Population Dynamics of Meloidogyne arenaria and Pasteuria penetrans in a Long-Term Crop Rotation Study

The endospore-forming bacterium Pasteuria penetrans is an obligate parasite of root-knot nematodes (Meloidogyne spp.). The primary objective of this study was to determine the effect of crop sequence on abundance of P. penetrans. The experiment was conducted from 2000 to 2008 at a field site naturally infested with both the bacterium and its host Meloidogyne arenaria and included the following crop sequences: continuous peanut (Arachis hypogaea) (P-P-P) and peanut rotated with either 2 years of corn (Zea mays) (C-C-P), 1 year each of cotton (Gossypium hirsutum) and corn (Ct-C-P), or 1 year each of corn and a vegetable (V-C-P). The vegetable was a double crop of sweet corn and eggplant (Solanum melongena). A bioassay with second-stage juveniles (J2) of M. arenaria from a greenhouse (GH) population was used to estimate endospore abundance under the different crop sequences. A greater numerical increase in endospore densities was expected in the P-P-P and V-C-P sequences than in the other sequences because both peanut and eggplant are good hosts for M. arenaria. However, endospore densities, as determined by bioassay, did not substantially increase in any of the sequences during the 9-year experiment. To determine whether the nematode population had developed resistance to the resident P. penetrans, five single egg-mass (SEM) lines from the field population of M. arenaria were tested alongside the GH population for acquisition of endospores from the field soil. Four of the five SEM lines acquired 9 to 14 spores/J2 whereas the GH population and one of the SEM lines acquired 3.5 and 1.8 spores/J2, respectively. Endospore densities estimated with the four receptive SEM lines were highest in the P-P-P plots (14-20 spores/J2), intermediate in the V-C-P plots (6-7 spores/J2), and lowest in the Ct-C-P plots (< 1 spore/J2). These results indicate that the field population of M. arenaria is heterogeneous for attachment of P. penetrans endospores. Moreover, spore densities increased under intensive cropping of hosts for M. arenaria, but the GH population of the nematode was not receptive to spore attachment. However, previously, the GH population was very receptive to spore acquisition from this field site. One explanation for this inconsistency is that the M. arenaria population in the field became resistant to the dominant subpopulation of P. penetrans that had been present, and this led to the selection of a different subpopulation of the bacterium that is incompatible with the GH population.     

Life history and laboratory host range of Stenopelmus rufinasus, a natural enemy for Azolla filiculoides in South Africa

The frond-feeding weevil, Stenopelmus rufinasus Gyllenhal, was imported into quarantine for testing as a potential natural enemy for the invasive fern Azolla filiculoides Lamarck in South Africa. Adult S. rufinasus lived for approximately 55 days during which the females produced on average 325 offspring. The developmental period for the immature stages (egg, three larval instars and pupation) was about 20 days indicating the potential for several overlapping generations per year. Both the adults and the larvae caused severe damage to A. filiculoides in the laboratory. Host specificity of this insect was determined by adult no-choice oviposition and larval starvation tests on 31 plant species in 19 families. Adult feeding, oviposition and larval development was only recorded on the Azolla species tested (A. filiculoides, A. pinnata subsp. poss. asiatica R.K.M. Saunders and K. Fowler, A. pinnata subsp. africana (Desv.) R.K.M. Saunders and K. Fowler and A. nilotica De Caisne Ex Mett.). A. filiculoides proved to be significantly the most suitable host for the weevil. The low adult emergence from A. nilotica and A. pinnata subsp. africana would most probably prevent the weevil from establishing on them in the field. A. pinnata subsp. poss. asiatica which supported greater development, is thought to be introduced and has a weedy phenology in South Africa and is thus of low conservation value. Therefore, any damage inflicted on this plant in the field may be an acceptable trade-off for the predicted impact of S. rufinasus on the aggressive exotic weed, A. filiculoides.     

Biology and host range of Platyphora semiviridis, a leaf beetle evaluated as a potential biological control agent for Solanum mauritianum in South Africa

