‘巴斗杏’组培快繁体系建立与耐盐植株筛选

以淮北黄里‘巴斗杏’茎段为外植体,MS为基本培养基,通过茎段诱导植株再生及进行耐盐筛选。结果表明:采用4、5月‘巴斗杏’茎段用0.1%升汞灭菌8 min较适宜;在含有1.0 mg/L 6-BA和0.1 mg/L IBA的MS培养基上茎段增殖较快,生长旺盛;较理想的生根培养基为MS+NAA 0.1 mg/L+IBA 0.2 mg/L,生根率达46.3%;‘巴斗杏’组培苗进行耐盐筛选的适宜盐浓度为0.4%～0.8%,筛选植株与对照相比差异显著。     

高质量浓度IBA和插条木质化程度对兔眼蓝浆果扦插生根的影响

采用速蘸法研究了高质量浓度(3 000、5 000、7 000和9 000 mg.L-1)IBA对兔眼蓝浆果(Vaccinium asheiReade)品种‘粉蓝’(‘Powderblue’)、‘梯芙蓝’(‘Tifblue’)和‘芭尔德温’(‘Baldwin’)插条生根率和开始生根时间的影响,并对木质化程度不同的‘顶峰’(‘Climax’)和‘雾达德’(‘Woodard’)插条的生根率和开始生根时间进行了比较。结果表明:经3 000~9 000 mg.L-1IBA溶液速蘸3 s,3个品种插条的生根率均高于或显著高于对照(P<0.05),开始生根时间则较对照明显提前,但不同品种有明显差异;随IBA质量浓度的提高,‘粉蓝’和‘芭尔德温’的生根率逐渐增加,开始生根时间逐渐提早,以9 000 mg.L-1IBA处理组的生根率最高(分别为93.98%和86.46%),开始生根时间最早(分别为15和21 d);品种‘梯芙蓝’插条的生根率则以5 000 mg.L-1IBA处理组最高(88.02%),开始生根时间最早(18 d)。木质化程度不同的插条生根率有显著差异(P<0.05),木质化程度越低,插条的生根率越高且开始生根时间越早;采用木质化程度低的插条,品种‘顶峰’和‘雾达德’插条的生根率分别达到88.54%和77.60%,开始生根时间分别为25和29 d。研究结果显示:高浓度IBA速蘸法是兔眼蓝浆果扦插生根的有效方法,在实际生产中应根据不同品种的特性选择适宜的IBA浓度;从木质化程度及插条可取数量两方面考虑宜选用半木质化枝条作为插条。     

Use of the temporary immersion bioreactor system (RITA<Superscript>®</Superscript>) for production of commercial <Emphasis Type="Italic">Eucalyptus</Emphasis> clones in Mondi Forests (SA)

In order to optimise tissue culture systems and to meet production targets, Mondi Forests biotechnology programme has in the last 2years concentrated efforts on the use of the RITA^® temporary immersion bioreactor system. Protocols have been established for six Eucalyptus clones. Results indicate a four- to six-fold increase in yield, in half the time, with the RITA^® system when compared with axillary bud proliferation on semi-solid media. Furthermore, plants produced from the RITA^® system are hardier and acclimatize better, giving higher yields of hardened-off plants. The establishment of aseptic axillary shoots into the RITA^® system is from shoots in the semi-solid system. Highest multiplication was achieved using 30-second flushes of medium every 10min, starting with 50 shoots per vessel. The multiplication cycles in RITA^® are between 14 and 18days, compared with 25–28days in a semi-solid system. There is minimal callus evident on the leaves and bases of the stems of plants in the RITA^® system and, in addition, cold-tolerant plants have a greater rooting competence when compared with plants coming from the semi-solid system. Ex vitro rooting of RITA^® – derived plantlets is substantially better than the plants from the semi-solid media.     

