Seed bank longevity and age to reproductive maturity of Calotropis procera (Aiton) W.T. Aiton in the dry tropics of northern Queensland

Understanding the reproductive biology of Calotropis procera (Aiton) W.T. Aiton, an invasive weed of northern Australia, is critical for development of effective management strategies. Two experiments are reported on. In Experiment 1 seed longevity of C. procera seeds, exposed to different soil type (clay and river loam), pasture cover (present and absent) and burial depth (0, 2.5, 10 and 20 cm) treatments were examined. In Experiment 2 time to reach reproductive maturity was studied. The latter experiment included its sister species, C. gigantea (L.) W.T. Aiton, for comparison and two separate seed lots were tested in 2009 and 2012 to determine if exposure to different environmental conditions would influence persistence. Both seed lots demonstrated a rapid decline in viability over the first 3 months and declined to zero between 15 and 24 months after burial. In Experiment 1, longevity appeared to be most influenced by rainfall patterns and associated soil moisture, burial depth and soil type, but not the level of pasture cover. Experiment 2 showed that both C. procera and C. gigantea plants could flower once they had reached an average height of 85 cm. However, they differed significantly in terms of basal diameter at first flowering with C. gigantea significantly smaller (31 mm) than C. procera (45 mm). On average, C. gigantea flowered earlier (125 days vs 190 days) and set seed earlier (359 days vs 412 days) than C. procera. These results suggest that, under similar conditions to those that prevailed in the present studies, land managers could potentially achieve effective control of patches of C. procera in 2 years if they are able to kill all original plants and treat seedling regrowth frequently enough to prevent it reaching reproductive maturity. This suggested control strategy is based on the proviso that replenishment of the seed bank is not occurring from external sources (e.g. wind and water dispersal).

Diagnostic Molecular Markers for Phosphine Resistance in U.S. Populations of Tribolium castaneum and Rhyzopertha dominica

Stored product beetles that are resistant to the fumigant pesticide phosphine (hydrogen phosphide) gas have been reported for more than 40 years in many places worldwide. Traditionally, determination of phosphine resistance in stored product beetles is based on a discriminating dose bioassay that can take up to two weeks to evaluate. We developed a diagnostic cleaved amplified polymorphic sequence method, CAPS, to detect individuals with alleles for strong resistance to phosphine in populations of the red flour beetle, Tribolium castaneum, and the lesser grain borer, Rhyzopertha dominica, according to a single nucleotide mutation in the dihydrolipoamide dehydrogenase (DLD) gene. We initially isolated and sequenced the DLD genes from susceptible and strongly resistant populations of both species. The corresponding amino acid sequences were then deduced. A single amino acid mutation in DLD in populations of T.castaneum and R.dominica with strong resistance was identified as P45S in T.castaneum and P49S in R.dominica, both collected from northern Oklahoma, USA. PCR products containing these mutations were digested by the restriction enzymes MboI and BstNI, which revealed presence or absence, respectively of the resistant (R) allele and allowed inference of genotypes with that allele. Seven populations of T.castaneum from Kansas were subjected to discriminating dose bioassays for the weak and strong resistance phenotypes. Application of CAPS to these seven populations confirmed the R allele was in high frequency in the strongly resistant populations, and was absent or at a lower frequency in populations with weak resistance, which suggests that these populations with a low frequency of the R allele have the potential for selection of the strong resistance phenotype. CAPS markers for strong phosphine resistance will help to detect and confirm resistant beetles and can facilitate resistance management actions against a given pest population.