Monomorphism of isoenzyme loci in natural and cultivated amaranth (Amaranthus L.) populations

Starch gel electrophoresis was used for isozyme analysis of ADH, GDH, MDH, IDH, and ME in populations of amaranth. Experiments were performed with 93 populations and 4 cultivars. Some populations proved to be polymorphic, and this fact allowed analysis of the genetic control of the enzymes listed. The populations examined showed poor allozyme variability. Monomorphism for all loci studied was observed in 73 populations and 4 varieties. Starch gel electrophoresis was used for isozyme analysis ofADH, GDH, MDH, IDH, and ME in populations of amaranth. Experiments were performed with 93 populations and 4 cultivars. Some populations proved to be polymorphic, and this fact allowed analysis of the genetic control of the enzymes listed. The populations examined showed poor allozyme variability. Monomorphism for all loci studied was observed in 73 populations and 4 varieties. Only some populations demonstrated rare polymorphism for a single locus each: Adh, Mdh 2, Gdh, Idh 1, Idh 2, or Mod 2. The results demonstrate genetic monomorphism of amaranth for the studied loci.     

Estimation of the genetic variability of an amaranth collection (Amaranthus L.) by RAPD analysis

The genetic variability of an amaranth collection was studied by RAPD analysis. A high level of polymorphism of the studied samples was detected. It was about 85% on average. Unique fragments characteristic of only definite accessions and 18 monomorphic loci proper for all amaranth accessions were detected using several primers. The genetic distances of Nei and Li were calculated. This index varied from 0.0009 to 0.0141. A cluster analysis was carried out. The amaranth accessions were classified into two clusters according to the species affiliation.     

In vitro culture of lavenders (Lavandula spp.) and the production of secondary metabolites

Lavenders (Lavandula spp., Lamiaceae) are aromatic ornamental plants that are used widely in the food, perfume and pharmaceutical industries. The large-scale production of lavenders requires efficient in vitro propagation techniques to avoid the overexploitation of natural populations and to allow the application of biotechnology-based approaches for plant improvement and the production of valuable secondary metabolites. In this review we discuss micropropagation methods that have been developed in several lavender species, mainly based on meristem proliferation and organogenesis. Specific requirements during stages of micropropagation (establishment, shoot multiplication, root induction and acclimatization) and requisites for plant regeneration trough organogenesis, as an important step for the implementation of plant improvement programs, were revised. We also discuss different methods for the in vitro production of valuable secondary metabolites, focusing on the prospects for highly scalable cultures to meet the market demand for lavender-derived products.     

Genetic variation in outcrossing rate and correlated floral traits in a population of grain amaranth (Amaranthus cruentus L.)

The genetic control of breeding system in a population of grain amaranth is described here by the estimates of outcrossing rate variation among individuals and their response to mass selection. In the first generation (S₀), a single morphological marker locus(R/r) controlling plant pigmentation was used for the estimation of outcrossing rate. A mixture of genotypes RR (red) and rr (green) mixed in equal proportions was grown and all rr plants were progeny-tested for estimating the outcrossing rate (t), using the relationship t=H/p, where H = proportion of heterozygotes (Rr) in the progeny, and p = frequency of alleleR. A full range (0 to 100%) of outcrossing rates was found in the first generation, % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaGabeiDayaara% Gaeyypa0JaaGimaiaac6cacaaIZaGaaGymaiaacYcacqGHGaaicqGH% GaaicuaHdpWCgaqcamaaBaaaleaacaqG0baabeaakiabg2da9iaaic% dacaGGUaGaaGOmaiaaiwdaaaa!441B!\[{\text{\bar t}} = 0.31, \hat \sigma _{\text{t}} = 0.25\], and the distribution was significantly skewed toward high outcrossing.A bidirectional mass selection experiment was initiated for high and low outcrossing rates. In the Hi and Lo lines (Generation S₁), the rate of outcrossing was estimated using the R locus in addition to two other completely dominant morphological markers(B, G1). Relative sex ratios in the monoecious inflorescences of selected lines were estimated by the number of male flowers per glomerule. Outcrossing rate was significantly different between the Hi and Lo lines, and Lo line exhibited much higher male fertility than the Hi line. Such a response to selection indicated a polygenic control of sex expression, and consequently, of the potential for outcrossing in this population. In the second generation (S₂), selection was based on the number of male flowers/glomerule as an index of outcrossing ability. Differences in sex expression between the Hi and Lo lines in S₂ generation had 1.0 male flower/glomerule, while the Hi selection individuals had more or less rectangular distribution from 0.1 to 1.0 male flowers/glomerule.Thus, breeding system in amaranth, as represented by the sex ratios within glomerules and the outcrossing rate parameters, responded to bidirectional selection. Such a genetic component of variation in breeding system has significant implications in the evolution of amaranth species and landraces under domestication.     

