RÉSUMÉ
En el presente trabajo, es estudiada la diversidad genética de tres poblaciones atribuidas a ecotipos de aguaymanto, Physalis peruaviana. Las tres poblaciones eran atribuidas a los ecotipos Agroandino (provincia de San Pablo), Celendino (provincia de Celendín) y Cajabamba (provincia de Cajabamba) del departamento de Cajamarca. Se realizó la cuantificación proteica y evaluó el polimorfismo de las proteínas de reserva seminal (SSPs) mediante electroforesis en gel de poliacrilamida denaturante (SDS-PAGE). Además, se identificaron características bioquímicas de las proteínas seminales en esta especie. No se hallaron diferencias entre las tres poblaciones basados en la cuantificación proteica. Las globulinas (82.4%) fueron la fracción mayoritaria seguida por las albuminas (13.9%), glutelinas (3.7%) y prolaminas (0.7%). Sólo las albuminas mostraron polimorfismo, hallándose 21 proteínas entre ~ 6.5 a ~45 kDa y tres perfiles electroforéticos diferentes, los cuales fueron compartidos entre las poblaciones. Se identificaron las leguminas y vicilinas en la fracción globulina. Las glutelinas mostraron proteínas de mismo peso molecular (PM) a las leguminas; y las prolaminas sólo una banda de bajo PM. La población de San Pablo fue completamente homogénea a diferencia de la población de Cajabamba que mostró la mayor diversidad genética seguida de Celendín. No fue posible diferenciar las poblaciones designadas como ecotipos Agroandino, Cajabamba y Celendino basados en el análisis de proteínas seminales.
The genetic diversity of three populations designated as ecotypes of golden berry (Physalis peruaviana) is studied using protein quantification and polymorphism of seed storage proteins (SSPs) by denaturating polyacrylamide gel electrophoresis (SDS-PAGE). As well, biochemical characteristics of seed proteins were identified. The populations were from San Pablo province (Agroandino ecotype), Celendín province (Celendino ecotype) and Cajabamba province (Cajabamba ecotype), all from Cajamarca Department. There was not difference among the three populations based on protein quantification. Globulins (82.4%) were the majority fraction followed for albumins (13.9%), glutelins (3.7%) and prolamins (0.7%). Only albumins showed polymorphism, showing 21 proteins between ~6.5 to ~45 kDa and three different electrophoretic profiles, which were share among the three populations. Legumins and vicilins were identified in globulin fraction. Glutelins showed proteins of same molecular weight (MW) to legumins; and prolamins only a band of low MW. San Pablo province population (Agroandino ecotype) was completely uniform, while Cajabamba population showed higher genetic diversity followed by Celendin population. Our results shows that, based on seed proteins analyses is not possible to distinguish the three populations designated as Agroandino, Cajabamba and Celendino ecotypes.
RÉSUMÉ
Objective: To establish a stable, reproducible, and suitable reaction system for ISSR analysis of genetic differences in Illicium difengpi. Methods: The ISSR-PCR amplification system on I. difengpi in five factors (Mg2+, dNTPs, primers, Taq DNA polymerase, and DNA template) was optimized by orthogonal design, and the PCR result was analyzed by SPSS. Then based on the optimal ISSR-PCR amplification system, the annealing temperature and cycle times in PCR were proposed by gradient determenation. Results: Most of the factors in different levels had the significant effects on the result of PCR, and the most remarkable factor was the quantity of Taq DNA polymerase. The optimized ISSR-PCR reaction system (20 μL) for I. difengpi was constructed of Mg2+ (1.60 mmol/L), dNTP (0.22 mmol/L), primer (0.90 μmol/L), Taq polymerase (0.50 U), and DNA template (70.00 ng). The optimized annealing temperature and cycle times were 51.8 °C and 40 cycles, respectively. Thirteen ISSR primers with stable amplification and abundant polymorphism were selected from 62 ISSR primers. Conclusion: The established and optimized ISSR reaction system is stable and credible according to the testing results of 16 samples of I. difengpi, and provides the basis for the genetic analysis of I. difengpi.