L-lactate dehydrogenase from leaves of Capsella bursa-pastoris (L.) Med.

By ammonium sulfate fractionation and gel filtration an enzyme preparation which catalyzed NAD⁺-dependent L-lactate oxidation (10^-4 kat kg^-1 protein), as well as NADH-dependent pyruvate reduction (10^-3 kat kg^-1 protein), was obtained from leaves of Capsella bursa-pastoris. This lactate dehydrogenase activity was not due to an unspecific activity of either glycolate oxidase, glycolate dehydrogenase, hydroxypyruvate reductase, alcohol dehydrogenase, or a malate oxidizing enzyme. These enzymes could be separated from the protein displaying lactate dehydrogenase activity by gel filtration and electrophoresis and distinguished from it by their known properties. The enzyme under consideration does not oxidize D-lactate, and reduces pyruvate to L-lactate (the configuration of which was determined using highly specific animal L-lactate dehydrogenase). Based on these results the studied Capsella leaf enzyme is classified as L-lactate dehydrogenase (EC 1.1.1.27). It has a K_m value of 0.25 mmol l^-1 (pH 7.0, 0.3 mmol l^-1 NADH) for pyruvate and of 13 mmol l^-1 (pH 7.8, 3 mmol l^-1 NAD⁺) for L-lactate. Lactate dehydrogenase activity was also detected in the leaves of several other plants.Abbreviation FMN flavin adenine mononucleotide     

Somatic embryogenesis from cell suspension and protoplast cultures ofCapsella bursa-pastoris (L.) Medic

Summary Rapidly growing cell suspension cultures of shepherd’s purse (Capsella bursa-pastoris L. Medic.) were established from leaf-derived calli. These suspensions remained unorganized in the presence of 2,4-D, but underwent extensive root organogenesis in a growth regulator-free liquid medium. Attempts to induce direct embryogenesis in liquid cultures were unsuccessful, but numerous embryos were obtained from cells plated onto growth-regulator-free solid medium. These embryos were frequently abnormal, and secondary embryogenesis was problematic for plant recovery but fertile plants were recovered. Viable protoplasts could readily be isolated from these cell suspensions. After 1 wk of culture, protoplast viability was 62%, and 7% of the cells had divided. Embryogenesis was observed from protoplast-derived microcolonies, plated on growth-regulator-free medium. Although these somatic embryos were difficult to root, plants were recovered. New cell suspensions were more recently established, which were only 4 to 6 mo. old when plant regeneration was attempted. Numerous shoots were obtained when these cells were plated onto growth-regulator-free solid media. However, these shoots differed from the embryos previously obtained in that they readily rooted and rapidly developed into plantlets. This system may allow the use of shepherd’s purse as a gene source for introgression of agronomically interesting traits intoBrassica crop species through protoplast manipulation and somatic hybridization.     

Origin of the quiescent center in the root of Capsella bursa-pastoris (L.) Medik

The origin of the quiescent center in the embryonic radicle of Capsella bursa-pastoris was investigated by in-situ hybridization to cellular polyadenylic-acid-containing RNA using [³H]polyuridylic acid as a probe. In the globular embryo, autoradiographic silver grains were localized in all cells of the presumptive root apex except in the hypophysis. As the inner cell formed by a transverse division of the hypophysis cut off new cells toward the central procambial cylinder of the embryo, these cells remained characteristically unlabeled, in contrast to the labeled cells of the rest of the embryo. In the embryonic radicles of mature seeds and of seedlings, cells derived from the hypophysis appeared as a nonmeristematic, unlabeled, hemispherical group, bounded by the procambium to the inside and the root epidermis to the outside. When root tips excised from 2-d-old seedlings were incubated in [methyl-³H]thymidine, sectioned, and autoradiographed, cells derived from the inner cell of the hypophysis were found to be unlabeled, thus showing that they constitute the specific cells of the quiescent center. These results present evidence for the single-cell origin of the quiescent center in an angiosperm root and a role for the hypophysis in it.Abbreviations poly(A)⁺RNA polyadenylicacid-containing RNA - [³H]poly(U) [³H]polyuridylic acid - QC quiescent center This work was supported in part by National Science Foundation grants PCM-7902898 and DCB-8709092.     

