CN113615649A - Purification method and application of new soybean cyst nematode microspecies X12 - Google Patents
Purification method and application of new soybean cyst nematode microspecies X12 Download PDFInfo
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- 241000498254 Heterodera glycines Species 0.000 title claims abstract description 62
- 238000000034 method Methods 0.000 title claims abstract description 36
- 238000000746 purification Methods 0.000 title claims abstract description 28
- 208000031513 cyst Diseases 0.000 claims abstract description 28
- 235000010469 Glycine max Nutrition 0.000 claims abstract description 27
- 244000068988 Glycine max Species 0.000 claims abstract description 27
- 201000010099 disease Diseases 0.000 claims abstract description 16
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 16
- 238000012216 screening Methods 0.000 claims abstract description 12
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 11
- 230000000644 propagated effect Effects 0.000 claims abstract description 7
- 238000009395 breeding Methods 0.000 claims abstract description 3
- 230000001488 breeding effect Effects 0.000 claims abstract description 3
- 239000002689 soil Substances 0.000 claims description 37
- 238000012360 testing method Methods 0.000 claims description 18
- 238000012258 culturing Methods 0.000 claims description 15
- 241000196324 Embryophyta Species 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 6
- 230000001717 pathogenic effect Effects 0.000 claims description 3
- 238000004321 preservation Methods 0.000 claims description 3
- 239000004576 sand Substances 0.000 claims description 3
- 230000004807 localization Effects 0.000 claims 1
- 238000012163 sequencing technique Methods 0.000 claims 1
- 238000011160 research Methods 0.000 abstract description 15
- 230000007918 pathogenicity Effects 0.000 abstract description 7
- 206010011732 Cyst Diseases 0.000 abstract description 6
- 238000012268 genome sequencing Methods 0.000 abstract description 4
- 208000035473 Communicable disease Diseases 0.000 abstract 1
- 208000015181 infectious disease Diseases 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 16
- 238000011081 inoculation Methods 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 235000003276 Apios tuberosa Nutrition 0.000 description 3
- 241001300423 Strophostyles Species 0.000 description 3
- 235000013601 eggs Nutrition 0.000 description 3
- 241001260012 Bursa Species 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 241000223199 Humicola grisea Species 0.000 description 1
- 108010064851 Plant Proteins Proteins 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 239000010773 plant oil Substances 0.000 description 1
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Abstract
The invention provides a purification method and application of a new race X12 of soybean cyst nematode, which is characterized in that a soybean cyst nematode group identified as a soybean cyst nematode X12 physiological race is continuously propagated for 4 generations on domestic excellent Source-resistant county grey bark branch (number: ZDD2315) by adopting a single cell cyst propagation method, then is continuously purified and propagated for 3-4 generations on the county grey bark branch, and then is recovered for 1 generation by utilizing an infectious disease variety Lee, so that the purification and propagation of the soybean cyst group are realized. The method can separate and purify X12 race with super pathogenicity from soybean cyst nematode colony mixed in nature, and the purified X12 race can be continuously used for genome sequencing research, soybean cyst nematode pathogenicity gene research, soybean resistance resource screening, soybean cyst nematode resistance gene positioning and phenotype identification screening research in the soybean disease-resistant variety breeding process for multiple generations, and has important application value in the aspect of soybean cyst nematode related research.
Description
Technical Field
The invention relates to a purification method and application of a new soybean cyst nematode microspecies X12, belonging to the technical field of biology.
Background
Soybeans are an important source for people to take plant proteins and plant oils, so that soybeans play a significant role in agricultural production. However, Soybean Cyst Nematodes (SCNs) are widely distributed in major Soybean producing areas worldwide, and cause serious economic losses in Soybean production. The Huang-Huai region is one of two major soybean production regions in China, and early research results show that the soybean cyst nematodes in the region are common, the No. 2 is a dominant physiological race, and the No. 5 is a second physiological race. In the investigation research, a new soybean cyst nematode microspecies X12 with super pathogenicity is also detected, the microspecies can successfully parasitize the excellent Humicola grisea (ZDD2315) in the county of resisting source and all other published resisting sources in China at present, and belong to the new soybean cyst nematode microspecies with super pathogenicity, and the discovery of the physiological microspecies shows that the soybean cyst nematode further threatens the soybean production.
