Isolation and characterization of novel microsatellite markers and their application for diversity assessment in cultivated groundnut (Arachis hypogaea)

Background: Cultivated peanut or groundnut (Arachis hypogaea L.) is the fourth most important oilseed crop in the world, grown mainly in tropical, subtropical and warm temperate climates. Due to its origin through a single and recent polyploidization event, followed by successive selection during breeding efforts, cultivated groundnut has a limited genetic background. In such species, microsatellite or simple sequence repeat (SSR) markers are very informative and useful for breeding applications. The low level of polymorphism in cultivated germplasm, however, warrants a need of larger number of polymorphic microsatellite markers for cultivated groundnut. Results: A microsatellite- enriched library was constructed from the genotype TMV2. Sequencing of 720 putative SSR-positive clones from a total of 3,072 provided 490 SSRs. 71.2% of these SSRs were perfect type, 13.1% were imperfect and 15.7% were compound. Among these SSRs, the GT/CA repeat motifs were the most common (37.6%) followed by GA/CT repeat motifs (25.9%). The primer pairs could be designed for a total of 170 SSRs and were optimized initially on two genotypes. 104 (61.2%) primer pairs yielded scorable amplicon and 46 (44.2%) primers showed polymorphism among 32 cultivated groundnut genotypes. The polymorphic SSR markers detected 2 to 5 alleles with an average of 2.44 per locus. The polymorphic information content (PIC) value for these markers varied from 0.12 to 0.75 with an average of 0.46. Based on 112 alleles obtained by 46 markers, a phenogram was constructed to understand the relationships among the 32 genotypes. Majority of the genotypes representing subspecies hypogaea were grouped together in one cluster, while the genotypes belonging to subspecies fastigiata were grouped mainly under two clusters. Conclusion. Newly developed set of 104 markers extends the repertoire of SSR markers for cultivated groundnut. These markers showed a good level of PIC value in cultivated germplasm and therefore would be very useful for germplasm analysis, linkage mapping, diversity studies and phylogenetic relationships in cultivated groundnut as well as related Arachis species.

Incorporating dugong habitats into the marine protected area design for the Great Barrier Reef Marine Park, Queensland, Australia.

Dugong habitats were considered in the design for the new zoning network for the Great Barrier Reef Marine Park as part of the Representative Areas Program. One of the specific design guidelines developed as part of the biophysical operational principles recommended that 50% of all high priority dugong habitats should be incorporated in the network of no-take areas. The high priority dugong habitat incorporated in no-take protection increased from 1396 to 3476 km2 (or 16.9-42.0% of all identified sites). Although this increase in protection fell short of the recommended 50%, overall the level of protection afforded by the Great Barrier Reef Marine Park Zoning Plan 2003 increased for all the locations identified.

SSR analysis of cultivated groundnut (Arachis hypogaea L.) germplasm resistant to rust and late leaf spot diseases

Cultivated groundnut (Arachis hypogaea L.) is an agronomically and economically important oilseed crop grown extensively throughout the semi-arid tropics of Asia, Africa and Latin America. Rust (Puccinia arachidis) and late leaf spot (LLS, Phaseoisariopsis personata) are among the major diseases causing significant yield loss in groundnut. The development of varieties with high levels of resistance has been constrained by adaptation of disease isolates to resistance sources and incomplete resistance in resistant sources. Despite the wide range of morphological diversity observed in the cultivated groundnut gene pool, molecular marker analyses have thus far been unable to detect a parallel level of genetic diversity. However, the recent development of simple sequence repeat (SSR) markers presents new opportunities for molecular diversity analysis of cultivate groundnut. The current study was conducted to identify diverse disease resistant germplasm for the development of mapping populations and for their introduction into breeding programs. Twenty-three SSRs were screened across 22 groundnut genotypes with differing levels of resistance to rust and LLS. Overall, 135 alleles across 23 loci were observed in the 22 genotypes screened. Twelve of the 23 SSRs (52%) showed a high level of polymorphism, with PIC values ≥0.5. This is the first report detecting such high levels of genetic polymorphism in cultivated groundnut. Multi-dimensional scaling and cluster analyses revealed three well-separated groups of genotypes. Locus by locus AMOVA and Kruskal-Wallis one-way ANOVA identified candidate SSR loci that may be valuable for mapping rust and LLS resistance. The molecular diversity analysis presented here provides valuable information for groundnut breeders designing strategies for incorporating and pyramiding rust and late leaf spot resistances and for molecular biologists wishing to create recombinant inbred line populations to map these traits.

