Note: Descriptions are shown in the official language in which they were submitted.
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FIELD OF INVENTION
The present invention relates to a culture vessel for plant
tissue culture and is particularly useful for in vitro
restoration and mass propagation of plantlets.
PRIOR ART
It is presently known to the art to use solid as well as
liquid culture mediums for such in-vitro regeneration and
mass propagation of plantlets. However, it has been found
that the use of liquid culture medium in conventional
processes of in-vitro regeneration of plantlets is more
effective since it facilitates more oxygen and nutrient
uptake by the cells/tissues which improves production
levels. Culture vessels known in the art are normally found
to be more suitable for use in solid culture medium since
this suffers from certain inherent limitations in their
construction for having the advance of the use of liquid
culture medium. It is essential that for plant tissue
culture with liquid culture medium, the plant tissue during
culture be free of any contamination either from the
atmosphere or in improper handling of the tissue while
carrying out various stages of the culture. Further, in
use of liquid culture medium, it is necessary to
periodically monitor the culture medium to ensure proper
quantity of the liquid medium to facilitate proper uptake
of the medium by the tissue for the purpose of its culture.
Moreover, to carry out the various stages of culture of
tissue, the liquid culture media requires to be changed
from one formulation to another during the culture process.
While changing the liquid culture media, it is necessary
that the tissue is not subjected to contamination. The
conventional culture vessels presently used in the art
are not suitable for plant tissue culture using liquid
culture medium.
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OBJECTS OF THE INVENTION
:; .
An object of the present invention is to provide a culture
vessel for plant tissue culture which can be advantageously
utilised in carrying out the different shapes of plant
tissue culture with liquid culture medium by changing the
medium in the same vessel and thereby reducing the chances
of contamination.
,
BRIEF DESCRIPTION OF THE INVENTION
According to the present invention, there is provided a
culture vessel comprising :
a transparent container having a releasably fitted cap;
atleast one side tube having a releasably fitted cap and
being in flow communication with the lower part of said
container through atleast one passage;
said side tube disposed at an inclination with respect
to the wall of said container;
atleast one float member disposed within said container
and having a porous support sheet to support the tissue
for culture.
The culture vessel of the present invention allows a tissue
culture without subjecting the tissue to contamination.
The releasable cap of the container restricts inflow of
contaminants within the vessel during culture. Further,
the side tube provides for necessary monitoring change
of liquid culture media and/or maintaining particular
predetermined level of the liquid media in the vessel and
drain in/out to and from the vessel as and when required.
The requirement of the tissue to be transferred from one
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vessel to the other for subjecting the same to the culture
stages is also carried out in the same vessel by introducing
the appropriate liquid medium through the side tube into
the vessel at each stage of culture. The side tube is also
provided with a releasable cap at its open end to restrict
inflow of any contaminants within the vessel through the
tube.
DESCRIPTION WITH REFERENCE TO THE ACCOMPANYING DRAWINGS
Fig.1 is a schematic representation of the culture vessel
according to the present invention showing the vessel fitted
with the cap and housing the float member with the supported
porous sheet inside the float;
Fig.2 is a sectional view of the float member illustrating
the provision of the flange near its lower end to support
the porous sheet; and
Fig.3 is a further sectional view of the complete culture
vessel along line AA' of Fig.1 with the cap fitted thereon.
Reference is made to Fig.1 wherein the culture vessel (V)
comprises a container (CO) which may be cylindrical or
of any other shape having a screw cap (Ca). The container
(CO) is provided with side tube (ST) disposed at an
inclination with respect to the side wall (WS) of container
(CO). Side tube (ST) has a cap (Cp) removably held thereto.
As shown in Fig.3, cap (Cp) has a membrane made of any
suitable resilient or puncturable material so as to allow
an introduction or withdrawal of a needle of a syringe.
The side tube (ST) communicates with the inside of the
container (CO) through passages (01) and (02) provided
in side wall (WS). Passages (01) and (02) are disposed
in a spaced relationship to each other, and such that
passage (01) allows an introduction, addition or withdrawl
of the culture medium into container (CO) without the
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removal of the cap (Ca). Passage (02) is disposed at a
height above passage (01) and breaks the vacuum within
side tube (ST). The container (C0) has a float member (F)
which may also be cylindrically shaped or any other shape.
