Genetic engineering of insect viruses for insect biological control

Insect viruses can be exploited for the biological control of insect pests and serve as well as vehicles for the introduction of ‘foreign genes’ into insect cells and insects. The present work describes some of the molecular events that take place during the replication of theGalleria mellonella densonucleosis virus in the insect host. Viral RNA and protein synthesis were monitored. An understanding of the virus biology is essential for further implementation of these viruses for biological control purposes.     

Indol-3-ylacetic acid- and calmodulin-regulated Ca2+-ATPase: A target for the phytotoxic action of hexachlorocyclohexane

In rice seedlings, the organochlorine insecticide hexachlorocyclohexane (HCH) and its four major isomers—α,β, γ, and δ—have been shown to elicit their phytotoxic action by interacting with indol-3-ylacetic acid (IAA)-regulated growth and Ca²⁺-ATPase activity. When rice seedlings were grown in the presence of 0.34 mM HCH, seedling vigour was reduced to 33% of the control. A similar effect was observed when seedlings were grown in the presence of the γ and δ isomers, but not with the α and δ isomers of HCH. This reduced vigour could be restored by treating the seedlings with 100 nM IAA, suggesting that HCH and its isomers (γ and δ) limit either IAA synthesis or action or both. In a microsomal fraction from rice seedlings grown in the presence of either HCH or its isomers, the calmodulin-regulated Ca²⁺-ATPase activity was inhibited as follows: technical HCH = 46%, α isomer = 80%, β isomer = 72%, γ isomer = 65% and δ isomer = 62% of the control value, respectively. TLC analysis suggested that the various isomers of HCH are metabolised by the plant, except for the δ isotner, which accumulated. This isomer, along with the degraded products of other isomers, may be responsible for the phytotoxic action of HCH.     

Development of glasshouse pests of cucumber under different conditions,in relation to possibilities for their biological control1

Experiments were performed under controlled conditions on the development of four cucumber pests (Trialeurodes uaporariorum, Tetranychus urticae, Aphis gossypii and Thrips tabaci). The main variables were temperature, photoperiod and light intensity. The effect of these factors on the natural enemies Encarsia formosa and Amblyseius mckenziei was also assessed. Comparison of the results allows recommendations on manipulation of the glasshouse environment to favour biological control.     

Variation for pathogenicity among isolates of bean common mosaic virus in Africa and a reinterpretation of the genetic relationship between cultivars of Phaseolus vulgaris and pathotypes of BCMV

Bean common mosaic virus (BCMV) isolates were collected from crops of Phaseolus vulgaris (bean) and from wild legume species in 13 African countries. Isolates of pathotype VIa from both beans and wild legume species were predominant in central, eastern and southern Africa. Isolates of pathotypes I, III, IVa, IVb and Va were also found. Some isolates did not conform to previously published pathotypes, and therefore represent records of novel pathotypes. The susceptibility of various wild legume species to BCMV was investigated and isolates of the virus obtained from Crotalaria incana, Rhynchosia sp., Macroptilium atropurpureum and Cassia occidentalis (synonym Senna occidentalis) were aphid-transmitted both from P. vulgaris to their original host species and to P. vulgaris. Isolates of BCMV from wild legume species were seed-transmitted in bean and in several other legume species. The natural occurrence of BCMV in wild legume species in Africa is probably a significant factor in the ecology and epidemiology of the virus and possibly the evolution of isolates of the 'A' serotype which induce necrotic reactions in cultivars carrying the I gene for resistance. The occurrence of viruses other than BCMV from P. vulgaris and other legume hosts is also reported. The gene-for-gene model described by Drijfhout (1978) is reinterpreted to explain the variation for pathogenicity, and it is proposed that there may be genes which control the temperature sensitivity of necrosis in combination with the I gene.     

Losses in grain yield of winter crops from Lolium rigidum competition depend on crop species, cultivar and season

The competitive abilities of eight winter crops were compared against Lolium rigidum Gaud, (annual ryegrass), an important weed of southern Australia, as a potential strategy to suppress weeds and reduce dependence on herbicides. Two cultivars of each species were chosen to represent the range of competitive ability within each crop and grown in field experiments in 1992 and 1993. The order of decreasing competitive ability (with the ranges of percentage yield reduction from L. rigidum at 300 plants m^?2 in parenthesis) was as follows: oats (Avena sativa L.), 2–14%; cereal rye (Secale cereale L.), 14–20%; and triticale (×Triticosecale), 5–24%; followed by oilseed rape, (Brassica napus L.), 9–30%; spring wheat (Triticum aestivum L.), 22–40%; spring barley (Hordeum vulgare L.), 10–55%; and, lastly, field pea (Pisum sativum L.), 100%, and lupin (Lupinus angustifolius L.), 100%. Differences in competitive ability of cultivars within each species were identified, but competition was strongly influenced by seasonal conditions. Competition for nutrients (N, P and K) and light was demonstrated. L. rigidum dry matter and seed production were negatively correlated with grain yield of the weedy crops. More competitive crops offer the potential to suppress grass weeds while maintaining acceptable grain yields. Ways of improving the competitive abilities of grain legume crops are discussed.     

