Eremophila glabra is an Australian plant that reduces lactic acid accumulation in an in vitro glucose challenge designed to simulate lactic acidosis in ruminants

Lactic acidosis is a major welfare issue affecting animal health and production systems such as dairy and feedlot beef. We used two bioassays to identify bioactive plants of Australia with the potential to prevent acidosis in ruminants. In the first bioassay, a potentially acidotic environment was induced by adding glucose to rumen fluid and pH and gas production were used to estimate the effect on acid production and microbial fermentation after 5-h incubation. Australian plants (n = 104) were screened for their ability to prevent a decline in the pH without inhibiting normal gas production, and five plants namely Eremophila glabra, Kennedia eximia, Acacia saligna, Acacia decurrens and Kennedia prorepens with such properties were identified. We investigated further the two top ranking plants, E. glabra and K. prorepens, in the second bioassay to determine the extent of their effect in vitro, by extending the incubation to 24 h and measuring d-lactate, and volatile fatty acids (VFA) in addition to pH and gas production. These were measured at 0, 5, 10, 16 and 24 h after inoculation. Eremophila glabra maintained pH values that were higher and d-lactate concentrations that were lower than the control (P < 0.001), and comparable to the antibiotic-protected environment (AB; 12 μg of virginiamycin/ml). Eremophila glabra and AB treatments did not restrict fermentation, as judged by gas production and VFA. Kennedia prorepens slowed the decline in pH and reduced the accumulation of lactate but inhibited gas production. We concluded that, in vitro, E. glabra was effective at controlling events that can lead to acidosis and the effect was comparable to that of virginiamycin, while K. prorepens was less effective than E. glabra and also inhibited fermentation.     

Allozyme diversity in the endangered pitcher plant Sarracenia rubra ssp. Alabamensis (Sarraceniaceae) and its close relative S. rubra ssp. rubra

Genetic variability in the federally endangered pitcher plant Sarracenia rubra ssp. alabamensis was assessed in eight Alabama populations using starch gel electrophoresis. Ten populations of the more widespread Sarracenia rubra ssp. rubra were sampled in the southeastern United States for comparison. Fifteen allozyme loci representing 13 enzyme systems were scored for each species. In contrast to S. oreophila and S. jonesii, two previously analyzed endangered pitcher plants, genetic diversity was high for both S. rubra subspecies. Within ssp. alabamensis the percentage polymorphic loci (Ps) was 80.0, the mean number of alleles per polymorphic locus was (APs) = 2.58, and expected heterozygosity (Hes) was 0.209. Genetic diversity was slightly lower for ssp. rubra (Ps = 73.3, APs = 2.91, and Hes = 0.177). The proportion of total genetic diversity found among populations was fairly low for both species (GST = 0.09 for ssp. alabamensis and 0.14 for ssp. rubra). Little genetic divergence has occurred between the two subspecies as indicated by the lack of diagnostic alleles, the proportion of total genetic diversity between taxa (GST = 0.09), and the genetic identity estimate (I = 0.90). The relatively high genetic diversity found for ssp. alabamensis indicates that the maintenance of its evolutionary potential is possible if population sizes are maintained or increased. Low levels of genetic diversity found within small Georgia ssp. rubra populations indicate that genetic erosion may increase extinction risks for these populations.     

Polymorphic microsatellite markers for inferring diversity in wild and domesticated sugar beet (Beta vulgaris)

Eight microsatellite loci were characterized within two cultivated beet (Beta vulgaris ssp. vulgaris) accessions and one accession of the wild progenitor of domesticated sugar beet, Beta vulgaris ssp. maritima. Allele diversity was high, yielding two to 11 alleles per locus. Polymorphism information content (PIC) values obtained for these eight loci where also high and indicate the highly informative nature of the microsatellites presented here. These described markers add to a small set of publicly available microsatellite markers for beet and will be instrumental in identifying patterns of genetic diversity and origins of domestication.     

Characterization of polymorphic microsatellite DNA markers for Acacia saligna (Labill.) H.L.Wendl. (Mimosaceae)

A genomic library was constructed and 10 novel polymorphic microsatellite markers developed for Acacia saligna, a species complex being developed for agroforestry across southern Australia. Polymorphism was investigated for these markers in 30 individuals from two subspecies. Alleles per locus averaged 4.0 for ssp. saligna and 5.2 for ssp. lindleyi. Population specific alleles were detected for all loci. The microsatellite markers will be used for diagnostic assessment of the four main subspecies of the species complex.     

