CN1227961C - Screening method of late bolting character of radish - Google Patents

Abstract

The present invention discloses a screening method of a late bolting character of radishes, which comprises a stage of vernalization and a photostage. Seeds to be identified are sterilized and are soaked in bacteria-free water for accelerating germination, seeds which are in a sprouting state are inoculated in an MS culture medium and placed in a refrigerator, artificial vernalization treatment is carried out in different time periods, the seeds are placed in a tissue culture room after being treated at a low temperature to realize the growth culture of radish seedlings. In the photostage, the radish seedlings are transplanted in seedling-raising plates with substrates and placed in the tissue culture room, and flower bud bolting conditions of different materials after different treatment are observed. In the present invention, the property that radishes need a low temperature and long sunshine during the phasic development is utilized, radish seeds in a sprouting state are artificially vernalized at a low temperature, a long sunshine condition in the tissue culture room is utilized to realize the light irradiation, bolting properties of different materials are judged by observing flower bud bolting conditions of plants, and the process of late bolting breeding of the radishes is accelerated.

Description

The screening technique of radish bolting proterties in evening

Technical field:

The present invention relates to the screening technique of a kind of radish bolting proterties in evening, be used for introducing a fine variety and breeding of biennial vegetable spring such as radish and winter-spring season cultivation specific breed, belong to the crop breeding technical field.

Background technology:

Radish (Raphanus sativus L.) is one of main vegetables of widely cultivating of Southeast Asia, and is not only nutritious, and also has very high medical value.Though the history of China cultivation radish is very long, variety source is also very abundant, and the research of variety source is compared the also very big gap of existence with utilization with external advanced level.

Current, the radish varieties that is suitable for off-season cultivation is few, and of low quality be one of subject matter of existing during radish produces.Radish is 2 years living vegetable crops, and normal cultivation season is autumn and autumn and winter.The winter-spring season radish and spring the radish off-season cultivation success or not depend primarily on breed selection, otherwise may be because of bolting (do not form product organ-fleshy root, promptly bolting is bloomed) in advance and have no harvest.And the success or not of carrying out winter-spring season and off-season cultivation radish specific breed seed selection in spring depends primarily on the selection to the bolting characteristic.In addition, radish can only open once under field conditions (factors) in 1 year spends, this limited greatly the radish breeding process (Chen Huoying etc., CPPU is to the influence of the spring booth radish chaff heart, bolting and output, Shanghai Agricultural College's journal, 1999,17 (1), 48-51), (Wang Longzhi, Chen Huoying etc., the high-quality and efficient four seasons cultivation of radish, scientific and technical literature publishing house, 2001).

Summary of the invention:

The objective of the invention is to the deficiency that exists in the bolting breeding evening at existing radish, research provides a kind of screening technique of new radish bolting proterties in evening, shortens the time of radish bolting breeding in evening, and screening process is controlled and The selection result is more reliable.

For realizing such purpose, the present invention utilizes radish to need low temperature, long-day to satisfy this characteristic of its phasic development, radish seed to the state of sprouting under cryogenic conditions carries out artificial vernalization, utilize tissue culture room long-day condition to carry out illumination again, pass judgment on the bolting characteristic of different materials by the bolting situation of buddingging of observing plant.

Method of the present invention comprises vernalization stage and photostage, will use the sterile water presoaking and germinating after the seed disinfection of expert evidence, the seed that will be in the state of sprouting is inoculated in MS (Murashige and Skoog, development in 1962) in the macroelement medium, and place refrigerator, time segment is carried out artificial vernalization, places tissue culture room to carry out the grown cultures of Turnip Sprouts after the low temperature treatment again, utilizes the long-day condition to carry out illumination.Again Turnip Sprouts are transplanted and in seedling-cultivating tray, carry out open culturing and place tissue culture room, observe the bolting situation of buddingging at last, thereby pass judgment on the bolting characteristic of different materials, accelerate the process of radish bolting breeding in evening simultaneously through the plant of different disposal.

In the technical scheme of the present invention, the key problem in technology of each step comprises:

1. vernalization stage: seed that will expert evidence was soaked seed 24 hours vernalization 2 days with sterile water again with saturated bleaching fluid sterilization 20 minutes under 25 ± 1 ℃ of conditions under aseptic condition; The seed (the long 1-2mm of radicle) that will be in the state of sprouting is inoculated in the MS medium, and then the refrigerator that places temperature to be controlled at 3 ± 1 ℃ culture carries out artificial vernalization, and that handles the duration is set at 0 day (contrast), 5 days, 10 days, 15 days, 20 days; After low temperature treatment finishes, place temperature to be controlled at the grown cultures that 25 ± 1 ℃, 16 hours tissue culture room of illumination every day carry out Turnip Sprouts culture.

