CN105613290B

CN105613290B - A kind of inducing culture of Southeast Asia common wild-rice self-mating system embryo callus

Info

Publication number: CN105613290B
Application number: CN201511018196.8A
Authority: CN
Inventors: 潘英华; 梁云涛; 徐志健; 黄娟
Original assignee: Rice Research Institute Guangxi Academy Of Agricultural Sciences
Current assignee: Rice Research Institute Guangxi Academy Of Agricultural Sciences
Priority date: 2015-12-29
Filing date: 2015-12-29
Publication date: 2017-11-14
Anticipated expiration: 2035-12-29
Also published as: CN105613290A

Abstract

The invention provides a kind of inducing culture of Southeast Asia common wild-rice self-mating system embryo callus, the inducing culture is that the specific hormone that certain concentration is added on the basis of N6D culture mediums obtains, Fiber differentiation can improve healing rate on the culture medium, increase callus diameter, improve callus quality.

Description

A kind of inducing culture of Southeast Asia common wild-rice self-mating system embryo callus

Technical field

The present invention relates to a kind of inducing culture and abductive approach, and in particular to a kind of Southeast Asia common wild-rice self-mating system The inducing culture and abductive approach of embryo callus.

Background technology

Plant dedifferentiation and differentiation it is affected by various factors, as genotype, explant selection, type of culture medium and Interaction between them, but up to now, it can not establish a kind of " omnipotent culture medium " for adapting to all genotype (Bolibok, 2006).But the genome of wild rice and cultivated rice genome difference are larger, therefore suitable for the training of cultivated rice Foster system can not produce same positive role to all wild rices.By genotype if the cultured in vitro of oryza officinalis Limitation, it is more difficult to obtain the embryo callus (Ding Yumei, 2003) of oryza officinalis；The effect of high―strong nurserystock leaf callus induction Rate is relatively low (Ni Wenjin, 2007)；But ripe plant can be divided into from the callus of the freezen protective of common wild-rice (Zeliang,2010)。

The content of the invention

It is commonly wild that a kind of Southeast Asia is provided it is an object of the invention to overcome in place of the shortcomings of the prior art The inducing culture and abductive approach of rice self-mating system embryo callus.

To achieve the above object, the technical scheme taken：A kind of Southeast Asia common wild-rice embryo callus lures Culture medium is led, the inducing culture includes the component of following concentration：

The pH of the inducing culture is 5.4.

Inducing culture of the present invention is commonly wild to Southeast Asia by comparing hormon on the basis of N6D culture mediums Obtained after the influence of rice self-mating system embryo callus.

The invention provides a kind of abductive approach of Southeast Asia common wild-rice self-mating system embryo callus, methods described It is that the Southeast Asia common wild-rice self-mating system seed of shelling is inoculated on inducing culture described above to carry out Fiber differentiation 's.

Preferably, the Fiber differentiation temperature is 25~28 DEG C.

Preferably, the Fiber differentiation time is 12~18 days.

Preferably, the Fiber differentiation is carried out under light illumination.

Preferably, the intensity of illumination is 30000lx, light application time 16h/d.

Preferably, before the Southeast Asia common wild-rice seed of shelling is inoculated in into the inducing culture, to described Southeast Asia common wild-rice seed carries out disinfection processing.

Preferably, the Processing for removing comprises the following steps：The Southeast Asia common wild-rice self-mating system seed is first used 75% alcohol solution dipping 2min, then vibrate sterilization 50min with 20%NaClO solution, then with aseptic water washing 6~7 times, most After dry up the Southeast Asia common wild-rice self-mating system seed.75% ethanol solution refers to the ethanol that mass concentration is 75% The aqueous solution, the 20%NaClO solution refer to the NaClO aqueous solution that mass concentration is 20%.

The beneficial effects of the present invention are：The invention provides a kind of Southeast Asia common wild-rice self-mating system embryo callus subculture group The inducing culture knitted, Fiber differentiation can improve healing rate on the culture medium, increase callus diameter, improve callus quality.

Brief description of the drawings

Fig. 1 is to induce Southeast Asia common wild-rice self-mating system to produce embryo on inducing culture described in the embodiment of the present invention 1 The aspect graph of property callus.

Embodiment

To better illustrate the object, technical solutions and advantages of the present invention, below in conjunction with specific embodiment to the present invention It is described further.