Seasonal population dynamics of dusky cotton bug (Oxycarenus spp.) in transgenic cotton varieties under field conditions

Dusky cotton bug (Oxycarenus spp.) has become a major insect pest for cotton crop in Pakistan. Transgenic cotton varieties provided resistance to a variety of insects pests. But, these are not safe for this emerging potential threat. In present study, nine transgenic cotton varieties (IUB-222, MNH-886, FH-142, CIM-599, A-555, CIM-602, NIAB-777, MNH-786 and Bt-666) were assessed for seasonal population dynamics of dusky cotton bug (DCB) under field conditions. All transgenic varieties showed a differential DCB population over the months and no transgenic variety was free from DCB population throughout the crop duration. DCB population appeared during 3rd week of July and crossed the economic threshold level (10–15 nymph/ adults or both per plant) during August. A substantial increase in DCB population was noted during September-November with its peak population during October, 2014. Among all varieties, three varieties (CIM-599, CIM-602 & IUB-222) showed a significantly lower mean population per plant (37.76, 37.87, 43.84) and two varieties (FH-142, MNH-886) gave highest population (44.71, 46.81), respectively. Correlation matrix revealed that low temperature and high humidity were promoting the DCB population. Cluster analysis revealed interesting findings that IUB-222 with least population fall in a cluster where other two varieties (FH-142 & MNH-886) possessed highest population. Moreover, two varieties (CIM-599 & CIM-602) with least population fall in second cluster regarding DCB population. These findings would be helpful for the farmers to select the varieties that showed relatively higher resistance towards DCB population and to adopt proper management strategies keeping in view the trend of DCB population during the crop season.     

Susceptibility of willow clones (Salix spp.) to herbivory by Phyllodecta vulgatissima (L.) and Galerucella lineola (Fab.) (Coleoptera, Chrysomelidae)

Levels of damage by mixed natural infestations of the leaf-feeding chrysomelid beetles, Phyllodecta vulgatissima (L.) (the blue willow beetle) and Galerucella lineola (Fab.) (the brown willow beetle), were determined in replicated field plots of 24 Salix clones at Long Ashton (Bristol, UK) during 1993–94. Over the same period, the host plant preferences of both chrysomelids were investigated in a standard multiple-choice laboratory procedure, where beetles were enclosed in Petri dishes with leaf discs cut from young pot-grown trees propagated from shoot cuttings taken from 20 of the 24 willow clones represented in the field study. The laboratory experiments indicated that P. vulgatissima and G. lineola had similar host plant preferences in the range of willows examined (r >0.85). In both field and laboratory, the least preferred Salix clones and hybrids were those of 5. eriocephala, followed by S. purpurea, S. burjatica, S. dasyclados and S. triandra. Clones of S. eriocephala and S. purpurea were frequently rejected altogether in laboratory tests. Most preferred were clones of S. viminalis and several hybrids of S. viminalis, S. aurita, S. caprea and S. cinerea. These results substantiate the reports that P. vulgatissima and G. lineola are deterred from feeding on willows which have relatively high concentrations of phenolic (salicylate) glucosides in the leaves. The least preferred willows, particularly S. eriocephala, S. purpurea and S. burjatica, could be of great potential value in plant breeding for resistance to these willow beetle pests.     

Needle nematodes (Longidorus spp.) and stubby-root nematodes (Trichodorus spp.) harmful to sugar beet and other field crops in England

Longidorus attenuatus produces galls at the tips of roots of field crops, including sugar beet, growing in alkaline, sandy soils in eastern England. L. elongatus produces similar, but often larger, galls on the roots of sugar beet and other crops in sandy soils in the W. Midlands and in Fen peats. Trichodorus spp. cause ‘stubby root’ of sugar beet and can feed on many field crops. Seven species of Trichodorus were found in sandy soils in eastern England. L. attenuatus, L. elongatus and Trichodorus spp. aggregate around roots and stunt sugar beet and other crop plants. L. attenuatus is commoner below plough depth than in the topsoil, whereas T. cylindricus, T. pachy-dermus and T. anemones are more abundant in the topsoil. These nematodes cause some forms of ‘Docking disorder’.     

