Beta-amylase in germinating millet seeds

Beta-amylase (EC 3.2.1.2) was isolated from germinating millet (Panicum miliaceum L.) seeds by a procedure that included ammonium sulfate fractionation, chromatography on DEAE-cellulofine and CM-cellulofine, and preparative isoelectric focusing. The enzyme was homogeneous by SDS-PAGE. The M(r) of the enzyme was estimated to be 58,000 based on its mobility on SDS-PAGE and gel filtration with TSKgel G4000SW(XL), which showed that it is composed of a single unit. The isoelectric point of the enzyme was 4.62. The enzyme hydrolyzed malto-oligosaccharides more readily as their degree of polymerization increased, this being strongest for malto-oligosaccharides larger than 13 glucose residues and very weakly for maltotriose. Amylose, amylopectin and soluble starch were the most suitable substrates for the enzyme. While the enzyme showed some activity against native starch by itself, starch digestion was accelerated 2.5-fold using alpha-amylase, pullulanase and alpha-glucosidase. This enzyme appears to be very important for the germination of millet seeds.     

克隆哺乳动物的一些进展

克隆哺乳动物的一些进展陈秀兰（中国科学院遗传研究所,北京１００１０１）ＲｅｃｅｎｔＰｒｏｇｒｅｓｉｎＭａｍｍａｌｉａｎＣｌｏｎｉｎｇＣｈｅｎＸｉｕｌａｎ（ＩｎｓｔｉｔｕｔｅｏｆＧｅｎｅｔｉｃｓ,ＣｈｉｎｅｓｅＡｃａｄｅｍｙｏｆＳｃｉｅｎｃｅｓ,Ｂｅｉ...     

Somatic hybridization in the gramineae: Pennisetum americanum (L.) K. Schum. (Pearl millet) +Panicum maximum Jacq. (Guinea grass)

Summary Protoplasts from Pennisetum americanum resistant to S-2-amino-ethyl-l-cysteine (AEC) were fused with protoplasts of Panicum maximum utilizing polyethylene glycol-dimethylsulfoxide after inactivation of the Pennisetum protoplasts with 1 mM iodoacetic acid. The iodoacetate treatment prevented division of Pennisetum protoplasts; therefore, only Panicum protoplasts and heterokaryons potentially could give rise to colonies. A second level of selection was imposed by plating 3–4-week-old colonies on AEC medium. Putative somatic hybrid calli were analyzed for alcohol dehydrogenase, 6-phosphogluconate dehydrogenase, aminopeptidase, and shikimate dehydrogenase isozymes. Three somatic hybrid cell lines (lines 2, 3, and 67) were identified which showed two bands of alcohol dehydrogenase activity representing homodimers of P. maximum and P. americanum as well as a novel intermediate band of activity where Panicum-Pennisetum heterodimers would be expected. Aminopeptidase and shikimate dehydrogenase were useful for identifying presumptive hybrid calli but the isozyme patterns were additive-evidence which would not preclude the selection of chimeric callus. A more complex isozyme pattern which varied among the somatic hybrids was observed for 6-phosphogluconate dehydrogenase. In the hybrid calli, the presence of DNA sequences homologous to both P. maximum and P. americanum sequences was confirmed by hybridization of a maize ribosomal DNA probe to XbaI and EcoRI restriction fragments. Growth of hybrid lines on various concentrations of AEC was either similar to the AEC-resistant parent (hybrid line 2) or intermediate between the resistant and sensitive parents (hybrid lines 3, 67).     

Impact of the fungus Balansia henningsiana on Panicum agrostoides: frequency of infection,plant growth and reproduction,and resistance to pests

Summary The impact of the systemic fungus Balansia henningsiana (Clavicipitaceae) on the grass Panicum agrostoides was examined in field and greenhouse studies comparing infected and uninfected plants. Approximately one-half of all plants in three populations located in southern Indiana were infected. In field samples and greenhouse studies infected plants were significantly heavier than uninfected plants and produced significantly more tillers. Infection tended to suppress flowering but occasional asymptomatic tillers on infected plants produced healthy inflorescences. Although infected plants produced fewer inflorescences than uninfected plants as a proportion of total tillers, absolute numbers of inflorescences were similar in the two groups. Because other grasses infected by different species of Balansia and related fungi often are more resistant to insect damage, pest damage was quantified in one population. No differences between infected and uninfected plants were detected in levels of herbivory but infected plants had significantly less damage by the common leaf spot fungus Alternaria triticina. The results suggest that there is no selective disadvantage for plants infected by B. henningsiana.     

