Comparative Transcriptomics of Non-Embryogenic and Embryogenic Callus in Semi-Recalcitrant and Non-Recalcitrant Upland Cotton Lines.

Somatic embryogenesis-mediated plant regeneration is essential for the genetic manipulation of agronomically important traits in upland cotton. Genotype specific recalcitrance to regeneration is a primary challenge in deploying genome editing and incorporating useful transgenes into elite cotton germplasm. In this study, transcriptomes of a semi-recalcitrant cotton (Gossypium hirsutum L.) genotype 'Coker312' were analyzed at two critical stages of somatic embryogenesis that include non-embryogenic callus (NEC) and embryogenic callus (EC) cells, and the results were compared to a non-recalcitrant genotype 'Jin668'. We discovered 305 differentially expressed genes in Coker312, whereas, in Jin668, about 6-fold more genes (2155) were differentially expressed. A total of 154 differentially expressed genes were common between the two genotypes. Gene enrichment analysis of the upregulated genes identified functional categories, such as lipid transport, embryo development, regulation of transcription, sugar transport, and vitamin biosynthesis, among others. In Coker312 EC cells, five major transcription factors were highly upregulated: LEAFY COTYLEDON 1 (LEC1), WUS-related homeobox 5 (WOX5), ABSCISIC ACID INSENSITIVE3 (ABI3), FUSCA3 (FUS3), and WRKY2. In Jin668, LEC1, BABY BOOM (BBM), FUS3, and AGAMOUS-LIKE15 (AGL15) were highly expressed in EC cells. We also found that gene expression of these embryogenesis genes was typically higher in Jin668 when compared to Coker312. We conclude that significant differences in the expression of the above genes between Coker312 and Jin668 may be a critical factor affecting the regenerative ability of these genotypes.

Evaluation of ultrasound diagnosis of fetal anomalies in women with pregestational diabetes: University of Florida experience.

Fetal congenital anomalies have become the major cause of perinatal morbidity and mortality in pregnancies complicated by insulin-dependent diabetes. We evaluated the use of level II ultrasound in predicting congenital anomalies, to determine if the management of pregnant women with insulin-dependent diabetes would be altered by these findings. We examined 43 insulin-dependent diabetic pregnancies. In this group, 10 newborns (23%) were diagnosed either at birth or later to have an abnormality. Of these, four (9%) were diagnosed by ultrasound. Of the seven cases that were undiagnosed, three women did not undergo the level II examination, two women had lesions undetectable by ultrasound, and in two women cardiac abnormalities were missed. Of the four congenital anomalies that were prenatally diagnosed, the findings influenced the management in three cases. The level II ultrasound used as a screening test had a 67% sensitivity and a 100% specificity, with a positive predictive value of 100% and a negative predictive value of 91%. We conclude that high-resolution ultrasound may be used as a screening tool for congenital anomalies in the insulin-dependent diabetic pregnancy. This study also suggests that the use of fetal echocardiography with evaluation of aortic and pulmonary outflow tracts, the arch of the aorta, and ventricular size may be helpful in the diagnosis of fetal cardiac anomalies in the pregnant woman with insulin-dependent diabetes.

Continuous-wave Doppler ultrasound and decreased amniotic fluid volume in pregnant women with intact or ruptured membranes.

The cause(s) of decreased amniotic fluid in the absence of fetal anomalies and intrauterine growth retardation is not clear. A prospective study was performed to evaluate umbilical and uterine artery Doppler velocimetric results in pregnancies complicated by decreased amniotic fluid. Three medically high-risk groups were studied: women with (1) normal fluid and intact membranes, (2) decreased fluid and intact membranes, and (3) decreased fluid and ruptured membranes. The decreased fluid/intact membranes group had a significantly increased incidence of abnormal uterine artery waveforms (diastolic notching or absence of end-diastolic velocity); however, uterine systolic/diastolic ratios were not significantly different. The umbilical systolic/diastolic ratios were marginally higher in the intact membranes/decreased fluid group when compared with the ruptured membranes group. This study suggests that problems with maternal blood supply to the placenta may be related to decreased amniotic fluid when membranes are intact.