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Method for quickly identifying genome size of sweet potatoes and application thereof

A genome and sweet potato technology, applied in the field of plant ploidy detection, can solve the problems of sweet potato genome size controversy, poor detection results, and failure to meet the accuracy requirements of sweet potato genome

Active Publication Date: 2019-04-09
XUZHOU INST OF AGRI SCI IN JIANGSU XUHUAI DISTRICT (JIANGSU XUZHOU SWEETPOTATO CENT)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2017, the genome of sweet potato (Ipomoea batatas (L.)) was predicted to be 4.4Gb according to the haplotype sequencing results, but the size of the sweet potato genome is still controversial in the academic circle
However, sweet potatoes are rich in polyphenols, which are not only effective health-care ingredients, but also cause the young sweet potato tissues to quickly brown, deteriorate and die once they are separated from the mother, resulting in poor detection results. Existing detection methods cannot meet the requirements of sweet potato. Accuracy Requirements for Genomic Testing

Method used

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  • Method for quickly identifying genome size of sweet potatoes and application thereof
  • Method for quickly identifying genome size of sweet potatoes and application thereof
  • Method for quickly identifying genome size of sweet potatoes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] The test sample is: mixed sample of Xu Zishu No. 8 and Trilobed Petunia-1

[0050] Sampling in the field: Take samples from the field and keep only 10 parts of the 10cm stem tip of the unexpanded leaves. Try to take the samples in the morning or evening. The cuttings are placed in an aeroponic box (100cm×60cm) prepared in advance with 30L of Hoagland’s culture medium ×40cm). Hoagland's culture solution is prepared with deionized water. When cutting, pay attention to rinse the sample with deionized water to remove browning wounds and tissues. Avoid mechanical damage to tender tissues when sampling. When sampling, ensure that the sample to be tested is virus-free, pest-free and robust.

[0051] Slow rooting of seedlings: immerse the 5 cm stem section of the cutting sample in the culture solution, and take 50 mg of young unexpanded leaves after 1 day for testing.

[0052] Cracking: Take 50 mg young stems and leaves, and the materials to be tested (when taking leaves, pa...

Embodiment 2

[0060] The test sample is: mixed sample of Xushu 18 and rice Nipponbare

[0061] Sampling in the field: Take samples from the field and keep only 10 parts of the 10cm stem tip of the unexpanded leaves. Try to take the samples in the morning or evening. The cuttings are placed in an aeroponic box (100cm×60cm) prepared in advance with 30L of Hoagland’s culture medium ×40cm). Hoagland's culture solution is prepared with deionized water. When cutting, pay attention to rinse the sample with deionized water to remove browning wounds and tissues. Avoid mechanical damage to tender tissues when sampling. When sampling, ensure that the sample to be tested is virus-free, pest-free and robust.

[0062] Slow rooting of seedlings: immerse the 5cm stem section of the cutting sample in the culture solution, and take 50mg of young young roots for testing after 1 day.

[0063] Lysis: take 50 mg of young young roots, the material to be tested, wash the surface dust with distilled water, blot ...

Embodiment 3

[0070] Test sample: Xushu 18

[0071] Field sampling: only keep 8 parts of the 10cm stem tips of unexpanded leaves from the field sampling, try to take the samples in the morning or evening, and put the sample cuttings in the aeroponic box (100cm × 60cm) prepared in advance with 30L of Hoagland culture medium ×40cm). Hoagland's culture solution is prepared with deionized water. When cutting, pay attention to rinse the sample with deionized water to remove browning wounds and tissues. Avoid mechanical damage to tender tissues when sampling. When sampling, ensure that the sample to be tested is virus-free, pest-free and robust.

[0072] Slow rooting of seedlings: immerse the 4cm stem section of the cutting sample in the culture solution, and take 50mg of new root tips after 1 day for testing.

[0073] Lysis: Take 50 mg of newborn root tips, wash the dust on the surface with distilled water, blot the surface moisture with filter paper, put it into a pre-cooled sterile petri di...

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Abstract

The invention provides a method for quickly identifying the genome size of sweet potatoes, belonging to the technical field of plant ploidy detection. The method comprises the steps of 1) taking sweetpotato tips from a field and performing cottage on the sweet potato tips in aHoagland nutrient solution for culturing for 20-28h; 2) mixing unexpanded leaves or new root tips with a lysis solution special for sweet potatoes for tissue lysis to obtain liquor produced after lysis; 3) filtering the liquor produced after lysis, collecting the filtrate, centrifuging, and collecting the centrifugationproduct as lysed cells; 4) mixing the lysed cells with a dye solution for dyeing, and obtaining a to-be-detected cell solution by vortex oscillation; 5) detecting the FL2-A value of the to-be-detectedcell by a flow cytometer; and 6) calculating to obtain the 1C value of the genome of the to-be-detected cell and the genome size of the to-be-detected cell according to the FL2-A value of the to-be-detected cell. The method can also identify the genome size of other species of ipomoea, and provide a fast, efficient and low-cost determination method for identifying the genome of sweet potatoes andrelated wild species.

Description

technical field [0001] The invention belongs to the technical field of plant ploidy detection, and in particular relates to a method for quickly identifying the genome size of sweet potato and its application. Background technique [0002] In recent years, with the vigorous development of flow cytometry (FCM), it can be used for immunophenotyping, cell counting, cytokine analysis, microsphere-based immunoassay, platelets, stem cells, apoptosis, cell cycle , DNA damage and proliferation, cancer research, cell signaling, calcium efflux, gene expression, kinetics, microbial enumeration and activity, water quality analysis, aquatic biology research, microbial species identification, bioprocessing, biofuels, plant biology, food science , veterinary medicine and other fields. In botany, flow cytometry is mainly used for the determination of nuclear DNA content, DNA ploidy identification, and cell cycle analysis. [0003] Flow cytometry (Flow cytometry, FCM) has been applied to t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N15/14G01N1/28
CPCG01N1/28G01N15/14
Inventor 苏一钧曹清河戴习彬赵路宽王珧唐君周志林赵冬兰张安
Owner XUZHOU INST OF AGRI SCI IN JIANGSU XUHUAI DISTRICT (JIANGSU XUZHOU SWEETPOTATO CENT)
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