Mapping a floral trait in Shepherds purse – ‘Stamenoid petals’ in natural populations of Capsella bursa-pastoris (L.) Medik

Striking feature of angiosperm diversity is the huge number of variations in corolla morphology including complex innovations like variations in symmetry or the identity and number of floral organs. Throughout the Brassicaceae, the disymmetric flower structure is highly conserved. Still, quite a few floral alterations occur like a variant of Common Shepherds purse (Capsella bursa-pastoris), in which all petals have been transformed into additional stamens. This “decandric” phenotype has been reported for the first time about 200 years ago. In some of the original locations the variant has been recovered recently. The long term persistence indicates the establishment of an evolutionary novelty in wild populations in sympatric occurrence with wild-type plants. Due to this fact the floral variant has become an interesting model for evolutionary studies. The phenotype is heritable and just a single locus, termed “Stamenoid petals” (Spe), is assumed to be involved in the molecular origin. To unravel the chromosomal localization of this locus, a linkage map analysis was carried out using molecular markers (AFLPs, RAPDs). The final map includes 15 linkage groups and the floral trait was integrated on linkage group 12 (CBP12) including six AFLP markers. Out of these, five markers were successfully sequenced and revealed sequence identities with chromosome IV of the A. thaliana genome. Interestingly, AGAMOUS is located on this chromosome, the only class C floral organ identity gene in the A. thaliana genome, which is compatible with the assumption that Spe is an allele of AGAMOUS rather than a regulator of that gene.     

Flowering Ecotypes of Capsella bursa-pastoris(L.) Medik. (Brassicaceae) Analysed by a Cosegregation of Phenotypic Characters (QTL) and Molecular Markers

Capsella bursa-pastoris(Brassicaceae) is an annual to biennialpredominantly autogamous species distributed worldwide. Usinga linkage map with RAPDs and isozymes we studied quantitativetrait loci (QTL) controlling phenotypic traits in this invasivespecies. To obtain a mapping population we crossed two plantsoccurring in different climatic regions in California, USA (CentralValley and Sierra Nevada) with the most diverse ecotypes (phenotypicparameters) and genotypes (isozyme multilocus genotypes). Ahundred and thirteen F₂individuals were raised and analysedfor segregation at 107 RAPDs, six isozyme loci, and one locusdetermining leaf type. The number, location and magnitude ofgenes underlying 13 traits were determined by using both intervaland composite interval mapping. Two to five QTL affecting onecharacter have been detected. Altogether the 13 quantitativetraits produced 48 QTL. The inheritance patterns of traits rangedfrom those controlled by one QTL with a major effect to thosecontrolled by several QTL with only minor effects. Closely linkedQTL, e.g. onset of flowering with rosette leaf number, wereinterpreted as pleiotropic. Three major QTL account for onsetof flowering. These loci were linked to at least three isozymeloci and several other QTL responsible for developmental traitslike rosette leaf number. Heritability of quantitative traits,segregation of the leaf type, and segregation of the allozymeswas tested in the F₃generation. We conclude that historicalevents alone are insufficient to explain the distribution patternof isozyme multilocus genotypes during the colonization of newregions and habitats. The present evidence indicates that ecotypicadaptation and genetic linkage of isozyme loci with adaptivecharacters are involved. Copyright 2001 Annals of Botany Company Capsella bursa-pastoris, Shepherd’s purse, Brassicaceae, plant invasions, RAPDs, allozymes, Mediterranean multilocus genotype, flowering ecotypes, leaf type, fruit dimensions, cosegregation analysis, QTL     

Micropropagation of hornbeam (Carpinus betulus L.) and ash (Fraxinus excelsior L.)

