Sade, N. ; Moshelion, M. ; Gebretsadik, M. ; Kaldenhoff, Ralf ; Wallach, R.
Hrsg.: Hebrew University Jerusalem (2011)
Transgenic crop plant useful for generating tissue culture comprises root cell and leaf cell transformed with DNA construct having polynucleotide encoding Nicotiana tabacum aquaporin-1.
Norm, Patent, Standard, Bibliographie

Kurzbeschreibung (Abstract)

Abstract: NOVELTY - A transgenic crop plant (T1) comprises at least one root cell and at least one leaf cell transformed with a DNA construct having a polynucleotide encoding Nicotiana tabacum aquaporin-1 (NtAQP1), where the plant has increase yield compared to a corresponding non-transgenic plant.

USE - As a transgenic crop plant (where the plant is selected from a plant producing fruit, flower and ornamental plant, grain producing plant (i.e. wheat, oats, barely, rye, rice, maize), legumes (i.e. peanuts, peas soybean lentil), plant producing forage, plant producing fiber (including cotton and flax), a tree for wood industry, plant producing tuber or root crop, sugar beet, sugar came, plant producing oil (i.e. canola, sunflower, sesame) and tomato plant useful for generating tissue culture (claimed).

ADVANTAGE - The transgenic crop plant is grown under optimal water availability conditions or abiotic stress conditions selected from water stress (drought) (i.e. water content of less than 70%), high soil salinity (i.e. above 4 dS/m), extreme temperatures, low oxygen levels or presence of heavy metals; has a yield increase of at least 60% compared to a plant grown from a seed of corresponding non-transgenic plant; and shows an enhanced tolerance to drought stress compared to unmodified plants.

DETAILED DESCRIPTION - INDEPENDENT CLAIMS are included for the following:

(1) a seed of the plant (T1), where a plant grown from the seed comprises at least one root cell and at least one leaf cell comprising a DNA construct containing polynucleotide encoding NtAQP1;

(2) a tissue culture (R1) comprising at least one transgenic cell of the plant (T1) or a protoplast derived from it;

(3) a plant regenerated from the tissue culture (R1);

(4) increasing (m1) the yield of a crop plant involving transforming a plant cell with a DNA construct comprising a polynucleotide encoding NtAPQ1, and regenerating the transformed cell into a transgenic plant comprising at least one root cell and at least one leaf cell expressing NtAQP1 having an increased yield compared to a corresponding non-transgenic plant;

(5) screening (m2) for a plant capable of producing high yield when grown under abiotic stress conditions involving obtaining several samples from several plant lines and a control sample from a reference plant, the samples comprising genetic material, measuring the expression level of a polynucleotide encoding NtAQP1 or its ortholog in the samples, comparing the expression level of the polynucleotide encoding NtAQP1 or its ortholog in the samples to the control sample, where a plant line overexpressing the polynucleotide encoding NtAQP1 or its ortholog is capable of producing high yield when grown under abiotic stress conditions.

Technology Focus/Extension Abstract: TECHNOLOGY FOCUS - BIOLOGY - Preferred Plant: The plant lines are selected from plants of the same species and plants of different species. The control plant is tobacco (Nicotiana tabacum). Preferred Method: The method (m2) further involves planting the plant line overexpressing the polynucleotide encoding NtAQP1 or its ortholog and a corresponding control plant having lower expression of the polynucleotide under abiotic stress conditions; comparing the crop yield of the plant line to the crop yield of the control plant; and selecting plant lines having increased crop yield compared to the control plant. In the method (m2), the expression level of the polynucleotide is measured using nucleic acid technology (NAT)-based assays selected from quantitative polymerase chain reaction (PCR), Quantitative real time PCR and Northern Blot. The NAT assay is PCR employing a primer pair having the nucleic acid sequence of 5'-tatccttcgcaagaccctcc-3' (SEQ ID NO: 3) and 5'-tgcctggtctgtgttgtagat-3' (SEQ ID NO: 4). Preferred Tissue Culture: The tissue culture (R1) regenerates a plant having at least one root cell and at least one leaf cell comprising a DNA construct containing polynucleotide encoding NtAQP1.

TECHNOLOGY FOCUS - BIOTECHNOLOGY - Preferred Components: The polynucleotide encodes an NtATQP1 comprising the amino acids sequence of SEQ ID NO: 1, not given in specification. The polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 2, not given in specification. The DNA construct further comprises a regulatory element selected from a promoter, an enhancer, a termination sequence and a polyadenylation signal. The promoter is a constitutive promoter. The promoter is a tissue specific promoter selected from root specific promoter and shoot specific promoter. The tissue specific promoter is selected from guard cell specific promoter (shoot); endodermis (root) and bundle sheath (shoot) scarecrow promoter; bundle sheath OSTMT1 promoter (shoot); and the green tissue Fbpase promoter (shoot). The polynucleotide encodes NtATQP1 having at least 75 (preferably greater than or equal to 85)% homology to the protein having the amino acids sequence of SEQ ID NO: 1. The polynucleotide comprises a nucleic acids sequence having at least 75 (preferably greater than or equal to 85)% homology to the nucleic acid sequence of SEQ ID NO: 2.

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