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  • Author or Editor: Pablo S. Jourdan x
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Before attempting transformation of Pelargonium spp., it is necessary to develop procedures for shoot regeneration under selective conditions (kanamycin resistance). Leaf disks of various cultivars of regal, zonal, ivy-leaved, and scented pelargoniums were compared for shoot regenerability and phenolic production. Disks were cultured on MS medium supplemented with 10.7 μM NAA and 8.9 μM BA. After three weeks in darkness, each disk was transferred to MS with 0.89 μM BA and evaluated for proportion of disks with callus, extent of disk development, and the rate and intensity of phenolic halo production. Phenolic production was not correlated with shoot regeneration ability. Regal pelargoniums had low phenolic production, while ivy-leaved pelargoniums had both the highest phenolic production and callus formation. Shoot regeneration from leaf disks was inhibited on media containing 100 μg/ml kanamycin.

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Abstract

More than 65 different genotypes, including cultivars and inbred lines, from five cruciferous species (Brassica oleracea L., B. campestris L., B. napus L., B. juncea L., and Raphanus sativus L.) were tested for their in vitro response of leaf protoplasts. Protoplasts were cultured in three liquid media and the resulting colonies were placed on seven test regeneration media. Significant differences among the species were found in plating efficiency in the frequency of shoot regeneration. Two broad response groups were identified: 1) Cultivars from B. oleracea and B. napus—these generally yielded protoplasts that were able to divide, form colonies at high frequencies, and regenerate shoots at variable frequencies; and 2) cultivars of the other species evaluated, which typically exhibited low plating efficiencies and little, if any, shoot regeneration. Evaluation for the effect of the cytoplasmic constitution of a few B. oleracea breeding lines on in vitro performance indicated that protoplasts carrying the Ogura (R1) male-sterile cytoplasm regenerated shoots at slightly lower frequencies than the corresponding alloplasmic-fertile lines. Genotypes exhibiting high frequency of shoot formation in one medium also had efficient shoot regeneration in other media as well, while genotypes with low shoot regeneration responded consistently in the different media used. This consistency in response indicates that genotype plays a critical role in determining the success of leaf protoplast culture in the crucifers.

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The genus Aesculus (buckeyes and/or horsechestnuts) is composed of 13 species and a number of interspecific hybrids. Pollen from 11 genotypes from five Aesculus species and the hybrid Aesculus ×carnea were used to develop an in-vitro germination test to evaluate pollen viability under various storage treatments. This test was optimized using samples of both fresh pollen and pollen that had been stored up to 1 year. The most effective medium contained 20% sucrose, 100 mg·L-1 H2BO3, 150 mg·L-1 Ca(NO3)2, and 1% agar. The highest germination percentage was observed at 15 °C across all storage treatments. Fresh pollen germinated in excess of 80% over a wide range of germination temperatures. Based on this, all specimens studied would be good pollen parents. The differences in pollen germination between storage at -20 and -80 °C were nonsignificant, but the duration of the storage period was highly significant. At 3 months, viability remained above 60% for four of the six species/hybrid tested. However, at 12 months, all pollen tested dropped below the threshold for good fruit set based on in-vitro pollen germination. Based on these observations, short-term pollen storage may permit crosses between parents with temporally separate flowering phenologies. However, conventional storage procedures are inadequate to maintain pollen collected from a male parent for crosses in subsequent growing seasons.

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As a prelude to interspecific hybridization, we compared the floral biology of bottlebrush buckeye (Aesculus parviflora) and red buckeye (A. pavia) by examining inflorescence morphology, pattern of floral anthesis, sex expression, and the effects of panicle decapitation on complete flower development. Inflorescences of both species (n = 1606) were randomly selected and analyzed for length, total number of flowers and complete flower number and location. The pattern of anthesis was observed in four genotypes using 10–30 inflorescences per plant. For each flower, its date of anthesis, position on both the rachis and cincinnus, and sex were recorded. For studies of panicle decapitation, sets of panicles were selected and one member was severed in half early in development in an attempt to increase the number of complete flowers. More than one-fourth of all panicles observed were completely staminate. For both species, the ratio of complete flowers to male flowers (C:M) within mixed panicles was about 5%. Complete flowers were observed in the basal portion of A. pavia inflorescences and in the apical portion of A. parviflora inflorescences. Anthesis progressed from base to tip over a period of 6–11 days. Complete flowers are present in A. pavia from the beginning of anthesis but do not appear in A. parviflora until the fifth day of anthesis. Staminate flowers are present throughout anthesis in both species. Severing panicles in half increased the potential for differentiating complete flowers. In conclusion, the frequency of complete flowers in both species was quite low, but could be increased by panicle decapitation to increase opportunities for controlled hybridization.

