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Seeds of Pulsatilla turczaninovii were categorized to full and empty seeds based on observations under a light microscope and on X-ray images. A germination test for full or empty seeds was evaluated as affected by the duration of gibberellin GA3 and the moist 5 °C cold stratification (CS) treatment. The morphological and elemental components of P. turczaninovii and P. cernua var. koreana seeds were analyzed using low-temperature scanning electron microscopy (LT-SEM) and energy dispersive X-ray diffraction analysis. The results showed that 64% of full and semifull P. turczaninovii seeds 10 weeks after harvesting germinated in 17 to 19 days; however, the germination rate, including empty and semiempty seeds, was lower (52.6%). Full seeds with damaged or dried vegetative organs (embryo or endosperm) and semifull seeds with severely damaged vegetative organs were observed, and this result could be related to low germination rates. Germination patterns of seeds stored dry at 5 °C for 44 weeks that showed a sigmoid pattern were increased by immersing seeds in a GA3 solution for 8 hours and treating seeds with 16 or 32 days of CS. More seeds germinated between 12 and 17 days (as compared with 17 to 29 days), especially when they were treated with GA3 and received 32 days of CS. Comparison of germination rates of visually full seeds upon harvest (52.6%) with those that had been stored dry for 44 weeks (26.3% to 29.7%) suggested that the viability of seeds may have decreased. Dormancy could be a factor that decreased germination and can be removed by low temperature and GA3 treatment. LT-SEM revealed a valley-like, sunken streak in empty seeds of P. cernua var. koreana. The nickel content in the trichome and seedcoat of full and empty seeds of both taxa ranged from 2.98 to 4.62 (weight %), as determined on X-ray images. Our study suggested that the low germination rate was due to either the presence of dormancy, damage to either embryo or endosperm, a loss in viability, or the presence of nickel in the seeds.
The eriophyid mite, Phyllocoptes fructiphilus, vectors the causal agent, Rose rosette virus (RRV), that results in rose rosette disease. Parts of the southeastern United States have remained free of the disease, except for infected plant material introductions that were eradicated. A survey of sampling points through Alabama, Georgia, and Mississippi (n = 204) revealed the southeastern border of RRV. The presence of RRV in symptomatic plant tissue samples (n = 39) was confirmed by TaqMan-quantitative reverse transcription polymerase chain reaction (RT-qPCR). Samples were also collected at every plot for detection of eriophyid mites, specifically for P. fructiphilus. Three different species of eriophyid mites were found to be generally distributed throughout Alabama, Georgia, and Mississippi. Most of these sites (n = 60) contained P. fructiphilus, found further south than previously thought, but in low populations (<10 mites/gram of tissue) south of the RRV line of incidence. Latitude was found to be significantly correlated with the probability of detecting RRV-positive plants, but plant hardiness zones were not. Plot factors such as plant size, wind barriers, and sun exposure were found to have no effect on P. fructiphilus or the presence of RRV. The reason for the absence of RRV and low populations of P. fructiphilus in this southeast region of the United States are unclear.