2. photostage: when Turnip Sprouts reach 3-4 sheet true leaf, open culturing is carried out in transplantation of seedlings in the seedling-cultivating tray that matrix is housed, during need 1 week, 4 extra-nutrition liquid, nutrient solution is 1/10MS macroelement and trace element; Seedling-cultivating tray still places above-mentioned tissue culture room, observes the bolting situation of buddingging through the different materials of different disposal when 30 days finish experiment, thereby filters out variety type or the individuality that is difficult for bolting.

Compare with the conventional screen selecting technology, the present invention has certain advantage: method is easy, and general conventional breeding worker is easy to grasp; Utilize aseptic condition to carry out artificial vernalization, avoided causing the lost possibility of part germplasm owing to seed goes mouldy; The Preliminary screening of lot of materials is carried out in utilization group training limited space, chamber, both can significantly reduce the field working time, save land resources, again can be so that screening is not subjected to the restriction in time, place, thus shortened the breeding time limit greatly; Whole screening process all under controlled condition, makes The selection result more reliable.

It is Shanghai Communications University's agricultural and the part variety source of characteristic radish breeding of new variety seminar of biological institute and the intermediate materials of breeding that the present invention selects test material for use.

Studies show that, utilize the present technique invention, can accelerate new varieties and identify speed, put into production as early as possible if be used to introduce a fine variety; Be used for breeding and can finish the stability screening of radish bolting variant in evening in the time of 3-4, traditional triage techniques then needs the time in 6 years at least, and also can avoid the problem of losing of contingent elite germplasm in screening process to a certain extent; In addition, utilize natural conditions to carry out the screening of late bolting material, may can not get seed then, thereby incured loss through delay its process from generation to generation the material that is difficult for bolting individually especially.

Embodiment:

Below by specific embodiment technical scheme of the present invention and implementation result are further described.

Embodiment 1

Old from white jade spring (from Korea S), day big of spring and little political affairs of the four seasons (from Japan) and domestic existing kind Wuhan spring, go up farming 936, Duanye 13,60 days and respectively get 75 seeds the mid-autumn red 8 parts of materials and (divide 5 parts, be 5 processing, 15 of every processing), with saturated bleaching fluid sterilization 20 minutes, under aseptic condition, under 25 ± 1 ℃ of conditions, soaked seed 24 hours vernalization 2 days again with sterile water; The seed (the long approximately 1-2mm of radicle) that will be in the state of sprouting thereafter is inoculated in the MS macroelement medium, then each 4 of 8 parts of materials are handled the refrigerator that cultures place temperature to be controlled at 3 ± 1 ℃ and carry out low temperature treatment, and after 5 days, 10 days, 15 days, 20 days culture dislocation after the low temperature treatment is controlled at 25 ± 1 ℃ in temperature respectively at low temperature treatment, carry out the grown cultures of Turnip Sprouts in 16 hours the tissue culture room of illumination every day.1 of each of other 8 parts of materials is handled culture and is then directly placed temperature to be controlled at 25 ± 1 ℃ after vernalization, and 16 hours tissue culture room of illumination every day does not carry out vernalization.

When Turnip Sprouts reach 3-4 sheet true leaf, with transplantation of seedlings in the matrix (composition of matrix: carry out open culturing in the seedling-cultivating tray 60% peat+20% perlite+20% vermiculite) is housed, finish test after 30 days, during 1 week 4 extra-nutrition liquid (1/10MS macroelement and trace element).Test shows that not old low temperature treatment of white jade spring and Wuhan spring can't be by phasic development when finishing test after 20 days; Little political affairs low temperature treatment of the four seasons was buddingged through 30 days after 20 days; It spring big low temperature treatment buddingged through 25 days after 20 days; Go up agricultural 936 low temperature treatment after 15 days, just buddingged through 25 days; 60 days and the mid-autumn, red low temperature treatment through 10 days was promptly buddingged in 20 days; Duanye 13 was promptly buddingged through 10 days low temperature treatment in 15 days.The all material that does not carry out low temperature treatment in the test all can not be by phasic development.

