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InDel primer for identifying radish leaf shapes and application of InDel primer

A radish leaf and radish technology, applied in the field of InDel primers for identifying radish leaf shape, can solve the problems of complex regulation mechanism, incomplete dominance or recessiveness, etc.

Active Publication Date: 2020-10-30
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The leaf shape of radish is mainly determined by genetic factors, but the regulatory mechanism is more complicated. In different materials, the mosaic leaves are dominant, incompletely dominant or recessive relative to the plate leaves.

Method used

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  • InDel primer for identifying radish leaf shapes and application of InDel primer
  • InDel primer for identifying radish leaf shapes and application of InDel primer
  • InDel primer for identifying radish leaf shapes and application of InDel primer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Constructing parental BC using Maer radish from southern China (slab leaves, no splits, early bolting) and Nienenzi from Japan (flower leaves, full splits, late bolting) 1 (89 and 87 strains) and F 2 population (183 strains).

[0044] They were sown in the farm of Beijing Vegetable Research Center in 2016, and the results of the survey of leaf rosette stage traits showed that F 1 Plants exhibited fully cleaved leaves (see figure 1 ), F 2 In the segregation population, there were 131 plants and 52 plants of floral leaf (full cleavage, AA or Aa) and plate leaf (no cleavage, aa), respectively, and the segregation ratio was 3:1 (χ 2 =1.14, P=0.286). in bc 1 -P 1 All in the group showed mosaic (AA or Aa), in BC 1 -P 2 The separation ratio of flower leaves (Aa) and plate leaves (aa) in the population is 1:1 (χ 2 =0.01, P=0.915, see Table 1).

[0045] Table 1 Generational population shape observations

[0046]

[0047] The results showed that the leaf shape trait...

Embodiment 2

[0049] IlluminaHiseq 2500 sequencing platform was used to re-sequence the whole genome of the parents "Ma Er" and "Niannianzi", and the polymorphisms of the parents were detected using the 'Aokubi DH' as the reference genome. By GATK software analysis, approximately 534,000 InDel variants were detected between the parents. According to the reference genome information, design a pair of InDel markers every 300-800kb, and consider the distribution on the scaffold as much as possible. 500 pairs of InDel markers were preliminarily designed. The principles of primer design included the annealing temperature at 58-60°C, the base difference between the parents at 3-8 bp, and the size of the amplified fragment at 100-200 bp.

[0050] 626 pairs of EST-SSR markers and 500 pairs of InDel markers were used to detect the polymorphism between the parents "Ma'er" and "Niannianzi", among which 34 pairs of EST-SSR primers and 97 pairs of InDel primers were polymorphic between the parents Sex,...

Embodiment 3

[0052] According to the results of preliminary positioning, further use of F 2 The large population (1656 strains) was fine-mapped, and the InDel32 and SSR80 mapping intervals were located at the R07:25,100,327-25,490,368 position of the "XYB36-2" reference genome. In the interval, 49 InDel markers (InDel548-InDel596) were designed at intervals of 50-100 Kb based on parental genome data, and a locally encrypted linkage map was constructed (see image 3 ). Genetic linkage analysis located the radish leaf shape gene LS in the 45kb interval between Indel593 (R07:25,182,138) and Indel594 (R07:25,227,193) (see image 3 ).

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Abstract

The invention belongs to the technical field of radish breeding, and particularly relates to an InDel primer for identifying radish leaf shapes and an application of the InDel primer. The invention provides the InDel primer for identifying the shape of a radish leaf. The InDel primer comprises an InDel593 primer and an InDel594 primer, wherein the InDel593 primer is used for identifying the shapeof the radish leaf; wherein an upstream primer of the InDel593 primer is a nucleotide sequence as shown in SEQ ID NO: 1, and a downstream primer of the InDel593 primer is a nucleotide sequence as shown in SEQ ID NO: 2; wherein an upstream primer of the InDel594 primer is a nucleotide sequence shown as SEQ ID NO: 3, and a downstream primer of the InDel594 primer is a nucleotide sequence shown as SEQ ID NO: 4. The primer provided by the invention has a linkage relationship with a gene for controlling the radish leaf shape, and can accurately identify the radish leaf shape.

Description

technical field [0001] The invention belongs to the technical field of radish breeding, in particular to an InDel primer for identifying radish leaf shape and its application. Background technique [0002] As an important agronomic trait of crops, leaf shape affects plant type, ventilation and light transmittance of the population, and light energy utilization efficiency, and is also related to crops' adaptation to adversity. For example, Cotton pawpaw and compound leaf tomato can significantly increase the photosynthetic efficiency of the population; leaves with deep leaf margins can more effectively resist high temperature damage through convection and heat dissipation, and can regulate water stress; Source relationship, above-ground and below-ground growth balance to increase crop yield and biomass. [0003] Radish (Raphanus sativus L.) is an important vegetable native to my country, with an annual cultivated area of ​​18 million mu, accounting for 6% of the vegetable ar...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/13C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 张丽王庆彪王艳萍
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES