CN103773885A - Primer sequence and method for detecting Ogura cytoplasmic sterility of brassica crops - Google Patents
Primer sequence and method for detecting Ogura cytoplasmic sterility of brassica crops Download PDFInfo
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- CN103773885A CN103773885A CN201410049460.3A CN201410049460A CN103773885A CN 103773885 A CN103773885 A CN 103773885A CN 201410049460 A CN201410049460 A CN 201410049460A CN 103773885 A CN103773885 A CN 103773885A
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Abstract
The invention discloses a primer sequence and a method for detecting Ogura cytoplasmic sterility of brassica crops. The primer sequence comprises an upstream primer sequence shown as SEQ ID No.1 and a downstream primer sequence shown as SEQ ID No.2. The method is rapid and simple and convenient, the traditional field identification method has long period and costs a lot of manpower and soil fertility, the method can finish a batch of identification only in 5-6 hours, is not limited by sample materials, can sample and identify any tissue of the plant in the whole growth duration, and is accurate and reliable; the method is used for identifying more than 100 genetic resources preserved in the institute like broccoli, cauliflower, cabbage, kohlrabi and the like, strips of the characteristic can be amplified in all the sterility materials, and the fertility identification result is the same as the field identification result.
Description
Technical field
The present invention relates to agricultural biological technical field, be specifically related to a kind of primer sequence and method for detection of Brassica Crops Ogura cytoplasmic sterility.
Background technology
Male sterile refers to that the male reproductive system of plant can not normal development produce stamen, pollen or the male gamete with normal function, and its female repro ductive system is grown and nourish and grow normal a kind of biological phenomena completely.Male sterile extensively exists in vegitabilia, is divided into two types of nuclear male sterility and cytoplasmic male sterilities.
Cruciferae Brassica Crops hybrid vigour is obvious.For a long time, in the breeding of Brassica Crops, utilizing selfing not affine is one of the main path of preparing hybrid kind, and this approach exists long-term selfing vitality and easily degenerates, parent needs artificial cabage-type rape, breeding cost high, and the hybrid rate of cross-fertilize seed is difficult to reach the shortcoming such as 100%.Utilizing male sterile is an effective way of preparing hybrid kind, and genie male sterile line is so-called dual-purpose system, has been used in and has produced a upper hybridization generation.But utilizing dual-purpose is that the production of hybrid seeds need be pulled out 50% fertile plant, has both increased cost, affect again hybrid rate, thereby its utilization is restricted.Application cytoplasm male sterility line is produced cross-fertilize seed seed, just can overcome above-mentioned difficulties, improves first generation of hybrid seed purity, reduces breeding cost, is a kind of Perfected process of producing cenospecies.
Cytoplasmic male sterilty is commonly considered as being done mutually by nucleus sterile gene and Cytoplasmic male sterile genes the inherited character of co-controlling, and its mode of inheritance is matrocliny.The sterile degree of cytoplasmic male sterilty is high, and pollen abortion is thorough, on producing, has application prospect very widely.At present, in cress, utilizing is the Ogura sterile cytoplasm that comes from radish the most widely, it has genetic stability, sterile degree and sterile plant rate advantages of higher, being more satisfactory male sterile source, successfully proceeding in the brassicaceous vegetables such as Cauliflower, broccoli, wild cabbage, Chinese cabbage, turnip and cabbage heart, is one of sterile source being most widely used in vegetable crop.
The method of the evaluation male sterile cytoplasm extensively adopting is at present to identify extensive guarantor's relation.The method need to utilize Testers to prepare a large amount of cross combination, observes in the coming year the fertility of the first generation of hybrid, more accurate, but much time power, determination rates is low.
Summary of the invention
The object of this invention is to provide the primer sequence for detection of Brassica Crops Ogura cytoplasmic sterility that a species specificity is good.
Second object of the present invention is to provide a kind of accurately and rapidly for detection of the method for Brassica Crops Ogura cytoplasmic sterility.
