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An optimized method for efficiently and rapidly constructing a library of homogeneous mutants with high saturation and high resistance in stepgrass

An optimization method and a step-grass technology, applied in the fields of botanical equipment and methods, horticultural methods, plant regeneration, etc., can solve the problem of difficult to quickly obtain genetically stable mutation-rich and directional mutation homogeneous mutants, difficult to saturate Targeted homogeneous mutant library and other issues, to achieve the effect of low cost, uniform distribution, and increased capacity

Active Publication Date: 2019-09-10
JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is difficult to quickly obtain genetically stable mutation-rich and directed mutation homoplasmic mutants in the past, and it is also difficult to efficiently and quickly construct a genetically stable saturated directed homoplasmic mutant library.

Method used

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  • An optimized method for efficiently and rapidly constructing a library of homogeneous mutants with high saturation and high resistance in stepgrass
  • An optimized method for efficiently and rapidly constructing a library of homogeneous mutants with high saturation and high resistance in stepgrass
  • An optimized method for efficiently and rapidly constructing a library of homogeneous mutants with high saturation and high resistance in stepgrass

Examples

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Embodiment 1

[0042] Example 1 illustrates the optimization method for the construction of a library of homogeneous mutants with high efficiency, rapid saturation and high resistance to C. argentina as an example. The steps are:

[0043] 1) High-frequency somatic embryo explant material preparation and early stage somatic embryo explant culture: select healthy and pest-free plants of A. figure 1 : Thin-leafed Grass; Dwarf Grass and Black Grass), peel off the old leaves, take its young stem base with 3-4 young leaves, put it into a sterile medium for cultivation after aseptic disinfection (1 / 2MS+2.0mg / L BA+0.5mg / L NAA+sucrose 20g / L+0.7% agar) 50-60d, subculture once in the middle, tissue culture into test-tube plantlets, take 0.5- 1.0cm size explants were placed in somatic embryogenesis induction medium (MS+BA(0.2mg / L)+NAA(0.5mg / L)+2.4D(0.5mg / L)+sucrose 30g / L+3.5g / L phytagel) for 50-56 days, subculture once in the middle, and transfer the resulting primary embryoid bodies to high-frequenc...

Embodiment 2

[0059] Example 2 illustrates the optimization method for the construction of a library of homogeneous mutants with high efficiency, rapid saturation and high resistance to C. nigella in C. nigella as an example. The steps are:

[0060] 1) Preparation of high-frequency somatic embryo explant material and cultivation of early somatic embryo explant: select healthy and pest-free plants of Nigella nigra, peel off old leaves, take its young stem base and bring 3-4 Young leaves, aseptically sterilized, placed in sterile medium for culture (1 / 2MS+2.0mg / L BA+0.5mg / L NAA+sucrose 20g / L+0.7% agar) for 50-60d, subcultured once in the middle, group Cultivate test-tube plantlets, get its test-tube plantlet leaves, stem base 0.5-1.0cm size explants, put on somatic embryogenesis induction medium (MS+BA(0.2mg / L)+NAA(0.5mg / L)+2.4 D (0.5mg / L)+sucrose 30g / L+3.5g / L phytagel) was cultured for 50-56d, subcultured once in the middle, and the primary embryoid bodies produced were transferred to high-...

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Abstract

The invention discloses an optimization method for efficiently and quickly building an ophiopogon japonicus saturation high resistance homogeneous mutant library. Ethylmethane sulfonate (EMS) and <60> Co-gamma ray are physical and chemical mutagenic agents which are most widely used, efficient and stable. The EMS can generate a plurality of point mutations on one genome, the generated mutations can be randomly distributed on the whole genome, the distribution is even, the density is high, the cost is low, and the obtaining of allelic mutants is facilitated, especially, a saturated mutant library can be obtained by a smaller mutant population; Gamma ray mutagenesis mainly causes the loss of DNA fragments and chromosomal rearrangement and is specially suitable for the building of a mutant library of which multiple family gene functions are lost; therefore, by using the two method simultaneously to make their respective advantages complementary to each other, the capacity of the mutant library is increased, the demands of the saturated mutant library are satisfied, the content of the saturated mutant library is enriched, and moreover, the obtained mutants do not involve a GM event, so the safety is high and the application is convenient.

Description

technical field [0001] The invention belongs to the field of plant breeding, and relates to a method for efficiently and rapidly constructing a plant mutant library, in particular to a method for efficiently and rapidly creating a mutant library through physical and chemical mutagen compound mutagenesis (r-ray and ethyl methanesulfonate (EMS)). Optimization method for a library of highly resistant homogeneous mutants saturated with cayenne grass. Background technique [0002] Ophiopogon japonicus, also known as Ophiopogon japonicus, has three species, Ophiopogon japonicus f.nanus and Ophiopogon japonicus (L.f.) Ker- Gawl.), all belong to the genus Liliaceae. Lemongrass is a unique perennial evergreen grass ground cover for foliage in my country. Because of its low plant, strong shade preference, good disease and insect resistance, and no pruning for life, it is an ideal foliage plant in many places. Ground cover plants or landscape plants in landscaping; step grass also has...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00A01H1/06
Inventor 梁慧敏周兴元刘南清郑凯谭晓燕
Owner JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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