Genetic differentiation in the Bladder campions, Silene vulgaris and S. uniflora (Caryophyllaceae), in Sweden

Allozyme variation was studied in Swedish populations of Silene vulgaris (a widespread weed), S. uniflora ssp. uniflora (restricted to coastal habitats) and S. uniflora ssp. petraea (endemic to Sweden and confined to open limestone habitats). The taxa are diploid, gynodioecious, perennial herbs and showed high levels of within-taxon and within-population gene diversity at four polymorphic loci. Within-taxon diversity was highest (H_tax=0.52) in S. vulgaris and lowest (H_tax=0.36) in S. uniflora ssp. uniflora. The weedy S. vulgaris has more alleles than either of the other two taxa and 5 out of a total of 27 alleles are unique to S. vulgaris. Most of the gene diversity within each of the taxa is accounted for by within-population diversity. The between-population component of diversity is 10% in S. vulgaris , and 24% and 5%, respectively, in S. uniflora ssp. uniflora and ssp. petraea. Hybrids may occur between S. vulgaris and S. uniflora , but introgression is limited by the species' ecology. Neither allozyme nor distributional data support the suggestion that ssp. petraea is a recent hybrid between S. vulgaris and S. uniflora ssp. uniflora , although an older hybrid origin for ssp. petraea is possible. Patterns of allele frequency variation suggest that there has been some historical gene flow between taxa, outside their present areas of sympatry. It is likely that the two subspecies of S. uniflora , which occur in naturally open habitats, were able to colonize Sweden during the Late Glacial or early post-glacial, whereas S. vulgaris followed the spread of agriculture into Sweden.     

Development of microsatellites from the rare ironstone endemic, Tetratheca paynterae ssp. paynterae and cross-species amplification

Nineteen microsatellite markers were developed from Tetratheca paynterae ssp. paynterae, a rare and endangered, leafless, perennial shrub. Twelve loci were polymorphic in T. paynterae ssp. paynterae with two to 14 alleles per locus and mean expected heterozygosity of 0.62. Primer pairs were tested on four other Tetratheca species from ironstone ranges in southern Western Australia. Ten loci were polymorphic in T. paynterae ssp. cremnobata and T. aphylla ssp. aphylla, three in T. harperi and four in T. erubescens. The level of polymorphism was adequate for studies of genetic structure and mating systems in three of the five taxa.     

Population Analysis of Licuala glabra Griff. var. glabra (Palmae) using RAPD Profiling

Random amplified polymorphic DNA (RAPD) profiling revealed geneticdiversity among three populations ofLicuala glabra var. glabra,an understorey palm growing in the dipterocarp forests of Terengganu,Malaysia. The Bukit Bauk population had individuals withoutaerial stems. The Bukit Besi individuals had stem heights to30 cm, while those of the Rasau Kerteh population had stemsup to 4 m in height. Southern blot analysis confirmed that anapparent monomorphic band generated by the primer OPU-17 amongthe three different populations corresponds to the same locus.Six primers were used to generate 87 RAPD bands that formed2-state character raw data for estimating genetic distancesusing the simple matching coefficient of similarity. The phenogramgenerated by UPGMA of the RAPD data grouped the individualsfrom the three populations under three clusters. This was inagreement with the morphological grouping. The infraspecificL. glabra var.selangorensis Becc. was grouped closer to theL. glabra var. glabra individuals than to the outgroupL. scortechiniiBecc. Principal component analysis supported the observationfrom the phenogram. The results show discrete genetic structuringin the populations studied and provide useful information forfuture conservation measures. Copyright 1999 Annals of BotanyCompany Licuala glabra Griff. var. glabra, RAPD, palm, AMOVA, population, genetic diversity, conservation.     

Isoenzymes of aspartate aminotransferase in perennial and annual rye and their hybrids.

