Seeds from Tabebuia roseo-alba lose viability very fast. Moreover, the seed germination rate is very low, reaching approximately 40%. This study aimed at the in vitro induction of embryogenic callus. This technology a...Seeds from Tabebuia roseo-alba lose viability very fast. Moreover, the seed germination rate is very low, reaching approximately 40%. This study aimed at the in vitro induction of embryogenic callus. This technology allows subsequent plant regeneration as an alternative for the production of T. roseo-alba seedlings. Seeds were germinated in vitro and after 20 days, cotyledonary leaves, hypocotyls and root segments excised from these seedlings were used as explants. They were inoculated on MS medium supplemented with sucrose （30 g/L）, agar （5.0 g/L） and different auxins. The effect of 2,4-D, picloram and NAA at concentrations 0.0, 0.5, 1.0, 2.0 and 4.0 mg/L was evaluated. For the analysis of callus with embryogenic characteristics, ultra-structural study by scanning electron microscopy and cytochemical test with carmine were performed. The results showed that the culture medium supplemented with 4 mg/L NAA presented induction of callus with embryogenic characteristics in all explants used, with cotyledonary leaves showing the highest percentage （70% of explants with embryogenic characteristics）. The use of 2, 4-D and picloram was efficient for callus formation in different explants, but no embryogenic characteristics were observed. From the ultra-structural analysis of callus with embryogenic characteristics, it was found that cells from different explant sources had isodiametric format. This format is similar to somatic embryos in globular stage. The cytochemical analysis confirmed the presence of pro-embryogenic cells in callus mass. Callus induced from cotyledonary leaves presented 46% positive reaction to carmine acetic.展开更多
AIM To investigate the contribution of polymorphisms in the CYP1A1, CYP2E1 and EPHX1 genes on sporadic colorectal cancer(SCRC) risk. METHODS Six hundred forty-one individuals(227 patients with SCRC and 400 controls) w...AIM To investigate the contribution of polymorphisms in the CYP1A1, CYP2E1 and EPHX1 genes on sporadic colorectal cancer(SCRC) risk. METHODS Six hundred forty-one individuals(227 patients with SCRC and 400 controls) were enrolled in the study. The variables analyzed were age, gender, tobacco and alcohol consumption, and clinical and histopathological tumor parameters. The CYP1A1 *2A, CYP1A1 *2C CYP2E1 *5B and CYP2E1 *6 polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). The EPHX1 Tyr113 His, EPHX1 His139 Arg and CYP1A1 *2C polymorphisms were detected by real-time PCR. Chisquared test and binary logistic regression were used in the statistical analysis. Haplotype analysis was conducted using the Haploview program, version 2.05.RESULTS Age over 6 2 years was a risk factor for SCRC development(OR = 7.54, 95%CI: 4.94-11.50, P < 0.01). Male individuals were less susceptible to SCRC(OR = 0.55, 95%CI: 0.35-0.85, P < 0.01). The CYP2E1*5B polymorphism was associated with SCRC in the codominant(heterozygous genotype: OR = 2.66, 95%CI: 1.64-4.32, P < 0.01), dominant(OR = 2.82, 95%CI: 1.74-4.55, P < 0.01), overdominant(OR = 2.58, 95%CI: 1.59-4.19, P < 0.01), and log-additive models(OR = 2.84, 95%CI: 1.78-4.52, P < 0.01). The CYP2E1*6 polymorphism was associated with an increased SCRC risk in codominant(heterozygous genotype: OR = 2.81, 95%CI: 1.84-4.28, P < 0.01; homozygous polymorphic : OR = 7. 3 2, 9 5 % C I : 1.85-28.96, P < 0.01), dominant(OR = 2.97, 95%CI: 1.97-4.50, P < 0.01), recessive(OR = 5.26, 95%CI: 1.35-20.50, P = 0.016), overdominant(OR = 2.64, 95%CI: 1.74-4.01, P < 0.01), and log-additive models(OR = 2.78, 95%CI: 1.91-4.06, P < 0.01). The haplotype formed by the minor alleles of the CYP2E1*5B(C) and CYP2E1*6(A) polymorphisms was associated with SCRC(P = 0.002). However, the CYP1A1 *2A, CYP1A1 *2C, EPHX1 Tyr113 His and EPHX1 His139 Arg polymorphisms were not associated with SCRC.CONCLUSION In conclusion, the results demonstrated that CYP2E1*5B and CYP2E1*6 minor alleles play a role in the development of SCRC.展开更多
