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Katrina J.M. Hodgson-Kratky, Olivier M. Stoffyn, and David J. Wolyn

environments ( Springer and Goldman, 2016 ). Of the numerous osmotica available (e.g., PEG, mannitol, sucrose, and sodium chloride), PEG has been identified as most effective because it is nontoxic and has large molecules excluded from the seedcoat pores ( Hohl

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Qi Zhang, Liqi Yang, and Kevin Rue

. Two hundred fifty milligrams of seeds of one cultivar were placed in one cell. Each plate was floated in PEG-6000 (VWR International, Batavia, IL) solutions at 0.0 MPa [0% PEG-6000 (control, nonstressed)], −0.3 MPa (15% PEG-6000), or −0.6 MPa (22

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Carlos A. Parera and Daniel J. Cantliffe

`Verina' leek (AIlium porrum L.) seed germination is normally reduced at temperatures > 25C. Leek seeds were primed in aerated solutions (1.5 MPa, 10 days at 15C) of d-mannitol (mannitol), polyethylene glycol-8000 (PEG), KNO, and a nonaerated solution of PEG-8000 (PEG). At high temperatures mannitol, PEG, and PEG significantly enhanced germination percentage relative to KNO, or the control. At constant 30C, the mannitol, PEG, and PEG treatments increased final germination almost 10 times and the coefficient of velocity (COV) was improved compared to KNO, and the control. 10 growth chambers with alternating day/night temperatures (38 to 28C or 32 to 22C, 10 to 14 hours, respectively), primed seeds had significantly higher emergence and a larger COV than the control. In a greenhouse study under good conditions for germination, total emergence of primed and nonprimed seeds was similar; however, mannitol, PEG, and PEG led to a significantly higher COV than the control or KNO, treatments. These controlled-environment results demonstrate that priming leek seeds via mannitol, PEG, and PEG may promote early emergence at high temperature and improve stand uniformity for container transplant production.

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Eucario Mancilla-Álvarez, Marco A. Ramírez-Mosqueda, Samantha Arano-Avalos, Rosalía Núñez-Pastrana, and Jericó J. Bello-Bello

retardants such as ancymidol ( El-Dawayati et al, 2012 ; Sarkar et al., 2001 ), and osmoregulators as polyethylene glycol (PEG-800) ( Seesangboon et al., 2018 ). In plant physiology, ancymidol produces a number of effects, including the inhibition of

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Yanxia Zhao, Guimei Qi, Fengshan Ren, Yongmei Wang, Pengfei Wang, and Xinying Wu

( Deng et al., 2014 ). The treatment methods of the samples described above were as follows: ‘Cabernet Sauvignon’ ( V. vinifera ) plants were treated with 120-m m salt [NaCl:CaCl, v/v (10:1)], polyethylene glycol (PEG), cold (5 °C), or unstressed

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Andreas Winkler, Stefanie Peschel, Kathleen Kohrs, and Moritz Knoche

in ‘Summit’ sweet cherry fruit. Microcracks were induced by incubating fruit for 24 h in isotonic PEG 6000 solution. Tonicity was established by water vapor pressure osmometry (VAPRO ® 5520 and 5600; Wescor, Logan, UT) of juice extracted from fruit

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Imed Dami and Harrison Hughes

Grape cv. Valiant was micropropagated in an MS medium with and without 2% (W/V) of polyethylene glycol (PEG, MW 8000). Leaf anatomy of control (in vitro, no PEG), treated (in vitro, PEG), field grown and greenhouse grown plants were compared under light microscopy. Cell size, palisade layer formation, relative intercellular air space and apparent chloroplast number varied between the leaves of control and PEG treated (high osmoticum) plantlets. These leaf characteristics in the high osmoticum medium appeared more similar to the leaves of the greenhouse and field grown plants. Leaves from control plantlets contained cells of larger size, lacked normal palisade layer formation, greater intercellular pore spaces and fewer chloroplasts. Leaves of PEG treated plantlets had smaller cells, a more defined palisade layer, reduced intercellular pore spaces and greater number of chloroplasts. Leaves of greenhouse and field grown plants had small cells, a well-defined palisade layer, least intercellular pore space and greatest number of chloroplasts. These results demonstrate that a high osmoticum medium may be used to induce more normal leaf development.

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Aaron J. Brown

Polyethylene glycol (PEG) was evaluated for its influence on hardening of in vitro-propagated `Fern' strawberries (Fragaria ×ananassa) when applied just before transplanting. Strawberries were micropropagated via shoot tips and grown in vitro until roots were well developed. Plantlets were then transferred onto filter paper bridges in liquid medium with 15% (w/v) of PEG-8000. After treatment in the medium for various periods, the plants were compared to the control (no PEG) for water loss from detached leaves, stomatal aperture, and survival rates after transplanting. Leaf epicuticular wax was also quantified. Overall, the in vitro PEG treatment was not successful in significantly increasing hardiness and survivability of the strawberry plants after transplanting from in vitro conditions to a soil medium. Osmotic stress was created, but apparently not for the time needed to increase survival. Further tests are needed to pinpoint the proper exposure time required to increase hardiness and survivability after transplanting plantlets. To increase survival, the time exposed to PEG should be 15, 18, or possibly 21 days.

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Farida Safadi and Harrison Hughes

Detached and intact leaves (first fully expanded leaf from the top) of tobacco (Nicotiana tabaccum L.) plantlets hardened in vitro with 2.0% polyethylene glycol (PEG) showed increased diffusive resistance (r) over those of nonhardened plantlets as measured by a steady state porometer. The leaves of the PEG hardened plants maintained a higher resistance throughout the one hour dessication period in approximately 30% relative humidity although both treatments showed an increase in diffusive resistance after 30 minutes. This indicates that the stomates are functioning in the in vitro tobacco plantlets. The higher (r) in the PEG treated plants may be due to more complete closure of stomates, higher cuticle wax content or a combination of both.

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Sandip Mukhopadhyay and Yves Desjardins

Transient expression of electroporation-mediated DNA uptake was monitored in callus-derived protoplasts of two asparagus (Asparagus offcinalis L.) genotypes by measuring the GUS activity. The level of expression and the viability of the protoplasts were influenced by the voltage and duration of the electric pulse. An increased plasmid DNA concentration and the presence of polyethylene glycol (PEG) in the electroporation medium enhanced the transient expression level. A considerable increase in GUS activity was observed in the presence of both PEG and heat-shock treatments than with PEG treatment alone. An optimal level of GUS activity was obtained after electroporation with a capacitive discharge of 500 V/cm and 94 ms duration. The two genotypes differed in their responses in vitro and also showed variable levels of transient expression. The present technique was suitable to obtain transgenic plants, as histochemical GUS assay revealed GUS activity in the protoplast-derived microcolonies as well as in callus tissues.