Method for improving plant drought hardiness

A drought tolerance and plant technology, applied in the directions of botanical equipment and methods, plant products, angiosperms/flowering plants, etc., can solve problems such as few candidate genes, and achieve improved drought resistance, improved drought tolerance, and improved drought resistance. performance effect

Inactive Publication Date: 2012-04-11
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few alternative genes for improving plant drought resistance, and they all focus on genes that can increase drought resistance by increasing expression. This area needs a new method for improving plant drought resistance

Method used

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  • Method for improving plant drought hardiness
  • Method for improving plant drought hardiness
  • Method for improving plant drought hardiness

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1. Cloning and vector construction of full-length UGT71C5 gene

[0026] 1. Primer design

[0027] The full-length UGT71C5 gene was queried using the NCBI database, primers were designed, and a BmaH I restriction site was added to the upstream primer, and a Sma I restriction site was added to the downstream primer.

[0028] Upstream primer (SEQ ID NO.3):

[0029] 5'-CGCGGATCCTAGATGAAGACAGCAGAGCTCATATTCG-3';

[0030] Downstream primer (SEQ ID NO.4):

[0031] 5'-TCCCCCGGGATTATCTTGCTCTGAAAGTGAAATGGTC-3'.

[0032] 2. Amplify the nucleotide sequence shown in SEQ ID NO.1 from Arabidopsis cDNA

[0033] PCR reaction system (50μl):

[0034] Template (cDNA) 5.0μl

[0035] 25mM MgCl2 4.0μl

[0036] 10×Taq Buffer 5.0μl

[0037] 2.5mM dNTPs 5.0μl

[0038] 20mM upstream primer (SEQ ID NO.3) 5.0μl

[0039] 20mM downstream primer (SEQ ID NO.4) 5.0μl

[0040] wxya 2 O 26.5 μl

[0041] Taq enzyme (5U / μl) 0.50μl

[0042] PCR reaction parameters: 95.0°C pre-denaturatio...

Embodiment 2

[0078] Example 2: Transferring the UGT71C5 gene into Arabidopsis thaliana

[0079] 1. Preparation of Agrobacterium competent cells (calcium chloride method)

[0080] (1) Pick a single colony of Agrobacterium EHA105 and inoculate it into LB medium containing 25 μg / ml streptomycin (Str), and culture and shake overnight at 28°C;

[0081] (2) Take 2ml of the overnight culture and add it to 50ml of LB+Str liquid medium, continue to shake and culture until OD600 reaches 0.5;

[0082] (3) Place the culture solution on ice for 10 minutes, and cool the culture to 0°C;

[0083] (4) 4°C, 8000rpm, centrifuge for 10min to collect bacteria;

[0084] (5) Suspend Agrobacterium with 1ml of ice-cold 10mM CaCl2 solution. Dispense 100 μl into 1.5ml EP tubes, freeze in liquid nitrogen and store at -80°C.

[0085] 2. Transformation of Agrobacterium

[0086] (1) Take out the competent Agrobacterium and thaw on ice;

[0087] (2) Add 2 μl of recombinant plasmid, freeze in liquid nitrogen for 5 m...

Embodiment 3

[0102] Embodiment three, the drought tolerance test of transgenic plant

[0103]Three overexpression lines (OE1, OE2, OE3) that were positively transformed into UGT71C5, and three (DN1, DN2, DN3) suppressed expression lines (reversely transformed into UGT71C5) were selected. Select the plants with relatively consistent growth conditions in the seedling trays with the same culture conditions for about 1 month among the above-mentioned strains, cut the seedling trays and divide each strain into two identical parts, one part is treated without watering, and the other part is used as a control. Still watering normally, another wild-type plant was selected as a control. Except for the factor of water, other culture conditions are strictly guaranteed to be consistent. Observe the growth of seedlings on time and make records.

[0104] Table 1 Drought tolerance test results of transferred UGT71C5 plants

[0105]

[0106] The experimental results showed that compared with normal ...

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Abstract

The invention belongs to the field of plant resistance improvement and particularly relates to a method for improving the plant drought hardiness, which aims at solving the technical problem to provide a novel effective method capable of improving the plant drought hardiness. In order to solve the technical problem, the method has the technical scheme that the method for improving the plant drought hardiness is provided, and the drought hardiness is improved through inhibiting the expression of uridine diphosphoglucose glycosyl transferase (UGT) genes in plants. The method provides a novel thought for the field of plant hardiness improvement, belongs to the novel efficient method capable of improving the plant hardiness and has good application prospects.

Description

technical field [0001] The invention belongs to the field of improving plant resistance, and in particular relates to a method for improving plant drought tolerance. Background technique [0002] Most proteins in organisms exist in the form of glycoproteins, which include enzymes, immunoglobulins, carrier proteins, hormones, toxins, lectins and structural proteins, and their functions involve cell recognition, information transmission, hormone regulation, fertilization, occurrence, development, Various aspects of differentiation, nervous system and immune system homeostasis maintenance. Moreover, the infection of bacteria and viruses, the proliferation and metastasis of cancer cells, and autoimmune diseases are all closely related to the sugar on the cell surface. [0003] Glycosyltransferases (Glycosyltransferases, GTs) are enzymes that catalyze the combination of sugar groups and specific receptor molecules to form glycosidic bonds. GTs widely exist in organisms and play...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63A01H5/00
Inventor 杨毅
Owner SICHUAN UNIV
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