The use of polymerase chain reaction to detect latent infection of Clavibacter michiganensis subsp. michiganensis in tomato seedlings

A method was developed for routine analysis to detect latent infection of Clavibacter michiganensis subsp. michiganensis in samples from tomato seedling lots, before transplant, using polymerase chain reaction. In samples of 300 stem segments 1 cm long, the sensitivity threshold of the method was estimated at around 1.1 × 10³ corynebacteria or 0.33% of latently infected stems. This method was shown to have a good specificity.     

Detection of Spongospora subterranea using monoclonal antibodies in ELISA

Five hybridoma cell lines secreting antibodies (MAbs) recognizing zoospores of S. subterranea were raised from splenocytes of mice. One MAb also weakly recognized plasmodia/zoosporangia and cystosori of S. subterranea , and another recognized only plasmodia/zoosporangia in plate-trapped antigen ELISA. Polymyxa graminis was recognized most strongly out of 26 micro-organisms other than S. subterranea against which the MAbs were tested. Most were recognized only weakly or not at all. The MAb that recognized zoospores of S. subterranea most strongly detected as few as three zoospores per microtitre plate well when 12 replicate wells per treatment were arranged randomly on plates and absorbance values subjected to analysis of variance. The sensitivity of detection was not improved by mixing antibodies, using a biotin-streptavidin amplification system, or by using a double antibody sandwich system. Zoospores of S. subterranea flushed from soil were detected only after unrealistically large numbers of cystosori had been added. They were not detected in samples of naturally infested soil removed from a field shortly after a severely scabbed potato crop had been harvested.     

The effect of timing of fungicide applications on control of frogeye leaf spot and grain yield of soybeans

Soybean cultivar Samsoy 1, and the breeding lines TGx 849-313D and TGx 996-26E, grown in a field with a heavy epidemic of frogeye leaf spot caused byCercospora sojina, were treated with double foliar applications of the fungicide benomyl. The treatments were made using four application schedules at six different growth stages, starting from V₃ (fully developed leaves, beginning with trifoliate nodes) to R₅ (beginning seed_, to determine the effect of the fungucide timing on frogeye leaf spot severity, soybean grain yield and grain quality. Generally, applications at R₁ (beginning bloom) and R₃ (beginning pod) significantly (P<-0.05) reduced disease severity in the 2 susceptible genotypes, Samsoy 1 and TGx 849-313D. Plot yields of these genotypes were also significantly greater than the untreated controls when the fungicide applications were made at R₁ and R₃. There was no significant difference in diseave severity or grain yield, between the untreated control and the different times of application, on the resistant genotype TGx 996-26E. Improved seed germination and lower levels of seed infection byC. sojina occurred for all fungicide timings in the susceptible genotypes. The results suggest that fungicide spraying initiated at R₁ and followed up at R₃ is most effective in frogeye leaf spot control and can also result in higher grain yields, than applications made earlier or later in the season. Control of frogeye leaf spot, however, is best achieved by growing resistant cultivars.     

Occurrence and viability of Septoria oenotherae in commercial seed samples of evening primrose

The antiphytoviral activity of 1-morpholinomethyl-tetrahydro-2(1H)-pyrimidinone (DD13) in a test system including protoplast cultures, surviving tissues and greenhouse plants was examined. The inhibitory effect was quantitatively investigated by immunofluorescence and enzyme linked immunosorbent assay. The antiviral action in vitro was 96%. The first 6 h after inoculation was the most sensitive period of the tomato mosaic virus (ToMV) reproduction cycle. DD13 possessed a protective effect in 97–100% plants infected with ToMV and cucumber mosaic virus (CMV).     

Persistence and fitness of carbendazim- and dicarboximide-resistant isolates of Monilinia fructicola (Wint.) Honey in flowers, shoots and fruit of stone fruit

Flowers on detached peach shoots and ripe fruit were inoculated under controlled conditions in order to estimate the competitive ability of Monilinia fructicola isolates sensitive and resistant to carbendazim and dicarboximide fungicides. Isolates varied considerably, but there was no consistent relationship between carbendazim resistance and competitive ability; there was, however, evidence of reduced competitive ability in the single dicarboximide-resistant and the dual dicarboximide/high carbendazim-resistant isolate examined. The ability to produce conidia on twig cankers inoculated in late spring 1989 was retained under field conditions by all sensitive and resistant isolates for at least 1 year. Dicarboximide-resistant isolates produced fewer conidia than the carbendazim-resistant and sensitive isolates on cankers. The production of conidia on mummified fruit inoculated in February 1990 decreased in the field during the winter and the following spring, but some conidia were produced by all isolates. Measures of pathogenicity, virulence and fitness for all isolates were similar to the original values after survival for 1 year. The evidence presented in this paper supports field observations that carbendazim-resistant isolates are likely to persist permanently in the M. fructicola population, whereas dicarboximide-resistant isolates are more likely to decline unless fungicide selection pressure is maintained.