Author: NANCY SANTANA BUZZY
NANCY SANTANA BUZZY (2015)
In order to determine the role of polyamines in the formation and development of the somatic embryos of Capsicum chinense, the effect of different concentrations (0, 0.01, 0.1, and 1.0 mM) of Putrescine, Spermidine and Spermine on the efficiency and morphology of the embryos was evaluated. The results show that none of the three polyamines evaluated had a significant effect on the number of embryos formed, except Spermidine (1 mM), which caused a significant reduction in their numbers, in comparison with the control treatment. However, the most noteworthy result was observed in the treatment containing 0.1 mM of Spermine. The embryos developed in this treatment showed harmonic apex-radicle development, pale-green coloration and the formation of two tiny cotyledonary leaves. Real-time PCR was used to analyze the differential expression of the WUS, WOX1 and WOX3 genes in somatic embryos treated with Spermine and untreated, including the zygotic embryo. The transcript levels of the genes analyzed were found to differ significantly between both types of embryos (somatic and zygotic), with the zygotic embryos presenting a higher level of transcripts; however, compared to the untreated somatic embryos, the somatic embryos treated with Spermine showed an increase in the transcript levels of the three genes analyzed (WUS, WOX1 and WOX3); the WOX1 gene in particular presented an accumulation pattern similar to that of the zygotic embryo of the species.
NANCY SANTANA BUZZY (2011)
The effect of NaCI salinity on growth and development of somatic embryos of Habanero pepper was examined. Addition of 75 and 100 mM NaCI into the medium greatly increased the growth and development of somatic embryos and both of these concentrations favored the proliferation of somatic embryos. However, supplementation of 200 and 300 mM NaCI to the medium showed a negative effect on the growth and development of somatic embryos. Concentration increases of NaCl provoked a significant reduction of the embryos survival rate with the average lethal dose (46%) being registered in the treatment of 100 mM. Furthermore, a lower tolerance to salt stress (NaCl) was observed in deformed somatic embryos. Concentrations of 200 and 300 mM NaCl significantly delayed development in the surviving embryos in both treatments. These embryos remained at the globular stage throughout culture time. At 75 mM NaCl, most of the embryos were observed in the torpedo stage. However, the embryos exposed to 100 mM NaCl were observed mainly in globular and cotiledonar stages. It is quite likely that the transition from one intermediate stage of development to another occurs rapidly. With the exception of the concentration at 300 mM NaCl, salt stress stimulated embryonic germination, particularly at 100 mM NaCl. The content of proline in somatic embryos increased substantially in response to salinization. The results suggest that somatic embryos of C. chinense can tolerate concentrations of NaCl up to 100 mM without their development being affected. Moreover, they have sufficient cellular mechanisms to tolerate salinity at relatively higher levels.
NANCY SANTANA BUZZY (2013)
Somatic embryo-like structureswere produced from the hypocotyls of aseptic plants of Capsicum Chinese.
Somatic embryogenesis is a powerful tool for the massive production of elite plant materials, as well as for molecular agricultural breeding through the use of biotechnological strategies. Although this technology can be applied to any plant species,
NANCY SANTANA BUZZY (2013)
Jatropha curcas is a small bush which has attracted atention as an energy crop for biodisel production.
NANCY SANTANA BUZZY (2010)
This article describes the performance of nodal segments from Habanero
pepper (Capsicum. chinense) during shoot induction and elongation under different
semisolid and liquid culture conditions with various degrees of ventilation in which they
were exposed to different levels of immersion and growth regulators. The ethylene
content in non-ventilated containers, the age of the explant donor plants as well as the
effect of thidiazuron and paclobutrazol on shoot induction and of gibberellic acid and
AgNO3 on shoot elongation were also evaluated. A temporary immersion bioreactor
(BioMINT) was used for the multiplication and elongation of isolated shoots with very
good results. We report an efﬁcient protocol for the in vitro propagation of Habanero
pepper that produces plants with a high survival rate when transplanted to soil.
NANCY SANTANA BUZZY (2014)
Intersimple sequence repeat (ISSR) markers were used to evaluate the effects of
in vitro culture on genetic variation in Habanero pepper (Capsicum chinense Jacq.)
regeneration protocols. A total of 219 ISSR clear and reproducible fragments were
generated with 13 ISSR primers in direct organogenesis, direct and indirect somatic
embryos, and the embryogenic callus system. A cluster analysis was performed to express
in the form of dendrogram the relationships among different regeneration systems and
the genetic variability detected. Genetic distance analysis indicated that our regeneration
protocols are inappropriate for micropropagation, conservation, or genetic transformation;
however, they may be applicable to breeding. This is the first report on the use of
molecular analysis to evaluate genetic variation of in vitro-regenerated plants of
Habanero pepper using ISSR markers.
