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Construction method of wild rice chromsome segment substitution line

The technology of chromosome fragment and replacement line is applied in the field of construction of chromosome fragment replacement line of wild rice, which can solve the problems of too large imported fragment, low efficiency of gene cloning, insufficient research and analysis, etc., and achieve the effect of improving efficiency.

Active Publication Date: 2016-12-07
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Claims
  • Application Information

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Problems solved by technology

However, the disadvantage of the introgression line is that the imported fragment is too large, the genome coverage of the population chromosome is not complete, and the fine mapping of the next step QTL is not efficient in gene cloning
Furuta et al. used 149 SNP molecular markers to construct 33 wild rice chromosome partial replacement lines, but this small population is not enough for a comprehensive study and analysis of wild rice genomics

Method used

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  • Construction method of wild rice chromsome segment substitution line
  • Construction method of wild rice chromsome segment substitution line
  • Construction method of wild rice chromsome segment substitution line

Examples

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Comparison scheme
Effect test

Embodiment 1

[0061] Example 1, Identification and Screening of Polymorphic SSR Molecular Markers

[0062] 1. Select 780 pairs of SSR primers and 175 pairs of InDel primers distributed on 12 chromosomes to screen the primers used to construct the replacement line. The genetic distance of all the primers (unit: cM centimolar root) comes from the website http: / / www .gramene.org.

[0063] 2. 369 pairs of SSR primers were detected in the reference "Eshed Y, Zamir D: A genomic library of Lycopersiconpennellii in L.esculentum: a tool for fine mapping of genes. Euphytica, 1994, 79(3): 175-179." There was polymorphism between the parents (wild rice and cultivated rice 9311), and finally 119 pairs of SSR primers and 62 pairs of InDel primers evenly distributed on 12 chromosomes were selected to construct the replacement line. The information and sequences of the 181 pairs of primers are shown in Table 1, and the average genetic distance between each pair of primers is 5.7cM.

[0064] Table 1. Info...

Embodiment 2

[0070] Embodiment 2, the construction method of wild rice chromosome segment replacement line

[0071] The method for constructing the wild rice chromosome segment replacement line of the present invention is to use continuous backcrossing combined with MAS to detect molecular markers from the BC3 generation, perform selective backcrossing according to the distribution of exogenous DNA fragments, and continuously track and detect up to the high-generation backcrossed offspring , to establish a chromosome segment replacement line of the whole genome series of wild rice. The construction flow diagram of the wild rice chromosome segment replacement line of the present invention is as follows figure 1 As shown, the construction method of the wild rice chromosome segment replacement line of the present invention is specifically as follows:

[0072] 1. Using the cultivated rice 9311 as the female parent and the common wild rice as the male parent, carry out hybridization to obtain ...

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Abstract

The invention discloses a construction method of a wild rice chromsome segment substitution line (CSSL). According to the construction method of the wild rice chromsome segment substitution line, a substitution line population capable of covering an entire chromosome genome is constructed, genetic backgrounds of substitution lines are very clear, for one substitution line, the genetic background of the substitution line and a receptor parent are approximately the same, and a difference with a donor parent exists only in one known marker segment; for a complete set of substitution line, all wild rice chromsome segments have corresponding marking segments, and duplications and deletions are avoided. The constructed wild rice chromsome segment substitution line provides a basis for building a platform for comprehensively conducting wild rice genomics research and long-term beneficial help is provided for rice genomics research and innovation germplasm resources.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for constructing a chromosome segment replacement line of wild rice. Background technique [0002] Common wild rice (Oryza.rufipogon Griff.) is the ancestor species of Asian cultivated rice (O.sativa L.), and it is an important germplasm resource for rice variety improvement. It has been found that wild rice has valuable genes such as resistance to diseases and insect pests, stress resistance, high yield, high quality, efficient phosphorus utilization, sterility and recovery. Therefore, excavating from wild rice the excellent genes that have been lost or weakened in cultivated rice, and finding and utilizing new favorable gene resources has become a new breakthrough in rice variety improvement. [0003] Many important agronomic traits such as crop yield, quality, and resistance are mostly complex quantitative genetic traits. Due to its complex genetic backgroun...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68A01H1/02A01H1/04
CPCA01H1/02A01H1/04C12Q1/6895C12Q2600/13
Inventor 乔卫华杨庆文郑晓明齐兰张丽芳程云连
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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