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Corylopsis seed germination tests were conducted to assess the influence of harvest date (seed maturity) and cold stratification (CS) at 5 °C. Corylopsis gotoana seeds harvested on 12 July, 2 and 22 Aug., 6 and 20 Sept., and 1 and 10 Oct. 2011 were immersed in water for 20 min to separate fully developed seeds (full seeds) from empty seeds by floatation, and by X-ray scanning to identify full from empty seeds (Expt. 1). Immersing seeds in water did not effectively separate full seeds from empty seeds as evaluated by seed germination tests. Seeds harvested on or around 6 Sept. that sank showed translucent X-ray images with fully developed internal structures composed of embryo, cotyledons, and endosperm, and were considered mature. Without CS, >12% seeds harvested on 20 Sept. germinated, regardless of whether seeds were full or empty. Seeds of C. coreana harvested on 5 and 15 Sept., and 5 and 18 Oct. were stored dry at 20 °C until 27 Dec. and germinated after 0, 3, 6, 9, and 12 weeks of CS (Expt. 2). Longer than 6 weeks of CS was required to accelerate and increase the germination of seeds harvested on or after 5 Sept. Germination percentage of full seeds harvested on Oct. 18 was increased to >72% as the duration of CS treatment increased to 12 weeks. In conclusion, fully developed seeds harvested on or after 6 Sept. were considered mature and 6 weeks of CS accelerated germination and increased the germination percentage. Further, dormancy of Corylopsis seeds appears to be shallow since germination occurred without any CS.

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Greenhouse growers must use water more efficiently. One way to achieve this goal is to monitor substrate moisture content to decrease leaching. A systems approach to irrigation management would include knowledge of substrate matric potentials and air-filled pore space (AS) in addition to substrate moisture content. To study the relationship between substrate moisture and plant growth, annual vinca (Catharanthus roseus L.) was subject to a 2 × 2 factorial combination of two irrigation treatments and two substrates with differing moisture characteristic curves (MCCs). A gravimetric on-demand irrigation system was used to return substrate moisture content to matric potentials of −2 or −10 kPa at each irrigation via injected drippers inserted into each container. Moisture characteristic curves were used to determine gravimetric water content (GWC), volumetric water content (VWC), and AS at target substrate matric potential values for a potting mix consisting of sphagnum moss and perlite and a potting mix consisting of sphagnum moss, pine bark, perlite, and vermiculite. At each irrigation event, irrigation automatically shut off when the substrate-specific weight of the potted plants associated with the target matric potential was reached. Irrigation was triggered when the associated weight for a given treatment dropped 10% from the target weight. VWC and AS differed between substrates at similar matric potential values. Irrigating substrates to −2 kPa increased the irrigation volume applied, evapotranspiration, plant size, leaf area, shoot and root dry weight, and flower number per plant relative to irrigating to −10 kPa. Fafard 3B had less AS than Sunshine LB2 at target matric potential values. Plants grown in Fafard 3B had greater leaf area, shoot dry weight, and root dry weight. Leachate fraction ranged from 0.05 to 0.08 and was similar across all treatment combinations. Using data from an MCC in conjunction with gravimetric monitoring of the container–substrate–plant system allowed AS to be determined in real time based on the current weight of the substrate. Closely managing substrate matric potential and AS in addition to substrate water content can reduce irrigation and leachate volume while maintaining plant quality and reducing the environmental impacts of greenhouse crop production.

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