Because the soybean cyst nematode propagation belongs to intraspecific hybridization, the cysts propagated in the diseased soil collected from the field infected with the soybean cyst nematode are mixed by a plurality of groups, and the mixed groups of the soybean cyst nematode without purification seriously obstruct the processes of the soybean cyst nematode in genome sequencing research, the soybean cyst nematode pathogenic gene research, the soybean resistance resource screening, the soybean cyst nematode resistance gene positioning and the soybean disease-resistant variety cultivation. By combining the agricultural green development guideline of 'paying attention to resource saving and environmental friendliness' in China, a disease-resistant variety is urgently needed to be cultivated for preventing the economic loss of soybean production caused by soybean cyst nematodes, which is the most economical, effective and green environmental protection measure for preventing the soybean cyst nematodes, and the development of the purification of the new soybean cyst nematode microspecies X12 for screening the disease-resistant resources is a precondition for cultivating new disease-resistant varieties.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a method for purifying a novel soybean cyst nematode microspecies X12 and application thereof, wherein the method can separate and purify the X12 microspecies with super pathogenicity from a natural mixed soybean cyst nematode population.
In order to achieve the purpose, the invention adopts the technical scheme that:
a method for purifying a new microspecies X12 of soybean cyst nematode comprises the following steps:
(1) collecting diseased soil infected by soybean cyst nematode X12, and mixing the diseased soil and sterile fine sand in a ratio of 3: uniformly mixing the materials in a volume ratio of 0.5-1.0 to prepare test disease soil, and subpackaging the test disease soil in a test cup for identifying the resistance of the soybean cyst nematodes;
(2) transplanting the excellent anti-source Xingxian grey bark branches of soybeans which germinate for 3-5 days and are in the cotyledon unearthing period into test cups filled with test disease soil, transplanting 1 plant in each test cup, and placing in a greenhouse for culturing;
(3) after culturing for 20-25 d, taking the single cell sacs from the root system by using forceps, inoculating the single cell sacs to the vicinity of the ash branch root system in Xingxian county growing for 3-5 d in sterile soil, inoculating single cell sacs in each cup, and culturing for 20-25 d in a greenhouse;
(4) repeating the step (3), after the single cell sacs are propagated for 4 times, completely inoculating the 4 th generation of the single cell sacs collected from a single laboratory cup to the vicinity of the root system of the ash bark branch in Xingxian county growing in sterile soil for 3-5 d, and culturing in a greenhouse for 20-25 d to obtain purified 5 th generation of the cell sacs;
(5) inoculating all 5 th generation cysts collected from a single laboratory cup to the vicinity of the root system of the ash bark branch in Xingxian county growing in sterile soil for 3-5 d, and culturing in a greenhouse for 20-25 d to obtain purified 6 th generation cysts;
(6) and (5) repeating the step (5) until purified 8 th generation or higher generation cysts are obtained, and completely inoculating the 8 th generation or higher generation cysts obtained from a single laboratory cup to the vicinity of the root system of the Lee of the susceptible variety growing in sterile soil for 3-5 d, so as to realize the group purification and generation-added preservation of the cysts.
The collection method of the disease soil comprises the following steps: collecting the soil 2-6 cm below the surface of the soil from the soybean field infected by the soybean cyst nematode X12 for later use.
The greenhouse culture conditions are as follows: alternating 16h in the daytime and 8h in the night; temperature: day at 27 +/-1 ℃ and night at 24 +/-1 ℃; humidity was 70% ± 5%.
The purification method is applied to the genome sequencing of the soybean cyst nematode.
The purification method is applied to the research of pathogenic genes of soybean cyst nematode and the gene location of soybean cyst nematode resistance.
The purification method is applied to screening of soybean resistance resources and phenotypic identification and screening in cultivation of soybean disease-resistant varieties.