The <Emphasis Type="Italic">rolB</Emphasis> gene promotes rooting <Emphasis Type="Italic">in vitro </Emphasis>and increases fresh root weight <Emphasis Type="Italic">in vivo</Emphasis> of transformed apple scion cultivar ‘Florina’

Transgenic apple (Malus × domestica Borkh.) Florina plants were obtained by Agrobacterium-mediated transformation. The efficiency of gene transfer was 7.9%, calculated as a number of explants producing at least one transgenic shoot, after co-cultivation of leaf explants from in vitro-grown shoots in a thin layer of the A. tumefaciens C58C1 strain with the binary vector pCMB-B:GUS. Polymerase chain reaction revealed that all the clones contained the nptII and rolB genes, while four of them did not contain the gus gene. Southern blot analysis confirmed the integration of the nptII and rolB genes, with one to three copies per genome being present. All independent rolB-transgenic lines were able to produce roots in vitro on the hormone free medium, while the plants, transformed with the vector pIB16.1, or untransformed control plants did not root, and only half of shoots of MM106 rootstock rooted on this medium. The average root number in the rolB-transgenic clones ranged from 4 to 7.7. Pretreatment with indole-3-butyric acid caused root formation in all transgenic and control plants and significantly increased root number in the rolB-transgenic lines, compared to untransformed plants. RolB-transgenic plants, grown in vivo in greenhouse for 2 years, did not differ phenotypically from the wild type line with the exception of root parts. All rolB-transformed plants produced altered root systems containing more fine roots leading to significantly increased fresh root weight in five plant lines.     

Ancient gene duplications and the root(s) of the tree of life

Summary. Tracing organismal histories on the timescale of the tree of life remains one of the challenging tasks in evolutionary biology. The hotly debated questions include the evolutionary relationship between the three domains of life (e.g., which of the three domains are sister domains, are the domains para-, poly-, or monophyletic) and the location of the root within the universal tree of life. For the latter, many different points of view have been considered but so far no consensus has been reached. The only widely accepted rationale to root the universal tree of life is to use anciently duplicated paralogous genes that are present in all three domains of life. To date only few anciently duplicated gene families useful for phylogenetic reconstruction have been identified. Here we present results from a systematic search for ancient gene duplications using twelve representative, completely sequenced, archaeal and bacterial genomes. Phylogenetic analyses of identified cases show that the majority of datasets support a root between Archaea and Bacteria; however, some datasets support alternative hypotheses, and all of them suffer from a lack of strong phylogenetic signal. The results are discussed with respect to the impact of horizontal gene transfer on the ability to reconstruct organismal evolution. The exchange of genetic information between divergent organisms gives rise to mosaic genomes, where different genes in a genome have different histories. Simulations show that even low rates of horizontal gene transfer dramatically complicate the reconstruction of organismal evolution, and that the different most recent common molecular ancestors likely existed at different times and in different lineages. Correspondence and reprints: Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269-3125, U.S.A. Present address: Genome Atlantic, Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, Nova Scotia, Canada.     

Effects of carbon source and concentration on development of lingonberry (<Emphasis Type="Italic">Vaccinium vitis-idaea</Emphasis> L.) shoots cultivated <Emphasis Type="Italic">in vitro</Emphasis> from nodal explants

Summary Carbohydrate type and concentration and their interactive effects on in vitro shoot proliferation of three lingonberry (Vaccinium vitis-idaea ssp. vitis-idaea L.) cultivars (‘Regal’, ‘Splendor’, and ‘Erntedank’) and two V. vitis-idaea ssp. minus (Lodd) clones (‘NL1’ and ‘NL2’) were studied. Nodal explants were grown in vitro on medium with 2 μM zeatin and either glucose, sorbitol, or sucrose at a concentration of 0, 10, 20, or 30 gl⁻¹. The interactive effects of carbohydrate type and concentration and genotype were important for shoot proliferation. The best response was afforded by sucrose at 20 gl⁻¹ both in terms of explant response and shoot developing potential, although glucose supported shoot growth equally well, and in ‘NL1’ at 10 gl⁻¹ it resulted in better in vitro growth than sucrose. Carbohydrate concentration had little effect on shoot vigor. The genotypes differed in terms of shoots per explant, length, and vigor, leaves per shoot, and callus formation at the base of explants; this was manifested with various types and concentrations of carbohydrate. Changing the positioning of explants on the medium from vertically upright to horizontal increased the shoot and callus size, but decreased shoot height and leaves per shoot. Proliferated shoots were rooted on a peat:perlite (1∶1, v/v) medium and the plantlets were acclimatized and eventually established in the greenhouse.     

