Molecular marker based taxonomy and phylogeny of Guinea yam (Dioscorea rotundata - D. cayenensis).

Four different molecular techniques were used to assess relationships among 21 accessions of Guinea yam (Dioscorea rotundata and Dioscorea cayenensis) and 21 accessions belonging to seven putative progenitor species. Random amplified polymorphic DNA (RAPD) and microsatellite-primed PCR (MP-PCR) analysis yielded 246 informative characters that were transformed into a matrix of pairwise distances and analyzed by neighbor joining or split decomposition. Both methods gave congruent results. Well-separated groups were formed that corresponded to their species designation. Dioscorea rotundata and D. cayenensis accessions were clearly separated from each other, supporting the concept that both are distinct species. Two morphological intermediates grouped together with D. rotundata. All investigated species fell into two main clusters, one comprising D. rotundata, D. cayenensis, Dioscorea abyssinica, Dioscorea liebrechtsiana, and Dioscorea praehensilis, the other comprising Dioscorea smilacifolia, Dioscorea minutiflora, Dioscorea burkilliana, and Dioscorea togoensis. The same grouping was also obtained by comparative sequence analysis of chloroplast DNA, which supports earlier studies of nuclear rDNA variation and chloroplast restriction fragment length polymorphisms. We also analyzed the same set of Dioscorea samples with the recently developed random amplified microsatellite polymorphism (RAMPO) technique. A series of diagnostic RAMPO bands was identified that clearly distinguished between D. rotundata and D. cayenensis. Some of these bands could also be traced back to the putative progenitors of both species. The evolutionary origin of Guinea yam is discussed in light of the present results.     

Use of cpSSRs for the characterisation of yam phylogeny in Benin.

The Dioscorea cayenensis - Dioscorea rotundata species complex is the most widely cultivated yam in West Africa. This species complex has been described as deriving from wild yams belonging to the Enanthiophyllum section through domestication by African farmers. To study patterns of yam evolution and to establish phylogenetic relationships existing between wild and cultivated species sampled in Benin, we investigated changes in chloroplast DNA simple sequence repeats (cpSSR) in 148 yam accessions selected to cover the wider possible genetic diversity existing in the country. Dioscorea cayenensis and D. rotundata share the same haplotype. The morphotype "abyssinica" appeared to be subdivided into 2 haplotypes. One of these haplotypes shares the same haplotype with the Dioscorea cayenensis - Dioscorea rotundata species complex and with morphotypes praehensilis, suggesting that they might belong to the same species. Relationships among sections Lasiophyton, Macrocapaea, Opsophyton, and Enanthiophyllum were clarified, and some taxonomic changes within the Enanthiophyllum section were suggested. Dioscorea minutiflora, D. smilacifolia, and D. burkilliana might be considered as 1 single genetic group, and they are suspected of belonging to the same species.     

A set of 100 chloroplast DNA primer pairs to study population genetics and phylogeny in monocotyledons

Chloroplast DNA sequences are of great interest for population genetics and phylogenetic studies. However, only a small set of markers are commonly used. Most of them have been designed for amplification in a large range of Angiosperms and are located in the Large Single Copy (LSC). Here we developed a new set of 100 primer pairs optimized for amplification in Monocotyledons. Primer pairs amplify coding (exon) and non-coding regions (intron and intergenic spacer). They span the different chloroplast regions: 72 are located in the LSC, 13 in the Small Single Copy (SSC) and 15 in the Inverted Repeat region (IR). Amplification and sequencing were tested in 13 species of Monocotyledons: Dioscorea abyssinica, D. praehensilis, D. rotundata, D. dumetorum, D. bulbifera, Trichopus sempervirens (Dioscoreaceae), Phoenix canariensis, P. dactylifera, Astrocaryum scopatum, A. murumuru, Ceroxylon echinulatum (Arecaceae), Digitaria excilis and Pennisetum glaucum (Poaceae). The diversity found in Dioscorea, Digitaria and Pennisetum mainly corresponded to Single Nucleotide Polymorphism (SNP) while the diversity found in Arecaceae also comprises Variable Number Tandem Repeat (VNTR). We observed that the most variable loci (rps15-ycf1, rpl32-ccsA, ndhF-rpl32, ndhG-ndhI and ccsA) are located in the SSC. Through the analysis of the genetic structure of a wild-cultivated species complex in Dioscorea, we demonstrated that this new set of primers is of great interest for population genetics and we anticipate that it will also be useful for phylogeny and bar-coding studies.     