Aspartate aminotransferase patterns were screened in a collection of rye genotypes that included 24 accessions of wild perennial rye (Secale montanum Guss.), 6 accessions of cultivated perennial Derzhavin and Tsitsin rye (Secale cereale x S. montanum), 15 accessions of winter and spring rye cultivars (S. cereale L.), and 9 accessions of perennial and annual rye genotypes bred from S. montanum ssp. kuprijanovii, Derzhavin rye, and winter rye for their resistance to fungal diseases. Aspartate aminotransferase is coded for by four loci. The data fit the model where AAT 1/4 is coded by Aat 1 and Aat 4, two duplicate loci, with null and two active alleles for each locus, alleles 1 and 3 for locus Aat 1 and alleles 2 and 4 for locus Aat 4; dimeric AAT 1/4 enzyme molecules are the products of both intralocus and interloci complementation. Allele 1 of Aat 1 was the most prominent in the isoenzyme patterns of the rye species. Alleles null and 2 of Aat 4 were twice as frequent in the perennial rye accessions, including Derzhavin and Tsitsin rye, than in winter and spring rye. In contrast, allele 4 of Aat 4 was characteristic of S. cereale. Within the screened collection, locus Aat 2 was monomorphic. Among three alleles of Aat 3, allele 2 dominated isoenzyme profiles of both rye species, whereas the other two alleles were species-specific: allele 1 was characteristic of S. montanum and allele 3 was found only in S. cereale. Key words : rye, Secale cereale, Secale derzhavinii, Secale montanum, aspartate aminotransferase, isoenzymes, perennial habit, polymorphism.     

中国特有小麦Gli-1、Gli-2和Glu-1位点的遗传多样性(英文)

运用APAGE和SDS_PAGE方法 ,研究了 32份中国特有小麦Gli_1、Gli_2和Glu_1位点的遗传多样性。在 1 4份云南铁壳麦 (Triticumaestivumssp .yunnaneseKing)中 ,共出现 8种醇溶蛋白带型和 3种高分子谷蛋白带型。在 9份西藏半野生小麦 (T .aestivumssp .tibetanumShao )中 ,发现 9种醇溶蛋白带型和 4种高分子谷蛋白带型。在 9份新疆稻麦 (T .petropavlovskyiUdacz.etMigusch .)中 ,观察到 9种醇溶蛋白带型和 5种高分子谷蛋白带型 ,其中 1份新疆稻麦 (稻麦 2 )具有Glu_D1编码的新亚基 2 .1 1 0 .1。在这 3种中国特有小麦群体中 ,Gli_1位点分别检测出 1 0、1 4和1 1个等位基因 ;Gli_2位点各具有 1 1、1 4和 1 2个等位基因 ;Glu_1位点也分别出现 5、6和 8个等位基因。云南铁壳麦、西藏半野生小麦和新疆稻麦群体内的Nei’s遗传变异系数分别为 0 .3798、0 .56 2 5和 0 .56 93。这些结果说明 ,与云南铁壳麦相比 ,西藏半野生小麦和新疆稻麦群体内的遗传变异相对较大。     

Systematics and phylogeography of the Dactylorhiza maculata complex (Orchidaceae) in Scandinavia: insights from cytological, morphological and molecular data

Flow cytometry, morphometry and molecular markers [plastid DNA and internal transcribed spacers (ITS) of nuclear ribosomal DNA] were used to determine taxonomic and phylogeographic patterns in Dactylorhiza maculata s.l. from Scandinavia. A total of 238 individuals from 27 populations from throughout all of Scandinavia, including the adjacent Kola Peninsula of Russia, were analyzed. Diploid D. maculata ssp. fuchsii and autotetraploid D. maculata ssp. maculata are morphologically differentiated. Fragment size variants from 10 plastid DNA loci (seven microsatellite loci and three loci with indel variation) were combined to give 43 haplotypes. Three major groups of haplotypes were found. Group I haplotypes were prevalent in the north and the northeast, whereas Group II haplotypes were prevalent in the south and the southwest. Group III was represented by only a single haplotype and appeared to be the result of introgression from D. incarnata s.l. Group I and Group II haplotypes did not correspond with cytologically and morphologically defined D. maculata ssp. fuchsii or D. maculata ssp. maculata. Past introgressive gene flow rather than recent hybridization is envisaged. Intermediate Group I haplotypes between Group II and the rest of Group I were detected in a zone of contact in central Sweden, which may suggest plastid DNA recombination. The six ITS alleles scored showed strong positive correlation with taxonomy. All data sets obtained for ssp. maculata were significantly correlated with geography. Three different autotetraploid lineages are hypothesized. One lineage may represent postglacial immigration from the south and the other two lineages may represent eastern immigration routes. Morphology and ITS data suggested that subarctic populations of ssp. maculata should be recognized as var. kolaënsis.     

