Methods and compositions for improving plant traits
Disclosed herein are methods of increasing nitrogen fixation in a non-leguminous plant. The methods can comprise exposing the plant to a plurality of bacteria. Each member of the plurality comprises one or more genetic variations introduced into one or more genes or non-coding polynucleotides of the bacteria's nitrogen fixation or assimilation genetic regulatory network, such that the bacteria are capable of fixing atmospheric nitrogen in the presence of exogenous nitrogen. The bacteria are not intergeneric microorganisms. Additionally, the bacteria, in planta, produce 1% or more of the fixed nitrogen in the plant.
1. A method of increasing an amount of atmospheric derived nitrogen in a corn plant in a field, the method comprising:
exposing a corn plant in a field to a plurality of engineered diazotrophs comprising at least one genetic variation introduced in a nitrogen fixation genetic regulatory network that increases expression or activity of nifH, wherein the at least one introduced genetic variation comprises genetic material that originates from at least one organism of the same genus as said plurality of engineered diazotrophs,
whereby the corn plant exposed to the engineered diazotrophs comprises an increased amount of atmospheric derived nitrogen relative to a corn plant exposed to the same amount of non-engineered diazotrophs of the same species as the engineered diazotrophs.
2. The method of claim 1 , wherein the at least one introduced genetic variation comprises an insertion of genetic material from a diazotroph of the same species as the engineered diazotrophs.
3. The method of claim 1 , wherein said genetic material that originates from diazotrophs of the same genus as said plurality of engineered diazotrophs comprises a regulatory element.
4. The method of claim 3 , wherein said regulatory element is a heterologous regulatory element.
5. The method of claim 3 , wherein said regulatory element is a promoter.
6. The method of claim 5 , wherein said promoter is a promoter that is active in the presence of environmental nitrogen.
7. The method of claim 1 , wherein said plurality of engineered diazotrophs further comprise a modification within a nitrogen assimilation genetic regulatory pathway.
8. The method of claim 7 , wherein said plurality of engineered diazotrophs comprise a modification which alters the activity or expression of a gene selected from the group consisting of: GlnE, AmtB, ntrB, ntrC, polynucleotide encoding glutamine synthetase, glnA, glnB, glnK, draT, polynucleotide encoding glutaminase, and glnD.
9. The method of claim 1 , wherein said at least one genetic variation introduced in a nitrogen fixation genetic regulatory network that increases expression or activity of nifH increases the expression or activity of nifH under non-nitrogen limiting conditions.
10. The method of claim 1 , wherein said plurality of engineered diazotrophs have increased expression or activity of nifH when in the field compared to a plurality of non-engineered diazotrophs of the same species in the same field.
11. The method of claim 1 , wherein said field comprises at least 0.01 mM of fixed nitrogen.
12. The method of claim 1 , wherein said field comprises at least 0.1 mM of fixed nitrogen.
13. The method of claim 1 , wherein said field comprises at least 0.5 mM of fixed nitrogen.
14. The method of claim 1 , wherein the plurality of engineered diazotrophs comprises epiphytes.
15. The method of claim 1 , wherein the plurality of engineered diazotrophs comprises endophytes.
16. The method of claim 1 , wherein the plurality of engineered diazotrophs comprises rhizophytes.
17. The method of claim 1 , wherein the genetic material of the plurality of engineered diazotrophs consists essentially of genetic material that originates from diazotrophs of the same species as said plurality of engineered diazotrophs.