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Molecular marker assisted selection method for female line breeding of cucumber
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A molecular marker-assisted and molecular marker technology, applied in the field of vegetable molecular genetics and breeding, can solve the problems of heavy workload, high cost and long cycle, and achieve the effects of improving selection efficiency, simple operation and short cycle.
Active Publication Date: 2014-12-03
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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[0004] The purpose of the present invention is to: address the defects of large workload, long cycle, easy to be affected by environmental conditions, low breeding efficiency, high cost, etc. , to provide a molecular marker-assisted selection method for accurate, fast and efficient selection of female line materials for early segregation generation and seedling female line identification in the process of cucumber female line breeding
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Embodiment 1
[0028] Example 1: (Detection of resistance to hybrid progeny materials using molecular marker-assisted selection method 1)
[0029] (1) Collection of cucumber leaf samples to be tested: sow the cucumber material H174-1-2 to be tested, and when 1-2 true leaves are reached, take 0.5g of relatively young leaves at the top of the plant, and after collection, Use or freeze at -20°C;
[0030] (2) Collection of leaves of cucumber control samples: all female cucumber variety 240-1-2-2-3-1 inbred line, weak female variety 3-5-1-3-2-1-1-1-1 -2 Inbred line materials are sown, and when 1-2 true leaves are in stage, take the relatively young leaves at the top of the plant, 0.5 per share, and use them immediately after collection or freeze them at -20°C;
[0031] (3) Genomic DNA extraction of samples: DNA of each leaf was extracted by CTAB method. Weigh 150 mg of fresh leaves, pre-cool in liquid nitrogen together with adapters and grinding balls, and use MM301 cell disruptor to grind into...
Embodiment 2
[0035] Example 2: (Detection of resistance to hybrid progeny materials using molecular marker-assisted selection method 2)
[0036] (1) Collection of cucumber leaf samples to be tested: Sow the cucumber material H158-1-1-2-2 to be tested, and when 1-2 true leaves are reached, take the young leaves at the top of the plant, 0.5g per portion , used immediately after collection or frozen at -20°C;
[0037] (2) Collection of leaves of cucumber control samples: all female cucumber variety 240-1-2-2-3-1 inbred line, weak female variety 3-5-1-3-2-1-1-1-1 -2 Inbred line materials are sown, and at the stage of 1-2 true leaves, take the relatively young leaves at the top of the plant, 0.5g per portion, and use them immediately after collection or freeze them at -20°C;
[0038] (3) Genomic DNA extraction of samples: DNA of each leaf was extracted by CTAB method. Weigh 150 mg of fresh leaves, pre-cool in liquid nitrogen together with adapters and grinding balls, and use MM301 cell disrup...
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Abstract
The invention discloses a molecular marker assisted selection method for female line breeding of a cucumber. The method includes screening out a simple sequence repeat (SSR) labeled primer which is in close linkage with all-female genes of the cucumber through an all-female variety 230-1-2-2-3-1 inbred line, a weak female variety 3-5-1-3-2-1-1-1-1-2 inbred line and F2-generation segregation population of the cucumber; through extraction of DNA of a control variety and seedling genome of a seed to be detected, obtaining the gel maps of the control variety and the sample to be detected through the screened SSR labeled primer and after polyacrylamide gel electrophoresis and silver staining; and detecting the existence of the total-female genes of the cucumber rapidly and accurately according to a relative position of a stripe on the gel. According to the method, the molecular marker assisted breeding of the female line of the cucumber can be carried on effectively through SSR molecular marker screening of the all-female variety 230-1-2-2-3-1 inbred line and the weak female variety 3-5-1-3-2-1-1-1-1-2 inbred line, the selection efficiency and the standardization degree are improved, and the breeding process is accelerated.
Description
technical field [0001] The invention relates to a molecular marker-assisted selection method for cucumber female line breeding, and belongs to the technical field of vegetable molecular genetic breeding. Background technique [0002] Cucumber is one of the main vegetable crops in my country and has extremely high economic and social value. Cucumber plants have various sex types, including pure male plants, hermaphrodite plants, monoecious plants, androsexual flower plants and all female plants. The primary factor limiting the economic yield of cucumber is the ratio of male and female flowers, because except for all female plants, other sexual types of plants are characterized by luxuriant branches and leaves, large source and small pool, and low economic coefficient. Female-line cucumbers refer to lines in which the plants only grow female flowers but not male flowers or grow very few male flowers. This strain often shows the advantages of early maturity, dense melon, conc...
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