analysed in the same products in the context of a previous study.