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A method for rapidly identifying the genome size of sweet potato and its application

A genome and sweet potato technology, applied in the field of plant ploidy detection, can solve the disputes over the size of the sweet potato genome, the inability to meet the accuracy requirements of the sweet potato genome, browning, deterioration and death, etc.

Active Publication Date: 2021-07-23
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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  • A method for rapidly identifying the genome size of sweet potato and its application
  • A method for rapidly identifying the genome size of sweet potato and its application
  • A method for rapidly identifying the genome size of sweet potato and its application

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 size of the sweet potato genome, which belongs to the technical field of plant ploidy detection, and comprises the following steps: 1) taking sweet potato stem tip tissue cuttings from the field and culturing them in Hoagland culture solution for 20 to 28 hours; 2) taking Unexpanded leaves or new root tips are mixed with sweet potato special lysate for tissue lysis; 3) filtering the lysed feed liquid, collecting the filtrate, centrifuging, and collecting centrifuged products as lysed cells; 4) lysing the lysed cells Mix the cells with the dye solution for staining, and vortex to obtain the cell solution to be tested; 5) detect the FL2-A value of the cell to be tested by flow cytometry on the cell solution to be tested; 6) The FL2-A value of the cell to be tested is calculated to obtain the 1C value of the genome of the cell to be tested and its genome size. The method of the invention can also identify the genome size of other species of the genus Ipomoea, providing a fast, efficient and low-cost determination method for genome identification of sweet potato and its close relative 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 Patents(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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