文摘Seeds from Tabebuia roseo-alba lose viability very fast. Moreover, the seed germination rate is very low, reaching approximately 40%. This study aimed at the in vitro induction of embryogenic callus. This technology allows subsequent plant regeneration as an alternative for the production of T. roseo-alba seedlings. Seeds were germinated in vitro and after 20 days, cotyledonary leaves, hypocotyls and root segments excised from these seedlings were used as explants. They were inoculated on MS medium supplemented with sucrose （30 g/L）, agar （5.0 g/L） and different auxins. The effect of 2,4-D, picloram and NAA at concentrations 0.0, 0.5, 1.0, 2.0 and 4.0 mg/L was evaluated. For the analysis of callus with embryogenic characteristics, ultra-structural study by scanning electron microscopy and cytochemical test with carmine were performed. The results showed that the culture medium supplemented with 4 mg/L NAA presented induction of callus with embryogenic characteristics in all explants used, with cotyledonary leaves showing the highest percentage （70% of explants with embryogenic characteristics）. The use of 2, 4-D and picloram was efficient for callus formation in different explants, but no embryogenic characteristics were observed. From the ultra-structural analysis of callus with embryogenic characteristics, it was found that cells from different explant sources had isodiametric format. This format is similar to somatic embryos in globular stage. The cytochemical analysis confirmed the presence of pro-embryogenic cells in callus mass. Callus induced from cotyledonary leaves presented 46% positive reaction to carmine acetic.
基金Supported by Sao Paulo Research Foundation(FAPESP),No.2011/23969-1 and No.2012/02473-0Coordination for the Improvement of Higher Education Personnel(CAPES)(Master grant)and National Council of Technological and Scientific Development(CNPq),No.310582/2014-8
文摘AIM To investigate the contribution of polymorphisms in the CYP1A1, CYP2E1 and EPHX1 genes on sporadic colorectal cancer(SCRC) risk. METHODS Six hundred forty-one individuals(227 patients with SCRC and 400 controls) were enrolled in the study. The variables analyzed were age, gender, tobacco and alcohol consumption, and clinical and histopathological tumor parameters. The CYP1A1 *2A, CYP1A1 *2C CYP2E1 *5B and CYP2E1 *6 polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). The EPHX1 Tyr113 His, EPHX1 His139 Arg and CYP1A1 *2C polymorphisms were detected by real-time PCR. Chisquared test and binary logistic regression were used in the statistical analysis. Haplotype analysis was conducted using the Haploview program, version 2.05.RESULTS Age over 6 2 years was a risk factor for SCRC development(OR = 7.54, 95%CI: 4.94-11.50, P < 0.01). Male individuals were less susceptible to SCRC(OR = 0.55, 95%CI: 0.35-0.85, P < 0.01). The CYP2E1*5B polymorphism was associated with SCRC in the codominant(heterozygous genotype: OR = 2.66, 95%CI: 1.64-4.32, P < 0.01), dominant(OR = 2.82, 95%CI: 1.74-4.55, P < 0.01), overdominant(OR = 2.58, 95%CI: 1.59-4.19, P < 0.01), and log-additive models(OR = 2.84, 95%CI: 1.78-4.52, P < 0.01). The CYP2E1*6 polymorphism was associated with an increased SCRC risk in codominant(heterozygous genotype: OR = 2.81, 95%CI: 1.84-4.28, P < 0.01; homozygous polymorphic : OR = 7. 3 2, 9 5 % C I : 1.85-28.96, P < 0.01), dominant(OR = 2.97, 95%CI: 1.97-4.50, P < 0.01), recessive(OR = 5.26, 95%CI: 1.35-20.50, P = 0.016), overdominant(OR = 2.64, 95%CI: 1.74-4.01, P < 0.01), and log-additive models(OR = 2.78, 95%CI: 1.91-4.06, P < 0.01). The haplotype formed by the minor alleles of the CYP2E1*5B(C) and CYP2E1*6(A) polymorphisms was associated with SCRC(P = 0.002). However, the CYP1A1 *2A, CYP1A1 *2C, EPHX1 Tyr113 His and EPHX1 His139 Arg polymorphisms were not associated with SCRC.CONCLUSION In conclusion, the results demonstrated that CYP2E1*5B and CYP2E1*6 minor alleles play a role in the development of SCRC.
