Production of biomass in ultra high density plantations
The present application relates to methods for the production of plant material suitable for the extraction and purification of saponins, through plantations of clone plants of defined chemotype, including ultrahigh density plantations of Quillaja saponaria Molina, and methods to increase the recovery of saponins by solid/liquid extraction of the harvested plant material.
1. A method of making an enriched saponin compositions, comprising:
(i) selecting Quillaja sp. plants based on chromatographic analysis of aqueous extracts of plant biomass for chemotype or total R series of X series saponin content, wherein said plant biomass is selected from bark, trunk, leaves, stems, roots, seeds, flowers, fruits or a combination thereof; and wherein the selection comprises sampling Quillaja plant biomass, conducting an aqueous extraction of the plant biomass, analyzing the aqueous extracts of the sampled plant biomass by reversed phase HPLC or UHPLC chromatography, and selecting trees of a defined chemotype, X or R;
(ii) removing axillary buds from said Quillaja sp. plants and inducing shoots of said axillary buds;
(iii) growing apical meristems of the induced shoots to provide clone plantlets;
(iv) growing said clone plantlets under controlled conditions to provide young plants;
(v) transferring said young plants to an ultrahigh-density plantation;
(vi) providing irrigation and nutrients to the young plants to provide young trees;
(vii) harvesting the aerial biomass from the young trees;
(viii) drying, grinding, and sieving said harvested biomass to provide biomass suitable for extraction:
(ix) extracting said suitable biomass to provide an enriched saponin composition relative to that obtained from non-plantation biomass; and
(x) repeating steps (vi)-(ix) about every year to about every 6 years.
2. The method of claim 1 , wherein the Quillaja ,sp. is Quillaja saponaria Molina.
3. The method of any one of claims 1 - 2 , wherein the ultrahigh-density plantation comprises from about 20,000 plants per hectare to about 100,000 plants per hectare.
4. A method of increasing the saponin content of Quillaja sp. plants, comprising:
(i) selecting Quillaja sp. plants based on chromatographic analysis of aqueous extracts of plant biomass for chemotype or total R series or X series saponin content, wherein said plant biomass is selected from bark, trunk, leaves, stems, roots, seeds, flowers, fruits or a combination thereof; and wherein the selection comprises sampling Quillaja plant biomass, conducting an aqueous extraction of the plant biomass, analyzing the aqueous extracts of the sampled plant biomass by reversed phase HPLC or UHPLC chromatography, and selecting trees of a defined chemotype, X or R;
(ii) removing axillary buds from said Quillaja sp. plants and inducing shoots of said axillary buds;
(iii) growing apical meristems of the induced shoots to provide clone plantlets;
(iv) growing said clone plantlets under controlled conditions to provide young plants;
(v) transferring said young plants to an ultrahigh-density plantation;
(vi) providing irrigation and nutrients to the young plants to provide young trees; and
(vii) harvesting biomass from the young trees,
wherein the biomass from said young trees has a higher saponin content than in non-plantation Quillaja sp. plants, and wherein the higher saponin content is selected from X-series saponins or R-series saponins.