大豆体细胞胚增殖保存与萌发植株体系的建立(英文)

以大豆(Glycine max(L.)Merr.)未成熟子叶为外植体,用高浓度生长素诱导东北地区主栽大豆的18个基因型体细胞胚胎发生,诱导率为0.29％～77.62％。在此基础上成功地诱导10个大豆基因型产生可继代增殖的体细胞胚,诱导率在5.2％～22.1％之间。经在固体培养基上多次继代增殖,首次建立了可在固体培养基上继代增殖的大豆体细胞胚萌发再生体系,继代一年以上的体细胞胚仍具有萌发能力和正常育性,得到了结荚植株。此体系的建立为大豆的遗传转化提供了新的、更为有效的受体体系。     

大豆体细胞胚增殖保存与萌发植株体系的建立

以大豆(Glycine max(L.)Merr.)未成熟子叶为外植体,用高浓度生长素诱导东北地区主栽大豆的18个基因型体细胞胚胎发生,诱导率为0.29%～77.62%.在此基础上成功地诱导10个大豆基因型产生可继代增殖的体细胞胚,诱导率在5.2%～22.1%之间.经在固体培养基上多次继代增殖,首次建立了可在固体培养基上继代增殖的大豆体细胞胚萌发再生体系,继代一年以上的体细胞胚仍具有萌发能力和正常育性,得到了结荚植株.此体系的建立为大豆的遗传转化提供了新的、更为有效的受体体系.     

苏云金芽孢杆菌杀虫晶体蛋白基因导入大豆的研究

用苏云金芽孢杆菌(BacillusthuringiensisBerliner)杀虫晶体蛋白(Bt)基因和葡糖苷酸酶(GUS)基因通过基因枪轰击和根癌土壤杆菌(Agrobacteriumtumefaciens(SmithetTownsend)Conn)介导转入大豆(Glycinemax(L.)Merr.),诱导大豆转基因植株再生。大豆主栽品种“中黄4号”和品系8502未成熟子叶有较强体细胞胚分化能力。体细胞胚的脱水处理显著促进“中黄4号”体细胞胚的萌发。未成熟子叶的预培养有利于根癌土壤杆菌感染子叶外植体体细胞胚的分化。基因型和受体的选择,转基因体系的改进,体细胞胚的脱水处理等是提高大豆转基因效率的重要因素。     

花生体细胞胚的诱导及其植株再生

采用不同成熟度的花生胚轴为外植体进行体细胞胚诱导及植株再生研究,结果表明,成熟胚轴在高浓度2,4－D的MS培养基中,经过30d左右的培养,可直接诱导产生出大量的体细胞胚,含40mgL~－12,4－D的培养基中体细胞胚的诱导率达100％,平均每个外植体产生11．58个体细胞胚．体细胞胚的继代培养需降低2,4－D的浓度（1－20mgL~－1）．未成熟胚轴的体细胞胚诱导及继代培养的2,4－D浓度宜为10mgL~－1．将诱导的体细胞胚转接到合5－10mgL~－1BA的MS培养基中,体细胞胚能够萌发再生成无根小植株,将其转接到生根培养基中可获得完整小植株．     

几种生长素对木薯体细胞胚发生和植株再生的作用

木薯（ＭａｎｉｈｏｔｅｓｃｕｌｅｎｔａＣｒａｎｔｚ）嫩叶外植体在含２,４－Ｄ（１－１６ｍｇＬ－１）或ＮＡＡ（４０ｍｇＬ－１）的诱导培养基上能直接诱导初生体细胞胚胎发生,而低活性的生长素ＩＢＡ或ＩＡＡ（４０ｍｇＬ－１）或低浓度的２,４－Ｄ（０．１ｍｇＬ－１）则不能。而以木薯初生体细胞胚切段为外植体时,次生体细胞胚的诱导对生长素的活性或浓度的要求降低。降低生长素浓度或活性能缩短体细胞胚诱导时间并促进根的形成,有利于提高体细胞胚的再生频率。体细胞胚外植体在诱导培养基上的培养时间对下一步体细胞胚胎发生的诱导产生影响。通过石腊切片观察,在含２,４－Ｄ诱导培养基上,木薯体细胞胚不能形成芽分生组织。结果表明,２,４－Ｄ等生长素类物质对诱导木薯体细胞胚胎发生是关键因子,但对体细胞胚的进一步发育和植株再生起抑制作用。     

