High-density linkage maps and loci for berry color and flower sex in muscadine grape (Vitis rotundifolia).

KEY MESSAGE: Linkage maps of muscadine grape generated using genotyping-by-sequencing (GBS) provide insight into genome collinearity between Muscadinia and Euvitis subgenera and genetic control of flower sex and berry color. The muscadine grape, Vitis rotundifolia, is a specialty crop native to the southeastern USA. Muscadine vines can be male, female, or perfect-flowered, and berry color ranges from bronze to black. Genetic linkage maps were constructed using genotyping-by-sequencing in two F1 populations segregating for flower sex and berry color. The linkage maps consisted of 1244 and 2069 markers assigned to 20 linkage groups (LG) for the 'Black Beauty' × 'Nesbitt' and 'Supreme' × 'Nesbitt' populations, respectively. Data from both populations were used to generate a consensus map with 2346 markers across 20 LGs. A high degree of collinearity was observed between the genetic maps and the Vitis vinifera physical map. The higher chromosome number in muscadine (2n = 40) compared to V. vinifera (2n = 38) was accounted for by the behavior of V. vinifera chromosome 7 as two independently segregating LGs in muscadine. The muscadine sex locus mapped to an interval that aligned to 4.64-5.09 Mb on V. vinifera chromosome 2, a region which includes the previously described V. vinifera subsp. sylvestris sex locus. While the MYB transcription factor genes controlling fruit color in V. vinifera are located on chromosome 2, the muscadine berry color locus mapped to an interval aligning to 11.09-11.88 Mb on V. vinifera chromosome 4, suggesting that a mutation in a different gene in the anthocyanin biosynthesis pathway determines berry color in muscadine. These linkage maps lay the groundwork for marker-assisted breeding in muscadine and provide insight into the evolution of Vitis species.

Recombination within the apospory specific genomic region leads to the uncoupling of apomixis components in Cenchrus ciliaris.

Apomixis enables the clonal propagation of maternal genotypes through seed. If apomixis could be harnessed via genetic engineering or introgression, it would have a major economic impact for agricultural crops. In the grass species Pennisetum squamulatum and Cenchrus ciliaris (syn. P. ciliare), apomixis is controlled by a single dominant "locus", the apospory-specific genomic region (ASGR). For P. squamulatum, 18 published sequenced characterized amplified region (SCAR) markers have been identified which always co-segregate with apospory. Six of these markers are conserved SCARs in the closely related species, C. ciliaris and co-segregate with the trait. A screen of progeny from a cross of sexual × apomictic C. ciliaris genotypes identified a plant, A8, retaining two of the six ASGR-linked SCAR markers. Additional and newly identified ASGR-linked markers were generated to help identify the extent of recombination within the ASGR. Based on analysis of missing markers, the A8 recombinant plant has lost a significant portion of the ASGR but continues to form aposporous embryo sacs. Seedlings produced from aposporous embryo sacs are 6× in ploidy level and hence the A8 recombinant does not express parthenogenesis. The recombinant A8 plant represents a step forward in reducing the complexity of the ASGR locus to determine the factor(s) required for aposporous embryo sac formation and documents the separation of expression of the two components of apomixis in C. ciliaris.

Insect-attracting and antimicrobial properties of antifreeze for monitoring insect pests and natural enemies in stored corn.

Insect infestations in stored grain cause extensive damage worldwide. Storage insect pests, including the Indianmeal moth, Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae); Sitophilus spp. (Coleoptera: Curculionidae); and their natural enemies [e.g., Cephalonomia tarsalis (Ashmead) (Hymenoptera: Bethylidae), and Anisopteromalus calandrae (Howard) (Hymenoptera: Pteromalidae)] inhabit a temporary, but stable ecosystem with constant environmental conditions. The objective of the present experiment was to assess the efficacy of using ethylene glycol antifreeze in combination with nutrient solutions to monitor storage insect pest and natural enemy populations in three bins of corn, Zea mays L. The treatments were deionized water, a diluted (1:5 antifreeze:water) antifreeze solution, 10% honey, 10% honey in the diluted antifreeze solution, 10% beer in the diluted antifreeze solution, 10% sucrose in the diluted antifreeze solution, and a commercial pheromone trap suspended in a 3.8-liter container filled with 300-ml of diluted antifreeze solution. The seven treatments captured storage insect pests and their natural enemies in the bins at 33-36 degrees C and 51-55% RH. The pheromone trap in the container with the diluted antifreeze captured significantly more P. interpunctella than the other treatments, but a lower percentage (7.6%) of these captures were females compared with the rest of the treatments (> 40% females). All trapping solutions also captured Sitophilus spp. and other beetle species, but the captures of the coleopteran pests were not significantly different among the seven treatments (P > 0.05). Two parasitoid wasps also were captured in the study. The number of A. calandrae was different among the seven treatments (P < 0.05), whereas the number of C. tarsalis was not different among the treatments (P > 0.05). Most A. calandrae adults were captured by the 10% honey in the diluted antifreeze, whereas the fewest were captured in the deionized water. Microbial growth was observed in the 10% honey solution, but no microbial growth occurred in the rest of the treatments, including 10% honey in the diluted antifreeze solution. The results of insect captures and microbial growth demonstrated that antifreeze could be used as a part of storage insect monitoring and/or control programs.

Sequence analysis of bacterial artificial chromosome clones from the apospory-specific genomic region of Pennisetum and Cenchrus.

Apomixis, asexual reproduction through seed, is widespread among angiosperm families. Gametophytic apomixis in Pennisetum squamulatum and Cenchrus ciliaris is controlled by the apospory-specific genomic region (ASGR), which is highly conserved and macrosyntenic between these species. Thirty-two ASGR bacterial artificial chromosomes (BACs) isolated from both species and one ASGR-recombining BAC from P. squamulatum, which together cover approximately 2.7 Mb of DNA, were used to investigate the genomic structure of this region. Phrap assembly of 4,521 high-quality reads generated 1,341 contiguous sequences (contigs; 730 from the ASGR and 30 from the ASGR-recombining BAC in P. squamulatum, plus 580 from the C. ciliaris ASGR). Contigs containing putative protein-coding regions unrelated to transposable elements were identified based on protein similarity after Basic Local Alignment Search Tool X analysis. These putative coding regions were further analyzed in silico with reference to the rice (Oryza sativa) and sorghum (Sorghum bicolor) genomes using the resources at Gramene (www.gramene.org) and Phytozome (www.phytozome.net) and by hybridization against sorghum BAC filters. The ASGR sequences reveal that the ASGR (1) contains both gene-rich and gene-poor segments, (2) contains several genes that may play a role in apomictic development, (3) has many classes of transposable elements, and (4) does not exhibit large-scale synteny with either rice or sorghum genomes but does contain multiple regions of microsynteny with these species.