GENERAL ANDROGENESIS PROCEDURES
COLLECTION
, DISINFESTATION
, EXCISION
,
AND
CULTURE
Floral buds may be collected from plants grown in the field, greenhouse, or growth chamber. Entire
inflorescences or individual buds are harvested and kept moist until ready for culturing. If buds are
to be pretreated (i.e., kept at low temperature), they should be wrapped in a moistened paper tissue
and placed into a small zipper-type plastic bag.
Flower buds are typically disinfested using a 5% sodium or calcium hypochlorite solution for
5 to 10 min, and then are rinsed thoroughly in sterile distilled water. Anthers are aseptically excised
in a laminar flow hood, taking care not to cause injury. If the anther is still attached to the filament,
the filament is carefully removed.
If a solid medium is used, the anthers are gently pressed onto the surface of the medium (just
enough to adhere to the medium), but should not be deeply embedded. When using a liquid medium
for intact anthers, the anthers are floated on the surface. Care must be taken when moving float
cultures so as not to cause the anthers to sink below the surface.
For most species, disposable petri dishes are utilized for anther cultures. For a species with
large anthers, such as tobacco, the anthers from 4 to 5 buds (20–25 anthers) may be cultured
together on one 100 x 15 mm diameter petri dish. For species with smaller anthers, or for certain
experimental designs, smaller petri dishes or other containers may be more useful. Petri dishes are
usually sealed and placed into an incubator; the specific temperature and light requirements of the
incubator depends on the species being cultured.
While many of the steps involved in microspore culture are similar to those of anther culture,
microspore culture also requires the separation of the microspores from the surrounding anther
tissue. Microspores may be squeezed out of anthers using a pestle or similar device, or a microblending
procedure may be used. See Dunwell (1996) for a review of literature pertaining to
microspore culture
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GENERAL ANDROGENESIS PROCEDURESCOLLECTION, DISINFESTATION, EXCISION,ANDCULTUREFloral buds may be collected from plants grown in the field, greenhouse, or growth chamber. Entireinflorescences or individual buds are harvested and kept moist until ready for culturing. If buds areto be pretreated (i.e., kept at low temperature), they should be wrapped in a moistened paper tissueand placed into a small zipper-type plastic bag.Flower buds are typically disinfested using a 5% sodium or calcium hypochlorite solution for5 to 10 min, and then are rinsed thoroughly in sterile distilled water. Anthers are aseptically excisedin a laminar flow hood, taking care not to cause injury. If the anther is still attached to the filament,the filament is carefully removed.If a solid medium is used, the anthers are gently pressed onto the surface of the medium (justenough to adhere to the medium), but should not be deeply embedded. When using a liquid mediumfor intact anthers, the anthers are floated on the surface. Care must be taken when moving floatcultures so as not to cause the anthers to sink below the surface.For most species, disposable petri dishes are utilized for anther cultures. For a species withlarge anthers, such as tobacco, the anthers from 4 to 5 buds (20–25 anthers) may be culturedtogether on one 100 x 15 mm diameter petri dish. For species with smaller anthers, or for certainexperimental designs, smaller petri dishes or other containers may be more useful. Petri dishes areعادة ما تكون مختومة وتوضع في حاضنة؛ على درجة حرارة معينة ومتطلبات الخفيفةحاضنة يعتمد على الأنواع التي يجري مثقف.بينما العديد من الخطوات التي تنطوي عليها الثقافة المئبر مماثلة لتلك الثقافة العضو الذكرى،يتطلب ثقافة المئبر أيضا فصل ميكروسبوريس عن العضو الذكرى المحيطةالأنسجة. وقد تقلص microspores الخروج من أنثيرس باستخدام جهاز مماثل، مدقة أو ميكروبليندينجيمكن استخدام الإجراء. انظر دونويل (1996) لإجراء استعراض للمؤلفات المتعلقةالثقافة المئبر
يجري ترجمتها، يرجى الانتظار ..
