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The NAM founders, the two NAM RIL subpopulations, and the B73 · NAM founder F1 crosses were all planted in 10·20 racks of tubes 1 inch wide and 8 inches deep. Every third row of 10 in each rack was left empty to allow for even air flow and light intensity and to reduce edge effects. The soil used was a 1:1 mixture of black soil and SunGro potting soil, blended with two teaspoons per square foot of Oscmocote Plus fertilizer. Plants were grown in growth chambers for 14 d (25 during 16-hr days and 20 during the nights). The 27 NAM parental lines (Table S1) (Yu et al. 2008) were grown in three replications of 10 plants per line, with lines randomly distributed throughout the growth chamber. All healthy plants were sampled for histology, and 10 to 27 images were measured per line. For the NAM subpopulations B73 · CML277 (Z005) and B73 · P39 (Z024), 140 and 137 lines, respectively, were examined (Table S1). These lines were grown in five replications of one plant per line, all in one chamber, with lines randomly distributed. In addition to the 140 and 137 RILs, all parental lines (B73, Mo17, P39, CML277) as well as 10 representative inbred lines (Table S1) from the IBMRIL population (selected to encompass the ranges of observed SAM height and SNP diversity) were included in each set for data standardization with the IBM RIL analyses in Thompson et al. 2014. Based on previous experiments, row–column variation for SAM architecture traits is trivial when compared with plant-to-plant and chamber-to-chamber variation, hence the decision to use five full replications of each genotype with adequate checks. All healthy plants were sampled for histology, and 1 to 5 images (average 3.8) were measured per line for RILs, 6 to 10 (average 8.3) were measured for each of the IBM controls, and 14 to 19 (average 16) were measured per parental line. Eight NAM founders were crossed to both B73 and Mo17, and these 16 F1 crosses (Table S1) as well as the 10 parental inbreds were grown in 20 replications of one plant per replication, scattered randomly throughout each replication. All healthy plants were sampled, and 14 to 18 images were measured per line, with the exception of Hp301 and its crosses. These lines had very poor germination, and only nine, three, and one plants were measured for Hp301, Hp301 · B73, and Hp301 · Mo17, respectively. The time course experiment included 18 lines, with five plants per line per time point. These were grown in the greenhouse in St. Paul, Minnesota in April/May 2014 in 6-inch square pots, 5 plants per pot, with soil mixture as described above. Greenhouse growth settings were 18-hr days, 27 during the days, and 21 during the nights. |
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