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Morphogenesis and Totipotency
A chlamydomonas mother cell gives rise to two daughter cells by a simple division. But a leaf cell cannot give rise to a new plant except in cases like Bryophyllum or Kalanchoe. What makes a daughter cell of a zygote to loose the 'potential' to develop into a whole plant? Or if all the daughter cells possess this potential- i.e. if they are totipotent-why it does not express? These questions troubled Haberlandt-hundred years ago. He postulated that any living plant cell should be able to grow into a full plant. He experimented with mesophyll cells at a time when tissue culture had not been unheard for. His experiments failed because we now know that isolated mesophyll cells are not really easy to grow to unleash their morphogenetic potential.
F.C. Steward-who took up this work sixty years later-successfully cultured whole carrot plants from carrot phloem parenchyma cell. He published the papers in the American Journal of Botany in the late 1950's. Steward and his coworkers took 2 mg. tissues of secondary phloem of carrot roots and grew them in special flasks with nipples or tubes called tumble tubes. The medium was whites' medium with coconut water. These tubes/flasks were mounted on a wheel that was rotated on a shaft at the rate of 1 r.p.m. So that the pieces were alternately aerated and bathed in the liquid medium. There was an enormous increase in size of the explants. A callus ensued. A few peripheral cells slaughtered off into the medium and started dividing and gave rise to clumps,-occasionally with roots these could be transferred to semi-solid medium in (still) tubes. Shoots arose opposite the roots to yield full plants. Subsequently other parts were also used to, demonstrate cellular totipotency.
Grasping instruments -Plier or clamp forceps -put piece of rubber in the plier -applied on the buccal and lingual surface -make pressure to
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