SSR (simple sequence repeats) markers, linked with LRGPP (loss rate of total grains per panicle) related Aphelenchoides besseyi Christie resistant QTL (quantitative trait locus), on chromosome 5 and application thereof
A technology for resistance to D. elegans in rice, which is applied to the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of limiting the effective utilization of resistance resources of D. elegans, and improve the breeding efficiency. , the effect of speeding up the breeding process and simplifying the selection method
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Embodiment 1
[0019] Example 1, Acquisition of Molecular Markers Linked to the Resistance QTL of D.
[0020] 1. Plant material
[0021] In 2008, Huaidao 5 and Tetep were planted in the field of Jiangsu Academy of Agricultural Sciences and crossed to obtain the hybrid F 1 , next year F 1 F 2 Seeds, propagated F in Hainan in 2009 2 group, as a mapping group. f 2 Individual plant number, parent and hybrid F at tillering stage 1 and F 2 Some leaves of each individual plant in the population were stored in a -70°C refrigerator for SSR analysis, F 2 Harvest the individual plants of the population for phenotypic identification.
[0022] 2. Cultivation and isolation of nematodes
[0023] Inoculate Botrytis cinerea (Botrytis cinerea) bacterium block on Potato Dextrose Agar (PDA) medium and cultivate at 25 ℃, after Botrytis cinerea is overgrown with medium, use 3% hydrogen peroxide to disinfect the surface of rice dry point nematode for 10min, After washing with sterilized ultrapure water f...
Embodiment 2
[0060] Example 2, the application of molecular markers on chromosome 5 linked to the resistance QTL of rice stem nematode resistance to rice 5 and Tetep in the offspring of the cross combination with Huaidao 5 and Tetep as parents
[0061] The SSR marker linked to the LRGPP-related rice stem nematode resistance QTL obtained on chromosome 5, the F 2∶3 Part of the individual plants of the family were predicted for the resistance of the rice stem nematode, and the DNA of each individual plant was extracted, and then PCR amplification analysis was performed with the primers of the SSR markers RM18632 and RM163, and the band type analysis was used to determine whether there were corresponding markers. The mark indicates that the line has reached the level of resistance to the rice stem nematode, and if it does not exist, it is susceptible. Subsequently, the actual resistance of the tested lines to rice A. spp. was determined by using the artificial inoculation identification method...
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