Drought tolerant transgenic plants and method of making same
Embodiments of the present invention relate generally to drought tolerant transgenic plants and methods of creating the drought tolerant transgenic plants. In one embodiment, plants are transformed such that a PYL polypeptide is overexpressed in the plant. In an embodiment, the plant is transformed with a cassette or vector that comprises a polynucleotide encoding for one or more of the PYL polypeptides, which may be under the control of an inducible or constitutive promoter. In embodiments, overexpression of PYL13 results in plants having increased drought tolerance, such as a decreased transpiration rate, a decreased stomatal conductance, an increased photosynthetic rate, accelerated stress responsive gene expression, and increased water use efficiency, and/or an increased survival rate compared to a control plant. In some embodiments, other PYL proteins having the single point mutations identified in PYL13 are expressed in the plant.
1. A transgenic plant having increased drought tolerance compared to a control plant, wherein the transgenic plant is transformed with a recombinant DNA construct comprising a polynucleotide sequence that is expressed to obtain a drought tolerant plant, wherein the polynucleotide sequence has a sequence similarity of E value <1e-90 to PYL13, wherein PYL13 has an amino acid sequence consisting of SEQ ID NO:1 and comprising CL1, CL2 and CL4 conserved regions, wherein the polynucleotide sequence expresses a mutant polypeptide having an amino acid sequence selected from the group consisting of:
SEQ ID NO:149 having at least one homologous point mutation selected from the group consisting of K59Q, L87F and N151T,
SEQ ID NO:150 having at least one homologous point mutation selected from the group consisting of K86Q, L114F and N181T,
SEQ ID NO:151 having at least one homologous point mutation selected from the group consisting of K64Q, L91F and N157T,
SEQ ID NO:152 having at least one homologous point mutation selected from the group consisting of K70Q, L111F and N180T,
SEQ ID NO:153 having at least one homologous point mutation selected from the group consisting of K81Q, L109F and N170T,
SEQ ID NO:154 having at least one homologous point mutation selected from the group consisting of K87Q, L115F and N176T,
SEQ ID NO:155 having at least one homologous point mutation selected from the group consisting of K90Q, L118F and N184T,
SEQ ID NO:156 having at least one homologous point mutation selected from the group consisting of K65Q, L91F and N155T,
SEQ ID NO:157 having at least one homologous point mutation selected from the group consisting of K61Q, L87F and N151T,
SEQ ID NO:158 having at least one homologous point mutation selected from the group consisting of K63Q, L89F and N153T,
SEQ ID NO:159 having at least one homologous point mutation selected from the group consisting of K56Q, L83F and N147T,
SEQ ID NO:160 having at least one homologous point mutation selected from the group consisting of K39Q, L66F and N129T, and
SEQ ID NO:161 having at least one homologous point mutation selected from the group consisting of K39Q, L66F and N128T.
2. The transgenic plant of claim 1 , wherein the mutant polypeptide is overexpressed as compared to a control plant.
3. The transgenic plant of claim 1 , wherein the polynucleotide is operably linked to a promoter selected from the group consisting of a constitutive promoter, an inducible promoter, a tissue specific promoter, and a stress-responsive promoter.
4. The transgenic plant of claim 1 , wherein the transgenic plant exhibits an increased survival rate after drought as compared to a control plant.
5. The transgenic plant of claim 1 , wherein the transgenic plant comprises at least one characteristic selected from the group consisting of a decreased relative transpiration rate, a decreased relative stomatal conductance, an increased relative photosynthetic rate, and an increased relative water use efficiency as compared to a control plant.
6. The transgenic plant of claim 1 , wherein the transgenic plant exhibits accelerated stress responsive gene expression of a stress gene selected from the group consisting of RD29A, KIN1, COR15A, and RAB18 as compared to a control plant.
7. A transgenic seed obtained from the transgenic plant of claim 1 , and wherein the transgenic seed comprises said recombinant DNA construct.
8. A method of increasing drought tolerance of a plant as compared to a control plant, the method comprising:
(a) providing a recombinant DNA construct comprising a polynucleotide sequence that is expressed to obtain a drought tolerant plant, wherein the polynucleotide sequence has a sequence similarity of E value <1e-90 to PYL13, wherein PYL13 has an amino acid sequence consisting of SEQ ID NO:1 and comprising CL1, CL2 and CL4 conserved regions, wherein the polynucleotide sequence expresses a mutant polypeptide having an amino acid sequence selected from the group consisting of:
SEQ ID NO:149 having at least one homologous point mutation selected from the group consisting of K59Q, L87F and N151T,
SEQ ID NO:150 having at least one homologous point mutation selected from the group consisting of K86Q, L114F and N181T,
SEQ ID NO:151 having at least one homologous point mutation selected from the group consisting of K64Q, L91F and N157T,
SEQ ID NO:152 having at least one homologous point mutation selected from the group consisting of K70Q, L111F and N180T,
SEQ ID NO:153 having at least one homologous point mutation selected from the group consisting of K81Q, L109F and N170T,
SEQ ID NO:154 having at least one homologous point mutation selected from the group consisting of K87Q, L115F and N176T,
SEQ ID NO:155 having at least one homologous point mutation selected from the group consisting of K90Q, L118F and N184T,
SEQ ID NO:156 having at least one homologous point mutation selected from the group consisting of K65Q, L91F and N155T,
SEQ ID NO:157 having at least one homologous point mutation selected from the group consisting of K61Q, L87F and N151T,
SEQ ID NO:158 having at least one homologous point mutation selected from the group consisting of K63Q, L89F and N153T,
SEQ ID NO:159 having at least one homologous point mutation selected from the group consisting of K56Q, L83F and N147T,
SEQ ID NO:160 having at least one homologous point mutation selected from the group consisting of K39Q, L66F and N129T, and
SEQ ID NO:161 having at least one homologous point mutation selected from the group consisting of K39Q, L66F and N128T; and
(b) introducing the recombinant DNA construct into a plant to produce a transgenic plant, wherein the transgenic plant exhibits increased drought tolerance.
9. The method of claim 8 , wherein the mutant polypeptide is overexpressed as compared to a control plant.
10. The method of claim 8 , wherein the polynucleotide sequence is operably linked to a promoter is selected from the group consisting of a constitutive promoter, an inducible promoter, a tissue specific promoter, and a stress-responsive promoter.
11. The method of claim 8 , wherein the transgenic plant comprises at least one characteristic selected from the group consisting of a decreased relative transpiration rate, a decreased relative stomatal conductance, an increased relative photosynthetic rate, and an increased relative water use efficiency as compared to a control plant.
12. The method of claim 8 , wherein the transgenic plant exhibits accelerated stress responsive gene expression of a stress gene selected from the group consisting of RD29A, KIN1, COR15A, and RAB18 as compared to a control plant.
13. The method of claim 8 , further comprising crossing the transgenic plant with itself or crossing the transgenic plant with another plant to produce a transgenic seed.