Shoot multiplication of hornbeam was stimulated onWPM, QL andDKW medium supplemented with a low concentration of BAP or BPA (0.1-0.2 mg I -1) andIBA (0.1 mg I -1). Low concentration of thidiazuron promoted axillary bud formation, higher concentration inhibited shoot elongation. Microshoots were rooted onWPM supplemented with a low auxin concentration (IBA or NAA 0.2-0.5 mg I -1). High rooting percentages were obtained. Shoot proliferation of ash was stimulated on MS andDKW medium supplemented withBAP orBPA (2.0-5.0 mg I -1) andIBA (0.1 mg I -1). Root formation was promoted onWPM containing a low auxin concentration. Rooted plantlets were transplanted into soil and after hardening off the micropropagated trees were planted in the field. The planted trees grew normally without showing signs of abnormality.     

扫帚菜、蕨菜营养成分分析

对扫帚菜、蕨菜两种山野菜的粗蛋白、粗脂肪、膳食纤维、胡萝卜素、氨基酸、维生素及微量元素等营养成分进行了分析.结果表明,两种山野菜营养丰富,各种营养成分较为齐全,为开发食用山野菜等绿色食品提供科学依据和参考.     

Karyotypes of Bembidion quadrimaculatum (L.) and Clivina fossor (L.) (Coleoptera: Carabidae)

Bembidion quadrimaculatum possesses 24 chromosomes: 2n male = 22 + XY, 2n female = 22 + XX; their structure is meta- and submetacentric and differences in length between them are slight. Achiasmatic meiosis has been identified in spermatogenesis. The diploid chromosome number in Clivina fossor is 44; 2n male = 42 + XY, 2n female = 42 + XX. The chromosome structure is meta-, submeta-, and subtelocentric and X is the longest element in the set. 1 to 2 chiasmata per bivalent occur in meiosis.     

Antimalarial constituents from Nauclea orientalis (L.) L

Bioassay-guided fractionation of the antimalarial-active CHCl3 extract of the dried stem of Nauclea orientalis (L.) L. (Rubiaceae) has resulted in the isolation of two novel tetrahydro-beta-carboline monoterpene alkaloid glucosides, naucleaorine (= (16alpha,17beta)-3,14:15,20-tetradehydro-16-ethenyl-17-(beta-D-glucopyranosyloxy)-19alpha-methoxyoxayohimban-21-one; 1) and epimethoxynaucleaorine (2), as well as the known compounds, strictosidine lactam (= (15beta,16alpha,17beta)-19,20-didehydro-16-ethenyl-17-(beta-D-glucopyranosyloxy)oxayohimban-21-one; 3), 3,4,5-trimethoxyphenol (4), 3alpha-hydroxyurs-12-en-28-oic acid methyl ester (5), 3alpha,23-dihydroxyurs-12-en-28-oic acid (6), 3alpha,19alpha,23-trihydroxyurs-12-en-28-oic acid methyl ester (7), and oleanolic acid (8). Compounds 1, 2, 6, and 8 showed moderate in vitro activities against Plasmodium falciparum. Their structures and configurations were elucidated by spectroscopic methods including 1D- and 2D-NMR analyses.     

Synchronization of protoplasts from Glycine max (L.) Merr. and Brassica napus (L.)

Cells of Glycine max originating in a suspension culture and cells of Brassica napus prepared from hypocotyls were synchronized. Synchronization was achieved by preparing protoplasts in the usual way and subsequently letting the protoplasts regenerate into cells by removing the cell-wall-digesting enzymes. More than 70% of the cells had divided synchronously at the end of the first cycle as determined by the mitotic index. The high frequency of mitosis critically depended on the osmolality of the medium. The duration of the S-phase was estimated by measuring the activity of thymidylate kinase as well as incorporation of [³H]deoxythymidine into acid-insoluble material. The data indicate that synchronization is induced by resetting the cell cycle.Abbreviations dTMP deoxythymidine 5-monophosphate - TCA trichloroacetic acid