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Sheri B. Crabtree*, Kirk W. Pomper, and Robert L. Geneve

The pawpaw (Asimina triloba) is the largest fruit native to the U.S. and has potential as a new fruit crop. Few methods are available to clonally propagate pawpaw, with grafting or budding onto a seedling rootstock being the only currently feasible method. Developing new options for clonal propagation of pawpaw could help advance this growing industry. Layering has been used to clonally propagate other difficult to root tree species. The objective of this study was to examine trench layering as a method to clonally propagate pawpaw. A randomized factorial experiment was implemented to examine the roles of plant juvenility and auxin concentration on rooting in a greenhouse trench layering system. Seedlings were defoliated, tipped, and transplanted into trench layering beds at 3, 6, and 12 weeks after emergence. Shoots were etiolated, then girdled and treated with three levels of IBA (0, 5000, and 10,000 ppm). The main effects of age and IBA concentration significantly affected the percentage of shoots producing roots. Juvenility enhanced rooting, with 15% of the shoots of the 3-week-old pawpaw seedlings producing roots, compared to only about 5% of the 12-week-old seedlings rooting. Auxin application to shoots also promoted rooting, with 16% of IBA-treated shoots producing roots, compared to the untreated control, with only 2% of shoots producing roots. There was no significant difference in rooting percentage between the two concentrations of IBA. The treatment combination most successful at promoting root initiation was 10,000 ppm IBA applied to shoots of 3-week-old seedlings, with 31% of shoots rooting.

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Robert L. Geneve, Sharon T. Kester, and Terriawkia A. Woods

Ethephon and ethylene gas applied to intact Eastern redbud seed induced germination in 44 or 53% of dormant seed. However, endogenous ethylene production was not found to be correlated with the release from dormancy during chilling stratification (5°C). Seeds stratified in the presence of 6000 ppm 2.5-norbomadicne germinated at the same percentage as control seeds. Isolated embryos treated with 100 to 500 μM AOA or 1000 μM silver thiosulfate germinated at a slower rate than control seeds, but the release from dormancy during stratification was unaffected by either ethylene inhibitor. Ethylene evolution, ACC and MACC content remained at a low level throughout stratification. EFE activity was not detectable in hydrated dormant or non-dormant seed. All ethylene parameters measured increased sharply during germination with peak activity correlated with radicle emergence. These data indicated that ethylene production was linked to germination, but unrelated to dormancy release in Eastern redbud seed.

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Susmitha Nambuthiri, Ethan Hagen, Amy Fulcher, and Robert Geneve

Two sensor-based irrigation scheduling systems were compared for water use and plant growth in container-grown Green Velvet boxwood (Buxus sempervirens L. × B. microphylla Siebold & Zucc. var. koreana ‘Green Velvet’) and slender deutzia (Deutzia gracilis Siebold & Zucc). These crops were chosen because they have different water requirements during production. The two sensor-based irrigation systems included a physiological-based on-demand (OD) irrigation system where the set point was derived from the relationship between substrate moisture and photosynthetic rate. The second system was a daily water use (DWU) method where the amount of water used by the crop was replaced each day. The objective of the study was to evaluate and compare water use and growth metrics using the OD and DWU irrigation scheduling regimes for two container-grown woody plants that differed in their water consumption. There were no differences in root and shoot biomass or growth index due to the irrigation schedule employed for either boxwood or deutzia. For boxwood plants, OD irrigation reduced water consumption by 35.5% and enhanced water use efficiency (WUE) by 54.5% compared with DWU. Total water use of deutzia in OD zones was reduced by 26.5% compared with DWU. DWU offers the labor scheduling advantage of irrigation occurring at a set time of day, and OD offers the advantage of watering as required, potentially reducing water stress as the season progresses and as the plant size and atmospheric demand increase.

