A device for the controlled cooling and freezing of excised plant specimens during magnetic resonance imaging.

BACKGROUND: Investigating plant mechanisms to tolerate freezing temperatures is critical to developing crops with superior cold hardiness. However, the lack of imaging methods that allow the visualization of freezing events in complex plant tissues remains a key limitation. Magnetic resonance imaging (MRI) has been successfully used to study many different plant models, including the study of in vivo changes during freezing. However, despite its benefits and past successes, the use of MRI in plant sciences remains low, likely due to limited access, high costs, and associated engineering challenges, such as keeping samples frozen for cold hardiness studies. To address this latter need, a novel device for keeping plant specimens at freezing temperatures during MRI is described. RESULTS: The device consists of commercial and custom parts. All custom parts were 3D printed and made available as open source to increase accessibility to research groups who wish to reproduce or iterate on this work. Calibration tests documented that, upon temperature equilibration for a given experimental temperature, conditions between the circulating coolant bath and inside the device seated within the bore of the magnet varied by less than 0.1 °C. The device was tested on plant material by imaging buds from Vaccinium macrocarpon in a small animal MRI system, at four temperatures, 20 °C, - 7 °C, - 14 °C, and - 21 °C. Results were compared to those obtained by independent controlled freezing test (CFT) evaluations. Non-damaging freezing events in inner bud structures were detected from the imaging data collected using this device, phenomena that are undetectable using CFT. CONCLUSIONS: The use of this novel cooling and freezing device in conjunction with MRI facilitated the detection of freezing events in intact plant tissues through the observation of the presence and absence of water in liquid state. The device represents an important addition to plant imaging tools currently available to researchers. Furthermore, its open-source and customizable design ensures that it will be accessible to a wide range of researchers and applications.

Responses of condensed tannins in poplar roots to fertilization and gypsy moth defoliation.

We examined the effects of fertilization and gypsy moth defoliation on condensed tannin concentration (%CT) of hybrid poplar (Populus x canadensis cv 'Eugeneii') fine roots in the summers of 1997 and 1998. This factorial experiment included two defoliation treatments (defoliated and a foliated control) and fertilization treatments (100 kg nitrogen (N) ha(-1) and an unfertilized control). Gypsy moth (Lymantria dispar L.) populations were experimentally increased to obtain defoliation in the summers of 1996, 1997 and 1998; fertilization subplots were supplemented with NH4NO3 (100 kg N ha(-1)) in the spring of each year. Despite the severity of defoliation, the effects were small, and significant on only two sampling dates: in May 1997, when fine root %CT was 23% lower in the defoliated trees, and in November 1997, when trees in the defoliated unfertilized plots had 35% higher root %CT than trees in all other plots. Defoliation effects on root %CT did not follow the same seasonal pattern as defoliation effects on root starch content, N uptake capacity or leaf %CT. Regulation of root condensed tannin concentration appeared to be partially uncoupled from these traits. The small transient effects on root defense reflect the resilience of this early successional tree to severe early season defoliation.

Cottonwood growth rate and fine root condensed tannin concentration.

We examined the relationship between trunk diameter and diameter relative growth rate (RGR) and fine root condensed tannin concentration in 12 genotypes of eastern cottonwood (Populus deltoides Bartr. ex Marsh.) planted in three locations across the north central United States. Across genotypes, trunk diameter, diameter RGR and root condensed tannin concentration were negatively correlated at one location (Wisconsin), but showed no significant correlation at the other locations (Iowa and Michigan). The factors responsible for this difference among sites remain unidentified, but may be related to soil fertility.

Inoculation of cranberry (Vaccinium macrocarpon) with the ericoid mycorrhizal fungus Rhizoscyphus ericae increases nitrate influx.

Despite the ubiquitous presence of ericoid mycorrhizal (ERM) fungi in cranberry (Vaccinium macrocarpon), no prior studies have examined the effect of ERM colonization on NO(3)(-) influx kinetics. Here, (15)NO(3)(-) influx was measured in nonmycorrhizal and mycorrhizal cranberry in hydroponics. Mycorrhizal cranberry were inoculated with the ERM fungus Rhizoscyphus (syn. Hymenoscyphus) ericae. (15)NO(3)(-) influx by R. ericae in solution culture was also measured. Rhizoscyphus ericae NO(3)(-) influx kinetics were linear when mycelium was exposed for 24 h to 3.8 mm NH(4)(+), and saturable when pretreated with 3.8 mm NO(3)(-), 50 microm NO(3)(-), or 50 microm NH(4)(+). Both low-N pretreatments induced greater NO(3)(-) influx than either of the high-N pretreatments. Nonmycorrhizal cranberry exhibited linear NO(3)(-) influx kinetics. By contrast, mycorrhizal cranberry had saturable NO(3)(-) influx kinetics, with c. eightfold greater NO(3)(-) influx than nonmycorrhizal cranberry at NO(3)(-) concentrations from 20 microm to 2 mm. There was no influence of pretreatments on cranberry NO(3)(-) influx kinetics, regardless of mycorrhizal status. Inoculation with R. ericae increased the capacity of cranberry to utilize NO(3)(-)-N. This finding is significant both for understanding the potential nutrient niche breadth of cranberry and for management of cultivated cranberry when irrigation water sources contain nitrate.