马尾松幼胚体细胞胚胎发生研究

本论文首次报道了马尾松（Pinus massoniana Lamb.）幼胚体细胞胚胎发生的完整发育过程,并对影响马尾松胚性愈伤组织诱导的因素如球果采种期、球果冷藏处理时间、外植体处理方式等进行了探讨,统计胚性愈伤组织诱导率,进行增殖评价,探讨ABA浓度梯度对马尾松体细胞胚分化成熟的影响,试验数据用SPSS16统计分析软件进行方差分析、差异显著性检验。结果表明：1）2008~2009连续2年内15个采种期得到的幼胚,胚性愈伤组织诱导和增殖有显著性差异,最适宜的马尾松球果采种期是6月下旬至7月下旬,诱导率在9.66%~22.59%之间;2）球果冷藏处理时间,对胚性愈伤组织诱导有显著性差异,其中4℃冷藏球果15d有利于幼胚胚性愈伤组织诱导;3）雌配子体包含幼胚的接种处理方式是可取的;4）胚性愈伤组织经稳定增殖培养后,转入分化成熟培养基,得到体细胞胚状体＂爆发式＂分化成熟,数量多,质量好。适宜体胚成熟转化的培养基为：成熟LP培养基添加ABA5.0mg·L-1＋60.0g·L-1蔗糖,并附加L-谷氨酰胺和水解酪蛋白;5）成熟体细胞胚在无激素萌发型LP培养基上正常萌发,并转化为结构完整的小植株。本研究首次建立了马尾松幼胚体细胞胚胎发生技术平台,为马尾松遗传改良种质创新、缩短育种周期奠定了研究基础。     

农杆菌介导大豆子叶节影响因素的研究

为提高大豆遗传转化效率,采用农杆菌介导遗传转化的方法,以大豆子叶节为外植体,研究共培养阶段浸染液浓度及外界培养条件(共培养温度和天数)和培养基添加物对转化效率和芽诱导率的影响。结果显示,浸染液OD600=0.5的诱导率最高,共培养温度为24℃,共培养天数为10 d具有较高转化效率,共培养基中加入一定浓度的单壁碳纳米管(Single-wall carbon nanotube,NM)对诱导卡那霉素抗性不定芽有促进作用。PCR检测表明PHR1基因已整合到T1代大豆基因组中,初步证明可在大豆基因组中稳定遗传。     

影响大豆体细胞胚诱导因素的研究

体细胞胚的诱导是大豆体外再生的关键。基因型,诱导光周期,外植体的英位,蔗糖浓度等因素,可导致诱导频率及正常胚比例不同,影响植株再生。本研究选用黑龙江省主栽大豆基因型的未成熟子叶,在含高浓度生长素的MSB培养基上诱导体细胞胚产生。合丰25和东农7819为优选基因型,生育前期下部英位大小为2－4mm未成熟子叶体细胞胚发生效果最好;四种光周期下体细胞胚诱导频率相近,但连续弱光了正常胚比例高;NAA诱导优于2－4,D;10mg/1NAA与1．5％蔗糖配比组合最佳。     

极东锦鸡儿幼胚子叶的体细胞胚胎发生和植株再生

以极东锦鸡儿未成熟合子胚子叶为外植体进行其体细胞胚胎发生和植株再生研究。在添加不同BA与NAA或2,4-D,外加500mg·L~（-1）水解酪蛋白、30g·L~（-1）蔗糖和8g·L~（-1）琼脂的MS培养基上诱导产生了体细胞胚。在5mg·L~（-1）NAA＋5mg·L~（-1）BA和5mg·L~（-1）2,4-D＋1mg·L~（-1）BA处理中体胚诱导率分别为14%和10%;NAA处理每外植体上诱导出的体胚数量最多为4.3个,而2,4-D为10.5个。体细胞胚经成熟培养后,在添加0.01mg·L~（-1）NAA、20g·L~（-1）蔗糖和6g·L~（-1）琼脂的MS培养基上萌发率达到58.94%。萌发的体胚在MS培养基上长成正常小植株,再生率为87%。经炼苗后的体胚苗移植到草炭土：蛭石：珍珠岩=5：4：1（V/V/V）的栽培基质中,可以正常生长,移栽成活率为40%。     