The invention has the beneficial effects that:
the invention provides a method for purifying a new race X12 of soybean cyst nematode, which is to continuously propagate a soybean cyst nematode group identified as a soybean cyst nematode X12 physiological race on a domestic excellent Source-resistant ash bark branch (number: ZDD2315) for 4 generations by a cyst unicellular propagation method, then continuously purify and propagate the soybean cyst on the ash bark branch of Xingxing county for 3-4 generations, and recover 1 generation by utilizing an susceptible variety Lee, thereby realizing the group purification and propagation of the cyst. Compared with the existing purified population obtained by expanding propagation after 1 generation of propagation by adopting the monad bursa, the method continuously screens 4 generations of purified populations obtained by expanding propagation by continuously screening 4 generations of grey bark branches in Xingxian county after 4 generations of propagation by adopting the monad bursa, has high population purity, stable progeny and relative homozygosis in the 32 th generation.
The method provided by the invention is adopted to purify the new soybean cyst nematode microspecies X12, X12 microspecies with super pathogenicity can be separated and purified from a natural mixed soybean cyst nematode population, the purified X12 microspecies can be continuously used for genome sequencing research, soybean cyst nematode pathogenicity gene research, soybean resistance resource screening, soybean cyst nematode resistance gene positioning and phenotype identification screening research in the soybean disease-resistant variety cultivation process for multiple generations, and the method has important application value in the aspect of soybean cyst nematode related research.
Drawings
FIG. 1 is a schematic diagram of a purification scheme of a novel soybean cyst nematode microspecies X12;
FIG. 2 shows the average cyst numbers before and after purification of the novel soybean cyst nematode race X12.
Detailed Description
The following examples further illustrate the embodiments of the present invention in detail.
Example 1
A method for purifying a new microspecies X12 of soybean cyst nematode comprises the following steps:
(1) collecting diseased soil infected by soybean cyst nematode X12, uniformly mixing the diseased soil and sterile fine sand according to the volume ratio of 3: 0.5-1.0 to prepare test diseased soil, and subpackaging the test diseased soil in a disposable plastic test cup for identifying the resistance of the soybean cyst nematode;
the collection method of the diseased soil comprises the following steps: according to reported investigation and research results, collecting soil between 2cm and 6cm below the surface layer of the soil from a soybean field infected by soybean cyst nematode X12 for later use; the soybean cyst nematode has 9 pairs of chromosomes;
(2) transplanting 3-5 d germinated soybeans at the cotyledon emergence period into test cups containing test disease soil, transplanting 1 strain in each test cup, and culturing in a greenhouse;
the greenhouse culture conditions were: alternating 16h in the daytime and 8h in the night; temperature: day at 27 +/-1 ℃ and night at 24 +/-1 ℃; humidity is 70% +/-5%;
(3) after culturing for 20-25 d, taking the single cell sacs from the root system by using tweezers, inoculating the single cell sacs to the vicinity of the Xingxian gray skin branch root system growing for 3-5 d in sterile soil, inoculating single cell sacs/cup, and culturing for 20-25 d in a greenhouse;
(4) repeating the step (3), after the single cell sacs are propagated for 4 times, completely inoculating all the 4 th generation of the single cell sacs collected from a single laboratory cup (about 10-30 cell sacs) to the position near the root system of the ash bark branch of Xingxian county growing in sterile soil for 3-5 d, and culturing in a greenhouse for 20-25 d to obtain the purified 5 th generation of cell sacs;
(5) inoculating all 5 th generation cysts collected from a single laboratory cup (about 20-60 cysts) to the vicinity of the root system of the ash bark branch in Xingxian county growing for 3-5 d in sterile soil, and culturing in a greenhouse for 20-25 d to obtain purified 6 th generation cysts;
(6) and (5) repeating the step (5) until purified 8 th-generation or higher-generation cysts are obtained, and completely inoculating (about 150) 300 th-generation cysts obtained from a single laboratory cup to the vicinity (more than 60) of the root system of the Lee of the infected variety which grows for 3-5 d in the sterile soil, so as to realize the population purification and the generation-added preservation of the cysts (the purification process is shown in figure 1).
Application example 1
The new soybean cyst nematode microspecies X12 purified by the purification method of the embodiment 1 and the new soybean cyst nematode microspecies X12 which are not purified are respectively inoculated to the root system of the branch of the ash bark of Xingxian county, 2000 eggs/plant, the inoculation method before purification is repeated for 3 times, the inoculation method after purification is repeated for 4 times, and the number of cysts on the root system of each plant is respectively counted after inoculation for 20d-30d, and the result is shown in figure 2.
