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A protocol was developed for production of transgenic iris plants (Iris germanica L. `Skating Party') from regenerable suspension cultures via Agrobacterium-mediated transformation. We tested a series of selection agents, and identified hygromycin and geneticin as the most suitable for selecting transformed iris cells. Suspension cultures of iris were cocultured for 3 days with A. tumefaciens LBA 4404(pTOK233) carrying an intron-interrupted uidA (GUS) gene encoding β-glucuronidase, and hpt (hygromycin) and nptII (geneticin) selectable marker genes. Hygromycin- or geneticin-resistant calli having GUS enzyme activity were identified and used to induce plant regeneration. More than 300 morphologically normal transgenic iris plants were obtained in ≈6 months. About 80% of the transformants were GUS-positive and NPTII-positive (paromomycin-resistant). Integration of transgenes into the nuclear genome of iris plants was confirmed by Southern blot analysis. We have, therefore, developed an efficient A. tumefaciens-mediated transformation system for Iris germanica, which will allow future improvement of this horticulturally important ornamental monocot via genetic engineering.
A proportion of `d'Anjou' pear fruit (Pyrus communis L.) developed a disorder, “black speck” or “skin speckling”, after prolonged controlled atmosphere (CA) storage (1% O2, - 0.5 C). A comparative study of biochemical components revealed that there was no significant difference in succinic, citric, fumaric, and pyruvic acids between the speckled' and normal skin tissues. The content of malic acid in the affected tissue was almost three times lower than that in the normal tissue. The specific activity of NADP-malic enzyme (EC 1.1.1.40) in the affected tissue was also lower, but the total activities were similar. The affected tissue contained higher percentages of dry matter and soluble proteins than the normal tissue. Two-dimensional gel electrophoresis of proteins showed that two groups of novel polypeptides appeared only in the affected skin tissue. This study indicated that a certain proportion of `d'Anjou' pear fruit might have been exposed to unfavorable preharvest environmental stresses, and, therefore, could no longer tolerate the subsequent semi-anaerobic and chilling stresses during prolonged CA storage.
Hybridization and selection has been one of the methods used to generate turfgrass cultivars in buffalograss improvement. Three half-sib populations were developed by crossing three buffalograss female genotypes, NE 3296, NE 2768, and NE 2769, with NE 2871, a male genotype, to 1) investigate the pattern of genetic variability generated for turfgrass characteristics through hybridization; 2) assess the effect of parental change on the level of genetic variability generated in a buffalograss diploid population; and 3) predict the performance of a progeny generated from two heterozygous parents for turfgrass performance. The four parents and 20 random F1 progeny selected from each population were established in 2006 at the John Seaton Anderson Turfgrass Research Facility located near Mead, NE. A randomized complete block design (RCBD) was used with the progeny nested in the crosses. A visual rating scale of 1–9 was used to evaluate the population. Mean population lateral spread, genetic color, density, and turfgrass quality from early summer to fall ranged from 3.5 to 4.5, 7.1 to 7.9, 6.9 to 8.1, and 5.2 and 6.8, respectively. There were significant differences among the crosses and the parents for all the traits studied except quality in June and August. The progeny nested within crosses differed for turfgrass genetic color and quality. Best linear unbiased prediction (BLUP) indicated a high improvement potential for turfgrass lateral spread and spring density in NE 2768 × NE 2871 and for turfgrass genetic color in NE 3296 × NE 2871. From these findings, it can be concluded that hybridization breeding is a worthwhile approach for generating and identifying transgressive segregants for specific buffalograss traits.