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 June 2010 - 31 July 2010

This article documents the addition of 205 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Bagassa guianensis, Bulweria bulwerii, Camelus bactrianus, Chaenogobius annularis, Creontiades dilutus, Diachasmimorpha tryoni, Dioscorea alata, Euhrychiopsis lecontei, Gmelina arborea, Haliotis discus hannai, Hirtella physophora, Melanaphis sacchari, Munida isos, Thaumastocoris peregrinus and Tuberolachnus salignus. These loci were cross-tested on the following species: Halobaena caerulea, Procellaria aequinoctialis, Oceanodroma monteiroi, Camelus ferus, Creontiades pacificus, Dioscorea rotundata, Dioscorea praehensilis, Dioscorea abyssinica, Dioscorea nummularia, Dioscorea transversa, Dioscorea esculenta, Dioscorea pentaphylla, Dioscorea trifida, Hirtella bicornis, Hirtella glandulosa, Licania alba, Licania canescens, Licania membranaceae, Couepia guianensis and 7 undescribed Thaumastocoris species.     

Monitoring microbiological and biochemical changes in fermented yam

The microbiological and biochemical changes occurring in yam (Dioscorea rotundata L.) tissues fermented in 2% brine under anaerobic conditions at ambient temperature were studied. The ash. fibre and fat contents of the yam tissues were reduced at an average rate of 1.98, 1.72 and 0.42 mg g⁻¹ d⁻¹, respectively.Lactobacillus andLeuconostoc spp., as well as other fermentative microorganisms, showed a tendency to increased growth. The decrease in the total aerobic count after a 4-d fermentation was due to depletion of available nutrients, increased acidity of the medium and complete anaerobiosis of the environment. The fermenting microbes showed a higher amylolytic activity, releasing between 52 and 73 mg glucose per mL, than proteolytic activity, where between 120 and 250 μg amino acids per mL was released.     

Influence of minisett sizes on dry matter accumulation and fresh tuber yield of white Guinea yam (Dioscorea rotundata)

Studies were conducted in 2005 and 2006 at Irrua, Nigeria, to determine the performance of various sizes of minisetts of Dioscorea rotundata cv. 'Obiaoturugo' evaluated for size and yield of yield matter and seed yams. These trials involved a macrosett (250 g) and six minisett sizes (25, 30, 35, 40, 45 and 50 g) in a randomised block design with four replicates. The results indicated that larger minisetts had higher degree of field establishment, foliation, leaf area index, total dry weight, percentage of leaf dry matter and net assimilation rate. These growth parameters influenced higher crop growth rate, which was responsible for higher fresh tuber yield. The macrosett's fresh tuber yield was the greatest (20.48 t ha⁻¹) but had the least multiplication ratio (8.19). The greatest multiplication ratio was obtained from 30-g setts.     

Multiple components of alpha-amylase in germinating tubers of a yam, Dioscorea dumetorum

alpha-Amylase from germinating tubers of a yam Dioscorea dumetorum was extracted and purified by four steps of purification. A total yield of 23.1% was obtained with over 1,600-fold increase in specific activity. Three distinct amylolytically active protein forms were resolved upon treatment of the preparation on DEAE-cellulose ion exchange chromatography at pH 8.3. All the partially purified alpha-amylase fractions have similar physical properties with respect to pH optimum, Km values, molecular weights, and energies of activation. Qualitative paper chromatographic analysis of the alpha-amylase-amylose digest revealed variable product specificity for the three alpha-amylase fractions. One form exhibited a dual product specificity for the formation of maltose and maltohexaose, while another form produced exclusively maltopentaose from polysaccharide substrates. The third amylase fraction showed usual action pattern characteristic of most alpha-amylases.     

