CN101280300A

CN101280300A - Method for transferring zizania latifolia gene to rice seed

Info

Publication number: CN101280300A
Application number: CNA2007100346724A
Authority: CN
Inventors: 臧曙
Original assignee: Individual
Current assignee: Individual
Priority date: 2007-04-03
Filing date: 2007-04-03
Publication date: 2008-10-08

Abstract

The invention relates to a hybridization method for rice and wild rice. The invention provides a method for introducing wild rice genes into rice seeds. The wild rice is adopted as the donator, and the rice seeds are adopted as the acceptor. The invention is characterized in that the method comprises the following steps: firstly, the stems or leaves of the wild rice in the booting stage are selected, cut into pieces after being cleaned, and then are disinfected; secondly, the wild rice solution accomplishing the above step is mixed with the disinfected rice seeds, added with water for agitation, and then is positioned into a dry and clean container after the water is cooled, and finally, the container is sealed; thirdly, five days later, the rice seeds in the container is agitated once, and is agitated once after another three days, and then is agitated once after two days; fourthly, the seeds through the steps are conventionally cultivated; fifthly, during the maturing stage, the wild rice seeds of the D1 generation are harvested. The method has the advantages that the method is simple and feasible for popularization, the genetic stability of the obtained wild rice is excellent, and the method is adapted to various gramineae.

Description

A kind of method that the wild rice gene is imported rice paddy seed

Technical field

The present invention relates to a kind of molecular breeding method, the hybridizing method of particularly a kind of paddy rice and wild rice.

Background technology

Plant molecular breeding is the breeding new way of starting modern age.Plant molecular breeding can be summarized as the engineering of two levels: the dna fragmentation that 1, will have the donor of purpose character gene imports the plant recipient cell of desire improvement, its offspring is morphed, therefrom filter out offspring who obtains the purpose proterties or the valuable new variety that suit the requirements; 2, separate goal gene, make up recombination, import the plant recipient cell that needs improvement,, filter out and obtained the purpose proterties through cultivating, and the comprehensive good offspring of proterties, bring out new variety.The present invention relates to a kind of method in ground.

Through 10 years of researches, the introduction method of plant foreign DNA has had significant progress, for example: particle bombardment, electrization, injection etc.As by the having or do not have of medium type, can be divided into carrier mediated method, direct guiding method and germplasm system mediated method.Carrier mediated method is that target DNA is changed in purulence bacillus or the virus, and be carrier, along with they to the infection of plant and target DNA is imported in the vegetable cell; Direct guiding method is directly to import vegetable cell with exposed foreign DNA by chemistry or physical method; And germplasm system mediated method, then be to realize the method that foreign DNA imports by germ cell and the medium (as pollen, ovum, ovary, pollen tube channel, rataria etc.) of plant self, it comprises that pollen tube introductory technique, ovary, young fringe, rataria injection, seed soak damp method etc.

So-called pickling process (imbibition) is meant seed, embryo, ovule, ovary, pollen granule, young fringe, the suspension cell of plant even has mausoleum etc. directly to be immersed in the foreign DNA a kind of method for transformation that utilizes osmosis that foreign gene is imported recipient cell and integrated and express.

Pickling process mainly is to utilize the substance operation system of vegetable cell self that exogenous DNA molecule is directly imported recipient cell.About the structure of vegetable cell haulage system and the existing a lot of research of function, proved that now vegetable cell can suck foreign DNA in the cell by following several approach at least: 1, foreign DNA can be transported in each cell by the haulage system of intercellular substance with the networking of plasmodesma composition; 2, vegetable cell also can be taken in foreign DNA in the cell by the endocytosis of similar animal; 3, the change of the membrane permeability of transfer cell also can be for the macromolecular substance permeate through cell membranes offers an opportunity in the vegetable cell, and especially in sexual cell, embryonic cell and meristematic cell, the chance that foreign DNA enters in the cell is bigger.

Wild rice (Zizania) is the perennial phreatophyte that the Gramineae wild rice belongs to, and is the relative genus of paddy rice, and it is distributed widely in various places, China north and south, grow in marsh, the pond etc., base portion is grown fat thickly owing to fungi autoeciousness, and fibrous root is sturdy, stalk is upright, high 90～180cm, flower unisexuality homophyletic, line up tectorum big panicle, male small ear is born on the branch of inflorescence bottom, and female small ear then is born in the top of inflorescence, caryopsis is cylindrical, autumn and winter in flowering fruit bearing stage summer, seed is tiny, easily shattering.So there is not tame wild rice all the time.

To experience all one's life of paddy rice and nourish and grow and two periods of reproductive growth, wherein, mainly comprise seeding stage and tillering phase vegetative growth phase.The paddy rice reproductive stage comprises jointing-booting stage, full heading time and grouting productive phase.Jointing-booting stage is meant that ear differentiation begins to comprise differentiation phase, formation phase and the phase of finishing till grow fringe, generally needs about one month; The productive phase of earing, is meant after spike of rice is bloomed and can be divided into milk stage, dough stage and full ripe stage again sophisticated period to grain.The growth characteristic of paddy rice reproductive stage is long shoot Zhang Sui, bloom, solid, formation and substantial seed, this is a main phase of capturing high yield, especially to pay attention to fertilizer, water in the cultivation, be so incensed that to coordinate, prolong the functional period of root system and blade, improve the substance accumulation transformation efficiency, reach the spike number foot, the fringe type is big, thousand seed weight and setting percentage height.

