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competing interests. Authors’ contributions GSL designed and performed most of the experiments, analyzed data and wrote the manuscript. STH designed, supervised all the experiments, analyzed data and wrote the manuscript. KFC provided ColE7 for colicin assay and gave suggestions. PHL provided the antibodies against BtuB, TolQ, TolR, TolA, TolB, Pal, and OmpF for this research. WJS and WSH gave suggestions and analyzed data for this research. All the authors have read and approved the final manuscript.”
“Background Metarhizium acridum is a haploid entomopathogenic

fungus (Hypocreales: Clavicipitaceae). M. acridum isolates have been used as biocontrol agents for crop pests, including sugar cane grubs, termites, cockroaches, and rhinoceros much beetles [1]. M. acridum was commercialized and used for locust control in Australia, West Africa [2], and China [3]. Insecticide resistance, pest resurgence, and concerns over environmental impact have made the search for alternative means of biological pest control more urgent. Unfortunately, large-scale use of fungal biocontrol agents is partially limited by the failure of conidia to retain virulence during long-term storage, transportation, and use under stressful conditions, such as high temperature, low humidity, and sunlight exposure [4–6]. Manipulation of culture conditions could optimize the concentration of spore polyols and sugars, including trehalose, and consequently increase tolerance to low relative humidity [7, 8]. However, genetic manipulations of these polyols and sugars to enhance environmental tolerance have not been explored in entomopathogenic fungi. To genetically engineer more robust entomopathogenic fungi, we focused on the trehalose pathways involved in stress response. Trehalose is a storage carbohydrate as trehalose concentrations are high when nutrients are limited in resting cells.

2) 118.4 (3.9) Sex (%) Male 7,121 (100.0) 49.6 Female   50.4 Height (cm) 7,047 (99.0) 139.5 (6.3) Weight (kg) 7,105 (99.8) 33.2 (29.4–38.4)a TBLH

BMC (g) 6,775 (95.1) 893.8 (184.0) TBLH BA (cm2) 6,775 (95.1) 1139.5 (164.3) TBLH BMD (g/cm2) 6,775 (95.1) 0.78 (0.05) TBLH ABMC 6,775 (95.1) 894.6 (39.8) Spine BMC (g) 5,487 (77.1) 78.4 (15.7) Spine BA (cm2) 5,487 (77.1) 100.7 (12.0) Spine BMD (g/cm2) 5,487 (77.1) 0.77 (0.08) Spine ABMC (g) 5,487 (77.1) 78.4 (7.1) Pubertal stage (%) Boys Tanner 1 2,365 (67.0) check details 82.9 Tanner 2   16.5 Tanner 3+   0.6 Girls Tanner 1 2,836 (79.0) 81.5 Tanner 2   15.0 Tanner 3+   3.5 Age at menarche for girls (years) (%) Up to 10 3,107 (86.5) 4.7 11+   95.3 Gestational age (weeks) 7,121 (100.0) 39.5 (1.8) Birth weight (kg) 7,035 (98.8) 3.4 (0.5) Household social class (%) I 6,544 (91.9) 15.5 II   45.1 III NM   24.8 III M   10.3 IV/V   4.3 Mother Age at delivery (years) 7121 (100.0) 29.0 (4.6) Height (cm) 6753 (94.8) 164.1 (6.6) Pre-pregnancy BMI (kg/m2) 6429 (90.3) 22.2 (20.5–24.4)a No. of previous births (%) 0 6879 (96.6) 45.8 1   35.5 2   13.7 3   3.8 4 or more   1.2 Smoking during pregnancy (%) Never 6379 (89.6) 78.7 1 or 2 trimesters   9.5 All trimesters   11.8 Education (%) None/CSE 6860 (96.3) 13.8 Vocational   8.5 O Levels   35.2 A Levels   26.6 Degree   15.8 Father STA-9090 Age at child’s

birth (years) 5106 (71.7) 31.4 (5.2) Height (cm) 4931 (69.2) 176.3 (6.9) BMI (kg/m2) 4887 (68.6) 24.8 (22.9–26.9)a Regular smoker (%) No 6679 (93.8) 65.3 Yes   34.7 Education (%) None/CSE 6467 (90.8) 19.3

Vocational   8.2 O Levels   21.7 A Levels   28.5 Degree   22.2 ABMC area-adjusted bone mineral content, BA bone area, BMC bone mineral content, BMD bone mineral density, BMI body mass index, IQR interquartile Farnesyltransferase range, TBLH total body less head aMedian and interquartile range are shown for skewed variables Pairwise correlations of total body and spinal bone measures are given in ESM Web Table 3, and correlations of these measures with child and parental characteristics are shown in ESM Web Table 4. The child’s height and weight were strongly positively correlated with TBLH and spine BMC and BA and moderately with TBLH and spine BMD. Higher birth weight, longer gestation and greater age at DXA scan were all associated with increased TBLH BMC, BA and BMD. Multiple imputation analysis of maternal and paternal smoking in relation to TBLH bone outcomes is shown in Table 2 and the analysis of spinal outcomes shown in Table 3.