Physiological and behavioral variation in estivation among mud turtles (Kinosternon spp.)

Kinosternid mud turtles, a primarily aquatic group, exhibit variable degrees of terrestrial activity in the Sonoran and Chihuahuan Deserts. We compared behavioral and physiological responses to dry conditions in four populations representing three species, Kinosternon sonoriense, Kinosternon flavescens, and Kinosternon hirtipes. All four groups were subjected to simulated dry season conditions in the laboratory, during which activity was monitored and physiological responses (blood chemistry and rates of resting metabolism and evaporative water loss) were measured. Kinosternon flavescens and K. hirtipes represented extremes in apparent ability to estivate, based on activity and rate of increase of plasma osmolality. Two populations of K. sonoriense exhibited intraspecific differences in behavioral and physiological measures that were related to extant environmental conditions. Large numbers of K. sonoriense from Arizona and K. hirtipes, the poorest estivators, had to be rehydrated after only 30 d out of water. Kinosteron flavescens had the lowest metabolic rates, but no evidence of metabolic depression during dehydration was found for any of the four populations. We conclude that the differences in capacity for estivation among populations are primarily linked to variable behavioral responses to dry conditions, though high rates of evaporative water loss in K. hirtipes represent a probable physiological constraint.     

Inter-population variability of secondary metabolites in Hypericum spp. (Hypericaceae) of the Northern Apennines, Italy

Hypericum spp. flowers were collected from different sites of Northern Apennines in Italy and were analysed by HPTLC technique to determine their contents in flavonoids (rutin, hyperosid, quercitrin, isoquercitrin and quercetin) and hypericins (pseudohypericin and hypericin). The H. perforatum and H. richeri composition was evaluated in order to investigate the influence of ecological conditions on secondary metabolites production. The species studied showed some differences in the amounts of investigated metabolites and high variability was observed in populations gathered in different places. No significant effects of environmental factors on active compound production were observed, except for the rutin content which is in positive correlation with the altitude of the growing site. Interesting informations are also given about the morphology of secretory structures found in Hypericum richeri.     

Extreme divergence in floral scent among woodland star species (Lithophragma spp.) pollinated by floral parasites

Backgrounds and Aims

A current challenge in coevolutionary biology is to understand how suites of traits vary as coevolving lineages diverge. Floral scent is often a complex, variable trait that attracts a suite of generalized pollinators, but may be highly specific in plants specialized on attracting coevolved pollinating floral parasites. In this study, floral scent variation was investigated in four species of woodland stars (Lithophragma spp.) that share the same major pollinator (the moth Greya politella, a floral parasite). Three specific hypotheses were tested: (1) sharing the same specific major pollinator favours conservation of floral scent among close relatives; (2) selection favours ‘private channels’ of rare compounds particularly aimed at the specialist pollinator; or (3) selection from rare, less-specialized co-pollinators mitigates the conservation of floral scent and occurrence of private channels.

Methods

Dynamic headspace sampling and solid-phase microextraction were applied to greenhouse-grown plants from a common garden as well as to field samples from natural populations in a series of experiments aiming to disentangle the genetic and environmental basis of floral scent variation.

Key Results

Striking floral scent divergence was discovered among species. Only one of 69 compounds was shared among all four species. Scent variation was largely genetically based, because it was consistent across field and greenhouse treatments, and was not affected by visits from the pollinating floral parasite.

Conclusions

The strong divergence in floral scents among Lithophragma species contrasts with the pattern of conserved floral scent composition found in other plant genera involved in mutualisms with pollinating floral parasites. Unlike some of these other obligate pollination mutualisms, Lithophragma plants in some populations are occasionally visited by generalist pollinators from other insect taxa. This additional complexity may contribute to the diversification in floral scent found among the Lithophragma species pollinated by Greya moths.     

High-resolution gas-chromatographic analysis of the secondary metabolites obtained by subcritical-fluid extraction from Colombian rue (Ruta graveolens L.)