Because wild rice is the relative genus of paddy rice, so its vegetative period and paddy rice are similar.

The existing report of prior art discloses the technology that the DNA of wild rice is imported paddy rice.For example, Zang Jian number is disclosed in the application of 200510032147.X " a kind of solid wild rice DNA is imported the method for paddy rice and the reproductive material of acquisition thereof " in Chinese patent application.

Summary of the invention

At the defective of prior art, the present invention aims to provide the method that a kind of just wild rice gene imports rice paddy seed, and the wild rice dna solution is passed through pressure, infiltration, combination mutually, this method simply should be promoted, and the stable gene of gained paddy rice wild rice is good, adapts to multiple Gramineae.

For achieving the above object, the invention provides a kind of method, adopt wild rice to make donor, make acceptor, it is characterized in that, comprise the steps: with rice paddy seed with wild rice gene importing rice paddy seed

A, select the stem or the leaf of the wild rice in boot stage, after the cleaning, chopping, sterilization again;

B, the wild rice solution that will finish after the above-mentioned steps mix with the rice paddy seed that disinfects, and add entry and stir, and treat to put into a dry and clean container after the water cooling, at last with container sealing;

C, after five days, the rice paddy seed in the container with stir once, after three days, once stir, once stir again after two days;

D, will carry out conventional cultivation through the seed of above-mentioned steps gained;

E, treat seed maturity after, results D1 for rice wild rice seed.

The wild rice solution in the above-mentioned B step and the blending ratio of rice paddy seed are 10: 1

Adding the water temperature that stirs in the above-mentioned B step is 50-70 ℃.

Sterilization in the above-mentioned A step adopts TCCA (Trichloroisocyanuric acid) to carry out disinfection.

A kind of method provided by the invention with wild rice gene importing rice paddy seed, by pressure, infiltration is made up mutually the wild rice dna solution, and this method simply should be promoted, and the stable gene of gained paddy rice wild rice is good, adapts to multiple Gramineae.

Embodiment

Embodiment one:

Adopt the glutinous acceptor of doing of native breed swimming in the present embodiment, the solid wild rice of Dongting Lake is made donor, carries out following steps:

A, select the stem or the leaf 5000g of the wild rice in boot stage, after the cleaning, the TCCA (Trichloroisocyanuric acid) sterilization is adopted in chopping again.

B, the wild rice solution that will finish after the above-mentioned steps mix with the 500g rice paddy seed that disinfects, and add 50-70 ℃ of warm water and stir, and treat to put into a dry and clean container after the water cooling, at last with container sealing.

C, after five days, the rice paddy seed in the container with stir once, after three days, once stir, once stir again after two days.

D, will carry out conventional cultivation through the seed of above-mentioned steps gained; After treating seed maturity, D1 is for rice wild rice seed for results.

March 11 in 2006, the daily container introductory technique carried out above-mentioned processing, vernalization on March 24 with ten day time, percentage of germination was sowed on the 20%, 26, seedling age 25 days, rice transplanting on April 20, the vegetative period-to pull out stem phase-boot stage be 96 days (26-July 1 March), fringe phase beginning is July 10 (13), in July 24 by academy of agricultural sciences's inspection, confirm the rice wild rice, its performance shows the peculiar property of wild rice, fragrant evening, glutinous peculiar property had both community, in the leaflet phase, be the leaflet pattern of wild rice, leaflet is more, along with growing tall of strain stem, produced the singularity of wild rice, the bright and clean nothing hair of cane, but the stipes wooden fork is locating to accompany the stem racemosus, the reality of setting seeds, belong to the effective leaflet of cane, effective leaflet that every wooden fork place has the 2-3 branch not wait, the stem height has 1.42m, the blade broad, light and effect are stronger, and root system is longer, for round or round partially, root system knot portion is closeer, be about 84cm, there are countless radiculas on the side, the about 12cm of radicula, the effect storage moisture synthesizing amino acid of root, this is the characteristic of wild rice.Seed is pale, and grain is carefully grown, and 0.89～0.98mm is arranged, and fringe portion is solid closeer, has 300 to 400 not wait, by solid two parts of the master of stem fringe and wooden fork portion.In addition, the characteristic of spike of rice is living all the year round, summer flood fighting water arid.Winter is cold-resistant, ties paddy for many batches for many years, and output is higher, and nutrition is very abundant, full grains.

Claims

1. the method with wild rice gene importing rice paddy seed adopts wild rice to make donor, makes acceptor with rice paddy seed, it is characterized in that, comprises the steps:

E, treat seed maturity after, results D1 for rice wild rice seed.

2. a kind of method with wild rice gene importing rice paddy seed according to claim 1 is characterized in that the wild rice solution in the above-mentioned B step and the blending ratio of rice paddy seed are 10: 1

3. a kind of method with wild rice gene importing rice paddy seed according to claim 1 and 2 is characterized in that adding the water temperature that stirs in the above-mentioned B step is 50-70 ℃.

4. a kind of method with wild rice gene importing rice paddy seed according to claim 1 and 2 is characterized in that the sterilization in the above-mentioned A step adopts TCCA (Trichloroisocyanuric acid) to carry out disinfection.