The Activity and Isoform Complement of Glutamine Synthetase in Panicum Species Differing in Photosynthetic Pathway

Anion exchange chromatography and immunoprecipitation have been used to demonstrate the presence of two forms (GS₁, and GS₂) of glutamine synthetase in the leaves of nine species of Panicum representative of C₃, C₄ and C₃-C₄ intermediate-type photosynthesis. GS₂ from the Panicum species, P. miliaceum and P. maximum was more thermostable than GS₁, GS₁, and GS₂ from P. laxum were equally thermostable but GS₂ from all the Panicum species examined was more sensitive to inhibition by N-ethylmaleimide than GS₁. GS₁, and GS₂ were characterised as being cytoplasmic and chloroplastic isoforms respectively by their reaction with N-ethylmaleimide and by immunoprecipitation with antibodies raised against the cytosolic isoform in barley and the chloroplastic form in tobacco. C₃ species were found to have higher activity of the chloroplastic isoform of glutamine synthetase than C₄ species. C₃-C₄ intermediate species had total leaf glutamine synthetase activities similar to those in C₃ species but were found to have a lower chloroplastic isoform content. The results are consistent with the reassimilation of photorespiratory ammonia by chloroplastic glutamine synthetase.     

Vegetation, substrate and hydrology in floating marshes in the Mississippi river delta plain wetlands, USA

In the 1940s extensive floating marshes (locally called flotant) were reported and mapped in coastal wetlands of the Mississippi River Delta Plain. These floating marshes included large areas of Panicum hemitomon-dominated freshwater marshes, and Spartina patens/Scirpus olneyi brackish marshes. Today these marshes appear to be quite different in extent and type. We describe five floating habitats and one non-floating, quaking habitat based on differences in buoyancy dynamics (timing and degree of floating), substrate characteristics, and dominant vegetation. All floating marshes have low bulk density, organic substrates. Nearly all are fresh marshes. Panicum hemitomon floating marshes presently occur within the general regions that were reported in the 1940's by O'Neil, but are reduced in extent. Some of the former Panicum hemitomon marshes have been replaced by seasonally or variably floating marshes dominated, or co-dominated by Sagittaria lancifolia or Eleocharis baldwinii.     

糜子EST-SSR分子标记的开发及种质资源遗传多样性分析

基于前期高通量测序结果设计EST-SSR引物, 用于评估国内外不同生态区144份糜子(Panicum miliaceum)种质资源的遗传差异。结果表明, 200对引物中80对呈多态性, 开发效率为40%; 引物分辨率(Rp)为0.67-4.67 (平均值为2.00), 扩增产物大小为50-500 bp。144份材料在80个位点共检测到206个等位变异, 每个位点为2-3个; 多样性指数(I)为0.659 3 (RYW108)-1.087 2 (RYW124), 平均为0.859 9; 多态性信息含量(PIC)为0.222 9 (RYW98)-0.717 2 (RYW124), 平均为0.457 3。基于UPGMA将144份资源划分为3个群组, 其中2个群组主要为北方春糜子区材料, 另一个群组主要为黄土高原春夏糜子区材料。基于Structure (K=4)将材料划分为4个类群, 即2个代表北方资源基因库以及代表黄土高原和国外资源基因库各1个。基于主成分分析将材料聚为7个类群, 划分结果与材料的地理来源一致。     

Chromosomal and embryological analyses in sexual x apomictic hybrids ofPanicum maximum Jacq.

Summary Cytological analyses in series of crosses between 7 sexual pistillate and 8 apomictic staminate parents of speciesPanicum maximum (Gramineae) are reported. Although these 15 progenitors were tetraploid (2n = 32), 2 dihaploids (2n = 16), 45 hexaploids (2n = 48) and 5 octoploids (2n = 64) were observed among 333 progeny plants. The role of unreduced gametes as the originators of polyploidy is discussed in relation to the so-called elements of apomixis. The 2 dihaploids appeared to be sexual while the hexaploids and octoploids were all apomictic. At the tetraploid level sexual and apomictic hybrids segregated in a ratio close to 11. These results were then compared to those already obtained from studies on other tropical grasses and indicate a simple genetic determinism for gametophytic apomixis.     