Subcritical (CO(2)) extraction, carried out in a J&W Scientific High Pressure Soxhlet Extractor, was used to isolate secondary metabolites from leaves, flowers, stems and roots of Colombian rue (Ruta graveolens L.). The various extracts were analyzed by capillary chromatography, on an HP-5 (30 m) column, using nitrogen-phosphorus, flame ionization, and mass selective detection systems. Kováts indexes and mass spectra (electron impact, 70 eV) were employed for compound identification. The extracts from the various parts of rue studied had different compositions. The number of compounds detected at concentrations above 0.01% (w/w) in the extracts from leaves, flowers, stems and roots, was 78, 45, 25 and 24, respectively. 2-Nonanone (8.9%), 2-undecanone (13.4%), chalepensin (13.0%), and geijerene (19.3%) were the main constituents found in the extracts from rue leaves, flowers, stems and roots, respectively. Furanocoumarins, furoquinolines, hydrocarbons and benzodioxol derivatives were the main compound families found in all extracts, at total concentrations between 3.7 and 33.9%, depending on the part of the plant. The extraction method used has low environmental impact and produced solvent-free extracts in good yield with no pigments, waxes, resins, or high-molecular weight compounds which may interfere with the isolation and analysis of the alkaloids responsible for rue's biological activity, which were extracted in relatively high yield.     

Differences in root and shoot development of tomato (Lycopersicon esculentum L.) varieties across contrasting soil environments

Summary The root and shoot growth of three tomato (Lycopersicon esculentum L.) varieties, VF-10, VF-1908 and Paste 56 were compared in either a river-washed sand or an alluvial silty loam, with or without added nutrients, to determine factors influencing root growth and development. VF-10 had consistently higher shoot weights than the other two varieties, particularly by 45 days from germination. Although the root-to-shoot ratio varied with soil treatment, no significant differences in this ratio occurred among varieties. The roots were characterised by a taproot, thickened in the uppermost 3 to 5 cm, with 86 to 190 secondary roots when harvested 30 days after germination. The variety VF-1908 had a higher proportion of fine roots than the other two varieties over all treatments. On the basis of root weight and length, VF-1908 was also more stable across the imposed environmental gradient than the varieties VF-10 or Paste 56. This arose from less variability across soil treatments in secondary branching in VF-1908; over the four soil treatments the number of secondary roots varied from 104 to 131 in VF-1908 compared to 86 to 139 in VF-10 and 95 to 190 in Paste 56. VF-1908 and Paste 56 also had less variation in tertiary branching than VF-10, which had a consistently greater number of major tertiary roots than the other two varieties over all soil treatments. With the exception of Paste 56 in the fertilized silt, shoot growth was correlated with root length.     

Chitinolytic microbes confer Meloidogyne incognita resistance and augment secondary metabolites in Bacopa monnieri (L.) Pennell

Meloidogyne incognita (Kofoid and White) Chitwood a calamitous plant pathogen affects almost all the commercial crops, especially medicinal plants viz. Bacopa monnieri L. In this study, the efficacy of indigenous chitinolytic microbes against M. incognita in B. monnieri was evaluated under sterilised and natural soil conditions. The plant growth was observed for various parameters viz. nutrient uptake, phytochemical activities and secondary metabolites (bacoside). In the natural soil under green house conditions, the isolates significantly (P ≤ 0.05) reduced M. incognita infestation (1.38–2.78-folds) with significant increase in secondary metabolites (1.28–1.88-fold) over the untreated control. The potential strains were identified through 16S-rRNA as Streptomyces sp., Microbacterium arabinogalactanolyticum, Cellulosimicrobium cellulans, Actinomycetales bacterium, Chitiniphilus sp. and Flavobacterium johnsoniae. These findings thus highlight the potential of indigenous chitinolytic microbes that can be utilised for overall fitness of the plant along with the reduced root-knot nematode infestation in B. monnieri.     

Recurrent introgression of mitochondrial DNA among hares (Lepus spp.) revealed by species-tree inference and coalescent simulations

