葱属Amerallium亚属（石蒜科）的系统发生与性状进化

运用贝叶斯和简约法对葱属（Allium）Amerallium亚属的核糖体DNA内转录间隔区（ITS）进行了分析,对该亚属的系统发生进行了推测。系统分析证实 Amerallium是单系的,并表明该亚属由三个隔离的地理群组成：北美Ameralliums,地中海区Ameralliums和东亚Ameralliums。性状进化的重建表明鳞茎是原始或祖先状态,根状茎和肉质增粗的根是衍生状态且在Amerallium这个亚属的类群中独立进化发生了几次。重建也表明该亚属的原始染色体基数x=7,其它染色体基数（x=8, 9, 10, 11）是由它转化而来的。在北美类群中,异基数性相当罕见,而多倍性似乎是一个相对频繁的进化事件。在地中海区类群和东亚类群中,异基数性和多倍性是染色体进化的两个主要驱动力。     

中国葱属根茎组植物15种25居群的核型研究

采用常规压片法,对15种25居群葱属根茎组植物进行核型研究,首次报道荒漠韭Allium tekesicola、蓝花韭A.beesianum的染色体数目和核型,并增补了一些根茎组植物的细胞学资料。结果表明：供试类群为二倍体或四倍体,核型类型为1A、2A或2B;高山韭A.sikkimense壤塘居群和荒漠韭具短臂随体,宽苞韭A.platyspathum的和布克赛尔居群、奇台居群具居间随体;野黄韭A.rude雅江居群和荒漠韭中各存在1条B染色体。结合前人研究结果,我们讨论了中国根茎组植物的染色体基数、随体染色体的多型性、B染色体的进化意义和该组植物的进化方式,探讨了天蓝韭A.cyaneum、齿丝山韭A.nutans的地理分布成因,得到了如下推论：（1）中国根茎组植物的染色体基数x=8;（2）中国根茎组植物的核型进化趋势为：1A→2A→2B→2C;（3）中国根茎组植物的随体染色体具多型性;（4）多倍化和结构变异是中国根茎组植物进化的两种重要方式;（5）天蓝韭和齿丝山韭以多倍化和无性生殖来克服扩大新的生存空间遇到的困难。     

Allium ursinum L. in Germany – surprisingly low genetic variability

Allium ursinum s.l. is a widely spread species of the herb layer in beech forests throughout Europe. Little is known about its phylogenetic origin and its biogeographic history. Molecular genetic analyses of eleven populations from Germany were used to clarify the relationship between populations of A. ursinum s.l. and its relationship to several other species of the genus Allium. The study focused mainly on the Teutoburg Forest in Lower Saxony and the Franconian mountain area in Bavaria. Sequences of the nuclear internal transcribed spacer ITS, and the external transcribed spacer ETS, as well as the plastidic trn L‐rpl 32 and the trn L‐trn F spacer regions were compared. No variation was detected within the species. Even sequences of populations from Belfast, Ireland did not differ from populations of Germany. The closest relative to Allium ursinum s.l. turned out to be Allium moly or Allium scorzonerifolium from the section Molium. Random amplified polymorphic DNA fingerprinting was performed and revealed 29% polymorphic bands. Genetic distances of the populations within the Teutoburg Forest coincided with geographical distances. Three populations (Osnabrück Westerberg, Osnabrück Honeburg and Leer, East Frisia) out of eleven analysed populations were identified as garden escapes. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Host-derived media used as a predictor for low abundant, in planta metabolite production from necrotrophic fungi