The South American tree Solanum mauritianum is a major environmental weed in the high-rainfall regions of South Africa and has been targeted for biological control. Potential agents included five species of the genus Platyphora, which were imported from South America in 1994. Platyphora species associated with Solanaceae reputedly have very specific habitat requirements and host plant preferences in the field. Despite this, host-specificity tests on one species, Platyphora semiviridis, revealed a broad physiological host range. Although laboratory tests showed that P. semiviridis is confined to Solanum species and cannot survive on solanaceous crops outside that genus, it developed on potato and cultivated eggplant (aubergine) as well as on 10 native South African Solanum species. With few exceptions, there were no consistent differences in survival and duration of development on these compared with S. mauritianum. Furthermore, at least six of these non-target species, including potato and eggplant, supported breeding colonies of the beetles in cages. During choice tests in both small and larger cages, P. semiviridis avoided potato but did not consistently discriminate between S. mauritianum, eggplant and six native solanums for larviposition. Despite these findings, P. semiviridis has never been recorded on either potato or eggplant in South America, where it was only observed to feed on S. mauritianum. Although there are several reasons why P. semiviridis is unlikely to attack non-target Solanum species in the field, it will not be released in South Africa because there are other imported agents which have displayed narrower physiological host ranges and which may be more effective.     

Host range and risk assessment of Zygogramma bicolorata,a defoliating agent released in South Africa for the biological control of Parthenium hysterophorus

Parthenium hysterophorus (parthenium) is a weed of international importance and is spreading rapidly in sub-Saharan Africa. Consequently, it has been targeted for biocontrol in South Africa since 2003. Based on precedents elsewhere in the world, the defoliating beetle Zygogramma bicolorata was prioritised as a candidate agent. Although no-choice tests, involving some 48 test plant species, indicated a significant preference for parthenium, significantly reduced feeding and oviposition was recorded on some species. Multiple-choice tests resolved many of these non-target results; however, Helianthus annuus (sunflower) was still selected for oviposition and feeding. Of the 12 sunflower cultivars tested, four were selected for oviposition, while two were selected for oviposition and feeding. These six cultivars were then subjected to larval development trials, together with three native and two weed species (in the Asteraceae). These trials showed high levels of complete development on parthenium, significantly reduced development on sunflower cultivars, and partial development on only one of the weed species. Finally, a risk assessment was conducted on the six sunflower cultivars to quantify Z. bicolorata feeding and reproductive performance. Feeding risk calculations revealed these cultivars to have an extremely low risk (<0.2%) of supporting Z. bicolorata feeding and development. Similarly, reproductive risk calculations showed a very low risk (<0.16%) of supporting viable Z. bicolorata populations. These data are supported by findings from both the native (Mexico) and introduced ranges (Australia, India) of Z. bicolorata, where it has never been recorded as a pest of sunflower. These considerations were accepted by the regulatory authorities and in August 2013, Z. bicolorata became the second insect agent to be released in South Africa for the biocontrol of parthenium.     

The host specificity of Anomalococcus indicus Ayyar (Hemiptera: Lecanodiaspididae), a potential biological control agent for prickly acacia (Vachellia nilotica ssp. indica) in Australia

ABSTRACT

Prickly acacia, Vachellia nilotica ssp. indica (Benth.) Kyal. & Boatwr, is a significant weed of northern Australia and has been a target of weed biological control in Australia since the 1980s. Following native range surveys in India, the scale insect Anomalococcus indicus Ayyar was identified as the most promising agent and was imported into Australia for further research. A. indicus is a major pest of prickly acacia on the Indian subcontinent, where it causes shoot tip dieback and plant death. Despite field observations suggesting the species was specific to V. nilotica, A. indicus completed development on 17 of the 84 non-target plant species tested during no-choice host specificity trials under quarantine conditions. Of these, Acacia falcata, V. bidwillii, V. sutherlandii and Neptunia major supported high numbers of mature females in all replicates. All of these species were utilised in choice trials. Combined risk scores indicate that V. sutherlandii, N. major and A. falcata may be attacked in the field. Due to the limited ability of scale insects to disperse, only those non-target species that occur on the Mitchell grass downs (i.e. V. sutherlandii) are considered to be at risk. Nevertheless, in view of the disparity between quarantine test results and the observed field host specificity of A. indicus in India, field trials are currently being conducted in India using Australian native species on which complete development has occurred. The future of A. indicus as a biological control agent for prickly acacia in Australia will be determined once results from these field trials are known.     

Stain-free detection as loading control alternative to Ponceau and housekeeping protein immunodetection in Western blotting

It is currently a routine practice to require a measurement of a housekeeping reference, including actin, glyceraldehyde-3-phosphate dehydrogenase, β-tubulin, among others, in Western blots, as it is the rule in RNA blots. Reversible Ponceau staining has been applied successfully to check equal loading of gels. Here we test a new technique, with the Stain-Free gels from Bio-Rad, against both Ponceau staining and housekeeping protein immunodetection under different conditions. Our results show that Stain-Free gels outperform Ponceau staining and that both are more consistent than housekeeping proteins as a loading control.     

