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Erianthus fulvus calmodulin-like gene ErCML30 under low-temperature stress expression in erianthus fulvus wild species

A low temperature stress, wild species technology, applied in genetic engineering, plant genetic improvement, microbial assay/inspection, etc. problem, to achieve the effect of high sensitivity, high expression stability, and strong specificity

Inactive Publication Date: 2018-05-18
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the mechanism of action of calmodulin-like genes on plants under low temperature stress, and no calmodulin-like genes have been cloned in Saccharum.

Method used

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  • Erianthus fulvus calmodulin-like gene ErCML30 under low-temperature stress expression in erianthus fulvus wild species
  • Erianthus fulvus calmodulin-like gene ErCML30 under low-temperature stress expression in erianthus fulvus wild species
  • Erianthus fulvus calmodulin-like gene ErCML30 under low-temperature stress expression in erianthus fulvus wild species

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Acquisition of the Calmodulin Gene ErCML30 Expressed by Low Temperature Stress in the Wild Species of Canegrass

[0027] 1. Design and amplify the special primers for amplifying the full-length sequence of the calmodulin gene ErCML30 expressed in the wild species of Pseudomonas under low temperature stress.

[0028] In the early stage, the differently treated wild species of Canegrass were sent for high-throughput transcriptome sequencing, and the returned transcriptome sequencing results were analyzed and screened by bioinformatics to obtain the spliced ​​Unigene sequence of the transcriptome data of the gene. According to the spliced ​​Unigene Sequence Utilize biological software Primer5.0, Oligo 7 to design and amplify the special primer of succulent class calmodulin gene ErCML30, this special primer is made up of upstream primer GP-F and downstream primer GP-R, described upstream primer GP-F The base sequence is shown in SEQ ID NO: 2, the base sequence of th...

Embodiment 2

[0064] Embodiment 2 A method for detecting the differential expression of the calmodulin gene ErCML30 of the succulata class provided by the present invention in the wild species of the succulent grass under low temperature stress, comprising the following steps:

[0065] 1. Sample processing:

[0066] Take 6 pots of wild species of Canegrass with basically the same growth state at the seedling stage, and use the light incubator for experimental treatment. Set up the control group and the experimental group. (ER-2), 2 pots of low temperature treatment for 72h (ER-3), low temperature treatment temperature is 4 ℃.

[0067] 2. Extraction of total RNA

[0068] (1) Place the treated leaves in a mortar and add liquid nitrogen to grind quickly, add 1ml TRIzol extract per 100mg sample, and vortex for 15s. The sampling in each period is a treated sample, a total of 6 treated samples. Place each sample at 15-30°C for 5 minutes; add chloroform, the amount of chloroform is 200 μl of chl...

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Abstract

The invention discloses an erianthus fulvus calmodulin-like gene ErCML30 under low-temperature stress expression in erianthus fulvus wild species, a special primer for amplifying the erianthus fulvuscalmodulin-like gene ErCML30, a method for detecting difference expression of the gene in low-temperature stress of the erianthus fulvus wild species, and a specific primer. The expression quantity ofthe gene is constantly increased when the erianthus fulvus wild species are under low-temperature stress, and the gene directly participates in low-temperature stress response. The technical supportis provided for understanding the resistance mechanism of the gene as well as important germplasm resource development and utilization of the erianthus fulvus wild species serving as new cold-resistant sugarcane species.

Description

technical field [0001] The invention belongs to the technical field of biomolecular cloning, and in particular relates to a gene encoding a calmodulin-like (Calmodulin-like, CML) in the wild species of succulents under low temperature stress, and primers are designed based on the sequence of the gene, and detected by relative quantitative RT-PCR The method for the differential expression of the calcium-binding protein gene in the wild species of succulent grass. Background technique [0002] E.fulvus is a wild species of Erianthus, which is closely related to Saccharum. It is mainly distributed in Yunnan, Sichuan and Tibet in my country. It grows at an altitude of 500-2700m and has cold and drought resistance. , resistance to barrenness, strong perennial roots, anti-embroidery disease and other excellent characteristics, and easy to bloom, with a large amount of pollen. The Sugarcane Research Institute of Yunnan Agricultural University collected more than 150 clones of diffe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12Q1/6895C12N15/11
CPCC07K14/415C12Q1/6895C12Q2600/13
Inventor 李富生孟玉陈疏影何丽莲徐荣曹哲群王先宏刘鲁峰狄义宁肖芙荣
Owner YUNNAN AGRICULTURAL UNIVERSITY
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