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Laura A. Wood, Sharon Kester, and Robert L. Geneve

Echinacea are North American members of the Asteraceae, and all can show some degree of endogenous physiological seed dormancy that is alleviated by chilling stratification. In some species, ethephon has been shown to substitute for chilling stratification to relieve dormancy. The objective of this research was to investigate the effect of ACC on dormancy and germination in five Echinacea species. Germination for each species was 90%, 59%, 99%, 81%, and 21%, respectively. Germination on 5 mM ACC improved germination in E. tennesseensis, E. paradoxa, and E. simulata to 82%, 99%, and 82%, respectively, but there was no change for E. purpurea and E. angustifolia. Germination rate was dramatically accelerated in all species in the presence of ACC. On average, there were 57% more seeds germinated on ACC after 3 days compared to untreated seeds. Exposing E. purpurea and E. tennesseensis seeds to 1 or 2 days of 5 mM ACC before drying and subsequently re-hydrating the seeds did not have the same effect as continual exposure to ACC. Similarly, there was no clear enhancement of adding ACC during stratification over the improvement gained by chilling stratification alone. Seeds produced more ethylene upon germination following both stratification and ACC treatment. However, significantly more ethylene is produced during germination in ACC-treated seeds. It was clear that ACC-treated seeds showed improvement for enhanced germination speed and in some cases germination percentage. Unfortunately, this enhanced germination was not retained in seeds treated with ACC and dried prior to germination. Additional work is required to develop a commercially viable method of loading ACC into seeds for germination enhancement.

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S. Yusnita, Robert L. Geneve, and Sharon T. Kester

In vitro shoot multiplication of white Eastern redbud was successful using two-node mature explants from the initial spring flush on a woody plant medium (WPM) supplemented with benzylaminopurine (BAP). Optimal shoot proliferation was obtained at 10-15 μM BAP. Treatment with thidiazuron produced fasciated (stunted) adventitious shoots which failed to elongate. Successive subcultures increased the ability of explants to form shoots. However, shoot tip necrosis became a problem after 7-8 subcultures. Shoot tip necrosis is being studied by comparing shoot multiplication on bacto-agar vs. gelrite, increasing the Ca concentration in WPM and by trying to reduce the phenolic exudate by the explants with PVP or activated charcoal. Microshoots >3 cm long were rooted by pulse treatment on half strength WPM containing 300 μM IBA or NAA before being moved to hormone free WPM. There was a different morphology between IBA and NAA induced roots, although the number of roots were comparable. IBA treated microcuttings developed branched, fine roots, whereas NAA treated plants produced unbranched, coarse roots. Rooted microshoots were successfully acclimated to greenhouse condition.

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Gunching Siriwitayawan, A. Bruce Downie, and Robert L. Geneve

Sweet corn (Zea mays L.) and tomato (Lycopersicon esculentum Mill.) seeds were aged naturally for 18 months or artificially aged using saturated salt accelerated aging to provide seed lots that differed in seed vigor, but retained a high standard germination percentage. Seed vigor was confirmed using standard vigor tests, including time to radicle emergence, cold, and accelerated aging tests. Ethylene evolution from both sweet corn and tomato seeds during germination was positively correlated with seed quality. Differences in ethylene evolution between nonaged and aged seeds were greater in seeds germinated on exogenous 1-aminocyclopropane-1-carboxylic acid (ACC). After 36 hours, there was about a 15-fold increase in ethylene evolution from seeds treated with 5 mm ACC compared to untreated seeds. Naturally and artificially aged seeds responded similarly and showed reduced ethylene production compared to nonaged seeds. In contrast to ethylene production, endogenous ACC titers were less for nonaged compared to aged seeds. Exogenous application of ACC to artificially aged seeds reduced the time to radicle protrusion, but did not completely reverse age-related effects on vigor. The data indicate that the reduced ability to produce ethylene in aged seeds was related to ACC oxidase (ACCO) synthesis or activity. Using Northern blot analysis, ACCO mRNA was detected after 48 hours of imbibition in nonaged seeds, but was undetectable in aged seeds affirming the contention that ACCO synthesis was delayed or reduced by aging. The current study provides additional support for ethylene as a biochemical indicator of seed vigor in seed lots with reduced vigor but high germination capacity.