离体条件下几种不同的因子对芦笋诱导生根的研究

以芦笋的茎尖为外植体,分别从糖、无机盐、激素等方面研究了在离体条件下影响芦笋生根的因素。结果分析表明：1/2 MS、20 g·L^-1的糖有利于生根;10 g·L^-1糖、1/4 MS培养基上预处理12 d后,生根率可达到100%,根的质量明显优于生长在含有激素培养基上的根;激素预处理并不能提高生根率。因此,低盐及低糖预处理是提高芦笋生根率的一种比较好的方法。     

In vitro propagation of Cassia angustifolia through leaflet and cotyledon derived calli

High efficiency shoot regeneration was achieved through leaflet and cotyledon derived calli in Cassia angustifolia - an important medicinal plant. Dark brown compact callus was induced at the cut ends of the explants on Murashige and Skoog's (MS) medium augmented with 1 μM N⁶-benzyladenine (BA) + 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Such callus pieces on transfer to cytokinins (BA or kinetin) supplemented medium differentiated shoots within 10 – 15 d. Of the two cytokinins, 5 μM BA was optimum for eliciting morphogenic response in 83.33 and 70.83 % cultures with an average of 4.16 ± 0.47 and 3.70 ± 0.56 shoots in cotyledon and leaflet derived calli, respectively. The addition of 0.5 μM α-naphthaleneacetic acid (NAA) to MS + 5 μM BA further elevated the maximum average number of shoots to 12.08 ± 1.04 and 5.37 ± 0.52 for cotyledon and leaflet calli, respectively. The excised shoots were transferred to a rooting medium containing either IAA (indole-3-acetic acid), IBA (indole-3-butyric acid) or NAA. Nearly 95 % shoots developed an average of 5.4 ± 0.41 roots on half strength MS medium supplemented with 10 μM IBA.     

影响四季桂插穗生根若干因子研究

本文研究扦插基质、插穗枝梢成熟度和插穗留叶量对四季桂生根的影响。结果表明:枝梢成熟度和扦插基质对插穗生根有显著影响,留1片叶和留2片叶对插穗生根没有显著影响。因此,四季桂扦插宜选用半木质化成熟枝梢,并采用2/3泥炭土+1/3珍珠岩作为扦插基质,留1～2片叶效果最好。     

Distinct modes of adventitious rooting in Arabidopsis thaliana

The literature describes different rooting protocols for Arabidopsis thaliana as models to study adventitious rooting, and results are generally perceived as comparable. However, there is a lack of investigations focusing on the distinct features, advantages and limitations of each method in the study of adventitious rooting with both wild-type (WT) ecotypes and their respective mutants. This investigation was undertaken to evaluate the adventitious rooting process in three different experimental systems, all using A. thaliana, analysing the same rooting parameters after transient exposure to auxin (indole-3-acetic acid) and control conditions: excised leaves, de-rooted plants and etiolated seedlings. The founding tissues and sites of origin of roots differed depending on the system used, whereas all rooting patterns were of the direct type (i.e., without callus formation). None of the systems had an absolute requirement for exogenous auxin, although rooting was enhanced by this phytohormone, with the exception of de-rooted plants, which had adventitious rooting strongly inhibited by exogenous auxin. Root elongation was much favoured in isolated leaves. Auxin-overproducing mutants could not be used in the detached leaf system due to precocious senescence; in the de-rooted plant system, these mutants had a WT-like rooting response, whereas the expression of the 'rooty' phenotype was only evident in the etiolated seedling system. Adventitious rooting of etiolated WT seedlings in the presence of exogenous auxin was inhibited by exogenous flavonoids, which act as auxin transport inhibitors; surprisingly, the flavonoid-deficient mutant chs had a lower rooting response compared to WT. Although Arabidopsis is an excellent model system to study adventitious rooting, physiological and developmental responses differed significantly, underlining the importance of avoiding data generalisation on rooting responses derived from different experimental systems with this species.