Increasing energy efficiency in Australian protected cropping

Management of environmental emissions is a significant challenge and opportunity for all of horticulture, including the protected cropping sector. Energy is a significant input in controlled environment horticulture and an important source of environmental emissions. Energy underlies this industry’s capacity to provide a consistent supply of fresh, quality, safe food in a changing global climate.

Insect induced bud fall in cultivated hibiscus and aspects of the biology of Macroura concolor (Macleay) (Coleoptera: Nitidulidae)

The influence of insect attack on bud fall and subsequent poor flowering in cultivated hibiscus (Hibiscus rosa-sinensis) was studied in cages and in the field in southern Queensland. Three species of Hemiptera (most importantly Aulacosternum nigrorubrum but also Nezara viridula and Tectocoris diophthalmus) caused some bud fall in 2 plantations studied. Adults of Macroura concolor suppressed flowering for long periods in spring and summer. Data from white funnel traps and counts in flowers showed that M. concolor was most active in these seasons. Methiocarb (0.75 g a.i./litre) reduced beetle numbers and increased flowering. When 15 or more adults of M. concolor occurred per bud (or flower) most buds fell and few flowers were produced, but when beetles declined to 10 or fewer many buds survived and widespread flowering occurred. Larvae fed in fallen buds and flowers and the mean duration of development of the combined immature stages was 14 days at 26 deg C. The preference of adults of M. concolor for pale coloured flowers was examined. Hibiscus plants produced most buds from December to June with lower numbers in winter and spring (July to November). Bud production in spring and early summer (September-December) varied greatly and probably contributed to poor flowering, however, even when large numbers of buds occurred very few flowers were produced because of the activities of M. concolor.

Phyllosticta spp. on cultivated Citrus in Australia

The occurrence of pathogenic and endophytic species of Phyllosticta on cultivated Citrus in Australia was investigated by DNA sequence analysis of specimens held in plant pathology herbaria and culture collections. Sequences of the internal transcribed spacer region (ITS1, 5.8S, ITS2), and partial translation elongation factor 1-alpha (TEF) gene of 41 Phyllosticta-like isolates from Citrus were compared to those sequences from the type specimens of Phyllosticta recorded from around the world. Phylogenetic analysis resolved all the sequences of Australian accessions into two major clades. One clade corresponded to P. citricarpa, which causes citrus black spot disease. The other clade contained P. capitalensis, which is a known endophyte of Citrus and many other plant species. All included herbarium accessions previously designated as Guignardia mangiferae are now designated P. capitalensis. No Australian isolates were identified as the newly described pathogens of citrus P. citriasiana or P. citrichinaensis, or the endophytes Guignarida mangiferae, P. brazilianiae, or P. citribraziliensis. © 2013 Australasian Plant Pathology Society Inc.

An inordinate fondness for Fusarium: Phylogenetic diversity of fusaria cultivated by ambrosia beetles in the genus Euwallacea on avocado and other plant hosts