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Cynthia H. Finneseth, Desmond R. Layne, and Robert L. Geneve

The North American pawpaw [Asimina triloba (L.) Dunal], a temperate member of the Annonaceae, is a deciduous woody tree with ornamental value and has merit as a fruit crop. Anatomical studies of pawpaw seed revealed a small, linear embryo that does not change in length during cold or warm stratification. Radicle and cotyledon growth from planting until radicle protrusion was concurrent and at about the same rate. Cotyledons grew through a specialized channel of cells extending above the cotyledon tips, but never emerged from the seed. The extended period of time required for the development of the cotyledons delayed seedling emergence more than 50 days. The cotyledons appear to be haustorial and translocate storage material from the endosperm to the growing embryo. At the time of epicotyl elongation, the radicle and developing root system were well developed and comprised 81 % of the seedling biomass. Seedling development could be divided into four distinct stages, including radicle protrusion, hypocotyl emergence, epicotyl elongation, and seedcoat abscission.

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Kay Oakley, Robert Geneve, Sharon Kester, and Myra Stafford

Root and shoot development in Marigold `Little Devil Flame' was studied after being grown for varying lengths of time in 392-count plugs before transplanting to six-pack cells. Seedlings were grown for 0, 5, 10, 15, 20, and 25 days before transplanting to six-packs. All plants were measured at day 25. There was no significant difference in total root length, area and dry weight per plant or in leaf area and shoot dry weight per plant for seedlings transplanted from 0 to 15 days. Both total root dry weight and total shoot dry weight of seedlings transplanted on day 20 was reduced by 32% compared to seedlings that were not transplanted. Total root dry weight of seedlings transplanted at day 25 was reduced by 60% while total shoot dry weight of seedlings was reduced by 56% from those not transplanted. In a separate experiment, the growth rate of seedlings grown in plugs was sigmoidal (r 2 = 0.98). Growth rate was significantly reduced between 20 and 25 days in the plug. These results suggest that root restriction in the plug may be a factor in the reduction of seedling growth following transplanting.

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Kay Oakley, Robert Geneve, Sharon Kester, and Patchara Wonprasaid

Standardized seed vigor tests must be developed for greenhouse-grown flower species. Current vigor tests used to evaluate large-seeded agronomic crops are generally not useful for evaluating smaller-seeded flower species. One alternative is to use radicle length in seedlings grown under controlled environments as an indicator of seed vigor. For that purpose, a seed vigor test was developed that uses digital images taken using a flat bed scanner to measure radicle length in small-seeded flower species. A novel, cellulose substrate was used for germinating seeds. It provided similar moisture-holding properties to standard germination blotters used by commercial seed analysts, but is clear. This has allowed for quick image acquisition without removing seedlings from the petri dish. Correlations were made between seedling growth (radicle length, total seedling length, and total seedling area) with other vigor tests (saturated salts accelerated aging) and greenhouse plug flat emergence. For several seed lots of impatiens that varied in initial seed quality, radicle length after 4 days showed good correlations (>R 2 = 0.79) with other measures of seed vigor for describing seed quality. This system is an improvement over other attempts to use computer-aided assessment of digital images because it provides digital images that do not vary due to external lighting; it uses software that can evaluate radicle length in a petri dish assay that does not require a slant-board for straight radicle growth; it relies on standard germination technics used by every seed lab; it uses a clear substrate to replace the opaque blotter to allow digital images to be taken within the petri dish; and accurate measurements of seedling parts is performed in under 2 min per petri dish.

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Robert L. Geneve, Jack W. Buxton, and Myra Stafford

Capillary mat subirrigation provides uniform water in the growing medium to optimize seedling growth in plugs. It also offers a closed system that allows the grower to regulate the amount of water available to seedlings and to reduce water runoff. However, root outgrowth into the capillary mat can be a significant problem. Copper hydroxide (Spin Out) was painted on the bottom, outside surface of the plug container to control root outgrowth into the capillary mat. Three square and two octagonal plug sizes were treated with copper. Regardless of the plug size or shape, copper treatment was an effective treatment to control root outgrowth in marigold seedlings. Copper treatment reduced overall root outgrowth by 80% to 92%. Marigold and geranium seedlings in copper-treated square plug containers showed some reduced shoot and root development during plug production, but there were no differences in copper-treated plants compared to nontreated plants following transplanting to cell packs.