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Simple and efficient potato multi-R gene polymerization assisted breeding method
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A technology of assisted breeding and gene aggregation, applied in the field of plant genetic engineering, can solve problems that have not been reported, and achieve the effect of improving accuracy
Inactive Publication Date: 2020-10-16
SOUTHWEST UNIVERSITY
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Problems solved by technology
[0004] Problems in the prior art: There is no report in the prior art that can target the persistent resistance genes RB and R8 of late blight and the PVX resistance gene Rx1 and PVY resistance gene Ry in the source S.tuberosum ssp.andigena adg multiplex PCR report
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Embodiment 1
[0039] This example provides the late blight persistent resistance genes RB and R8, the PVX resistance gene Rx1 and the PVY resistance gene Ry adg sequence, including:
[0040] (1) The full-length CDS fragment sequence of RB is shown in SEQ ID NO: 1 in the sequence listing.
[0041] (2) The sequence of the full-length CDS fragment of R8 is shown in SEQ ID NO: 2 in the sequence listing.
Embodiment 2
[0043] This embodiment provides a late blight resistance screening primer sequence set, including:
[0044] (1) RB marker primers are shown in SEQ ID NO: 3 and SEQ ID NO: 4 in the sequence listing.
[0045] (2) R8 labeled primers are shown in SEQ ID NO: 5 and SEQ ID NO: 6 in the sequence listing.
[0046] (3) Rx1 marker primers are shown in SEQ ID NO: 7 and SEQ ID NO: 8 in the sequence listing.
[0047] (4)Ry adg The labeled primers are shown in SEQ ID NO: 9 and SEQ ID NO: 10 in the sequence listing.
[0048] (5) GBSS marker primers are shown in SEQ ID NO: 11 and SEQ ID NO: 12 in the sequence listing.
Embodiment 3
[0050] This example provides a potato multi-R gene aggregation assisted breeding method, using R8, RB, Rx1 and Ry adg The multiplex PCR detection is realized, as follows:
[0052] The DNA of 218 potato resource materials collected earlier was used for single-marker detection of Rx1 and Ryadg (Liu Xun et al., 2019). The high-generation lines of potato breeding were planted in Chongqing Beibei and Chongqing Wuxi Breeding Base of Chongqing Key Laboratory of Potato Biology and Genetic Breeding, Southwest University, respectively. After 4 weeks of emergence, 1-2 young leaves were taken for DNA extraction.
[0053] (2) Potato resource materials Rx1 and Ry adg marker detection
[0054] In the early stage, 218 potato resources were tested for late blight race-specific R1, R2 family and R3a / b, as well as the composition of persistent resistance gene RB and R8 markers (Liu Xun et al., 2019). To these resources Rx1 and Ry adg Resist...
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Abstract
The invention provides a simple and efficient potato multi-R genepolymerization assisted breeding method. The method specifically comprises the following steps of obtaining late blight resistance screening primer sequence groups RB, R8, Rx1, Ryadg and GBSS, constructing a multiple PCR reaction system by using the primer group, and carrying out pyramiding breeding selection on late blight lastingresistance and PVX and PVY virusdisease resistance genes. The method is high in detection specificity, has good tolerance to the quality and concentration of a DNA template, and is convenient in adaptation to screening and identification of molecular markers of large-scale breeding intermediate materials. The method provides a simple and efficient technology for late blight and virusdisease polyclonal antibodygene pyramiding breeding and provides a technical scheme for breeding of new polyclonal antibody varieties.
Description
technical field [0001] The invention relates to the field of plantgenetic engineering, in particular to a simple and efficient multi-R gene aggregation-assisted breeding method for potatoes. Background technique [0002] China is the world's largest potato producer, and its total output and per capita consumption have been growing steadily. The sustainable development of the potato industry is of great significance to ensuring food security and promoting regional economic development. However, potato production has always been threatened by diseases and insect pests, which seriously affect the safe production of potatoes. Especially the seed potato degeneration caused by the outbreak of late blight and virus accumulation is the most serious, and it is also the main bottleneck restricting the development of the potato industry. At present, chemical agents are mainly used to prevent and control late blight, but their use is limited due to the cost of chemical agents, drug re...
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