The results show that the average number of cysts on the root of the plant to be inoculated is 110.2 for the plant inoculated with the X12 races which have not been purified, and 250.2 for the plant inoculated with the X12 races which have been purified. The purity of the X12 physiological race purified from 2000 inoculated eggs is higher than that of the X12 physiological race not purified, and the purification method eliminates other physiological races and non-homozygous X12 races mixed in the population after purifying X12 by multi-generation unit cell sacs, so that the homozygous X12 eggs account for a large proportion, and can be effectively infected on the grey bark branch of Xingxian county, so that the number of the unit cell sacs is large, and the number of the unit cell sacs effectively infected on the grey bark branch root system of Xingxian county is large, and the repeatability is high.
Application example 2
The X12 physiological race purified by the purification method of the embodiment 1 of the invention is screened out the resistance source in the wild soybean resistance resource screening.
The X12 physiological race purified by the purification method of the embodiment 1 of the invention is utilized to carry out greenhouse inoculation and phenotype identification on 420 parts of wild soybean resources. The method is characterized in that single wild bean material plants which show disease resistance are propagated, repeated resistance identification is carried out after 2019.10 single plants are harvested (HG type identification mode + Riggs identification mode + excellent resistance source in China is adopted), resistance identification results show that one part of wild bean materials shows high resistance (FI is 2.2 and the material name is YSD56), and the resistance identification results of YSD56 wild bean resources in multiple soybean cyst nematode physiological races are shown in Table 1. YSD56 and susceptible parents are selected to construct a colony, which can be used for subsequent resistance gene positioning and breeding research of soybean cyst nematode disease-resistant varieties.
TABLE 1 identification of resistance of YSD56 and other major resistance sources in various soybean cyst nematode physiological races
aCyst Index, FI ═ Female Index;
blevel of resistance to soybean cyst nematode: FI ≦ 10(Resistant ═ R, Resistant); 10<FI ≦ 30 (modertel resistance ═ MR, medium resistance); 30<FI ≦ 60(Moderately susceptable, MS, Mizhongzhong), and FI>60 (stable. S, feeling); the resistance level is divided according to the FI value, the FI is equal to the average number of cysts on the root system of the material to be identified/the average number of cysts on the Lee root system 100;
caverage number of cysts (in) that grew on Lee;
drepresents a differential host in the Riggs pattern;
erepresenting a discriminative host in HG type mode;
frepresenting excellent resistance source in China.
Claims (6)
1. A method for purifying a new microspecies X12 of soybean cyst nematode is characterized by comprising the following steps:
(1) collecting diseased soil infected by soybean cyst nematode X12, and mixing the diseased soil and sterile fine sand in a ratio of 3: uniformly mixing the materials in a volume ratio of 0.5-1.0 to prepare test disease soil, and subpackaging the test disease soil in a test cup for identifying the resistance of the soybean cyst nematodes;
(2) transplanting the excellent anti-source Xingxian grey bark branches of soybeans which germinate for 3-5 days and are in the cotyledon unearthing period into test cups filled with test disease soil, transplanting 1 plant in each test cup, and placing in a greenhouse for culturing;
(3) after culturing for 20-25 d, taking the single cell sacs from the root system by using forceps, inoculating the single cell sacs to the vicinity of the ash branch root system in Xingxian county growing for 3-5 d in sterile soil, inoculating single cell sacs in each cup, and culturing for 20-25 d in a greenhouse;
(4) repeating the step (3), after the single cell sacs are propagated for 4 times, completely inoculating the 4 th generation of the single cell sacs collected from a single laboratory cup to the vicinity of the root system of the ash bark branch in Xingxian county growing in sterile soil for 3-5 d, and culturing in a greenhouse for 20-25 d to obtain purified 5 th generation of the cell sacs;
(5) inoculating all 5 th generation cysts collected from a single laboratory cup to the vicinity of the root system of the ash bark branch in Xingxian county growing in sterile soil for 3-5 d, and culturing in a greenhouse for 20-25 d to obtain purified 6 th generation cysts;
(6) and (5) repeating the step (5) until purified 8 th generation or higher generation cysts are obtained, and completely inoculating the 8 th generation or higher generation cysts obtained from a single laboratory cup to the vicinity of the root system of the Lee of the susceptible variety growing in sterile soil for 3-5 d, so as to realize the group purification and generation-added preservation of the cysts.
