Breeding method of low-temperature-resistant cotton variety

A cotton variety and low-temperature-resistant technology, applied in botany equipment and methods, biochemical equipment and methods, and plant products, can solve problems such as plant low-temperature chilling damage, prolong the growth period, increase farmers' income, and avoid yield and fiber damage. quality reduction effect

Active Publication Date: 2019-07-05
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problem of plant low temperature chilling injury, the object of the present invention is to provide the application of the spinach choline monooxygenase gene (AhCMO) in cultivating low temperature resistant plant varieties

Method used

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  • Breeding method of low-temperature-resistant cotton variety
  • Breeding method of low-temperature-resistant cotton variety
  • Breeding method of low-temperature-resistant cotton variety

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1A

[0058] The cloning of embodiment 1AhCMO gene and the construction of expression vector

[0059] (1) Materials and methods

[0060] 1) Cotton material: Cotton line R15 is a self-fertile line of the Institute of Biotechnology, Chinese Academy of Agricultural Sciences, and is preserved by the Crop Molecular Breeding Laboratory of the Institute of Biotechnology, Chinese Academy of Agricultural Sciences.

[0061] 2) Strains: Escherichia coli Transl-T1, Agrobacterium GV3101.

[0062] 3) Vectors: pEASY-Blunt, pEASY-T5, pUC19, pBI121.

[0063] 4) Tool enzymes and modification enzymes: various restriction enzymes and modification enzymes were purchased from TaKaRa Company, NEB Company and TIANGEN Company.

[0064] 5) Chemical reagents: All chemicals are analytically pure at home and abroad.

[0065] 6) Primer synthesis: Synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd.

[0066] 7) Sequencing: completed by BGI.

[0067] (2) Test steps

[0068] The AhCMO ge...

Embodiment 2

[0072] Example 2 AhCMO Gene Transformation and Identification and Screening of Transgenic Materials

[0073] 1) AhCMO expression vector transformed into Agrobacterium GV3101

[0074] Agrobacterium cultivation: Take 100uL of the prepared Agrobacterium GV3101 competent cells, add 10ug pBI121-AhCMO plasmid DNA, mix well, and place on ice for 5min to obtain a competent mixture. Add the competent mixed solution to a 2mm electric shock cup (try to avoid generating air bubbles when adding), and electric shock at 2500V; quickly add 800uL of YEB liquid medium after electric shock, and induce culture at 28°C and 180rpm for 5 hours; take out the induced bacterial liquid , coated on YEB plates containing 5Oug / mL each of kanamycin and rifampicin, and cultured at 28°C for 48 hours. Pick a single clone and inoculate it in 5mL YEB liquid medium containing kanamycin and rifampicin (both at a concentration of 50ug / mL), shake the bacteria at 180rpm for 36 hours, and extract a small amount of pl...

Embodiment 3

[0092] Embodiment 3AhCMO transgenic plant low temperature tolerance identification test

[0093] 1) The transgenic cotton CMO24 and wild-type cotton R15 were cultured continuously for 3 weeks under normal growth conditions (28°C), and the results (see Pic 4-1 and Figure 4-2 ) leaves of both transgenic and wild-type cotton plants appeared normal. Subsequently, the transgenic and wild-type cotton plants were transferred to 12°C for 24 hours, and it was found that the phenotype of transgenic cotton CMO24 leaves was still normal (see Figure 4-4 ), the leaves of non-transgenic plants were severely wilted or even died (see Figure 4-3 ). It shows that the AhCMO transgenic cotton CMO24 is not sensitive to low temperature, that is, it has strong low temperature tolerance.

[0094] 2) Placenta blue staining: Select 3 wild-type R15 and transgenic CMO24 cotton leaves of similar plant size before and after low-temperature treatment, put them into a 50ml centrifuge tube and write th...

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Abstract

The invention discloses application of an atriplex hortensis choline monooxygenase (AhCMO for short) gene in the culture of low-temperature-resistant plant varieties and application of expression vectors and plant cells containing the AhCMO gene in the culture of the low-temperature-resistant plant varieties. The invention further discloses a breeding method of a low-temperature-resistant cotton variety. The breeding method is characterized in that the AhCMO gene is transformed into cotton varieties, and the cotton variety with high AhCMO gene expression quantity is selected as the low-temperature-resistant cotton variety. The AhCMO gene is applied to low-temperature-resistant plant breeding, and a novel field is exploited for the application of the gene; the transgenic cotton modified bythe AhCMO gene overcomes the low-temperature sensitivity of cotton, cotton yield lowering and fiber quality lowering are avoided, cotton planting area can be expanded, and farmer income can be increased.

Description

technical field [0001] The invention belongs to the field of cotton breeding methods, and specifically relates to the use of spinach choline monooxygenase gene in cultivating low-temperature-resistant cotton varieties; and the use of expression vectors containing the gene or plant cells in cultivating low-temperature-resistant cotton varieties; The invention relates to a breeding method of a low temperature resistant cotton variety. Background technique [0002] Cotton (Gossypium hirsutum L.) originated in tropical and subtropical regions, so cotton is very sensitive to cold stress. Cold stress significantly inhibits cotton seed germination and seedling growth, and can also cause fertilization failure, cotton boll shedding and even plant death, which will result in a decline in cotton yield and fiber quality. Cotton planting in my country is mainly distributed in the northwest inland such as Xinjiang, as well as in cool areas such as high latitude and high altitude. Cold st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N9/02C12N15/82C12Q1/6895A01H1/02A01H5/00A01H6/60A01H6/46A01H6/54
CPCA01H1/02C12N9/0077C12N15/8273C12Q1/6895C12Q2600/13C12Y114/15007
Inventor 郭三堆张锐梁成真王雅楠孟志刚
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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