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DNA fragment related to drought tolerance and alkali resistance of plant and application thereof

A technology of DNA molecules and fragments, applied in the direction of DNA / RNA fragments, recombinant DNA technology, applications, etc., can solve the problems of difficult transgenic technology and long distances

Inactive Publication Date: 2013-11-06
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most genus plants are relatively low-level mosses and gymnosperms, and their genetic relationship is relatively far away from food crops, and their drought resistance mechanism is also relatively special, so there are great difficulties in transgenic technology

Method used

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  • DNA fragment related to drought tolerance and alkali resistance of plant and application thereof
  • DNA fragment related to drought tolerance and alkali resistance of plant and application thereof
  • DNA fragment related to drought tolerance and alkali resistance of plant and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Example 1. Utilizing DNA Fragment Bh1-4 to Cultivate Drought and Alkali Resistant Transgenic Plants

[0037] 1. Construction of plant expression vectors

[0038] Prepare the 6381bp DNA fragment shown in Sequence 1 of the Sequence Listing, digest it with the restriction endonuclease BamH I and connect it to the vector backbone of pCLD04541 digested with BamH I to obtain the recombinant plant expression vector pCLD04541-Bh1-4 , it was confirmed by sequencing that the recombinant plant expression vector was inserted into the BamH I restriction site of vector pCLD04541 with the DNA fragment shown in the 7th to 6375th nucleotide sequence of Sequence Listing Sequence 1 (named Bh1-4 fragment).

[0039] 2. Preparation of recombinant Agrobacterium tumefaciens

[0040] The recombinant plant expression vector pCLD04541-Bh1-4 prepared in step 1 was electroporated to transform Agrobacterium tumefaciens GV3101 at a voltage of 2.2kv, and it was confirmed by PCR and sequencing that A...

Embodiment 2

[0075] Embodiment 2, the discovery of DNA fragment Bh1-4

[0076] 1. Construction of Genome Library of Large Fragment Binary Bacterial Artificial Chromosome

[0077] Extract the nuclei of the leaf cells of the drought-tolerant plant Boea hygrometrica (Bunge) R.Br, purify the nuclear DNA, digest with BamH I, separate fragments of different lengths by pulse electrophoresis, and obtain a 50-80Kb large DNA by gel cutting and purification. The fragmented DNA fragments were digested with BamH I and dephosphorylated, and then purified to obtain a binary bacterial artificial chromosome (BIBAC) linear vector pCLD04541, and then transformed into Escherichia coli DH10B to construct a BIBAC library.

[0078] 2. Transformation of Arabidopsis

[0079] From the BIBAC library, pick a single clone, shake the bacteria to extract the plasmid, transform Agrobacterium tumefaciens GV3101 by electroporation, and then transform Arabidopsis thaliana. Harvest T 1 Progeny seeds, selected to T on MS m...

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Abstract

The invention discloses a DNA fragment related to drought tolerance and alkali resistance of a plant and application thereof. The DNA fragment is originated from Boea hygromettica(Bunge)R. Br and composed of a nucleotide sequence ranging from site 7 to site 6375 in a sequence 1 in a sequence table. Under the condition of osmotic stress of PEG with a concentration of 25%, the main root of a T3-generation homozygous transgenic Arabidopsis thaliana plant which transfers the DNA fragment is substantially longer than the main root of a wild Arabidopsis thaliana plant; and under the condition of alkaline stress of the pH value of 8.0, the number of lateral roots of the transgenic Arabidopsis thaliana plant is substantially more than the number of lateral roots of the wild Arabidopsis thaliana plant, and the overground part of the transgenic Arabidopsis thaliana plant is more thriving. The DNA fragment provided by the invention has critical theoretical and practical significance to breeding of novel species of drought-tolerant and alkali-resistant crops, forests and grass, is applicable to cultivation and identification of plant varieties with resistance desired in farming and animal husbandry and in ecological environment treatment and provides a specific anti-adversity resource gene which cannot be provided by model plants and crops for cultivation of plants with high stress tolerance.

Description

technical field [0001] The invention relates to a DNA segment related to plant drought and alkali resistance in the field of biotechnology and its application. Background technique [0002] "Having a harvest or not depends on water." Drought has become one of the main problems plaguing the world's food production. Drought damages plants in many ways, affecting various physiological processes of plants. In recent years, people have identified and located many genes related to drought resistance through various methods, and obtained some transgenic plants resistant to drought to a certain extent through genetic engineering methods. However, with the deterioration of the global climate, it is necessary to fundamentally To greatly improve the drought resistance of crops and avoid yield reduction or even crop failure caused by crop death, it is necessary to find more genes that can resist severe drought, determine their functions in resisting drought and dehydration, and analyze...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/63C12N5/10C12N1/21C12N7/01C12N15/11C12Q1/68C12N15/82C12N15/84A01H5/00
Inventor 邓馨赵燕
Owner INST OF BOTANY CHINESE ACAD OF SCI
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