香根草不同处植体诿导体细胞胚胎发生和器官发生

以禾本科植物香根草（Ｖｅｔｉｖｅｒｉａ ｚｉｚａｎｉｏｉｄｅｓ）的嫩叶鞘基部切段为外植体,再取其试管苗的基部切段诱导体细胞胚胎发生和芽的器官发生。结果表明生长素是诱导外植体脱分化和形成体细胞胚的关键因子,而芽的形成则源于体细胞胚的萌发,它们是由ＮＡＡ或低浓度的２,４－Ｄ所诱导的。通过研究建立了有效的香根草循环诱导再生体系和试管苗繁殖体系,为进一步研究香根草的生物技术奠定了良好的基础。     

Tuning of electronic properties of one-dimensional cyano-bridged Cu-Ni, Cu-Pd, and Cu-Pt bimetallic assemblies by stereochemistry of ligands

Preparations, XPS and electronic spectroscopy, and magnetism of seven new one-dimensional cyano-bridged coordination polymers, chiral [Cu(RR-chxn)₂][Pd(CN)₄] · 2H₂O (1), [Cu(trans-chxn)₂][M(CN)₄] · 2H₂O (2, 4, and 6 for M = Pd, Ni, and Pt), and [Cu(cis-chxn)₂][M(CN)₄] · 2H₂O (3, 5, and 7 for M = Pd, Ni, and Pt) (RR-chxn = cyclohexane-(1R,2R)-diamine, trans-chxn = racemic trans-cyclohexane-(1,2)-diamine, and cis-chxn = racemic cis-cyclohexane-(1,2)-diamine) have been reported in view of tuning of their electronic properties by stereochemistry of chxn ligands and metal-substitution. Comparison of Cu 2p_1/2 and 2p_3/2 peaks of XPS and broad d-d bands around 18 000 cm⁻¹ of electronic spectra are described systematically for 1-7. Variable-temperature magnetic measurement shows that complexes 1-7 indicate weak antiferromagnetic interactions via cyano-bridges. Because of semi-coordination coupled with pseudo Jahn-Teller elongation and electrostatic interaction for 1, the axial Cu-N coordination bond distances of 2.330(7) and 3.092(8) Å are considerably longer than those of equatorial ones in the range from 2.016(6) to 2.030(6) Å. The former bond distances of 1 are intermediate values among the related Ni (2.324(6) and 3.120(8) Å) and Pt (2.34(1) and 3.09(1) Å) complexes.     

Land,livestock, and livelihoods: Changing dynamics of gender,caste, and ethnicity in a Nepalese Village

Over the past 10 years, Ghusel VDC, Lalitpur District has moved from primarily subsistence agriculture into the wider cash economy aided by the Small Farmers' Development Program (SFDP), which provides credit to farmers mainly for the purchase of buffalo for milk production, and by the National Dairy Corporation, which supports local dairy cooperatives. Analysis reveals that buffalo-keeping and milk sales are increasing the well-being of many households, while at the same time creating new inequalities in gender roles and responsibilities, greater inequities between Brahmin and Tamang residents in Ghusel, and placing pressures on the ecosystem for increased supplies of fodder and fuelwood. Evidence suggests that there is critical, need for attention to the social, and particularly gender-based, implications of maintaining livestock for milk sales and to the ecological underpinnings of this livelihood system.     

Structure and function of S-adenosylhomocysteine hydrolase

In mammals, S-adenosylhomocysteine hydrolase (AdoHcyase) is the only known enzyme to catalyze the breakdown of S-adenosylhomocysteine (AdoHcy) to homocysteine and adenosine. AdoHcy is the product of all adenosylmethionine (AdoMet)-dependent biological transmethylations. These reactions have a wide range of products, and are common in all facets of biometabolism. As a product inhibitor, elevated levels of AdoHcy suppress AdoMet-dependent transmethylations. Thus, AdoHcyase is a regulator of biological transmethylation in general. The three-dimensional structure of AdoHcyase complexed with reduced nicotinamide adenine dinucleotide phosphate (NADH) and the inhibitor (1′R, 2′S, 3′R)-9-(2′,3′-dihyroxycyclopenten-1-yl)adenine (DHCeA) was solved by a combination of the crystallographic direct methods program, SnB, to determine the selenium atom substructure and by treating the multiwavelength anomalous diffraction data as a special case of multiple isomorphous replacement. The enzyme architecture resembles that observed for NAD-dependent dehydrogenases, with the catalytic domain and the cofactor binding domain each containing a modified Rossmann fold. The two domains form a deep active site cleft containing the cofactor and bound inhibitor molecule. A comparison of the inhibitor complex of the human enzyme and the structure of the rat enzyme, solved without inhibitor, suggests that a 17° rigid body movement of the catalytic domain occurs upon inhibitor/substrate binding.     