Chloroplast DNA phylogeography of Photinia glabra (Rosaceae) in Japan

Climate changes during glacial periods have had significant effects on the present geographic distribution of plant species. To elucidate the evolutionary history of a plant species with a disjunctive distribution, we investigated the geographic distribution patterns of cpDNA haplotypes in Photinia glabra (Rosaceae) growing in broadleaved evergreen forests in Japan. We examined cpDNA in 42 populations of P. glabra sampled over a geographic range that included Kinki and its surrounding areas and the disjunctive regions in the Amakusa Islands. Both areas had unique cpDNA haplotypes. Moreover, the AMOVA revealed that a large proportion of the total variance (51%, P < 0.001) could be explained by differences among these regions. These results suggest a past fragmentation of this plant species into two separate refugia: southwestern Kyushu and Kinki, including the surrounding area, during the Quaternary glacial periods. A particularly interesting result was that in the southern disjunct distribution in the Amakusa Islands, the genetic subdivision (Φ(CT) = 1.00, P < 0.001) appears to lie between the populations from nearly contiguous islands located across a fairway only approximately 80 to 150 m in width.     

Characterization of microsatellite loci in polyploid Lavatera assurgentiflora ssp. assurgentiflora and ssp. glabra (Malvaceae)

Lavatera assurgentiflora (Malvaceae) is one of four species of the genus Lavatera native to California and Baja California. Two geographically defined subspecies are recognized: L. a. assurgentiflora on the northern islands and L. a. glabra on the southern islands. We isolated nine polymorphic microsatellite loci that amplify in both subspecies of L. assurgentiflora. Substantial levels of polymorphism were observed at many of the loci. Four loci exhibited more than 10 alleles, polymorphism information content ranged from 0.4 to 0.8, and up to six alleles were found in some individuals, supporting reports that these taxa are hexaploid. All loci also amplified in Lavatera lindsayi from Guadalupe Island, and we anticipate that they will cross‐amplify in other California Lavatera species as well.     

Genetic Variation and the Reproduction of Cordylanthus maritimus ssp. maritimus to Sweetwater Marsh, California

This study evaluated the genetic consequences of a reintroduction of the endangered annual plant Cordylanthus maritimus ssp. maritimus to Sweetwater Marsh (San Diego County, California). A survey of 21 enzyme loci in natural populations revealed that genetic diversity is very low and is primarily found as rare alleles at a few loci, making this species especially susceptible to the loss of alleles and heterozygosity through genetic drift. The reintroduction was performed in 1991 and 1992 by sowing seeds (collected from Tijuana Estuary) in numerous small patches of suitable habitat. For this study, leaf tissue was collected from all plants in all patches during flowering in 1995 and surveyed for genotype at the three enzyme loci that are polymorphic at Tijuana Estuary. Rare alleles were absent in 27 out of 30 patches for Pgm-1, in 17 out of 30 patches for Pgm-2, and in 10 out of 11 patches for Mdh-1. In all, half of the patches lacked any rare allele. Rare alleles tended to occur in patches with few individuals. Overall rare allele frequency was lower than in the colonies from which seeds were collected at two of the three loci, and heterozygosity was reduced. The Sweetwater Marsh population is at risk of losing most of its genetic variation at enzyme loci through the extinction of patches with few individuals. Future reintroduction attempts should attempt to create contiguous sets of patches or to periodically reseed existing patches to reduce the loss of genetic variation.     

Development of microsatellite markers in a large perennial herb, Veratrum album ssp. oxysepalum

We developed 11 polymorphic microsatellite [simple sequence repeat (SSR)] loci from genomic DNA of Veratrum album ssp. oxysepalum using a dual-suppression polymerase chain reaction technique and an improved method. These markers, with four to 17 alleles per locus, identified 47 genotypes in 48 samples collected from a population in Hokkaido, Japan. The observed and expected heterozygosities ranged from 0.042 to 0.938 and from 0.448 to 0.909, respectively. These SSR markers will be available to identify genets and evaluate genetic diversity of V. album ssp. oxysepalum.     