The shape of container (C0) and float member (F) is not
of importance. Float (F) has a porous sheet (PS). In
accordance with one e~bodiment, float (F) is provided with
a radially inwardly extending flange (FL), as shown in
Fig.2, which has a porous sheet (PS) held thereto.
Alternatively, as shown in Fig.3, flange (FL) supports
porous sheet (PS). The tissue is placed on porous sheet
(PS), which simultaneously allows absorption of the nutrient ;~
from the liquid media by the tissue. Thus, the tissue for
culture is placed on the porous sheet (PS) and the float
(F) is then introduced through the open end of container
(C0). Thereafter, cap (Ca) closes container (C0) so as
to provide a contaminant free atmosphere to the tissue.
The fitting of cap (Ca) to container (C0) can be effected
either by means of threads which may be continuous or
discontinous or any press fit connection of the cap (Ca)
to coptainer (C0). The liquid media required for the tissue
culture is fed into container (C0) through side tube (ST)
and passage (01). As soon as the liquid media occupies
container (C0) under float (F), the latter rises up and
floats over the liquid media. The level of the liquid media
is maintained up to a maximum upper limit so that it is
lower than the height of second passage (02). The
transparency of container (C0) and side tube (ST) further
facilitates constant monitoring of the level of the liquid
media in vessel (C0). During tissue culture, the liquid
media is uptaken by the tissue and due to such uptake of
the liquid medium, the level of the liquid starts falling.
Such fall in level of the liquid media in vessel (C0) is
noted and the level is maintained by introduction of fresh
liquid media through side tube (ST). Likewise, while
carrying out various stages of the culture process, lf
,
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it is essential to change the liquid media, side tube (ST)
can be effectively utilised to drain out and/or introduce
the liquid media necessary for the growth of plant tissue.
Side tube (ST) is provided with a releasably fitted cap
(Cp). The cap (Cp) in accordance with a preferred
embodiment, has a membrane (M) of rubber or any puncturable
material to facilitate the draining of the introduction
of the culture media to the container (CO) through the
side tube (ST). The releasably fitted cap (Cp) can have
a threaded connection to the side tube or may be press
fitted onto the side tube's top end. Once the tissue is
supported on sheet (PS), and in order to maintain the
contAm;nAnt free atmosphere within container (CO), the
introduction, replenishment or withdrawal of a culture
medium is effected by puncturing membrane (M) with the
needle of a syringe.
As shown in Figs. 2 & 3, float (F) comprises an inner wall
(F), spaced from the outer wall (F2) so as to provide the
required strength to float (F).
The threaded press fit connection of caps (Ca) and (Cp)
may have continuous and/or discontinous threads to prevent
ingress of contAminAnts into culture vessel (CO).
Preferably, cap (Ca) is provided with a radially inwardly
and downwardly extending sloped bottom surface ~SL) as
illustrated in Fig.3. The inclined sloped surface cap
prevents cont~r;nAtion resulting from arrested condensed
droplets held along the cap wall joint and/or its dripping
down along the wall of the container.
It is obvious to mention that the culture vessel and its
side tube of the invention which is shown to be cylindrical
shaped as described above with reference to the accompanying
figures of the preferred embodiments can be of any other
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shape and configuration, triangular, circular, square,
rectangular, oval and polygonal. Also, the shape of the
float member and the perforated support structure (PS)
can also vary depending upon such shape and configuration
of the container. The container and the side tube can be
selectively obtained or autoclavable/sterilisable,
transparent, semi-transparent, non-corrosive metallic/non-
metallic composite material such as glass, polymeric
materials and stainless steel. The float can have side
walls either solid or hollow and can be obtained of non-
corrosive, metallic/non-metallic composite materials such
as stainless steel and polymeric materials. The central
support structure can be perforated/porous/woven mouled
and can be selectively obtained of non-corrosive
metallic/non-metallic composite material such as glass
wool, polymeric materials or stainless steel. The support
structure can be integral to the float matrix or may be
an independent structure adapted tobe supported on the
radially inwardly extended flange of the float and/or like
adaption provided in the float member.
Further, it is possible to make obvious modification of
the culture vessel of the invention described above and
the scope of the invention should be determined keeping
in view of the possibility of the obvious modifications
of the disclosure according to the present invention. It
may be noted in this respect that the culture vessel of
the invention can have more than one side tube instead
of the one side tube described in the illustrated preferred
embodiments.