Production of oxalic acid by some fungi infected tubers

Oxalic acid (as oxalate) was detected in four tubers commonly used for food in Nigeria-Dioscorea rotundata (White yam), Solanum tuberosum (Irish potato), Ipomoea batatas (Sweet potato), and Manihot esculenta (cassava). Whereas healthy I. batata had the highest oxalic acid content, healthy M. esculenta contained the lowest. When all tubers were artifically inoculated with four fungi-Penicillium oxalicum CURIE and THOM, Aspergillus niger VAN TIEGH, A. flavus and A. tamarii KITA, there was an increase in oxalate content/g of tuber tissue. The greatest amount of oxalate was produced by P. oxalicum in D. rotundata tuber. Consistently higher amounts of oxalate were produced by the four fungi in infected sweet potato tuber than in any other tuber and consistently lower amounts of oxalate were produced by the four fungi in Irish potato tuber. Differences in the carbohydrate type present in the tubers and in the biosynthesis pathway are thought to be responsible for variation in the production of oxalate in the different tubers by the four fungi used.     

A NEW SUBSPECIES OF ACROCEPHALUS BAETICATUS FROM LAKE CHAD AND A TAXONOMIC REAPPRAISAL OF ACROCEPHALUS DUMETORUM

A new population of African reed warblers, Acrocephalus baeticatus hopsoni , is described from Lake Chad. Its characters, together with those of unallocated material from Senegal, Eritrea and Tibesti, are intermediate between the Palaearctic A. dumetorum and other races of A. baeticatus . It is concluded that dumetorum is conspecific with A. baeticatus , and by priority becomes A. baeticatus dumetorum . The merits of A. palustris, A. scirpaceus and Hippolais pallida for inclusion in an A. baeticatus superspecies are considered, but allow no firm conclusion.     

A Quantitative Cytochemical Study of Glutamate and Glucose-6-Phosphate Dehydrogenase Activities during Chilling Injury in Tubers of Dioscorea rotundata Poir.

Onyia, G. O. C. and Gahan, P. B. 1985. A quantitative cytochemicalstudy of glutamate and glucose-6-phosphate dehydrogenase activitiesduring chilling injury in tubers of Dioscorea rotundala Poir.—J.exp. Bot. 36: 1249–1256. The response of glucose-6-phosphate dehydrogenase and glutamatedehydrogenase activities in healthy Jamaican Dioscorea rotundalatubers and those chilled at 3 ?C for 1,2,3,4, and 7 d at 70%r.h. were assessed by quantitative cytochemical assays. Bothenzymes in chill-damaged tuber tissues showed a substantiallyhigher activity than did those of the healthy tubers. An early,sharp increase in the response of the NADP-tetrazolium reductasesystem of damaged tuber tissue was significantly higher (P =0.001) than that of healthy tubers or those chilled but ableto recover. This response may be used as an early marker ofchilling injury in the yam tuber. Key words: Dioscorea rotundata Poir, quantitative cytochemistry, yam tuber, glucose-6-phosphate, dehydrogenase, glutamate dehydrogenase, NADPitetrazolium reductase     

Leaf Spot of Alternanthera sessilis and Its Implications for Food Crops

A leaf spot of Alternanthera sessilis caused by Fusarium pallidoroseum (syn. F. semitectum) is described. During pathogenicity tests, the fungus caused leaf spots of Amarantbus sp. Abelmoschus esculentus (okra), Celosia sp. and Sorghum bicolor. It also caused rots of tubers of Dioscorea rotundata, D. alata, Solanum tuberosum, corms of Colocasia esculenta, bulbs of Allium cepa and roots of Daucus carota when wounded. Unwounded tubers, corms, bulbs and roots of these named plants were not infected except roots of Daucus carota which disintegrated between 4 and 6 days after inoculation.     