Forticulcita glabra gen. et sp. n. (Digenea, Haploporidae) in a Red Sea Mullet

The genus Forticulcita was established solely for F. glabra sp. n. from Valamugil seheli (Forsskål) in the Gulf of Aqaba. Most closely related to Haploporus , it is diagnosed primarily by having a species with an acetabulum being conspicuously robust, prepharynx measuring shorter than the pharynx, ceca terminating at the testicular level, muscular hermaphroditic ejaculatory organ lacking accessory structures, and vitellarium consisting of a single mass.     

Microsatellite markers isolated from a polyploid saltbush, Atriplex nummularia Lindl. (Chenopodiaceae)

Atriplex nummularia is a polyploid Australian saltbush which has been identified as a suitable species for use in the rehabilitation of agricultural land affected by salinity. We isolated 12 polymorphic loci for a preliminary assessment of genetic variability and structure within the species as a basis for a breeding programme. Preliminary screening of loci in 40 individuals from two populations revealed multibanded genotypes consisting of up to seven alleles in a single individual, with up to 29 alleles observed at a single locus. The multibanded patterns are consistent with the polyploid status of this species.     

Different karyotype patterns among allopatric Pinus nigra (Pinaceae) populations revealed by molecular cytogenetics

To examine variation and taxonomic recognition of Pinus nigra (European black pine) at the intraspecific level, chromosomal distribution of 5S and 18S-5.8S-26S rDNA loci revealed by fluorescent in situ hybridisation (FISH) and fluorochrome banding with chromomycin A(3) and DAPI were analysed among allopatric populations belonging to different subspecies. Despite prevalent opinion on predominantly conserved and homogenous conifer karyotypes, several patterns were observed. Surprisingly, interstitial 18S rDNA loci and DAPI heterochromatin staining after FISH showed variations in distribution and localisation. Three subspecies shared a pattern with nine 18S rDNA loci (ssp. nigra, pallasiana and laricio) while ssp. dalmatica and salzmannii had eight rDNA loci. DAPI banding displayed two patterns, one with a high number of signals (ssp. nigra, pallasiana and dalmatica) and the other with a lower number of signals (ssp. salzmannii and laricio). We conclude that our results cannot provide proof for either classification scheme for the P. nigra complex, but rather demonstrate the variability of different heterochromatin fractions at the intraspecific level.     

小花棘豆(Oxytropis glabra DC.)内生真菌的培养与鉴定

小花棘豆(Oxytropis glabra DC.)是内蒙古草原上的重要毒草,实验结合微生物学和分子生物学手段进行了其内生真菌研究.结果表明:体外培养的小花棘豆内生真菌生长缓慢,呈圆形、隆起、边缘整齐、辐射状生长的白色菌落,后菌体分泌黑褐色的色素物质,分生孢子近圆柱形,有粗且比孢子壁厚的暗色横隔膜,隔膜数1～5个.10个菌株的5.8S rDNA/ITS序列与内生真菌Embellisia sp.L12株的序列高度相似.推测该内生真菌属于Embellisia.     

Characterization of genetic diversity in core collection accessions of wild barley,Hordeum vulgare ssp. spontaneum

Genetic variability in the 143 core accessions of wild barley, Hordeum vulgare ssp. spontaneum, was assessed by allozyme analysis. A total of 34 alleles were detected at ten isozyme loci. All loci were polymorphic except Pgd-1, which was monomorphic. Est-2 and Est-4 were the most diverse loci, with genetic diversity values of 0.747 and 0.686, respectively. The comparison of the results with those of previous studies indicates that all alleles occurring in cultivated and wild barley are observed in this set of the wild Barley Core Collection. Only one allele (Pgd-1 Tj) was absent. It is noteworthy that one new allele at the Ndh-2 locus and another new allele at Aco-2 locus were first detected in the present study. Nine of the 34 alleles were rare and detected only in one to four accessions. The genetic similarities among the 143 accessions ranged from 0.18 to 1.00. Data analysis based on clustering and principal coordinate analysis showed that a high level of genetic variability exists in this set of core accessions, and indicated that some duplication probably exists in this set core based on the present study.     

菝葜与土茯苓黄酮类化合物的比较研究

对菝葜和土茯苓根茎中黄酮类化合物进行了定量测定和定性分析.结果表明：在菝葜和土茯苓根茎中均含有黄酮,黄酮醇,二氢黄酮,二氢黄酮醇等多种黄酮类化合物.以芦丁为标准品,用分光光度法测知菝葜和土茯苓根茎中总黄酮含量分别为3.36%和2.22%.