Biology and host range of the stem-boring beetle Aphanasium australe, a promising agent for the biological control of Hakea sericea in South Africa

The invasive Australian shrubHakea sericea Shrader is a majorenvironmental weed in the Western and EasternCape Provinces of South Africa. Dense,impenetrable thickets severely threaten theunique endemic vegetation of the Cape FloristicKingdom, increase fire hazards and reduce wateryields in catchments. Biological control,initiated in the 1970s, is largely confined tothe use of seed-feeding insect agents. Becausenone of these agents reduce the density ofexisting hakea populations, a stem-boringbeetle, Aphanasium australe (Boisduval) (Cerambycidae),was imported into quarantine in South Africa in1975. During multichoice oviposition tests,involving 12 Australian and six South Africanproteaceous species, in a walk-in cage, A. australe only oviposited on four species ofHakea and two exotic species of Grevillea. However, culturing difficultiesresulted in the suspension of host-specificitytesting after three years. Testing was resumedfollowing re-importations in 1995, also becauseA. australe also attacks Hakeagibbosa (Sm.) Cav., which is not attacked byany of the existing biocontrol agents. Duringno-choice survival tests, involving 66test plant species from 15 families, A. australe only developed on H. sericea, H. gibbosa and one exoticspecies of Grevillea. The contention thatA. australe is confined to the genus Hakea was confirmed by host records andsurveys in Australia which provided no evidenceof attacks on crop, pasture or related plants.The regulatory authorities accepted theseresults and A. australe was cleared forrelease in South Africa during 2001.     

Biology and host specificity of Aulacobaris fallax (Coleoptera: Curculionidae), a potential biological control agent for dyer’s woad,Isatis tinctoria (Brassicaceae) in North America

Dyer’s woad, Isatis tinctoria, a plant of Eurasian origin is a problematic weed in western North America against which a classical biological weed control programme was initiated in 2004. Three European insect species were selected as candidate agents to control this invasive species, including the root‐mining weevil Aulacobaris fallax. To determine its suitability as an agent, the biology and host specificity of A. fallax were studied in outdoor plots and in the field between 2004 and 2006 in its native European range. Aulacobaris fallax is a univoltine species that lays its eggs from March to August into leaf stalks and roots of dyer’s woad. Larvae mine and pupate in the roots and adults emerge from August to October. Up to 62% of the dyer’s woad plants at the field sites investigated were attacked by this weevil. In no‐choice host‐specificity tests, A. fallax attacked 16 out of 39 species and varieties within the Family Brassicaceae. Twelve of these are native to North America. In subsequent multiple‐choice tests, seven species, all native to North America, suffered a similar level of attack as dyer’s woad, while none of the European species were attacked. Our results demonstrate the importance of including test plant species that have not co‐evolved with the respective candidate agent. In sum, we conclude that the risk of non‐target effects is too high for A. fallax to be considered as a biological control agent for dyer’s woad in the United States.     

Host specificity testing,release and establishment of Urophora solstitialis (L.) (Diptera: Tephritidae), a potential biological control agent for Carduus nutans L., in Australia

The seed‐fly Urophora solstitialis was imported into Australia from southern France. Previous workers had shown that this seed‐fly limited its attack to selected members of the genus Carduus. Host specificity studies revealed that no native Australian plant tested from the family Asteraceae was a suitable host plant. Permission to release the fly was granted, and it was released in three regions of the New South Wales tablelands. The fly underwent a second generation at two of the release sites, and reasons for its failure to do so at the third are suggested. It has successfully established itself in all three regions, having completed a full seasonal cycle in the field.     

Preliminary studies on Ramularia crupinae sp. nov. as a potential biological control agent for common crupina (Crupina vulgaris) in the USA

Common crupina, Crupina vulgaris (Asteraceae), is an alien weed of pastures and disturbed noncrop lands in the western USA. Research is underway in Europe to search for the natural enemies to control the weed. Among various natural enemies attacking C vulgaris, a new fungal species, Ramularia crupinae, has been found to cause serious damage to the weed. The fungus has been grown on various media and a modified V8 juice agar has proved to be suitable for both growth and sporulation. Greenhouse and field studies have shown that R. crupinae can kill infected leaves, thus reducing the plant growth, and in severe cases can kill the young plants. During preliminary host-range tests using various members of the Asteraceae, including artichoke and safflower, the fungus attacked only C. vulgaris.     