Chiral Separation of G‐type Chemical Warfare Nerve Agents via Analytical Supercritical Fluid Chromatography

Chemical warfare nerve agents (CWNAs) are extremely toxic organophosphorus compounds that contain a chiral phosphorus center. Undirected synthesis of G‐type CWNAs produces stereoisomers of tabun, sarin, soman, and cyclosarin (GA, GB, GD, and GF, respectively). Analytical‐scale methods were developed using a supercritical fluid chromatography (SFC) system in tandem with a mass spectrometer for the separation, quantitation, and isolation of individual stereoisomers of GA, GB, GD, and GF. Screening various chiral stationary phases (CSPs) for the capacity to provide full baseline separation of the CWNAs revealed that a Regis WhelkO1 (SS) column was capable of separating the enantiomers of GA, GB, and GF, with elution of the P(+) enantiomer preceding elution of the corresponding P(–) enantiomer; two WhelkO1 (SS) columns had to be connected in series to achieve complete baseline resolution. The four diastereomers of GD were also resolved using two tandem WhelkO1 (SS) columns, with complete baseline separation of the two P(+) epimers. A single WhelkO1 (RR) column with inverse stereochemistry resulted in baseline separation of the GD P(–) epimers. The analytical methods described can be scaled to allow isolation of individual stereoisomers to assist in screening and development of countermeasures to organophosphorus nerve agents. Chirality 26:817–824, 2014. © 2014 The Authors. Chirality published by John Wiley Periodicals, Inc.     

Influence of Fluoride on Rat Kidney Antioxidant System: Effects of Methionine and Vitamin E

The aim of the study has been to determine and compare the influence upon the kidney antioxidative system, exercised by administration of vitamin E, and vitamin E in combination with methionine, under conditions of oxidative stress induced by sodium fluoride. The experiment was carried out on Wistar FL rats (adult males) that, for 35 days, were administered water, NaF, NaF with vitamin E, or vitamin E with methionine (doses: 10 mg NaF/kg of body mass/24 h, 3 mg vitamin E per 10 μl per rat for 24 h, 2 mg methionine per rat for 24 h). The influence of administered sodium fluoride and antioxidants upon the antioxidative system in kidney was examined by analyzing the concentration of malondialdehyde (MDA) and the activity of the most important antioxidative enzymes (SOD, total and both its isoenzymes, GPX, GST, GR, and CAT). The studies carried out confirmed the disadvantageous effect of the administered dose of NaF upon the antixodiative system in rats (increase in the concentration MDA, decrease activity of all antioxidative enzymes). The administration of vitamin E increased the activity of studied enzymes with the exception of glutathione reductase GR; it also reduced the procesess of lipid peroxidation. It has been found that combined doses of vitamin E and methionine were most effective in inhibiting lipid peroxidation processes. The results confirmed the antioxidative properties of methionine.     

Reorientation of Microfibrils and Microtubules at the Outer Epidermal Wall of Maize Coleoptiles During Auxin-Mediated Growth

Auxin-mediated elongation growth of isolated subapical coleoptile segments of maize (Zea mays L.) is controlled by the extensibility of the outer cell wall of the outer epidermis (Kutschera et al., 1987). Here we investigate the hypothesis that auxin controls the extensibility of this wall by changing the orientation of newly deposited microfibrils through a corresponding change in the orientation of cortical microtubules. On the basis of electron micrographs it is shown that cessation of growth after removal of the endogenous source of auxin is correlated with a relative increase of longitudinally orientated microfibrils and microtubules at the inner wall surface. Conversely, reinduction of growth by exogenous auxin is correlated with a relative increase of transversely orientated microfibrils and microtubules at the inner wall surface. These changes can be detected 30–60 min after the removal and addition of auxin, respectively. The functional significance of directional changes of newly desposited wall microfibrils for the control of elongation growth is discussed.     