Understanding recent speciation history requires merging phylogenetic and population genetics approaches, taking into account the persistence of ancestral polymorphism and possible introgression. The emergence of a clear phylogeny of hares (genus Lepus) has been hampered by poor genomic sampling and possible occurrence of mitochondrial DNA (mtDNA) introgression from the arctic/boreal Lepus timidus into several European temperate and possibly American boreal species. However, no formal test of introgression, taking also incomplete lineage sorting into account, has been done. Here, to clarify the yet poorly resolved species phylogeny of hares and test hypotheses of mtDNA introgression, we sequenced 14 nuclear DNA and 2 mtDNA fragments (8205 and 1113 bp, respectively) in 50 specimens from 11 hare species from Eurasia, North America, and Africa. By applying an isolation-with-migration model to the nuclear data on subsets of species, we find evidence for very limited gene flow from L. timidus into most temperate European species, and not into the American boreal ones. Using a multilocus coalescent-based method, we infer the species phylogeny, which we find highly incongruent with mtDNA phylogeny using parametric bootstrap. Simulations of mtDNA evolution under the speciation history inferred from nuclear genes did not support the hypothesis of mtDNA introgression from L. timidus into the American L. townsendii but did suggest introgression from L. timidus into 4 temperate European species. One such event likely resulted in the complete replacement of the aboriginal mtDNA of L. castroviejoi and of its sister species L. corsicanus. It is remarkable that mtDNA introgression in hares is frequent, extensive, and always from the same donor arctic species. We discuss possible explanations for the phenomenon in relation to the dynamics of range expansions and species replacements during the climatic oscillations of the Pleistocene.     

Morphometric variation among river sturgeons (Scaphirhynchus spp.) of the Middle and Lower Mississippi River

Pallid sturgeon (Scaphirhynchus albus) captured in the Middle and Lower Mississippi River (i.e. below St. Louis, MO, USA) are morphologically very similar to shovelnose sturgeon (Scaphirhynchus platorynchus). Available empirical data are limited to a few studies based on low sample sizes from disjointed populations. Geneticists are currently searching for markers that will differentiate the two species, but the need for unequivocal species‐specific field characters remains. Continuation of commercial fishing for shovelnose sturgeon in some states necessitates an immediate means for accurate field identifications. Previous studies of lower basin river sturgeon classified individuals with simple morphometric character indices and interpreted intermediacy as interspecific hybridization. In this study, morphometric variation among Scaphirhynchus specimens from the Middle and Lower Mississippi River is examined for evidence of hybridization. Data are compared for large (>250‐mm standard length) hatchery‐reared and wild pallid specimens and wild shovelnose specimens. Specimens are compared using two morphometric character indices, two morphometric/meristic character indices and principal components analysis. Results indicate substantial morphological variation among pallid sturgeon below the mouth of the Missouri River. The amount of variation appears to decrease downstream in the Mississippi River. Sheared principal components analysis of morphometric data shows complete separation of shovelnose and pallid sturgeon specimens, whereas character indices indicate overlap. Both character indices and sheared principal components analysis demonstrate that pallid sturgeon in the Lower Mississippi River are morphologically more similar to shovelnose sturgeon than are pallids from the Upper Missouri River. This similarity, explained in previous studies as hybridization, may be the result of latitudinal morphometric variation and length‐at‐age differences between populations of the upper and lower extremes of the range.     

Upwind flight of tsetse (Glossina spp.) in response to natural and synthetic host odour in the field

Abstract. In Zimbabwe, studies were made of the flight responses of tsetse ( Glossina spp.) to synthetic and natural ox odour using arrangements of electric nets.Tsetse flying away from a target showed a significant upwind bias when a blend of carbon dioxide (2/1 min), acetone (500 mg/h), octenol (0.4 mg/h), 4-methylphenol (0.8 mg/h) and 3-n-propylphenol (0.1 ma) was dispensed 15 m upwind, with c. 35% flying upwind.Without carbon dioxide this percentage was significantly reduced to 15% which was not significantly different from that with no odour (8%).This pattern was not altered by reducing the doses of acetone, octenol and phenols by 10–100 times, to levels comparable to those produced by an ox.With natural ox odour or a synthetic equivalent of ox odour dispensed from a ventilated pit 8 m upwind of the target, c. 28% flew upwind.This was reduced significantly to 15% if carbon dioxide was removed.In studies using a 17 m line of nets arranged orthogonally across the prevailing wind line, c. 50% of the catch was caught on the downwind side in the absence of odour.This increased significantly to c.60% when acetone, octenol and phenols were dispensed 15 m upwind, with or without carbon dioxide.With a shorter line (9 m) or an incomplete one (16.5 m long with 5 times 1.5 m wide gaps along its length) there was no change in the proportion caught downwind.For all three lines, dispensing odour upwind increased the catch 2–5 times on both the up-and downwind sides of the nets.It is concluded that a stronger upwind response to host odour is elicited when carbon dioxide is present.It is suggested that in nature upwind flight is very imprecisely orientated, with tsetse making flights up and down an odour plume 'searching' for a host.