Systems and methods for cellular reprogramming of a plant cell
Plant cell fate and development is altered by treating cells with cellular reprogramming factors. Embryogenesis inducing morphogenic developmental genes are used as cellular reprogramming factors, specifically comprising polypeptides or polynucleotides encoding gene products for generating doubled haploids or haploid plants from gametes. Maize microspores treated by contacting the isolated cells with an exogenous purified, recombinant embryogenesis inducing morphogenic developmental gene polypeptide results in embryogenesis. The gametes of a maize plant develop into embryoids when transformed with a genetic construct including regulatory elements and structural genes capable of acting in a cascading fashion to alter cellular fate of plant cells. Developmental morphogenic proteins expressed from a genetic construct are used for ex situ treatment methods and for in planta cellular reprogramming.
1. A method of generating a haploid plant embryo comprising:
(a) providing a plant microspore with an embryogenesis inducing compound to promote microspore embryogenesis in the plant microspore, wherein the embryogenesis inducing compound is hemin or a kinase inhibitor selected from N-[(2R)-2,3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodoanilino)benzamide, anthra(1,9-cd)pyrazol-6(2H)-one:4-(4-Fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole, or N-benzyl-2-(pyrimidin-4-ylamino)-1,3-thiazole-4-carboxamide;
(b) isolating an embryogenic microspore from the plant microspore; and
(c) culturing the embryogenic microspore to generate the haploid plant embryo.
2. The method of claim 1 , wherein the embryogenesis inducing compound is present in a tissue culture media.
3. The method of claim 1 , further comprising culturing the haploid plant embryo.
4. The method of claim 3 , comprising contacting the haploid plant embryo with a chromosome doubling agent for a period sufficient to generate a doubled haploid plant embryo.
5. The method of claim 1 , wherein the plant microspore is obtained from maize, rice, sorghum, brassica , soybean, wheat, or cotton.