Limited knowledge currently exists regarding the roles of plant genes and proteins in the Agrobacterium tumefaciens-mediated transformation process. To understand the host contribution to transformation, we carried out root-based transformation assays to identify Arabidopsis mutants that are resistant to Agrobacterium transformation (rat mutants). To date, we have identified 126 rat mutants by screening libraries of T-DNA insertion mutants and by using various "reverse genetic" approaches. These mutants disrupt expression of genes of numerous categories, including chromatin structural and remodeling genes, and genes encoding proteins implicated in nuclear targeting, cell wall structure and metabolism, cytoskeleton structure and function, and signal transduction. Here, we present an update on the identification and characterization of these rat mutants.

ObjectivesTo determine the risk factors for retinal microvascular impairment on optical coherence tomography angiography (OCT-A) in type 2 diabetic patients without clinical diabetic retinopathy (DR).MethodsThis retrospective and cross-sectional study enrolled 74 diabetic patients without clinically evident DR for the study group and 34 healthy subjects for the control group. OCT-A parameters were measured to determine vascular density (VD) and the foveal avascular zone (FAZ) size in the superficial and deep capillary plexuses (SCP/DCP) of the retina. Clinical data were collected on sex, age, diabetes duration, hemoglobin A1c (HbA1c), hypertension, dyslipidemia, low-density lipoprotein cholesterol (LDL-C), estimated glomerular filtration rate (eGFR) and smoking status. Multiple linear regression analyses were performed to represent the associated clinical variables with OCT-A parameters in diabetic patients.ResultsIn comparison between the study and control groups, the VD in the SCP and DCP were significantly lower in diabetic patients compared to the controls (P = 0.022 and 0.003, respectively). The FAZ size in the SCP and DCP were significantly greater in diabetic patients compared to the controls (P = 0.035 and <0.001, respectively). In age- and sex-adjusted multiple regression analyses for the diabetic patients, dyslipidemia and hypertension were negatively associated with SCP-VD (β = -0.357, P = 0.002; β = -0.239, P = 0.039, respectively). Current smoking was correlated with lower DCP-VD (β = -0.255, P = 0.043). Greater SCP-FAZ size was associated with dyslipidemia and greater LDL-C (β = 0.254, P = 0.013; β = 0.232, P = 0.029, respectively), and greater DCP-FAZ size, with lower eGFR and greater LDL-C (β = -0.355, P = 0.004; β = 0.235, P = 0.037, respectively).ConclusionsDiabetic patients without clinical DR showed lower VD and greater FAZ size in the SCP and DCP compared to healthy controls. In diabetic patients without clinical DR, dyslipidemia and/or high LDL-C were important risk factors for retinal microvascular impairment. Hypertension, current smoking and lower eGFR also contributed to microvascular impairment.

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BackgroundTo describe the features of foveal microvasculature using optical coherence tomography angiography (OCTA) and to determine the related clinical factors in eyes with surgically closed macular hole (MH).MethodsA retrospective case series of 18 patients with unilateral MH was reviewed. The patients maintained complete hole closure after vitrectomy with inner limiting membrane (ILM) peeling for at least 12 months. The healthy fellow eyes were studied as controls. The foveal microvasculature of both eyes was examined by OCTA. The area of the foveal avascular zone (FAZ) and the vascular density (VD) ratio in the superficial and deep capillary plexuses (SCP and DCP) were determined after surgery. Several clinical factors including age, stage and dimensions of MH, papillofoveal distance, the extent of nasal displacement of the fovea after surgery, postoperative central foveal thickness, and outer-retina integrity were evaluated to determine any relationships with the OCTA parameters.ResultsThe mean FAZ area in both the SCP and DCP (0.29 ± 0.11 mm2 and 0.39 ± 0.14 mm2) was significantly smaller than those of the controls (0.45 ± 0.14 mm2 and 0.62 ± 0.22 mm2) (p = 0.001 and <0.001, respectively). The mean VD ratio in the SCP (0.270 ± 0.349) was similar to that of the controls (0.321 ± 0.189) (p = 0.231); however, that in the DCP (0.321 ± 0.189) was significantly lower than that of the controls (0.331 ± 0.119) (p = 0.025). Only the extent of nasal displacement of the fovea was correlated with the DCP FAZ-area difference values between the study group and the controls (correlation coefficient = 0.577; p = 0.012).ConclusionsAfter successful MH surgery, the FAZ area in both the SCP and DCP was smaller and the VD ratio of the DCP was lower, suggesting a possible DCP vulnerability to tractional stress. As the FAZ area of the DCP in closed-MH eyes became smaller than that in the controls, the fovea was less displaced toward the optic disc, possibly reflecting a lack of retinal redundancy caused by horizontal stretching accompanied by foveal displacement.Electronic supplementary materialThe online version of this article (10.1186/s12886-017-0607-z) contains supplementary material, which is available to authorized users.

Indole-3-acetic acid (IAA) markedly increased ethylene production by inducing the expression of three 1aminocyclopropane-1-carboxylate (ACC) synthase cDNAs (pVR-ACS1, pVR-ACS6 and pVR-ACS7) in mung bean hypocotyls. Results from nuclear run-on transcription assay and RNA gel blot studies revealed that all three genes were transcriptionally active displaying unique patterns of induction by IAA and various hormones in etiolated hypocotyls. Particularly, 24-epibrassinolide (BR), an active brassinosteroid, specifically enhanced the expression of VR-ACS7 by a distinct temporal induction mechanism compared to that of IAA. In addition, BR synergistically increased the IAA-induced VR-ACS6 and VR-ACS7 transcript levels, while it effectively abolished both the IAA- and kinetin-induced accumulation of VR-ACS1 mRNA. In light-grown plants, VR-ACS1 was induced by IAA in roots, and VR-ACS6 in epicotyls. IAA- and BR-treatments were not able to increase the VR-ACS7 transcript in the light-grown tissues. These results indicate that the expression of ACC synthase multigene family is regulated by complex hormonal and developmental networks in a gene- and tissue-specific manner in mung bean plants. The VR-ACS7 gene was isolated, and chimeric fusion between the 2.4 kb 5'-upstream region and the beta-glucuronidase (GUS) reporter gene was constructed and introduced into Nicotiana tabacum. Analysis of transgenic tobacco plants revealed the VR-ACS7 promoter-driven GUS activity at a highly localized region of the hypocotyl-root junction of control seedlings, while a marked induction of GUS activity was detected only in the hypocotyl region of the IAA-treated transgenic seedlings where rapid cell elongation occurs. Although there was a modest synergistic effect of BR on the IAA-induced GUS activity, BR alone failed to increase the GUS activity, suggesting that induction of VR-ACS7 occurs via separate signaling pathways in response to IAA and BR. A scheme of the multiple regulatory pathways for the expression of ACC synthase multigene family by auxin and BR is presented.