Under the natural conditions of winter-spring season, the planting seed of above-mentioned 8 parts of materials is carried out bolting CHARACTERISTICS IDENTIFICATION under the natural conditions in Shanghai Communications University's agricultural and biological institute farm, the performance of different materials bolting characteristic matches with above-mentioned indoor qualification result: bolting phenomenon does not in advance take place in white jade spring, day big of spring, little political affairs of the four seasons and Wuhan spring always; Bolting was not bloomed when upward agricultural 936 fleshy roots fully expanded; Duanye 13,60 days and the mid-autumn the red fleshy root that then do not form substantially be that bolting is bloomed.

Embodiment 2

According to the present invention, will have late bolting characteristic, and good Korea S's kind white jade spring of comprehensive agronomy proterties as one of object of spring radish kind production domesticization, carry out the selfing of flower bud phase and separate.And, the temperature of low temperature treatment being set at 3 ± 1 ℃ according to result of the present invention, the time set of processing is 30 days and 40 days, to guarantee receiving seed when the artificial vernalization.Specific embodiments:

Respectively on April 18th, 1998, April 28, get white jade spring 10 seeds with saturated bleaching fluid sterilization 20 minutes, under aseptic condition, under 25 ± 1 ℃ of conditions, soaked seed 24 hours vernalization 2 days again with sterile water; Be inoculated in the MS macroelement medium respectively at April 21, the seed (the long approximately 1mm of radicle) that will be in the state of sprouting on May 1, then the refrigerator that places temperature to be controlled at 3 ± 1 ℃ culture carries out low temperature treatment; In end of day low temperature treatment in Mays 31, and the culture dislocation is controlled at 25 ± 1 ℃ in temperature, carries out the grown cultures of Turnip Sprouts in 16 hours the tissue culture room of illumination every day.

When seedling reaches 3 true leaves, with transplantation of seedlings in be equipped with matrix (composition of matrix: carry out open culturing in the pot for growing seedlings 60% peat+20% perlite+20% vermiculite) (diameter is 20cm), during 1 the week 4 extra-nutrition liquid (1/10MS macroelement and trace element).Test shows, the white jade spring can budding through 30 days after handling 40 days under 3 ± 1 ℃ of conditions, to its bud pollination, and receives selfing 1 generation seed in by the end of July; By the end of December selfing 1 generation planting seed is screened in investigation and stability that plastic tunnel carries out economical character then received selfing 2 generation seed in by the end of June, 1999 in; At the beginning of 1999 7 months, selfing 2 generation seed is carried out artificial vernalization, obtain selfing 3 generation seed; In being seeded in the stability screening that booth carries out other economical character by the end of December, and, selecting to mix between the close different individual plants of Other Main Agronomic Characters and hand over for preventing the generation of selfing vitality decay, and receive in by the end of June, 2000 in the 4th generation seed; In July, 2000, with 4 generation seed carry out artificial vernalization, and selfing, obtain 5 generation seed, and divide individual plant to receive kind; In by the end of December, divide individual plant to be seeded in the stability screening that booth is proceeded economical character, and eliminate bad colony, in calendar year 2001 receive by the end of June the 6th generation seed, obtain 6 good colonies.August calendar year 2001 elected six clusters is seeded in open country, carries out the investigation of economical character regularity, and, plant respectively in 3 different booths to reserve seed for planting respectively with the radish kind strain of 3 colonies of neat and consistent.

Claims (1)

1, a kind of screening technique of radish bolting proterties in evening is characterized in that comprising:

1. vernalization stage: seed disinfection that will expert evidence 20 minutes, soaked seed 24 hours under 25 ± 1 ℃ of conditions with sterile water, vernalization 2 days, the seed that the long 1-2mm of radicle is in the state of sprouting is inoculated in the MS medium, the refrigerator that places temperature to be controlled at 3 ± 1 ℃ culture carries out artificial vernalization, that handles the duration is set at 0 day, 5 days, 10 days, 15 days, 20 days, after low temperature treatment finishes, place temperature to be controlled at the grown cultures that 25 ± 1 ℃, 16 hours tissue culture room of illumination every day carry out Turnip Sprouts culture;

2. photostage: when Turnip Sprouts reach 3-4 sheet true leaf, open culturing is carried out in transplantation of seedlings in the seedling-cultivating tray that matrix is housed, during need 1 week, 4 extra-nutrition liquid, nutrient solution is 1/10MS macroelement and trace element; Seedling-cultivating tray still places above-mentioned tissue culture room, observes the bolting situation of buddingging through the different materials of different disposal when 30 days finish experiment, thereby filters out variety type or the individuality that is difficult for bolting.