Embodiment 1

1st, material and method

1.1st, material

Southeast Asia common wild-rice self-mating system, now it is stored in national germplasm Nanning wild rice garden, takes pond to plant mode and enter Row dystopy Cultivate administration.

1.2nd, the inducing culture of Southeast Asia common wild-rice self-mating system embryo callus such as table 1

The inducing culture of table 1 forms

The pH of the inducing culture is 5.4.

Control medium：Culture medium after NAA, IAA and KT in above-mentioned culture medium is removed.

1.3rd, reagent

Chemical reagent used in IAA, NAA, 2,4-D, 6-BA, KT and various organization culture is that domestic analysis is pure.

1.4th, the method for inducing and cultivating of Southeast Asia common wild-rice self-mating system embryo callus subculture group

The Southeast Asia common wild-rice self-mating system seed newly harvested is put into 35 DEG C of drying 24h of baking oven, by the wild of drying Rice self-mating system seed shells, and then soaks 2min with 75% ethanol, then vibrate sterilization 50min, aseptic water washing with 20%NaClO 6~7 times, suck dry moisture on aseptic filter paper is subsequently placed in, 30min is dried up in super-clean bench and is seeded in callus induction respectively On culture medium (table 1) and control medium, 20 conducts, 1 repetition is placed per ware, every kind of culture medium sets 3 repetitions.In 28 Illumination cultivation, intensity of illumination 30000lx, light application time 16h/d are carried out under the conditions of DEG C.Healing rate is calculated after inoculation 15d, is gone out More seed number/inoculation seed number × 100% of rate=grow callus.

2nd, result

The callus rate induced by contrasting on different culture media, as a result find, inducing culture of the present invention goes out more Rate is 19.00%, and the healing rate than control group culture medium is high by 5%.

The diameter of the embryo callus subculture induced by comparing on different culture media, as a result find inducing culture of the present invention The callus diameter of upper growth reaches 0.25cm, and a diameter of 0.23cm of the callus of control medium.

In terms of embryo callus subculture quality, the callus of control medium culture, color and luster is milky white, loose to expand, and has graininess to dash forward Rise；The callus grown on inducing culture of the present invention, color and luster is partially yellow, and quality is close, and particle projection is more (such as Fig. 1 institutes Show).Illustrate that inducing culture of the present invention can improve callus health condition, improve callus quality.

In addition, present invention discover that the preferable temperature of Fiber differentiation is 25-28 DEG C, and culture just can be observed to be cured for 12 days Wound.

Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than the present invention is protected The limitation of scope is protected, although being explained in detail with reference to preferred embodiment to the present invention, one of ordinary skill in the art should Understand, technical scheme can be modified or equivalent substitution, without departing from the essence of technical solution of the present invention And scope.

Claims

A kind of 1. inducing culture of Southeast Asia common wild-rice self-mating system embryo callus, it is characterised in that the induction Culture medium includes the component of following concentration：

The pH of the inducing culture is 5.4.
2. a kind of abductive approach of Southeast Asia common wild-rice self-mating system embryo callus, it is characterised in that methods described is The Southeast Asia common wild-rice self-mating system seed of shelling is inoculated on inducing culture as claimed in claim 1 and induced Culture.
3. abductive approach according to claim 2, it is characterised in that the Fiber differentiation temperature is 25~28 DEG C.
4. abductive approach according to claim 2, it is characterised in that the Fiber differentiation time is 12~18 days.
5. abductive approach according to claim 2, it is characterised in that the Fiber differentiation is carried out under light illumination.
6. abductive approach according to claim 5, it is characterised in that the intensity of illumination is 30000lx, and light application time is 16h/d。
7. abductive approach according to claim 2, it is characterised in that by the Southeast Asia common wild-rice self-mating system of shelling Seed is inoculated in before the inducing culture, and carry out disinfection processing to the Southeast Asia common wild-rice seed.
8. abductive approach according to claim 7, it is characterised in that described disinfect comprises the following steps：By described in Southeast Asia common wild-rice self-mating system seed first with 75% alcohol solution dipping 2min, is then disappeared with the vibration of 20%NaClO solution Malicious 50min, then with aseptic water washing 6~7 times, finally dry up the Southeast Asia common wild-rice self-mating system seed.