2. The purification method according to claim 1, wherein the collection method of the diseased soil is: collecting the soil 2-6 cm below the surface of the soil from the soybean field infected by the soybean cyst nematode X12 for later use.
3. The purification method of claim 1, wherein the greenhouse culture conditions are: alternating 16h in the daytime and 8h in the night; temperature: day at 27 +/-1 ℃ and night at 24 +/-1 ℃; humidity was 70% ± 5%.
4. Use of a purification process according to any one of claims 1 to 3 for sequencing the genome of a soybean cyst nematode.
5. Use of the purification method according to any one of claims 1 to 3 for the study of pathogenic genes of soybean cyst nematode and for the localization of genes against soybean cyst nematode.
6. Use of the purification method according to any one of claims 1 to 3 in screening of soybean resistant resources and phenotypic identification and screening in breeding of soybean resistant varieties.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030126625A1 (en) * | 2001-01-18 | 2003-07-03 | Leo Liu | Screens and assays for agents useful in controlling parasitic nematodes |
| CN102487901A (en) * | 2011-12-06 | 2012-06-13 | 华南农业大学 | Method for breeding meloidogynes by utilizing single allosome of separated root system of non-exogenous cultured water spinach |
| CN105010245A (en) * | 2015-08-04 | 2015-11-04 | 山东省花生研究所 | Method for cultivating meloidogyne hapla by adopting peanut rooting |
| CN108719208A (en) * | 2018-05-15 | 2018-11-02 | 吉林省怡科农业生物科技有限公司 | Entomopathogenic nematode large-scale method for producing and entomopathogenic nematode |
| CN109496988A (en) * | 2018-11-01 | 2019-03-22 | 李小龙 | A kind of method and culture apparatus of entomopathogenic nematode pilot scale culture |
| CN110250113A (en) * | 2019-07-09 | 2019-09-20 | 云南农业大学 | A method of M hapla is cultivated using Eupatorium adenophorum |
| CN113179898A (en) * | 2021-05-28 | 2021-07-30 | 河南省农业科学院 | Resistance identification method for improving infection efficiency of soybean cyst nematodes |
-
2021
- 2021-09-24 CN CN202111121931.3A patent/CN113615649A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030126625A1 (en) * | 2001-01-18 | 2003-07-03 | Leo Liu | Screens and assays for agents useful in controlling parasitic nematodes |
| CN102487901A (en) * | 2011-12-06 | 2012-06-13 | 华南农业大学 | Method for breeding meloidogynes by utilizing single allosome of separated root system of non-exogenous cultured water spinach |
| CN105010245A (en) * | 2015-08-04 | 2015-11-04 | 山东省花生研究所 | Method for cultivating meloidogyne hapla by adopting peanut rooting |
| CN108719208A (en) * | 2018-05-15 | 2018-11-02 | 吉林省怡科农业生物科技有限公司 | Entomopathogenic nematode large-scale method for producing and entomopathogenic nematode |
| CN109496988A (en) * | 2018-11-01 | 2019-03-22 | 李小龙 | A kind of method and culture apparatus of entomopathogenic nematode pilot scale culture |
| CN110250113A (en) * | 2019-07-09 | 2019-09-20 | 云南农业大学 | A method of M hapla is cultivated using Eupatorium adenophorum |
| CN113179898A (en) * | 2021-05-28 | 2021-07-30 | 河南省农业科学院 | Resistance identification method for improving infection efficiency of soybean cyst nematodes |
Non-Patent Citations (1)
| Title |
|---|
| YUN LIAN 等: "Chromosome-level reference genome of X12，a highly virulent race of the soybean cyst nematode Heterodera glycines", 《MOLECULAR ECOLOGY RESOURCES》 * |
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