Estudio del grano de polen en especies del genero Heliconia L ...
Estudio del grano de polen en especies del genero Heliconia L ...
Estudio del grano de polen en especies del genero Heliconia L ...
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Sanabria et al.<br />
b. Tratami<strong>en</strong>to con hidróxido <strong>de</strong><br />
potasio (KOH) (2).<br />
Se incluyeron las anteras <strong>en</strong><br />
KOH al 10% y se colocaron <strong>en</strong> baño<br />
<strong>de</strong> María. La preparación se <strong>de</strong>jó <strong>en</strong>friar<br />
y se lavó con agua <strong>de</strong>stilada. El<br />
material vegetal se <strong>de</strong>shidrató pasándolo<br />
por una batería <strong>de</strong> alcohol etílico<br />
(70, 95 y 100%), cada paso <strong>de</strong> 5 minutos.<br />
Se c<strong>en</strong>trífugo a 2000 rpm tres veces<br />
y por 3 minutos cada vez y <strong>de</strong>cantando<br />
cuidadosam<strong>en</strong>te <strong>en</strong> cada paso.<br />
El material vegetal se colocó <strong>en</strong><br />
b<strong>en</strong>c<strong>en</strong>o por 5 minutos y se <strong>de</strong>cantó.<br />
Se cubrió con una mezcla <strong>de</strong><br />
b<strong>en</strong>c<strong>en</strong>o:silicón <strong>en</strong> proporción 1:1. Se<br />
<strong>de</strong>jó reposar por 24 horas o hasta que<br />
el b<strong>en</strong>c<strong>en</strong>o se evaporó. Para la observación<br />
al microscopio óptico (MO), se<br />
colocaron gotas <strong><strong>de</strong>l</strong> preparado <strong>en</strong> un<br />
portaobjetos, se cubrieron con un<br />
cubreobjeto y se esperó el tiempo necesario<br />
par que se ext<strong>en</strong>diera la muestra<br />
<strong>de</strong> <strong>grano</strong>s <strong>de</strong> <strong>pol<strong>en</strong></strong>.<br />
c. Sa<strong>en</strong>z (12) y Kress (6 y 7).<br />
Los <strong>grano</strong>s <strong>de</strong> <strong>pol<strong>en</strong></strong> fueron extraídos<br />
<strong>de</strong> las anteras con la ayuda<br />
<strong>de</strong> una aguja <strong>de</strong> disección, se colocaron<br />
<strong>en</strong> un portaobjeto y se les añadió<br />
una gota <strong>de</strong> xilol puro. Se cubrieron<br />
con un cubreobjeto y se sellaron las<br />
preparaciones con esmalte para uñas<br />
transpar<strong>en</strong>te. En caso <strong>de</strong> que se requirieran<br />
preparaciones perman<strong>en</strong>tes,<br />
el xilol fue sustituido por bálsamo<br />
<strong>de</strong> Canadá o Permount.<br />
d. Mello Filho (9).<br />
Esta metodología se pue<strong>de</strong> utilizar<br />
tanto para anteras frescas, fijadas<br />
o tomadas <strong>de</strong> material<br />
herborizado y se recomi<strong>en</strong>da para preparaciones<br />
rápidas. El procedimi<strong>en</strong>to<br />
consistió <strong>en</strong> extraer los <strong>grano</strong>s <strong>de</strong><br />
<strong>pol<strong>en</strong></strong> <strong>de</strong> las anteras, colocarlos <strong>en</strong> un<br />
preparation were placed in a<br />
microscope sli<strong>de</strong> and were covered<br />
with other one, and time required for<br />
grain sample being ext<strong>en</strong><strong>de</strong>d.<br />
c. Sa<strong>en</strong>z (12) and Kress (6, 7).<br />
Poll<strong>en</strong> grains were extracted<br />
from anthers with the aid of a<br />
dissection needle, it was placed in a<br />
microscope sli<strong>de</strong> and a pure xilol drop<br />
was ad<strong>de</strong>d. It was covered with an over<br />
microscope sli<strong>de</strong> and preparations<br />
were sailed with transpar<strong>en</strong>t nail. In<br />
case of requiring perman<strong>en</strong>t<br />
preparations, xilol was substituted by<br />
Canada balsam or Permount.<br />
D, Mello Filho (9).<br />
This methodology can be used<br />
for fresh anthers, fixed or tak<strong>en</strong> from<br />
dried material and it is recomm<strong>en</strong><strong>de</strong>d<br />
for rapid preparations. Procedure<br />
consisted in to extract poll<strong>en</strong> grains<br />
for anthers, place in a microscope sli<strong>de</strong><br />
and adds a drop of mix conformed by<br />
chloral foun<strong>de</strong>d hydrate, Lactoph<strong>en</strong>ol<br />
and glycerin 50% (1/3 each).<br />
For every methodology, poll<strong>en</strong><br />
grains were observed, photographed<br />
and measured with an optical<br />
microscope (OM) Olympus BX40 with<br />
50X and 100X augm<strong>en</strong>tations. For its<br />
characterization it was used in<strong>de</strong>x<br />
table established by Erdtman (2, 3)<br />
and 11. Size <strong>de</strong>termination was based<br />
on 25 measurem<strong>en</strong>ts of polar<br />
diameter (P) and equatorial (E) of<br />
these structures selection at random,<br />
in meridian optical cut and<br />
relationship P/E applying.<br />
Results and discussion<br />
After studying morphology of<br />
poll<strong>en</strong> grains of H. bihai, H.<br />
psittacorum, H. platystachys and H.<br />
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