Androgenic haploid plant development via embryogenesis with simultaneous determination of bioactive metabolites in Cambod tea (Camellia assamica ssp. lasiocalyx)

Abstract

This pioneering work reports successful androgenic plant development via embryogenesis from microspore calluses in anther cultures and estimation of bioactive metabolites in in vitro regenerants and parent plant (control) of Cambod tea, Camellia assamica ssp. lasiocalyx (Planch MS) cultivar TV19. Anthers bearing microspores at early-to-late uni-nucleate stage were selected to initiate androgenesis. A pre-treatment of 5 0 C for ve days in the dark was most effective to initiate profusely growing white callusing from microspores within 10 weeks of culture on MS medium (6% sucrose) supplemented with high cytokinins/ auxin ratio maintained by benzyl adenine (BAP) and 2,4-dichlorophenxyacetic acid (2, 4-D). Nodular structures on the callus surface differentiated into embryos. Further developement of the embryos occurred on embryogenesis medium but, with ten times reduced concentration of growth regulators and additives. Germination of embryos into complete plantlets was achieved when major salts in medium were reduced to half MS (½ MS) and augmented with BAP, GA 3 and IBA along with glutamine and serine. Cytological examination of the root-tip cells revealed that regenerated plantlets were haploids (2n=x=15), which was further con rmed through ow cytometry. The hot-water extracts from in vitro haploid calluses, embryos and eld-grown donor plant were utilized for quanti cation of (+)-catechin, (-)-epicatechin, (-)-epigallocatechin gallate, caffeine and theophylline. Our ndings revealed that the metabolite pro le of in vitro regenerated haploid cultures is comparable to that of the mother plant, thereby presenting them as potential source for genome duplication and development of genetically stable homozygous pure breeding lines. Key MessageThis is rst report on haploids in out-breeding tree, Cambod tea. Its a signi cant achievement towards generating homozygous lines, which is impossible using conventional methods. Haploids showed consistent metabolite production.

Plant Nutrition for Human Health: A Pictorial Review on Plant Bioactive Compounds for Sustainable Agriculture

“…Plant tissue culture is considered a promising tool and a perpetual source for producing plant bioactive compounds, as reported by several studies, such as Espinosa-Leal et al [228], Chandran et al [18], Arora et al [229], Fazili et al [88], and Mishra et al [230]. Many methods could be employed for producing plant bioactives on a large scale, such as using plant in vitro tissue culture through organogenesis (including both micropropagation and hairy root culture) or cell suspension culture using callogenesis [228].…”