Method for Producing Plant Seed Containing Endophytic Micro-Organisms

Abstract

The invention discloses a method for producing plant seed containing endophytic micro-organisms characterised in by the following steps: —contacting a flowering plant with a preparation of endophytic microorganisms, whereby the endophytic microorganisms enter the plant via the flowers and are conveyed to seed produced by the plant; and—obtaining the plant seed containing endophytic microorganisms from the plant.

1 - 27 . (canceled) 
     
     
         28 . A plant seed population produced by the process of i) providing an agriculturally-acceptable preparation comprising a population of isolated bacterial endophytes; ii) specifically contacting at least one flower of a flowering plant with the preparation under conditions such that the bacterial endophytes are localized to a plurality of seeds derived from the flowering plant, wherein the plurality of seeds individually contain at least about 100 bacterial endophytes per seed; and iii) collecting the plurality of derived seeds, thereby producing a plant seed population. 
     
     
         29 . The seed population of  claim 28 , wherein the isolated bacterial endophytes are heterologous to the flowering plant. 
     
     
         30 . The seed population of  claim 28 , wherein the isolated bacterial endophytes are localized to the seed interior. 
     
     
         31 . The seed population of  claim 28 , wherein the derived seeds have one or more improved traits as compared to an untreated control seed, wherein the one or more improved traits are individually selected from the group consisting of increased plant growth, increased yield, increased biomass, improved nutrient acquisition, increased resistance to environmental stresses, increased tolerance to drought, increased tolerance to heavy metals, increased tolerance to disease, and increased tolerance to herbivory, increased production of phytohormones, increased production of antibiotics, increased production of siderophores, increased production of pesticides, and increased promotion of nitrogen fixation. 
     
     
         32 . The population of  claim 28 , wherein the step of contacting the at least one flower is performed in a non-sterile environment. 
     
     
         33 . The seed population of  claim 28 , wherein the seeds are collected in a manner suitable for storage for at least about 1 month. 
     
     
         34 . A field of plants at least 1 acre in size derived from the seed population of  claim 28 . 
     
     
         35 . A method for producing a synthetic combination comprising a plant seed and endophytic microorganisms, comprising contacting at least one flower of a flowering plant with a preparation comprising a purified population of microorganisms, the population comprising an endophytic microorganism, wherein the endophytic microorganisms are present in the interior of the plant seed. 
     
     
         36 . The method of  claim 35 , further comprising obtaining the seed produced by the flowering plant. 
     
     
         37 . The method of  claim 35 , wherein contacting the flower of a plant with a preparation of endophytic microorganisms is performed by specifically spraying the population at the time of flowering. 
     
     
         38 . The method of  claim 36 , wherein the seed has an improved trait as compared to an untreated control seed, wherein the improved trait is selected from the group consisting of increased germination rate, increased germination index, increased germination energy, and more uniform germination as compared to an untreated control seed. 
     
     
         39 . The method of  claim 36 , further comprising storing the seed produced by the plant for at least 1 month. 
     
     
         40 . The method of  claim 39 , wherein the seed is stored for at least 6 months. 
     
     
         41 . The method of  claim 36 , further comprising growing the plant seed to a progeny plant, wherein the progeny plant has an improved trait as compared to an untreated control plant. 
     
     
         42 . The method of  claim 41 , wherein the improved trait is selected from the group consisting of increased plant growth, increased yield, increased biomass, improved nutrient acquisition, increased resistance to environmental stresses, increased tolerance to drought, increased tolerance to heavy metals, increased tolerance to disease, and increased tolerance to herbivory, increased production of phytohormones, increased production of antibiotics, increased production of siderophores, increased production of pesticides, and increased promotion of nitrogen fixation as compared to an untreated control plant. 
     
     
         43 . The method of  claim 35 , wherein the at least one flower of the flowering plant is contacted with a preparation of microorganisms in a suspension of at least 10 6  cfu/mL. 
     
     
         44 . A method of improving a trait in a seed or plant produced by the seed comprising:
 contacting at least one flower of a flowering plant with a preparation comprising a purified population of microorganisms, the population comprising an endophytic microorganism, and   obtaining the seed produced by the plant.   
     
     
         45 . The method of  claim 44 , wherein the improved trait is selected from the group consisting of increased germination rate, increased germination index, increased germination energy, more uniform germination increased plant growth, increased yield, increased biomass, improved nutrient acquisition, increased resistance to environmental stresses, increased tolerance to drought, increased tolerance to heavy metals, increased tolerance to disease, and increased tolerance to herbivory, increased production of phytohormones, increased production of antibiotics, increased production of siderophores, increased production of pesticides, and increased promotion of nitrogen fixation as compared to untreated control seed or plant. 
     
     
         46 . The method of  claim 44 , wherein the endophytic microorganisms are present in the interior of the plant seed. 
     
     
         47 . The synthetic combination produced by the method of  claim 35 .