Yam mosaic, a new potyvirus infecting Dioscorea cayenensis in the Ivory Coast

The major disease affecting Dioscorea cayenensis in the Ivory Coast is caused by a virus which was transmitted by mechanical inoculation to some Dioscorea spp. and Nicotiana benthamiana . In extracts of D. cayenensis leaves infectivity was lost after 10 min at 60 C but not 55 C and after dilution to 10^-3 but not 10^-2. A purification procedure is described. The virus particles are flexuous filaments c . 785 nm long. The virus was transmitted by four aphid species in the non-persistent manner, and is serologically related to four African potyviruses. The name yam mosaic virus is proposed; the present cryptogram is: */*:*/6:E/E:S/Ve/Ap, potyvirus group.     

A LEAFCUTTER BEE TRACE FOSSIL FROM THE MIDDLE EOCENE OF PATAGONIA, ARGENTINA, AND A REVIEW OF MEGACHILID (HYMENOPTERA) ICHNOLOGY

Abstract:  The ichnospecies Phagophytichnus pseudocircus isp. nov. is described to include trace fossils characterized by leaf-margin excisions showing eccentricity values of 0.35–0.65 and more than 270 degrees of an arc, a non cuspate margin and vein stringers or necrotic flaps of tissue along the margin. A method for determining ellipse eccentricity was performed on leaf discs obtained from the nests of the modern leafcutter bee Megachile rotundata (Hymenoptera: Megachilidae), which provided objectively obtained values comparable to the trace fossil from the middle Eocene of Argentina and other world-wide ichnological records, historically and subjectively considered to be 'circular' trace fossils and attributed to leafcutter bees. The material described herein represents the first evidence for fossil Megachilidae from the Southern Hemisphere.     

New microsatellite loci in the dwarf yams Dioscorea group Epipetrum (Dioscoreaceae)

? Premise of the study: Microsatellite loci were isolated and characterized from enriched genomic libraries of two taxa of the Chilean Epipetrum group of Dioscorea to assess their levels of genetic diversity and population differentiation. ? Methods and Results: Eleven microsatellite loci were identified. Six out of nine microsatellites from D. biloba amplified in D. humilis, and the two microsatellites from D. humilis amplified in both taxa. Two different sets of eight loci amplified in each of the two tested taxa, D. biloba and D. humilis. The average number of alleles was 5.75 and 5 for D. biloba and D. humilis, respectively. Higher levels of mean genetic diversity were found in D. biloba (H(E) = 0.639) than in D. humilis (H(E) = 0.414). ? Conclusions: These microsatellite primers will be useful in population genetic studies and to establish conservation strategies in the endangered taxa of the Epipetrum group of Dioscorea.     

菊叶薯蓣的快速繁殖研究

我国野生的薯蓣皂甙类植物由于盲目采集已日渐枯绝,甚至面临绝种的危险。原产于墨西哥的菊叶薯蓣不仅产量高、含薯蓣皂甙也高,因此在我国有很大的开发利用潜力。本文的工作是对菊叶薯蓣的外植体进行组织培养,筛选出了诱导芽和诱导根的优化培养基与培养条件,建立了无性快速繁殖培养系,获得了完整的培养植株并移栽成功。     

Biocontrol and other beneficial activities of Bacillus subtilis isolated from cowdung microflora

Bacillus subtilis strains isolated from cowdung (CD) had several beneficial attributes, which included biocontrol, plant growth promotion, sulphur (S) oxidation, phosphorus (P) solubilization and production of industrially important enzymes (amylase and cellulase). The B. subtilis strains from CD inhibited the in vitro growth of fungi, Fusarium oxysporum (25-34%) and Botryodiplodia theobromae (100%), isolated from the postharvest rots of yam (Dioscorea rotundata) tubers. Other than biocontrol, B. subtilis strains were able to promote root elongation in seedlings of Cicer arietinum up to 70-74% as compared to untreated seeds (control). B. subtilis strains had also the ability to oxidize elemental S to sulphate (2-15microgml(-1)) and showed distinct P-solubilization activity in vitro. In addition, the cultures showed cellulase activity in carboxy methyl cellulose medium (1.5-1.8mg of reducing sugar24h(-1)ml(-1)) and amylase activity in vitro.     

