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Application of maize secretory phospholipase A2 gene ZmsPLA2-2 to changes in plant drought resistance properties

A technology of secreted phospholipase and drought resistance, which is applied in the field of plant bioengineering breeding, and can solve problems such as the lack of understanding of the properties of low-molecular-weight secreted PLA2-encoding genes

Inactive Publication Date: 2016-03-23
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite the known physiological functions of PLA2 in plants, little is known about the nature of the low-molecular-weight secreted PLA2 (sPLA2) and the genes that encode it

Method used

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  • Application of maize secretory phospholipase A2 gene ZmsPLA2-2 to changes in plant drought resistance properties
  • Application of maize secretory phospholipase A2 gene ZmsPLA2-2 to changes in plant drought resistance properties

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Transforming the ZmsPLA2-2 gene to create a drought-resistant inbred line in maize

[0040] 1) Establishment of receptor system

[0041] The main inbred lines used in my country's agricultural production are used as materials, such as inbred seeds of Zheng 58 and Chang 7-2. First soak the seeds with 70% ethanol for 10 minutes, then soak them with 0.1% mercuric chloride for 10-15 minutes, and then wash them with sterile water for 3-5 times. After sterilization, put the seeds in a sterile Erlenmeyer flask to germinate, put a small amount (30-40ml / 250ml Erlenmeyer flask) of sterile water in the bottle, seal it and place it under dark conditions (23-30°C) for 1-2 days. sky. After germination (lubai), the seeds were placed on MS medium and continued to germinate under dark conditions. When the germ elongates to 3-4 cm, the coleoptile and 2-3 young leaves are peeled off to expose the stem tip growth cone for Agrobacterium-mediated maize genetic transformation. ...

Embodiment 2

[0063] Example 2: Transducing the ZmsPLA2-2 gene to create new drought-tolerant cotton materials

[0064] 1) Construction of ZmsPLA2-2 gene transformation vector and Agrobacterium transformation

[0065] The ZmsPLA2-2 gene coding frame was fused with the stress-inducible promoter Prd29B or the constitutive promoter PCaMV35S, and the fusion gene was inserted into the plant expression vector pCambia1300-PCaMV35S::bar by conventional gene recombination technology to generate the transformation vector pCambia1300- PCaMV35S::ZmsPLA2-2-PCaMV35S::bar and pCambia1300-Prd29B::ZmsPLA2-2-PCaMV35S::bar. Then the competent cells of Agrobacterium tumefaciens strain LBA4404 were prepared by calcium chloride method. Take 50 μl of competent cell suspension, add 1 μg of recombinant plasmid DNA at room temperature, mix well, and ice-bath for 30 minutes, place in liquid nitrogen for quick freezing for 1 minute, and after incubating at 37°C for 3 minutes, add 1ml of YEP medium and shake well, sha...

Embodiment 3

[0077] Example 3: Transducing the ZmsPLA2-2 gene to create a new drought-resistant material of Poa annua

[0078] Bluegrass (Poapratensis L.) is one of the most important turfgrass species in temperate regions. Because of its beautiful color, strong cold resistance, shade tolerance, and pruning resistance, it is widely used as a lawn grass in the northern and central regions of my country, as well as in some cool areas in the south. Bluegrass grass also has its own shortcomings, such as weak drought resistance, poor high temperature resistance, intolerance to heat, and susceptibility to pests and diseases. Its growth stops in midsummer, and even some of them die. These disadvantages have greatly restricted the widespread use of bluegrass in hot summer areas. Because Poa annua has the characteristic of parthenogenesis, it is difficult to improve these unfavorable traits by conventional breeding techniques, and the success rate is low. Therefore, the work of breeding new variet...

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Abstract

The invention discloses application of a maize secretory phospholipase A2 gene ZmsPLA2-2 to changes in plant drought resistance properties. According to the application, the maize secretory phospholipase A2 gene ZmsPLA2-2 is cloned from maize and recombined into a plan expression vector in the sense or anti-sense form; the obtained fusion genes are introduced into plants according to a transgenic technology; through measurement on the drought resistance properties of the transgenic plants, the transgenic plants of which the drought resistance properties obviously improved or reduced and progenies of such transgenic plants are screened out, so that new plant germplasms with application value are created. The cDNA sequence of the maize secretory phospholipase A2 gene ZmsPLA2-2 is show as SEQ ID No.1, and the amino acid sequence encoded by the maize secretory phospholipase A2 gene ZmsPLA2-2 is shown as SEQ ID No.2. The plants of which the drought resistance properties can be changed are cultivated herbage crops such as poa annua, herbage, maize and cotton. The application of the maize secretory phospholipase A2 gene ZmsPLA2-2 has an important meaning in improvement of the crop yield under drought stress.

Description

technical field [0001] The invention belongs to the field of plant bioengineering breeding, and specifically relates to the application of a corn secreted phospholipase A2 gene ZmsPLA2-2 in changing the drought resistance of plants. Background technique [0002] As the skeleton component of the cell membrane, phospholipids play a very important role in the process of the cell membrane sensing stimuli and transmitting information into the cell. When the receptors on the cell membrane receive external signals, the receptors activate related enzymes to convert phospholipids into signaling molecules, such as phosphoinositides (PIs), inositol triphosphates (IP3), and diacylglycerol (DAG) , phosphatidic acid (phosphatidic acid, PA), free fatty acid (freefatty acid, FFA), etc., and then trigger a series of reactions in the cell. Phospholipid signaling is involved in the regulation of photosynthesis, hormone signaling, etc., and affects plant growth and development and stress resis...

Claims

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Application Information

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IPC IPC(8): C12N15/55C12N9/18C12N15/82A01H5/00
CPCC12N9/18C12N15/8205C12N15/8273C12Y301/01004
Inventor 张举仁刘秀霞李朝霞
Owner SHANDONG UNIV
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