Ambrosia beetle fungiculture represents one of the most ecologically and evolutionarily successful symbioses, as evidenced by the 11 independent origins and 3500 species of ambrosia beetles. Here we document the evolution of a clade within Fusarium associated with ambrosia beetles in the genus Euwallacea (Coleoptera: Scolytinae). Ambrosia Fusarium Clade (AFC) symbionts are unusual in that some are plant pathogens that cause significant damage in naive natural and cultivated ecosystems, and currently threaten avocado production in the United States, Israel and Australia. Most AFC fusaria produce unusual clavate macroconidia that serve as a putative food source for their insect mutualists. AFC symbionts were abundant in the heads of four Euwallacea spp., which suggests that they are transported within and from the natal gallery in mandibular mycangia. In a four-locus phylogenetic analysis, the AFC was resolved in a strongly supported monophyletic group within the previously described Cade 3 of the Fusarium solani species complex (FSSC). Divergence-time estimates place the origin of the AFC in the early Miocene similar to 21.2 Mya, which coincides with the hypothesized adaptive radiation of the Xyleborini. Two strongly supported clades within the AFC (Clades A and B) were identified that include nine species lineages associated with ambrosia beetles, eight with Euwallacea spp. and one reportedly with Xyleborus ferrugineus, and two lineages with no known beetle association. More derived lineages within the AFC showed fixation of the clavate (club-shaped) macroconidial trait, while basal lineages showed a mix of clavate and more typical fusiform macroconidia. AFC lineages consisted mostly of genetically identical individuals associated with specific insect hosts in defined geographic locations, with at least three interspecific hybridization events inferred based on discordant placement in individual gene genealogies and detection of recombinant loci. Overall, these data are consistent with a strong evolutionary trend toward obligate symbiosis coupled with secondary contact and interspecific hybridization. (C) 2013 Elsevier Inc. All rights reserved.

Host specificity studies on Gynaikothrips (Thysanoptera: Phlaeothripidae) associated with leaf galls of cultivated Ficus (Rosales: Moraceae) trees

Host specificity tests on Gynaikothrips ficorum (Marchal) and Gynaikothrips uzeli (Zimmerman) (Thysanoptera: Phlaeothripidae) have shown that under experimental conditions, G. ficorum will induce leaf galls on both Ficus benjamina L. and Ficus microcarpa L. f. (Rosales: Moraceae), but G. uzeli will induce galls only on F. benjamina. A further interesting aspect of the results is that gall induction by G. uzeli on F. benjamina appears to have been suppressed in the presence of F. microcarpa plants in the same cage. Liothrips takahashii (Moulton) (Thysanoptera: Phlaeothripidae), an inquiline in the galls of these Gynaikothrips, is reported for the first time from Australia, mainland China, Malaysia, Costa Rica, and western USA.

Fungi associated with foliar diseases of wild and cultivated rice (Oryza spp.) in northern Queensland

During surveys of wild and cultivated rice in northern Queensland in 2014 and 2015, 92 fungal isolates were obtained from plants that were afflicted by foliar diseases, including the rice blast pathogen, Pyricularia oryzae, and the brown spot pathogen, Bipolaris oryzae. Seven species of Curvularia were found, viz. Curvularia aeria, C. alcornii, C. asianensis, C. clavata, C. lunata, C. muehlenbeckiae and an undescribed species. To remove uncertainty about the identity of the host plants from which the fungi were isolated, a DNA barcoding strategy was developed using regions of the chloroplast genome. Pathogenicity tests using wild rice isolates of P. oryzae indicated that many local rice varieties are susceptible to infection.

Protected cropping of strawberry plants in subtropical Queensland

The effect of protected cropping on the performance of two strawberry cultivars ('Festival' and 'Rubygem') and two breeding lines (Breeding Lines 1 and 2) was studied in subtropical Queensland, Australia over two years. Production in this area is affected by rain, with direct damage to the fruit and the development of fruit diseases before harvest. The main objective of the study was to determine whether plants grown under high plastic tunnels had less rain damage, less disease incidence, and higher yields than plants grown outdoors. Our studies show that marketable yields were up to 40% higher in the plants under the tunnels compared with yields of the plants outdoors. This was mainly because fruit from the plants grown under the tunnels had lower incidences of rain damage and/or grey mould. There were no consistent differences in the relative numbers of small and/or misshaped fruit in the two growing environments. This research highlights the potential of protected cropping for strawberry producers in subtropical areas that receive significant rainfall during the growing season.