Responses of antioxidant potentials in Dioscorea rotundata Poir. following paclobutrazol drenching

The effect of paclobutrazol (PBZ) treatments on the antioxidant metabolism of white yam (Dioscorea rotundata Poir.) was investigated in the present study. PBZ @ 15 mg l(-1) plant(-1) was given to plants by soil drenching, 30, 60, and 90 days after planting (DAP). The non-enzymatic antioxidant contents like ascorbic acid (AA), reduced glutathione (GSH) and alpha-tocopherol (alpha-toc), activities of antioxidant enzymes like superoxide dismutase (SOD), ascorbate peroxidase (APX), polyphenol oxidase (PPO) and catalase (CAT) were extracted and assayed on 100 DAP from leaf, stem and tubers of both control and PBZ treated plants. It was found that PBZ has a profound effect on the antioxidant metabolism and caused an enhancement in both non-enzymatic and enzymatic antioxidant potentials under treatments in white yam. Our results have good significance, as this increase the innate antioxidant potential of this food crop, which is helpful to satisfy the needs of antioxidants in diet and thereby make it an economically important food crop.     

苜蓿切叶蜂在我国的研究与应用现状

应用苜蓿切叶蜂MegachilerotundataFubricius为苜蓿授粉是苜蓿 (MedicagoL .)种植中不可缺少的措施之一 ,该文对苜蓿切叶蜂的生物学及羽化与环境因子的关系进行了阐述 ,指出影响苜蓿切叶蜂授粉及滞育的环境因子 ,分析了苜蓿切叶蜂繁育过程中病害及天敌的情况 ,介绍了繁育过程中的设备和管理技术。     

Low isozyme diversity in Nordic and central European populations of Vicia pisiformis and V. dumetorum (Fabaceae)

Vicia pisiformis and V. dumetorum are closely related perennial species occurring in small, disjunct populations throughout Europe. In Sweden, they are classified as "vulnerable" and "care-demanding", respectively. They possess low variation in RAPD (Random Amplified Polymorphic DNA) markers but relatively high morphological variation. We estimated the amount and distribution of isozyme diversity within and among marginal (Sweden, Norway) and central (Czech Republic) populations. Vicia dumetorum was monomorphic for all 14 isozyme loci in the 22 populations investigated, whereas V. pisiformis had 2.94 % polymorphic loci and a mean of 1.02 alleles per loci. The differentiation (F_sr) of populations of V pisiformis were 0.848, but no clear geographical pattern was found. Preliminary results from artificial pollination experiments suggests self-compatibility in V. dumetorum . The low overall polymorphism in Swedish and Czech localities in both species is explained in terms of common postglacial colonization events, recent anthropogenic activity, selfing and clonal reproduction. Finally, the implications of the data for conservation genetics is discussed.     

水杨酸对菊叶薯蓣中薯蓣皂素生物合成的影响

薯蓣皂素为甾体激素药物合成起始原料,主要来源于菊叶薯蓣等薯蓣属植物的块茎或根状茎,因而关于提高菊叶薯蓣中薯蓣皂素含量的研究有重要意义。利用水杨酸处理菊叶薯蓣的离体植株,研究其对薯蓣皂素生物合成的影响及作用机制。100μmol·L－1的水杨酸处理使薯蓣皂素积累量最大,且提高了叶绿素含量和可溶性糖含量,降低可溶性蛋白含量。半定量 RT-PCR 检测基因表达发现,除了法尼基二磷酸（FPP）基因,水杨酸增强菊叶薯蓣角鲨烯合酶（SQS）基因、甲基戊二酰辅酶 A 还原酶（HMGR）基因、环阿屯醇合成酶（CAS）基因的表达。研究结果为提高菊叶薯蓣中薯蓣皂苷的含量、揭示水杨酸促进薯蓣皂素生物合成的机制等研究提供了基础。