Abiotic stress tolerant plants and methods
A plant, drought-tolerant technology used in plant breeding and genetics to address issues such as insufficient water availability, limiting crop production, etc.
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Embodiment approach
[0138] Embodiments include isolated polynucleotides and polypeptides, recombinant DNA constructs that confer drought tolerance, compositions such as plants or seeds comprising the recombinant DNA constructs, and methods of utilizing these recombinant DNA constructs.
[0139] Isolated polynucleotides and polypeptides:
[0140] The present invention includes polynucleotides and polypeptides isolated as follows:
[0141] An isolated polynucleotide comprising (i) a nucleic acid sequence whose encoded polypeptide has an amino acid sequence of at least 50% compared to SEQ ID NO: 4, 7, 10, 13, 16, 19, 22 or 25 , 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67 %, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% or (ii) the full-length complementary sequence of the nucleic acid sequence (i), wherein the nucleic acid sequence (i) h...
Embodiment 1
[0251] Example 1. Cloning of drought-tolerant genes and construction of vectors
[0252] According to the preliminary screening results of the rice activation tag mutant library and the gene ID sequence information in Table 2, primers were designed and the rice drought tolerance genes OsICDH1, OsMtN3L, OsDN-DTP6, OsANKL1, OsMBD2, OsTP1, OsACOAT1 and OsDN-DTP7 were cloned. The primer sequences and the lengths of the amplified fragments are shown in Table 3.
[0253] The cDNAs of OsICDH1, OsACOAT1 and OsDN-DTP7 were cloned using the cDNA library of Zhonghua 11 rice leaves, stems and roots as a template; the cDNAs of OsMtN3L, OsDN-DTP6, OsMtN3L, OsDN-DTP6, gDNA of OsANKL1, OsMBD2 and OsTP1. The PCR reaction mix and PCR program are shown in Tables 4 and 5.
[0254] Table 2. Rice gene name, gene ID (TIGR) and vector ID
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[0257] Table 3. Primers for cloning rice drought tolerance genes
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[0259] Table 4. PCR reaction mixture for cloning droug...
Embodiment 2
[0266] Embodiment 2. transformation obtains transgenic rice
[0267] The overexpression vector and empty vector (DP0158) were transformed into Zhonghua 11 rice using the Agrobacterium-mediated method described by Lin Yongjun and Zhang Qifa ((2005) Plant Cell Rep. 23: 540-547). Zhonghua 11 rice is a variety cultivated by the Crop Research Institute of the Chinese Academy of Agricultural Sciences. The first batch of seeds was provided by Beijing Weiming Kaituo Agricultural Biological Company. Transgenic seedlings of the T0 generation obtained in the transformation laboratory were transplanted into paddy fields to obtain T1 seeds, and the T1 and T2 generation seeds were stored in a 4°C cold storage. The overexpression vector contains DsRED and HYG genes, and the seeds of T1 and T2 generations that emit red fluorescence under green fluorescent light are transgenic seeds, and are used for the following trait verification experiments.
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