AIMS: Penicillium ser. Corymbifera strains were assayed on a variety of media and from infected Allium cepa tissues to evaluate the stimulation and in planta prediction of low abundance metabolites. METHODS AND RESULTS: Stimulated production of corymbiferones and the corymbiferan lactones were observed for Penicillium albocoremium, Penicillium allii, Penicillium hirsutum, Penicillium hordei and Penicillium venetum strains cultured on tissue media. Target metabolites were sporadically detected from strains cultured on common laboratory media (CYA, MEA and YES). Up to a 376 times increase in corymbiferone and corymbiferan lactone production was observed when culture extracts from CYA and A. cepa agar were compared by high pressure liquid chromatography with ultraviolet and mass spectrometry (LC-UV-MS). The novel metabolite corymbiferone B was purified and structure elucidated from a P. allii/A. cepa tissue medium extract. In planta expression of low abundance, target metabolites were confirmed from infected A. cepa tissue extracts by LC-UV-MS. CONCLUSIONS: Secondary metabolite production was directly dependent and influenced by media conditions, resulting in the stimulated production of low abundance metabolites on host-derived media. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of macerated host tissue media can be applied in vitro to predict in planta expression of low abundance metabolites and aid in metabolite origin annotation during in planta metabolomic investigations at the host/pathogen interface.     

Tissue specific expression of potent insecticidal, Allium sativum leaf agglutinin (ASAL) in important pulse crop, chickpea (Cicer arietinum L.) to resist the phloem feeding Aphis craccivora

The phloem sap-sucking hemipteran insect, Aphis craccivora, commonly known as cowpea aphid, cause major yield loss of important food legume crop chickpea. Among different plant lectins Allium sativum leaf agglutinin (ASAL), a mannose binding lectin was found to be potent antifeedant for sap sucking insect A. craccivora. Present study describes expression of ASAL in chickpea through Agrobacterium-mediated transformation of “single cotyledon with half embryo” explant. ASAL was expressed under the control of CaMV35S promoter for constitutive expression and phloem specific rolC promoter for specifically targeting the toxin at feeding site, using pCAMBIA2301 vector containing plant selection marker nptII. Southern blot analysis demonstrated the integration and copy number of chimeric ASAL gene in chickpea and its inheritance in T₁ and T₂ progeny plants. Expression of ASAL in T₀ and T₁ plants was confirmed through northern and western blot analysis. The segregation pattern of ASAL transgene was observed in T₁ progenies, which followed the 3:1 Mendelian ratio. Enzyme linked immunosorbant assay (ELISA) determined the level of ASAL expression in different transgenic lines in the range of 0.08–0.38% of total soluble protein. The phloem tissue specific expression of ASAL gene driven by rolC promoter has been monitored by immunolocalization analysis of mature stem sections. Survival and fecundity of A. craccivora decreased to 11–26% and 22–42%, respectively when in planta bioassay conducted on T₁ plants compared to untransformed control plant which showed 85% survival. Thus, through unique approach of phloem specific expression of novel insecticidal lectin (ASAL), aphid resistance has been successfully achieved in chickpea. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Vesicle formation in the membrane of onion cells (Allium cepa) during rapid osmotic dehydration

Background and Aims

Optimization of osmotic dehydration in different plant cells has been investigated through the variation of parameters such as the nature of the sugar used, the concentration of osmotic solutions and the processing time. In micro-organisms such as the yeast, Saccharomyces cerevisiae, the exposure of a cell to a slow increase in osmotic pressure preserves cell viability after rehydration, while sudden dehydration involves a lower rate of cell viability, which could be due to membrane vesiculation. The aim of this work is to study cytoplasmic vesicle formation in onion epidermal cells (Allium cepa) as a function of the kinetics of osmotic pressure variation in the external medium.

Methods

Onion epidermal cells were submitted either to an osmotic shock or to a progressive osmotic shift from an osmotic pressure of 2 to 24 MPa to induce plasmolysis. After 30 min in the treatment solution, deplasmolysis was carried out. Cells were observed by microscopy during the whole cycle of dehydration–rehydration.

Key Results

The application of an osmotic shock to onion cells, from an initial osmotic pressure of 2 MPa to a final one of 24 MPa for <1 s, led to the formation of numerous exocytotic and osmocytic vesicles visualized through light and confocal microscopy. In contrast, after application of a progressive osmotic shift, from an initial osmotic pressure of 2 MPa to a final one of 24 MPa for 30 min, no vesicles were observed. Additionally, the absence of Hechtian strand connections led to the bursting of vesicles in the case of the osmotic shock.