Biology and host range of Ophiomyia camarae Spencer (Diptera: Agromyzidae), a potential biocontrol agent for Lantana spp. (Verbenaceae) in Australia

The life history and host range of the herringbone leaf-mining fly Ophiomyia camarae, a potential biological control agent for Lantana spp., were investigated. Eggs were deposited singly on the underside of leaves. Although several eggs can be laid on a single leaf and a maximum of three individual mines were seen on a single leaf, only one pupa per leaf ever developed. The generation time (egg to adult) was about 38 days. Females (mean 14 days) lived longer than males (mean 9 days) and produced about 61 mines. Oviposition and larval development occurred on all five lantana phenotypes tested. Eleven plant species representing six families were tested to determine the host range. Oviposition and larval development occurred on only lantana and another nonnative plant Lippia alba (Verbenaceae), with both species supporting populations over several generations. A CLIMEX model showed that most of the coastal areas of eastern Australia south to 30°16′ S (Coffs Harbour) would be suitable for O. camarae. O. camarae was approved for release in Australia in October 2007 and mines have been observed on plants at numerous field sites along the coast following releases.     

Observations on the host specificity and biology of Lixus salsolae (Col., Curculionidae), a potential biological control agent of Russian thistle, Salsola tragus (Chenopodiaceae) in North America

Abstract:  No-choice host specificity tests were performed on Lixus salsolae Becker (Col., Curculionidae) in a quarantine green house near Montpellier, France. Several varieties of seven species of economic and ornamental plants from six genera of Chenopodiaceae were tested. Adult feeding was observed on almost all test plants and larvae successfully developed on nine of the eleven species/varieties tested. Sex ratio of field-collected overwintering adults was close to 1 : 1. While no-choice tests may indicate a wider host range under field conditions, we no longer consider L. salsolae as a potential biological control agent of Salsola tragus L. (Chenopodiaceae) in North America.     

The biology and host specificity of the onion weed rust, Puccinia barbeyi, a potentially useful agent for biological control in Australia

Onion weed, Asphodelus fistulosus L., (Liliaceae) a weed of Mediterranean and Middle Eastern origin is widespread in southern Australia where it invades pastures making them unsuitable for grazing. A program of research is underway to discover natural enemies of this plant and to study their possible role in the biological control of onion weed. A rust fungus Puccinia barbeyi (Roum.) Magnus has been found to severely attack A. fistulosus . Observations on the biology of the rust confirmed that it is monoecious and microcyclic and multiplied essentially by aecial and telial stages, although occasionally urediniospores also appeared among teliospores. Several members of the Liliaceae exposed to the aeciospores of the rust remained unattacked indicating that it is most probably specific to Asphodelus spp. and thus its potential for the biological control of A. fistulosus in Australia should be studied further.     

Biology, host specificity and risk assessment ofGargaphia decoris, the first agent to bereleased in South Africa for the biological control ofthe invasive tree Solanum mauritianum

The South American tree Solanummauritianum is a major environmental weed in thehigh-rainfall regions of South Africa and has beentargeted for biological control since 1984. Althoughhost ranges of imported agents determined duringquarantine tests have resulted in the rejection ofeight of the 11 candidate agents tested so far, theleaf-sucking lace bug Gargaphia decoris,imported from Argentina in 1995, displayed anacceptably narrow host range in captivity. No-choicetests showed that G. decoris is confined to Solanum species and cannot survive on solanaceouscrops outside that genus. Although these tests alsoindicated that G. decoris colonies could surviveand reproduce on cultivated eggplant (aubergine) andat least five native South African Solanumspecies, all but one native species proved to beinferior hosts in terms of adult survival andoviposition potential. During paired choice andmultichoice tests in small and larger cagesrespectively, G. decoris displayed very strongfeeding and oviposition preferences for S. mauritianum. Analyses of the risk of attack onnon-target Solanum plants revealed that, withone possible exception, none were likely to suffermore than incidental damage in the field. Host recordsfrom South America have also indicated that G. decoris has not been recorded on any Solanumspecies other than S. mauritianum, providingfurther evidence of its host specificity. The resultsof this study were accepted by the regulatoryauthorities and in February 1999, G. decorisbecame the first agent to be released in South Africafor the biological control of S. mauritianum.     

Baculovirus-based strategies for the management of insect pests: a focus on development and application in South Africa

There is growing concern among governments, scientists, agricultural practitioners and the general public regarding the negative implications of widespread synthetic chemical pesticide application for the control of crop pests. As a result, baculovirus biopesticides are gaining popularity as components of integrated pest management (IPM) programmes in many countries despite several disadvantages related to slow speed of kill, limited host range and complex large scale production. In South Africa, baculoviruses are incorporated into IPM programmes for the control of crop pests in the field, and recent bioprospecting has led to the characterisation of several novel isolates with the potential to be formulated as commercial products. This contribution will provide an overview of the use of baculoviruses against insect pests in South Africa, as well as research and development efforts aimed at broadening their application as biocontrol agents. Challenges faced by researchers in developmental projects as well as potential users of baculoviruses as biopesticides in the field are also discussed.     