Technical scheme of the present invention is summarized as follows:
For detection of the primer sequence of Brassica Crops Ogura cytoplasmic sterility, described primer sequence is made up of the downstream primer sequence shown in the upstream primer sequence shown in SEQ ID No.1 and SEQ ID No.2.
Brassica Crops is preferably broccoli, Cauliflower, wild cabbage, cabbage mustard or kohlrabi.
For detection of the method for Brassica Crops Ogura cytoplasmic sterility, comprise the following steps:
(1) extract Brassica Crops genomic dna to be measured;
(2) take described Brassica Crops genomic dna to be measured as template, classify upstream primer as with nucleotides sequence shown in SEQ ID NO.1, classify downstream primer as with nucleotides sequence shown in SEQ ID NO.2, carry out pcr amplification;
(3) in amplified production, add sample-loading buffer, mix, electrophoretic separation in sepharose 1 × TAE of 2%, electrophoresis finishes rear gel imaging system result is analyzed;
(4) on gel, occur 333bp specific amplified band for Brassica Crops Ogura cytoplasmic sterility sample.
Brassica Crops is preferably broccoli, Cauliflower, wild cabbage, cabbage mustard or kohlrabi.
The present invention is fast and convenient: traditional field identification method cycle is long, expend a large amount of manpower soil fertilities, adopt present method only to need 5-6 hour just can complete the evaluation of batch, and be not subject to the restriction of sample material, whole breeding time, any organizing all of plant can be sampled and be identified; Accurately and reliably: utilize the germ plasm resources such as more than 100 parts of broccolis that present method preserves this, Cauliflower, wild cabbage, kohlrabi to identify, in all sterile materials, all can amplify this feature band; Fertility identification result is consistent with field test result.
The present invention is only applicable to the detection of Ogura type cytoplasmic sterility material for detection of the primer sequence of Brassica Crops Ogura cytoplasmic sterility and detection method.
Accompanying drawing explanation
Fig. 1 is that method of the present invention detects Brassica Crops result figure.
In figure, M is DNA Marker; Isosorbide-5-Nitrae, 8,10 respectively are not Cauliflower, broccoli, wild cabbage, the kohlrabi sample containing Ogura kytoplasm; 2,3 is the Cauliflower sample containing Ogura kytoplasm; 5,6 is the broccoli sample containing Ogura kytoplasm; 7,9 is the wild cabbage sample containing Ogura kytoplasm; 11,12 is the kohlrabi sample containing Ogura kytoplasm.
Embodiment
Below in conjunction with specific embodiment, the present invention is further illustrated.
For detection of the method for Brassica Crops Ogura cytoplasmic sterility, comprise the following steps:
(1) CTAB method is extracted the genomic dna of Brassica Crops Cauliflower to be measured, broccoli, wild cabbage, kohlrabi blade;
(2) carry out pcr amplification: the Brassica Crops DNA20ng that adds step (1) to extract in PCR pipe, add again the nucleotide sequence 15ng described in SEQ ID NO.1 in sequence table, nucleotide sequence 15ng in sequence table described in SEQ ID NO.2,1 times containing Mg
2+pCR damping fluid, dNTP2mM, Taq archaeal dna polymerase 0.5 unit, adds aseptic double-distilled water to 20 μ L; PCR program is: 94 ℃ of denaturation 120s; Then 94 ℃ of sex change 15s, 58 ℃ of annealing 20s, 72 ℃ are extended 30s, totally 30 circulations; Last 72 ℃ are extended 60s, and amplification completes;
(3) gel electrophoresis analysis of PCR product: add gel sample-loading buffer [30mM EDTA, 36% (v/v) glycerine, blue or green FF, 0.05% (g/mL) tetrabromophenol sulfonphthalein of 0.05% (g/mL) dimethylbenzene] in amplified production, mix, at 2%(g/mL) sepharose 1 × TAE in electrophoretic separation, 120V constant voltage electrophoresis to tetrabromophenol sulfonphthalein stops electrophoresis while reaching the other end of gel, EB dyeing, gel imaging system is observed, is taken a picture result, sees Fig. 1.
