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Method for using molecular marking to assist selecting rapid transfer broccoli male sterile lines

A molecular marker-assisted and molecular marker technology is applied in the direction of botany equipment and methods, applications, plant genetic improvement, etc. It can solve the problems of difficulty in guaranteeing purity and provenance, dependence on import of hybrid seeds, and high cost of seed production, so as to reduce generations Effects of population size, shortening backcross generation, and speeding up breeding

Inactive Publication Date: 2007-10-24
ZHEJIANG UNIV
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Problems solved by technology

[0005] The purpose of the present invention is to propose a method of using molecular marker-assisted selection to quickly transfer broccoli cells in view of the current situation that the cost of conventional seed production of broccoli hybrids is high, the hybrid seeds mainly rely on imports, are expensive, and the purity and provenance are difficult to guarantee. qualitative male sterile line method

Method used

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Embodiment

[0048] 1. Hybridization:

[0049] The broccoli male sterile line 725 (plant height about 58cm, spreading degree 70cm, curd diameter 16.5cm, single bulb weight 500g) is a cytoplasmic male sterile line that can be inherited stably and is not affected by environmental conditions such as light and temperature. , with normal stigma development and no pollen in male flowers) was the source of sterility and served as the female parent. The disease-resistant and high-yield 718 selected by the applicant was used as the reincarnation male parent, and the F1 was obtained and then crossed with the reincarnation parent to obtain the BC1 generation segregation population. Harvest the seeds and plant 100 individual plants.

[0050] 2. PCR molecular marker analysis

[0051] 1) DNA extraction Samples are numbered by individual plant, add 600 μL (preheated to 65°C) extraction buffer, mix with 40 μL SDS, and bathe in 65°C water bath for 30 minutes. Add 200μL of 5M KAc, ice bath for 60min. Ce...

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Abstract

The invention discloses a method for fast breeding male sterile broccoli by using molecular marking, comprising following steps: (1) taking cytoplasm male sterile heterozygote of broccoli as female parent and good auto-copulation of broccoli as male parent, carrying out sexual crossing; taking its auto-copulation as recurrent parent for continuous backcross; (2) choosing sterile deseription in sprout period for backcross generation by using PCR checking technology; (3) choosing the genetic background for said single plants by applying AFLP labeling; (4) choosing plant with its molecular marking reaction being positive for backcross; (5) repeating steps (2) and (3), getting stable sterile broccoli and its relevant maintenance line through 3-4 generation backcross. The invention applies molecular marking connected with male sterile gene for the first time to breed purified male sterile line. The bred male sterile line is free from influence from temperature change, and the sterile degree is 100%.

Description

technical field [0001] The invention belongs to the field of heterosis utilization. Specifically, the invention relates to a broccoli cytoplasmic male sterile line breeding technology, in particular to a method for quickly cultivating a broccoli male sterile line by using molecular marker-assisted selection. technical background [0002] Broccoli is a variety of Brassica oleracea in the family Cruciferae (Cruciferae), a variety with green or purple curds as the product, a one-year or two-year herbaceous plant. Scientific name: Brassica oleracea L. var. italica Planch. Chromosome number 2n=2x=18. Broccoli is rich in vitamin C, vitamin B1, B2, B5, etc., and contains anticancer substances, so it is very popular among domestic and foreign consumers. Broccoli is evolved from cabbage, and the center of evolution is the eastern Mediterranean coast. The cultivation history in our country is very short, but it develops very fast. It has become an important vegetable for export and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/02A01H1/04C12Q1/68
Inventor 卢钢朱祝军邹宜静邵泰良何勇
Owner ZHEJIANG UNIV
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