Variation of cutting rooting ability in cultivated and wild species of Macadamia

In Australia, macadamia trees are commonly propagated by germinating rootstock seed and grafting when seedlings reach a suitable size. The production of grafted trees is a protracted and complex process, however, propagation of macadamia via cuttings represents a simpler and faster method of multiplication. Macadamias have traditionally proven difficult to propagate from cuttings, and while recent developments in the process have improved success rates, substantial variation in rooting ability between cultivars and species has been reported. The cultivar 'Beaumont' (Macadamia integrifolia × M. tetraphylla) is commonly propagated by cutting for use as a rootstock, and is relatively easy to strike while other cultivars are more difficult. There is speculation that Hawaiian cultivars are more difficult to strike from cuttings than Australian cultivars due to species and genetic composition. In this experiment, cuttings of 32 genotypes were evaluated for rooting ability. Each genotype's species profile was estimated using historical data, and used to determine species effects on survival (percentage) and rooting ability (rating 0-2). M. jansenii (100%), M. tetraphylla (84%) and M. integrifolia/tetraphylla hybrids (79%) had the highest success rates while M. integrifolia (54%) and M. ternifolia (43%) had the lowest survival. Rooting ability of M. jansenii (1.75) was significantly higher than M. ternifolia (0.49) but not significantly higher than M. tetraphylla × M. integrifolia with (1.09), M. tetraphylla (1.03) or M. integrifolia (0.88).

Protected cropping for specialty melons in North Queensland

Protective cropping could be an effective system for growing specialty melons in the dry tropics of North Queensland. The growing system could reduce outdoor risks for production loss, improve fruit quality, increase yield per m2, allow production offseason, and used for supplying niche markets in a segment of the larger melon market in Australia. First evaluations in Giru, Queensland, included seven cultivars of fruit types 'Galia', 'Hami', 'Charentais', small 'Canary', and 'Rockmelon', transplanted July 25, 2013 under a high polyethylene-covered tunnel. Plants were grown at a density of 2.8 plants m-2 in containers filled with volcanic rock and irrigated with a complete nutrient solution. Pruning and trellising was done to a single vertical stem, keeping lateral shoots on the main stem after the 7th leaf node. After bearing small fruit, lateral shoots were cut off after their second or third leaf node. To facilitate insect pollination, a screen window in the tunnel was left partially opened. On November 20 the cultivars had combined marketable yields that ranged from 2.8 to 8.2 fruits m-2 and 3.1 to 7.8 kg m-2. Total soluble solids levels in fruit ranged from 6 to 13 °Brix. Cultivars 'Tempo' ('Galia'), 'Tikal' ('Canary') and 'Sultan' ('Charentais') had fruit yields that were up to 2.6 times greater than yields commonly achieved with field-grown rockmelon crops. Sugar levels in fruits and marketable yields may be increased with changes in fertigation management. Promising results in this first evaluation justify examination of a greater number of genetic materials, in addition to the development of economic feasibility studies and further adaptive research to refine crop recommendations for growing melons in protective cropping systems.

Genetic Options for Drought Management in Groundnut

Groundnut is one of the principal oilseeds in the world. It is cultivated on 24.8 million ha with a total production of 32.8 million t and an average productivity of 1.32 t ha-'. Developing countries account for 96.9% of the world groundnut area and 93.8% of total production. Production is concentrated in Asia (56.8% area and 66.5% production of the world) and Africa (38.0% area and 24.7% production). The groundnut productivity in Africa is only 0.86 t ha-' compared with 1.55 t hx1 of Asia. The world groundnut economy-facts, trends and outlook are desaibed in detail by Freeman et al., 1999. Briefly, in medium-term (i.e. up to 2010), 'groundnut production and consumption is likely to shift increasingly to developing countries; production will grow in all regions but most rapidly in Asia, slowly in sub-Saharan Africa and decline in Latin America; and utilizationwill continue to shift away from groundnut oil toward groundnut meal, specially confectionery products'.

First record of Nematospora coryli in Australia and its association with dry rot of Citrus

Nematospora (Eremothecium) coryli was isolated from Citrus and identified for the first time in Australia. This insect-transmitted yeast was associated with dry rot in cultivated and native Citrus fruits. Although N. coryli is known as a serious seed pathogen of many tropical and sub-tropical plants, evidence is presented that it has been present and undetected in Queensland for at least ninety years.