Effects of collembola (arthropoda) and relative germination date on competition between mycorrhizalPanicum virgatum (Poaceae) and non-mycorrhizalBrassica nigra (Brassicaceae)

In this experiment the separate and interactive effects of grazing of vesicular-arbuscular mycorrhizal hyphae by collembola and of relative germination date on competition between the mycorrhizal perennial grassPanicum virgatum and the non-mycorrhizal annual cruciferBrassica nigra were investigated. In the absence of competition,P. virgatum mass and P uptake were not affected by collembola grazing; grazing did reduce tissue N concentration and root: shoot ratio. Competition fromB. nigra plants of the same age/size (“simultaneous competition”) significantly reducedP. virgatum total, root, and shoot mass relative to control plants not subject to competition. In contrast, when in competition,B. nigra plants did not differ in biomass fromB. nigra controls grown without competition. Simultaneous competition also reduced N and P uptake byP. virgatum, but not byB. nigra. Grazing by collembola during simultaneous competition increased the differences in nutrient uptake and tended to shift the competitive balance further toward the non-mycorrhizalB. nigra. WhenP. virgatum plants were subjected to competition fromB. nigra plants which germinated three weeks later (“offset competition”) the situation was reversed: offsetB. nigra plants were negatively affected by competition while the larger, olderP. virgatum plants were not. Thus, relative germination date is important in determining the relative competitive ability of these two species. Grazing by collembola did not affect offset competition. The grazing of VAM hyphae by collembola appears to increase N availability in this experimental system. Under simultaneous competition, this N is taken up by the more extensive root system ofB. nigra; under offset conditions, the root system of the smallerB. nigra plants is insufficient to take advantage of the added resources. Thus, we suggest that grazing-induced transient changes in nutrient availability and the differential abilities of the two species to make use of these added resources constitute the mechanisms by which relative germination date and collembola grazing influence competition.     

Compartmental nitrate concentrations in barley root cells measured with nitrate-selective microelectrodes and by single-cell sap sampling

Nitrate-selective microelectrodes were used to measure intracellular nitrate concentrations (as activities) in epidermal and cortical cells of roots of 5-d-old barley (Hordeum vulgare L.) seedlings grown in nutrient solution containing 10 mol · m^–3 nitrate. Measurements in each cell type grouped into two populations with mean (±SE) values of 5.4 ± 0.5 mol · m^–3 (n=19) and 41.8 ± 2.6 mol · m^–3 (n = 35) in epidermal cells, and 3.2 ± 1.2 mol · m^–3 (n = 4) and 72.8 ± 8.4 mol · m^–3 (n = 13) in cortical cells. These could represent the cytoplasmic and vacuolar nitrate concentrations, respectively, in each cell type. To test this hypothesis, a single-cell sampling procedure was used to withdraw a vacuolar sap sample from individual epidermal and cortical cells. Measurement of the nitrate concentration in these samples by a fluorometric nitrate-reductase assay confirmed a mean vacuolar nitrate concentration of 52.6 ± 5.3 mol · m^–3 (n = 10) in epidermal cells and 101.2 ± 4.8 mol · m^–3 (n = 44) in cortical cells. The nitrate-reductase assay gave only a single population of measurements in each cell type, supporting the hypothesis that the higher of the two populations of electrode measurements in each cell type are vacuolar in origin. Differences in the absolute values obtained by these methods are probably related to the fact that the nitrate electrodes were calibrated against nitrate activity but the enzymic assay against concentration. Furthermore, a 28-h time course for the accumulation of nitrate measured with electrodes in epidermal cells showed the apparent cytoplasmic measurements remained constant at 5.0 ± 0.7 mol · m^–3, while the vacuole accumulated nitrate to 30–50 mol · m^–3. The implications of the data for mechanisms of nitrate transport at the plasma membrane and tonoplast are discussed.Symbol _n ² Chi-squared with n degrees of freedom R.-G.Z. was awarded a Sino-British Friendship Scholarship sponsored by the British Council and H.-W.K. was supported by an AFRC Linked Research Grant to A.D.T for collaboration with R.A.L. We wish to thank Dr. K. Goulding for advice on ion chromatography, Dr. K. Moore for assistance with statistical analysis and Dr. J.H. Williams for advice on the microsample analysis.