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The log-transformed counts of C. phlei spores produced per gram leaf also increased with C N (p < 0.001; adjusted R = 0.22; Figure 3).
The leaves were then ground into an extraction buffer (50 mM PBS, 1 mM PMSF, and pH 9.5), at a ratio of 3 mL per gram leaf material.
Twenty-five gram leaf tissue was ground into fine powder in liquid nitrogen following the slightly modified procedure described by Luo and Wing [ 28] to release their intact nuclei.
The ER-localized variant showed the highest azocarboxymethylcellulase (Azo-CMC) activity per gram leaf material (data not shown) and was selected for expression in transgenic plants and further analysis.
We found significant positive relationships between leaf C N and A. alternata and C. phlei spore production per gram leaf, although only A. alternata sporulation was associated with CO2 concentration level after accounting for changes in leaf C N.
The log-transformed counts of A. alternata spores produced per gram leaf were positively correlated with leaf C N (p < 0.001; adjusted R = 0.25; Figure 1), with an order-of-magnitude increase across the range of leaf C N of experimental plants.
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One-gram leaf disks from the one-month-old plants were homogenized in 10 mL of absolute ethyl alcohol and the homogenate was centrifuged at 3500 × g for 5 minutes.
The CC of leaf tissue (grams of glucose necessary to synthesize 1 g leaf tissue) was calculated as (5.39 × C − 1191)/1000 (Vertregt and Penning de Vries 1987).
Leaf carbon-to-nitrogen ratio was positively correlated with A. alternata spore production per gram of leaf but negatively correlated with antigenic protein content per spore.
In brief, a half gram of leaf was ground into fine powder in liquid nitrogen.
Fig. 5 Amount of soluble hydrogen peroxide per gram of leaf tissue from oilseed rape treated with nutrient solution containing IONs and challenged with drought for 5 days (n = 16, p value = 0.004).
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