Gamma-glutamylcysteine protects ergothioneine-deficient Mycobacterium tuberculosis mutants against oxidative and nitrosative stress

Mycobacterium tuberculosis (M.tb.), the causative agent of tuberculosis (TB), cannot synthesize GSH, but synthesizes two major low molecular weight thiols namely mycothiol (MSH) and ergothioneine (ERG). Gamma-glutamylcysteine (GGC), an intermediate in GSH synthesis, has been implicated in the protection of lactic acid bacteria from oxidative stress in the absence of GSH. In mycobacteria, GGC is an intermediate in ERG biosynthesis, and its formation is catalysed by EgtA (GshA). GGC is subsequently used by EgtB in the formation of hercynine-sulphoxide-GGC. In this study, M.tb. mutants harbouring unmarked, in-frame deletions in each of the fives genes involved in ERG biosynthesis (egtA, egtB, egtC, egtD and egtE) or a marked deletion of the mshA gene (required for MSH biosynthesis) were generated. Liquid chromatography tandem mass spectrometry analyses (LC-MS) revealed that the production of GGC was elevated in the MSH-deficient and the ERG-deficient mutants. The ERG-deficient ΔegtB mutant which accumulated GGC was more resistant to oxidative and nitrosative stress than the ERG-deficient, GGC-deficient ΔegtA mutant. This implicates GGC in the detoxification of reactive oxygen and nitrogen species in M.tb.     

In vitro studies of the rabbit immune system. IV. Differential mitogen responses of isolated T and B cells.

The mitogenic responses of separated rabbit lymphocyte populations functionally analogous to mouse T and B cells have been tested in vitro. Purified T cells were prepared by passage over nylon wool (NW) and purified B cells prepared by treatment with antithymocyte serum and complement (ATS + C). ATS + C kills 70% of peripheral blood lymphocytes (PBL's) and 50% of the spleen cells while passage over NW yields 40% of the applied PBL's and 5–23% of the applied spleen cells. NW-purified T cells from the spleen or PBL's respond fully to concanavalin A (Con A) but have a reduced response to phytohemaglutinin (PHA) and little or no response to goat anti-rabbit immunoglobulin (anti-Ig). PBL's that survive ATS + C (B cells) are stimulated by anti-Ig but not by Con A or PHA. B cells purified from spleen do not respond to Con A or PHA but will respond to anti-Ig under appropriate conditions. A full spleen B-cell response to anti-Ig required removal of Ig produced by the cultures that blocked anti-Ig stimulation. It is concluded that, for rabbit lymphocytes, Con A and PHA are primarily T-cell mitogens and that anti-Ig is primarily a B-cell mitogen. However, the mitogen response of unfractionated PBL or spleen cell populations indicates an overlap in reactivity. This could be due to cells sharing T and B properties, alteration of cell populations by the fractionation procedures used, or recruitment of one population in the presence of a mitogenic response of the other population.     

八种脑-肠肽侧脑室内注射对大鼠基础胃酸分泌的影响

用乌拉坦麻醉大鼠作急性实验,采用连续灌流胃并收集流出液的方法,观察向侧脑室内注射微量脑-肠肽对大鼠基础胃酸分泌的影响。实验结果如下:(1)雨蛙肽、八肽胆囊收缩素、促甲状腺素释放激素及四肽胃泌素均使总酸排出量增加;(2)生长抑素、胰多肽、P 物质、胰高血糖素则使总酸排出量减少;(3)上述肽类用侧脑室注射的剂量作肌肉注射,除四肽胃泌素也产生明显的刺激胃酸分泌作用外,对胃酸分泌均无明显影响。以上结果提示,脑内的一些肽类可能以神经递质或调制物的方式,参与中枢对胃酸分泌的调节。     

A chromium-tolerant plant growing in Cr-contaminated land

In this paper, a kind of Typhaceae plant species, Typha angustifolia L, with a high tolerance to Cr is described. Experiments were carried out to examine its ability to tolerant Cr and its physiological response. The results showed that there was no difference in growth, plant height, and biomass response to external Cr (VI) between the plants exposed to 100 microM Cr (VI) and control (0 microM), while increasing Cr levels to 200-800 microM induced a significant decrease in plant height and biomass, but no significant injury was detected, even for the plants exposed to 800 microM Cr. Chromium induced significant increases in superoxide dismutase (SOD) and peroxidase (POD) activities. Meanwhile, a significantly positive correlation was found between Cr and Mn or Cu in leaves and roots, respectively. The Cr tolerance of the plant appeared to be associated with the enhancement of SOD and POD activities and the improvement in uptake and translocation of the essential microelements.