CN1422524A

CN1422524A - Method for raising anther seedling greening-rate of rice

Info

Publication number: CN1422524A
Application number: CN 01138213
Authority: CN
Inventors: 冯双华; 谢小立; 汪汉林
Original assignee: CHANGSHA INST OF AGRICULTURAL
Current assignee: CHANGSHA INST OF AGRICULTURAL
Priority date: 2001-11-26
Filing date: 2001-11-26
Publication date: 2003-06-11

Abstract

The invention changes the perspecive of that agar only can be used as solidifying agent of culture medium and only has the action for supporting culture, provides an agar concentration index for obviously raising the indica rice anther induction culture and differentiation culture frequency, the agar concentration of inductino culture medium is 0.7-0.8%, and agar concentration of differetiation culture medium is 0.9-1.0%. Said invention includes: disinfecting the surface of rice ear, pretreating at 8 deg.C, selecting edge-approaching stage glumal flower for anther inoculation, after inoculation making dark culture, when the callus grows to 1.5-2 mm, it can be transferred onto differentiation culture medium.

Description

A kind of method that improves anther seedling greening-rate of rice

Technical field:

The present invention relates to a kind of raising paddy rice cultivation and plant, more specifically relate to a kind of method that improves anther seedling greening-rate of rice, is a kind of plant cell engineering practical approach of utilizing paddy rice method for breeding haploidy breeding rice new varieties approach.

Background technology:

Paddy rice is a kind of important crops, the people of many countries all with it as main food source.Because the increase of population is more and more to the demand of grain, thereby, also just more and more cause people's attention to improve its yield and quality to the improvement rice varieties.People place this task on the genetic manipulation of individual cells, tissue and organ level on, thereby wish to shorten breeding cycle, choose new rice variety disease-resistant, pest-resistant, best in quality and that patience is strong.And rice callus regeneration plant is one of effective ways of improvement rice quality.Because the long-grained nonglutinous rice anther culture is the crop that is difficult to induce high-frequency pollen regeneration plant in the important crops, the average healing rate of general long-grained nonglutinous rice is no more than 5%, often be lower than 1.0% and anther seedling greening is used for breeding, can't provide a certain amount of colony pollen plant seedling to use for breeding.

Summary of the invention:

The purpose of this invention is to provide a kind of method that improves anther seedling greening-rate of rice, propose a kind of suitable agar concentration, can significantly improve the differentiation frequency of the green seedling of long-grained nonglutinous rice callus.Concrete grammar of the present invention: 1. inducing culture period, by microscope inspection, to choose paddy rice and grow for the keep to the side flower pesticide of phase of microspore is connected on the inducing culture, inducing culture is medium (M8)+2mg/L dichlorphenoxyacetic acid (2,4-D), agar concentration is 0.7--0.8%; 2. differential medium: handle by the 25-30 days dark inducing culture down of 26 degree, callus grows 2 millimeters, changes on the differential medium and cultivates.The differentiation culture temperature is the 26-27 degree, and 12 hour photoperiod, intensity of illumination is 2000Lx.Differential medium is: medium (Ms)+2mg/L kinetin (KT)+0.5mg/L heteroauxin (IAA)+0.5mg/L a-methyl (NAA).Agar concentration is 0.9--1.0%.

The present invention is based on paddy rice training theory, according to the demand of Rice Anther microspore development to the medium nutriment, filter out suitable agar content concentration, make between medium and the culture and produce water stress, reduce the callus water content, improve ABA hormone-content and the level of proline content and the quality of callus differentiation in the callus, make the green seedling differentiation frequency of Rice Anther improve 10-20%.

The present invention has changed and has thought that agar is that the curing agent of medium plays a part to support culture, and has proposed a kind of a kind of agar concentration index that long-grained nonglutinous rice flower pesticide inducing culture and differentiation culture frequency are significantly improved, and is that flower is cultivated kind of a technology application.It is induced with differential medium agar concentration content and is respectively 0.8%, 1.0%, by the medium that this agar concentration is prepared respectively, can improve the green seedling differentiation frequency of long-grained nonglutinous rice 10-20%.

The present invention compared with prior art, have the following advantages: this method is than filter paper and silica gel method processing callus is easy, practical, effect is better, and be difficult for polluting culture, the most important thing is that it has changed the physiological situation of culture, help the growth of pollen mother cell and the growth and the differentiation of cell, organize green seedling differentiation frequency thereby improved rice callus significantly.

Concrete implementation step:

1, gather spike of rice at first according to a conventional method: the spike of rice of adopting back places 6--9 degree preliminary treatment Celsius after 3--5 hour after surface sterilization, checks by surface and microscope, chooses paddy rice and grows for the keep to the side flower pesticide of phase of microspore and be connected on the inducing culture; To be connected to flower pesticide on the inducing culture after the inoculation and place under the 26--27 degree Celsius dark culturing 25～30 days, when callus grows 1.5～2 millimeters, change on the differential medium and cultivate, the condition that callus breaks up green seedling is that temperature is 26～27 degree Celsius, 12 hour photoperiod, and intensity of illumination is 2000Lx.

2, the system of two kinds of medium is joined:

Inducing culture: (2,4-D), agar concentration is 0.7--0.8% to medium (M8)+2mg/L dichlorphenoxyacetic acid;

Differential medium: medium (Ms)+2mg/L kinetin (KT)+0.5mg/L heteroauxin (IAA)+0.5mg/L a-methyl (NAA).Agar concentration is 0.9--1.0%.

Claims

1, a kind of method that improves anther seedling greening-rate of rice, comprise the following steps: to adopt back spike of rice after surface sterilization according to conventional method, place 8～9 degree low temperature preliminary treatment Celsius 3～5 hours, inducing culture is chosen the keep to the side flower pesticide of phase of microspore period and is used for inducing culture, then flower pesticide is placed on the inducing culture of configuration, dark culturing is 25～30 days under 26 degree Celsius, when callus length to 1.5～2 millimeter, change on the differential medium and cultivate, the condition that callus breaks up green seedling is that temperature is 26～27 degree Celsius, 12 hour photoperiod, intensity of illumination is 2000Lx.

2, a kind of method that improves anther seedling greening-rate of rice according to claim 1 is characterized in that the prescription of described inducing culture is:

(2,4-D), agar concentration is 0.7--0.8% to medium (M8)+2mg/L dichlorphenoxyacetic acid.

3, a kind of method that improves anther seedling greening-rate of rice according to claim 1 is characterized in that the prescription of described differential medium is:

Medium (Ms)+2mg/L kinetin (KT)+0.5mg/L heteroauxin (IAA)+0.5mg/L a-methyl (NAA), agar concentration is 0.9--1.0%.