(4) on gel, occur 333bp specific amplified band for Brassica Crops Ogura cytoplasmic sterility sample.
Isosorbide-5-Nitrae in Fig. 1,8,10 respectively are not Cauliflower, broccoli, wild cabbage, the kohlrabi sample containing Ogura kytoplasm; 2,3 is the Cauliflower sample containing Ogura kytoplasm; 5,6 is the broccoli sample containing Ogura kytoplasm; 7,9 is the wild cabbage sample containing Ogura kytoplasm; 11,12 is the kohlrabi sample containing Ogura kytoplasm.
Having or not with the field investigation result of sample of specific band is in full accord.
Experiment showed, that method of the present invention is also suitable for other Brassica Crops.
Claims (4)
1. for detection of the primer sequence of Brassica Crops Ogura cytoplasmic sterility, it is characterized in that described primer sequence is made up of the downstream primer sequence shown in the upstream primer sequence shown in SEQ ID No.1 and SEQ ID No.2.
2. the primer sequence for detection of Brassica Crops Ogura cytoplasmic sterility according to claim 1, is characterized in that described Brassica Crops is broccoli, Cauliflower, wild cabbage, cabbage mustard or kohlrabi.
3. for detection of the method for Brassica Crops Ogura cytoplasmic sterility, its feature comprises the following steps:
(1) extract Brassica Crops genomic dna to be measured;
(2) take described Brassica Crops genomic dna to be measured as template, classify upstream primer as with nucleotides sequence shown in SEQ ID NO.1, classify downstream primer as with nucleotides sequence shown in SEQ ID NO.2, carry out pcr amplification;
(3) in amplified production, add gel sample-loading buffer, mix, electrophoretic separation in sepharose 1 × TAE of 2%, electrophoresis finishes rear gel imaging system result is analyzed;
(4) on gel, occur 333bp specific amplified band for Brassica Crops Ogura cytoplasmic sterility sample.
4. the method for detection of Brassica Crops Ogura cytoplasmic sterility according to claim 3, is characterized in that described Brassica Crops is broccoli, Cauliflower, wild cabbage, cabbage mustard or kohlrabi.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115644062A (en) * | 2022-11-03 | 2023-01-31 | 天津科润农业科技股份有限公司 | Culture medium and method for improving embryo induction rate of Chinese cabbage with embryos difficult to produce |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1913773A (en) * | 2004-01-30 | 2007-02-14 | 辛根塔参与股份公司 | Improved fertility restoration for ogura cytoplasmic male sterile brassica and method |
| CN102071250A (en) * | 2010-06-25 | 2011-05-25 | 温州科技职业学院 | Broccoli Ogu cytoplasm quick identification kit and detection method thereof |
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- 2014-02-13 CN CN201410049460.3A patent/CN103773885A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1913773A (en) * | 2004-01-30 | 2007-02-14 | 辛根塔参与股份公司 | Improved fertility restoration for ogura cytoplasmic male sterile brassica and method |
| CN102071250A (en) * | 2010-06-25 | 2011-05-25 | 温州科技职业学院 | Broccoli Ogu cytoplasm quick identification kit and detection method thereof |
Non-Patent Citations (3)
| Title |
|---|
| 张扬勇等: "两种甘蓝Ogura细胞质雄性不育源的分子鉴别", 《中国农业科学》 * |
| 白红涛等: "甘蓝型油菜胞质雄性不育育性恢复基因的分子标记", 《西北农业学报》 * |
| 罗延青等: "甘蓝型油菜Ogura胞质雄性不育系及恢复系聚类分析", 《南方农业学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115644062A (en) * | 2022-11-03 | 2023-01-31 | 天津科润农业科技股份有限公司 | Culture medium and method for improving embryo induction rate of Chinese cabbage with embryos difficult to produce |
| CN115644062B (en) * | 2022-11-03 | 2023-11-17 | 天津科润农业科技股份有限公司 | Culture medium and method for improving induction rate of embryo of difficult-to-embryo Chinese cabbage |
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