Embryo culture and endosperm culture

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Embryo culture and endosperm culture:

Embryo culture and endosperm culture By- Himanshu mandawat Reg no: 07/075

Defination :

Defination Embryo culture can be defined as the in vitro culture either of the polarized egg, zygote, proembryo or mature embryo.

Histroy:

Histroy The first embryo culture, although crude, was done by Hanning in 1904. he cultured nearly mature embryos of certain crucifers and grew them to maturity. The technique was utilised by Laibach in 1925 to recover hybrid progeny from an interspecific cross in Linum . Subsequently, contributions from several workers led to the refinement of this technigue .

Steps involve in embryo culture in cultivated papaya:

Steps involve in embryo culture in cultivated papaya 1.C. papaya var. “ Wahington ” was used as the female parent and C. Caullflora as the male parent. C. papaya C. Caullflora

Steps involve in embryo culture in cultivated papaya:

Steps involve in embryo culture in cultivated papaya Flower of female and male parents were covered with butter paper bags(8cm x 12cm) before anthesis of flower.

Steps involve in embryo culture in cultivated papaya:

Steps involve in embryo culture in cultivated papaya When the female flowers had fully opened, pollen from the male parent was collected in a watch glass(between 9 and 11a.m)and dusted on the stigma of the female flower with a fine brush. This was repeated 2-3 times a day for 2-3 days.After pollination, the female flower were covered again with butter-paper bags to avoid contamination.

Steps involve in embryo culture in cultivated papaya:

Steps involve in embryo culture in cultivated papaya The fruits were collected 89 days after fertilization and the seeds removed. The seeds were surface sterlized .

Steps involved in laboratory:

Steps involved in laboratory Dip the seeds in 70% alcohol for 1 min and then sterlize with 20% sodium hyprochloride solution for 10 min.

Steps involved in laboratory:

Steps involved in laboratory Rinse 3-5x with sterile distilled water.

Steps involved in laboratory:

Steps involved in laboratory Take capsule in depression slide Give cut at placenta it will divided into 2 halves.

Steps involved in laboratory:

Steps involved in laboratory With the help of forceps exposed the ovule Give small incision on ovule give pressure with needle

Steps involved in laboratory:

Steps involved in laboratory Transfer it with micropipette in 25ml of solid medium in petridish . Place 6-8 embryo in petridish .

Steps involved in laboratory:

Steps involved in laboratory Sealed with cello tape or petridish put on petridish . The globular stage embryos were inoculated on the culture medium.

Steps involved in laboratory:

Steps involved in laboratory The culture were incubated at 25+/-2 degree Celsius in dark for 30 days. After this rapid period, the cultures were transferred to a growth chamber with 16/8hr photoperiod(200lux) at 25 degree celcius .

Steps involved in laboratory:

Steps involved in laboratory The embryos developed into seedlings of about 5 cm height within 3-4 weeks.

Steps involved in laboratory:

Steps involved in laboratory After 4 week subculturing is going on.

Application:

Application To obtain hybrids. To study morphogenesis and nutritional requirements. To overcome seed dormancy and for shortening the breeding cycle of deciduous trees. For rapid seed viability test. Propagation of rare plants. Haploid production.

Endosperm culture:

Endosperm culture Tissue culture methods are also used for culturing endosperm, which is unique, firstly, in its function of supplying nutrition to the developing embryo and secondly, inbeing triploid in its chromosome constitution. Triploid plants are useful for production of seedless fruits (e.g. apple, banana, watermelon, etc.) and for production of trisomics for cytogenetic studies.

Steps involved in endospem culture:

Steps involved in endospem culture The technique of endosperm culture involves the following steps: ( i ) the immature seeds are dissected under aseptic conditions and endosperms along with embryos, are excited, sometimes mature seeds can also be used. (ii) the excised endosperms are cultured on a suitable medium and embryos are removed after initial growth. (iii) the initial callus phase is followed by ‘shoot bud differentiation’ or ‘embryogenesis’. (iv) the shoots and roots may subsequently develop, and complete triploid plants can be established for further use.

Endosperm transplant technique:

Endosperm transplant technique

Reference:

Reference Pratical manual of principal of plant biotechnology Google image http://www.molecular-plant-biotechnology Plant embryo culture by- proff . Springer Introduction to plant biotechnology By-H.S Chawala

Thanking you……:

Thanking you……

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