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- Author or Editor: Hazel Y. Wetzstein x
Tillandsia eizii is an epiphytic bromeliad that due to over-collection, habitat destruction, and physiological constraints has declined to near threatened status. This species exhibits high mortality in the wild, and seed are characterized by low percentages of germination. As a means to conserve this species, in vitro culture protocols were developed to enhance seed germination and seedling growth. A sterilization protocol using 70% ethanol for 2 minutes followed by 2.6% NaOCl for 40 minutes disinfested seed and promoted seedling growth. Sucrose incorporated into the culture medium had no effect on germination or growth, while NAA inhibited growth, but not germination. Cultures maintained under a 16-hour photoperiod at 22 °C exhibited greater growth than those grown at 30 °C. Seed that germinated in the dark remained etiolated and failed to develop even after transfer to light conditions. Plants grown in vitro were successfully acclimatized and transferred to the greenhouse. Over 86% survival and rapid growth were obtained with either an all-pine-bark medium, or a mixture of 2 redwood bark: 2 fir bark: 2 potting mix: 1 perlite. This demonstrated that in vitro culture of seed may be used to rapidly produce large numbers of T. eizii, and thus can be used for the conservation and reintroduction of this species.
Rabbiteye blueberry (Vaccinium ashei Reade) often exhibits problems with low fruit set. Little is known about the duration of flower receptivity in this species. The objective of this study was to investigate the effects of flower age at pollination on fruit set, seed number per fruit, and stigmatic receptivity. `Brightwell' and `Tifblue' rabbiteye blueberry plants were kept under controlled conditions in a growth chamber. Day/night temperatures during pollination were 23 °C/10 °C. Flowers were hand pollinated with self- or cross-pollen at 2-day intervals ranging from 0 to 8 days after anthesis (DAA). Flower age at pollination had a significant effect on both fruit set and seed number per fruit. Rabbiteye blueberry flowers were able to produce optimum fruit set during a period of at least five days. Fruit set was markedly reduced 6 to 8 DAA, depending on the cultivar. Flower age at pollination also had a significant effect on stigmatic receptivity, which was assessed as the number of germinated tetrads on the stigma 24 hours after pollination. Stigmas pollinated 0 DAA had a significantly lower number of germinated tetrads than those pollinated 8 DAA. Flower age at pollination and stigmatic receptivity were positively associated. To our knowledge, this is the first quantitative evidence of delayed stigma maturation in blueberry. Stigmatic receptivity and fruit set were not correlated. Overall, the data strongly suggest that stigmatic receptivity was not a limiting factor for fruit set of `Brightwell' and `Tifblue'. It is hypothesized that ovule longevity determines the duration of flower receptivity in these two rabbiteye blueberry cultivars.
Fruit set of rabbiteye blueberries (Vaccinium ashei Reade) can be pollen-limited under certain conditions. The objective of this study was to determine production, release, and viability of pollen, as well as pollen-ovule ratios in the rabbiteye blueberry cultivars Austin, Brightwell, Climax, and Tifblue. In vitro tetrad germination varied among genotypes, although, values were high (≥80%) in all cultivars. Pollen viability does not seem to contribute to reproductive failure in the cultivars studied. Total pollen production per flower averaged 8434 tetrads across all cultivars. On a per ovule basis, pollen production was very low relative to other xenogamous species. The low pollen-ovule ratio of rabbiteye blueberry (≈400) may be an indicator of the high efficiency of its pollen dispensing mechanism. Total pollen production varied among cultivars. Furthermore, a significant difference in pollen release was found between two cultivars with similar total pollen production per flower. The possible mechanism regulating pollen release in these cultivars is discussed.
Field evaluations were conducted of pecan [Carya illinoinensis (Wangenh.) C. Koch] trees regenerated via somatic embryogenesis to assess if the trees maintained clonal fidelity and exhibited true-to-type characteristics. Phenotypic and molecular comparisons were made of trees from two different tissue culture lines after 4 years in the field. Factors evaluated included shoot growth, leaf morphology, and susceptibility to fungal scab [Cladosporium caryigenum (Ellis & Langl.) Gottwald] and southern pecan leaf phylloxera (Phylloxera russellae Stoetzel). Genetic fidelity was examined using amplified fragment length polymorphism (AFLP) analysis. Statistically significant differences were observed between the culture lines in phenotypic leaf characteristics (i.e., specific leaf weight and leaf length-to-width ratio), number of shoots per 1-year-old branch, and in the frequency of scab lesions on leaves. No between-line differences were observed in trunk caliper, average and total shoot growth, shoot length per cross-sectional area, or presence of phylloxera galls. AFLP analysis readily detected differences between culture lines. Cluster analysis generally grouped trees together that were regenerated from the same line. Trees within a culture line usually exhibited similar leaf characteristics, but not shoot growth or tree height. A few trees exhibited more extreme leaf characteristics and differed from each other. However, they were statistically similar to most of the other trees in the population evaluated. AFLP data revealed that some trees exhibited greater divergence and less similarity than other trees from the same line. The nature and significance of such variation at this time are not related to any detectable phenotypic differences.
It is increasingly evident that the concentration and type of matrix used in nutrient media can profoundly influence the response of tissues cultured in vitro. Picea abies shoot cultures exhibited increased dry weight yields (4) and increased shoot growth (9) with decreasing agar concentrations. Singha (5) compared shoot proliferation of crabapple and pear in Phytagar concentrations ranging from 0% to 1.2%. Optimal proliferation and growth occurred in crabapple on 0.3% agar, with higher concentrations decreasing these growth characteristics. In pear, increasing agar concentration also decreased shoot growth, but shoot proliferation was greatest at 0.6% or higher agar concentrations. Rooting has been promoted without or with lower agar concentrations in grape (1), Norway spruce (9), and apple (10). However, vitrification (glassiness or waterlogging) has been shown to increase with lower agar concentrations (9) and limits the use of liquid culture in some species.
Tissue cultures of pecan [Carya illinoensis (Wangenh.) C. Koch] were initiated from embryo explants collected weekly from 12 weeks post-pollination until fruit maturity. Three cultures derived from immature embryos collected 14 weeks postpollination produced primary somatic embryos within 1 month following transfer from modified woody plant medium (WPM) with 2.0 mgliter−1 2,4-D and 0.25 mg liter−1 BA in the light to hormone-free medium in the dark. Scanning electron microscopy documented the development of secondary embryos, which followed a globular, heartshaped, and torpedo-stage developmental sequence. Chemical names used: (2,4-di-chlorophenoxy) acetic acid (2,4-D), N-(phenylmethyl)-1H-purin-6-amine (BA), and 1H-indole-3-butanoic acid (IBA).