This paper describes two in vitro regeneration systems through direct and indirect organogenesis in Pinus brutia using fascicles aseptic cultures as explants. Mechanical scarification and gibberellic acid (GA3) were evaluated on in vitro seed germination. Scarification was the treatment that allowed for in vitro seed germination. The highest direct organogenic response was obtained in Murashige and Skoog (MS) medium containing 4.5 µM thidiazuron, whereas the highest indirect organogenesis was obtained with 9.8 µM thidiazuron and 3.4 µM paclobutrazol. The isolated shoots were rooted on MS medium supplemented with 1.70 µM indoleacetic acid. A large variation in root ability was observed among plantlets. These results suggest that both regeneration systems can be applied to the micropropagation or genetic transformation of P. brutia.
Protein profile was studied during the development of Capsicum chinense somatic embryos. The total protein content and profile of polypeptides (by sodium dodecyl sulfate polyacrylamide gel electrophoresis) of somatic embryos at different developmental stages (globular, heart-shaped, torpedo and cotyledonary stages) were analyzed. The protein profile of zygotic embryos included nine exclusive bands with molecular weights of 4.0, 5.2, 8.1, 13.7, 20.9, 23.7, 41, 50 and 69.3 kDa; these bands were not observed in the protein profile of somatic embryos. Coincidently, five of these bands possessed similar molecular weights to those reported for storage proteins in other plant species. Protein content showed a clear decreasing tendency with increasing somatic embryo development. The lowest protein content was detected in somatic embryos at the cotyledonary stage (0.436 µg/mg fresh weight), and the highest content was found in somatic embryos at the globular stage (2.98 µg/mg fresh weight). Total proteins two-dimensional electrophoresis (2-DE) analysis of mature zygotic embryo (prior to the desiccation) and cotyledonal somatic embryo, showed significant differences in the protein profile of both types of embryos. Zygotic embryo showed the proteins expression of isoelectric point between 4 to 7 and 7 to 10, and molecular weights between 25 to 36 KDa, which were not expressed in the cotyledonal somatic embryo. The low protein content during the development of the somatic embryos, particularly at the cotyledonary stage, is a factor that could be related with the low rate of conversion to plantlets and the high frequency of deformed somatic embryos of C. chinense.
Hilda Eulalia Lee Espinosa ANTONIO LAGUNA CERDA Joaquín Murguía González PABLO ELORZA MARTINEZ Lourdes Georgina Iglesias Andreu BENJAMIN GARCIA ROSAS FELIPE ALONSO BARREDO POOL NANCY SANTANA BUZZY (2007)
Se germinaron in vitro semillas de Laelia anceps ssp. dawsonii, una orquídea silvestre amenazada, originaria de México y Mesoamérica, con alto potencial ornamental, utilizando el medio Murashige & Skoog (1962) suplementado con ácido 1-naftalén-acético (ANA), 6-benzyl-amino-purina (BAP), Kinetina (Kin) y ácido indol-3-acético (AIA), 2 mg L-1 de cada uno, el cual resultó óptimo para la inducción de callo bajo fotoperiodo de 16/8 h (20.2 µmol·m-2·s-1). El callo fue subcultivado a intervalos de 45 días en el mismo medio de cultivo, produciendo en promedio 524 embriones somáticos en el tercer subcultivo. Los embriones somáticos producidos se convirtieron en plantas completas con brotes y raíces en el mismo medio, y fueron transferidas al medio VW suplementado con BAP 2 mg L-1, AIA 1 mg L-1 y carbón activado 0.2 % para su desarrollo. Después de aproximadamente tres meses, las plántulas fueron aclimatizadas en el invernadero con un 100 % de tasa de sobrevivencia.
Most of the cycads contain high concentrations of essential oils, flavonoids, polyphenols, and polysaccharides that interfere with DNA extraction, causing erroneous or no PCR products. The optimization of DNA isolation, employing inter-simple sequence repeats (ISSRs) primers were investigated in Ceratozamia mexicana Brongn., an endangered Mexican cycad. The DNA obtained from fresh-leaf tissues with a modified cetyltrimethylammonium bromide buffer protocol gave a good quality of DNA with no colored pigments and contaminants. The main modification to the CTAB-based DNA extraction protocol was the one hour leaf tissue soaking pre-treatment with a 0.7 M NaCl solution, to facilitate the cell lysis. The DNA extracted was successfully amplified by PCR using six arbitrary ISSR primers. Reproducible amplifiable products were observed in all PCR reactions. Our results show a significant improvement in the DNA quality obtained using low primer concentration (25 pM). 23 strong bands were detected, 9 of which were polymorphic. The results indicated that the optimized protocol for DNA isolation and PCR system is suitable for further work in this specie. This work is the first DNA extraction and ISSR protocols reported for this ornamental and endangered species.