Biology, host specificity and risk assessment of the leaf-mining flea beetle, Acallepitrix sp. nov., a candidate agent for the biological control of the invasive tree Solanum mauritianum in South Africa

The South American tree Solanum mauritianum, a major environmentalweed in the high-rainfall regions of SouthAfrica, has proved to be a difficult target forbiological control. Artificially expanded hostranges of candidate agents during quarantinehost-specificity tests, have resulted in therelease of only one agent species which has sofar had a negligible impact on the weed. Thenecessity for additional agents resulted in theimportation of a leaf-mining flea beetle, anunidentified species of Acallepitrix,from Brazil in 1997. No-choice tests inquarantine showed that potential host plantsare confined to the genus Solanum.Although several non-target plants, includingcultivated eggplant (aubergine) and some nativeSouth African Solanum species, sustainedfeeding, oviposition and the development oflarval leaf mines, most of these were inferiorhosts. These results were confirmed by pairedchoice tests, where Acallepitrix sp. nov.displayed significant feeding and ovipositionpreferences for S. mauritianum and where,with few exceptions, more larval leaf mineswere initiated on S. mauritianum.Interpretation of the host-specificity testswas facilitated by a risk assessment matrixwhich suggested that the risk of feeding andoviposition on non-target Solanum plants,including eggplant cultivations, was relativelylow. These and other considerations, such asthe lack of evidence of damage inflicted oncultivated Solanaceae in South America, suggestthat Acallepitrix sp. nov. could beconsidered for release against S. mauritianum in South Africa. However, the results of the host-specificity tests remain ambiguous and until more compelling evidence is obtained from field surveys and open-field trials in Brazil, an application for permission to release the flea beetle in South Africa will not be considered.     

Use of a cyanine dye as a probe for albumin and collagen in the extracellular matrix

The aim of this work was to develop a quick method for analysis of macromolecules of the extracellular matrix. Of great interest are soluble components of the extracellular matrix, in particular, carrier proteins, whose variation dynamics can characterize the studied tissue in its development, adult stage, and aging. We suggest the method of analysis of the extracellular matrix to reveal the presence of albumin and collagen by using an anionic cyanine dye as a spectral and fluorescence probe. The method was applied for the analysis of the human vitreous body in the course of its development. Albumin was detected by the appearance of the trans monomer absorption and fluorescence bands in the dye spectra, and collagen was detected by the absorption and fluorescence bands of J aggregates. Hyaluronic acid present in the vitreous body does not interfere with the results of the analysis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis confirmed the presence of albumin in the vitreous body. We suppose that albumin as a protein carrying biologically active macromolecules plays an important role in the processes of differentiation and functional establishment of ocular tissues in the course of their prenatal development.     

The host range and biology of <Emphasis Type="Italic">Cometaster pyrula</Emphasis>; a biocontrol agent for <Emphasis Type="Italic">Acacia nilotica</Emphasis> subsp. <Emphasis Type="Italic">indica</Emphasis> in Australia

Prickly acacia, Acacia nilotica subsp. indica (Benth.) Brenan, a major weed of the Mitchell Grass Downs of northern Queensland, Australia, has been the target of biological control projects since the 1980s. The leaf-feeding caterpillar Cometaster pyrula (Hopffer) was collected from Acacia nilotica subsp. kraussiana (Benth.) Brenan during surveys in South Africa to find suitable biological control agents, recognised as a potential agent, and shipped into a quarantine facility in Australia. Cometaster pyrula has a life cycle of approximately 2 months during which time the larvae feed voraciously and reach 6 cm in length. Female moths oviposit a mean of 339 eggs. When presented with cut foliage of 77 plant species, unfed neonates survived for 7 days on only Acacia nilotica subsp. indica and Acacia nilotica subsp. kraussiana. When unfed neonates were placed on potted plants of 14 plant species, all larvae except those on Acacia nilotica subsp. indica and Acacia nilotica subsp. kraussiana died within 10 days of placement. Cometaster pyrula was considered to be highly host specific and safe to release in Australia. Permission to release C. pyrula in Australia was obtained and the insect was first released in north Queensland in October 2004. The ecoclimatic model CLIMEX indicated that coastal Queensland was climatically suitable for this insect but that inland areas were only marginally suitable.