Conclusions

It is concluded that the kinetics of osmotic dehydration strongly influence vesicle formation in onion cells, and that Hechtian strand connections between protoplasts and exocytotic vesicles are a prerequisite for successful deplasmolysis. These results suggest that a decrease in the area-to-volume ratio of a cell could cause cell death following an osmotic shock.     

Real-time PCR detection of Holophagae (Acidobacteria) and Verrucomicrobia subdivision 1 groups in bulk and leek (Allium porrum) rhizosphere soils

In the light of the poor culturability of Acidobacteria and Verrucomicrobia species, group-specific real-time (qPCR) systems were developed based on the 16S rRNA gene sequences from culturable representatives of both groups. The number of DNA targets from three different groups, i.e. Holophagae (Acidobacteria group 8) and Luteolibacter/Prosthecobacter and unclassified Verrucomicrobiaceae subdivision 1, was determined in DNA extracts from different leek (Allium porrum) rhizosphere soil compartments and from bulk soil with the aim to determine the distribution of the three bacterial groups in the plant-soil ecosystem. The specificity of the designed primers was evaluated in three steps. First, in silico tests were performed which demonstrated that all designed primers 100% matched with database sequences of their respective groups, whereas lower matches with other non-target bacterial groups were found. Second, PCR amplification with the different primer sets was performed on genomic DNA extracts from target and from non-target bacteria. This test demonstrated specificity of the designed primers for the target groups, as single amplicons of expected sizes were found only for the target bacteria. Third, the qPCR systems were tested for specific amplifications from soil DNA extracts and 48 amplicons from each primer system were sequenced. All sequences were > 97% similar to database sequences of the respective target groups. Estimated cell numbers based on Holophagae-, Luteolibacter/Prosthecobacter- and unclassified Verrucomicrobiaceae subdivision 1-specific qPCRs from leek rhizosphere compartments and bulk soils demonstrated higher preference for one or both rhizosphere compartments above bulk soil for all three bacterial groups.     

Ultrastructural Observation on the Intra- and Intercellular Microtrabecular Network of the Pollen Mother Cells in Onion (Allium cepa)

Using DGD embedment-free electron microscopy, ultrastructural observation on the intra- and intercellular microtrabecular network (MN) of the pollen mother cells (PMC) of the whole meiotic prophase Ⅰ in onion ( Allium cepa L.) was performed. Complex nuclear MN was observed in the nucleus of PMCs, spreading throughout the nuclear region. The nucleolus and chromosomes were connected with the MN filament network. The uniformity of nuclear MN changed with the development of the PMCs. A lamina-like structure surrounded the nucleus and joined the MN in nucleus and in cytoplasm, but it disassembled at the end of prophase Ⅰ. There was also a complex cytoplasmic MN in PMCs, without obvious variation during the prophase Ⅰ. Furthermore, MN in cytoplasmic connections (plasmodesmata and cytoplasmic channels) was noticed to link the frameworks in two neighboring PMCs into one entity. Cytomixis was observed at synizesis of prophase Ⅰ in this experiment, and MN in cytoplasm and in nucleus was noticed to distribute in these granules which migrated from one PMC into its neighboring cell. At this time the nucleus moved aside from center of the PMC, but the rest of the cell was still fulfilled with MN filaments. The relationships of nuclear MN with nucleolus and chromosomes, lamina with nucleus, as well as intra- and intercellular MN with cytomixis are discussed in this paper.     

共聚焦荧光镜检术揭示大蒜鳞片细胞间期核中的F肌动蛋白网络

采用免疫荧光和荧光分子探针技术与共聚焦激光扫描显微镜观察相结合,对大蒜（Ａｌｌｉｕｍ ｓａｔｉｖｕｍ Ｌ．）鳞片细胞间期核中是否存在Ｆ肌动蛋白进行了研究。结果表明,以兔抗肌动蛋折抗体为一抗、ＦＴＴＧ－羊抗兔ＩｇＧ抗体为二抗进行免疫荧光标记实验,在荧光镜下观察到蒜瓣薄壁组织的细胞核及表皮细胞核均发出明亮的黄绿色荧光经共聚焦激光扫描显微镜进一步检查,整个细胞核呈黄绿色荧光,说明其中含有肌动蛋白。经ＴＲ