CN103609453A

CN103609453A - Construction method of in vitro regeneration system of tea tree

Info

Publication number: CN103609453A
Application number: CN201310639259.6A
Authority: CN
Inventors: 孙威江; 吴婉婉; 陈志丹; 何碧珠
Original assignee: Fujian Agriculture and Forestry University
Current assignee: Fujian Agriculture and Forestry University
Priority date: 2013-12-04
Filing date: 2013-12-04
Publication date: 2014-03-05
Anticipated expiration: 2033-12-04
Also published as: CN103609453B

Abstract

The invention discloses a construction method of an in vitro regeneration system of a tea tree. According to the construction method, the construction of the in vitro regeneration system takes a stem with an axillary bud as an explant, the stem is cultured, the axillary bud is germinated to grow leaves upwards and roots downwards, and then a seedling is directly formed without callus induction. According to the invention, the stem of Fuding Dabaicha is used as the explant and the construction method of the regeneration system is researched; the hereditary stability of tea tree varieties is kept and the storage of germplasm resources of tea trees can also be promoted.

Description

A kind of construction method of tea tree vitro Regeneration System

Technical field

The invention belongs to tea tree tissue culture technique field, be specifically related to a kind of construction method of tea tree vitro Regeneration System.

Background technology

Tea tree ( camellia sinensisl. Kuntze.) belong to Theaceae, Camellia, is a kind of perennial woody, evergreen plant.Tea tree tissue is cultivated and to be started from eighties of last century sixties, the research that China cultivates about tea tree tissue is started late, but under scientific research personnel's joint efforts, also obtained in recent years the progress of significance, by the seed of tea tree, stem section, axillalry bud, flower pesticide etc. are cultivated and obtained complete regeneration plant.But there are some problems in cultivating in tea tree tissue, the application of tissue culture technique in tea tree is limited by very large, its subject matter shows the following aspects: 1. quick propagating technology is applied to tea Culture and grows seedlings, while making explant with the indefinite bud of test-tube plantlet or axillalry bud, become tip rate low, growth is slow, and cultivation cycle is long, the unrooted tea shoot relative difficult of taking root, transplants land for growing field crops survival rate low; 2. for Different Tea Varieties, organize cultivation results to differ greatly, comparativity is little, and the cultivation results of organizing of same tea tree breed is also difficult to repetition even, and less stable cannot form a set of complete reliable method system; 3. gene engineering has become the powerful measure of genetic modification of plants, and the regeneration of in vitro plant is depended in the success or failure of genetic manipulation, but the regeneration techniques of tea tree is ripe not enough, cannot set up high-frequency, stable regenerating system.Fuding white tea is a tea tree breed that Comprehensive Traits is good, area maximum is promoted and introduced a fine variety in the whole nation, it is a backbone parent in Tea Breeding work, it is explant that Fuding white tea stem section is take in the present invention, study the method for building up of its regenerating system, not only be conducive to keep the genetic stability of tea tree breed, also can promote the preservation of Tea Germplasm.

Summary of the invention

The object of the present invention is to provide a kind of construction method of tea tree vitro Regeneration System, it is explant that this method be take the stem section of the good Fuding white tea of proterties, set up its regenerating system, be not only conducive to keep the genetic stability of tea tree breed, also can promote the preservation of Tea Germplasm.

For achieving the above object, the present invention adopts following technical scheme:

A construction method for tea tree vitro Regeneration System, is characterized in that: the structure of described tea tree vitro Regeneration System is to adopt stem segment with axillary bud to cultivate as explant, and without the direct seedling of evoked callus;

Its method for building up comprises that step is as follows:

1), by after the Stem of Tea section sterilizing with full axillalry bud, on MS+1.5 mg/L 6-BA+0.3mg/L NAA medium, cultivate 30-35d;

2), when bud grows to 3-5cm, be cut on stem section access MS+1.0mg/L 6-BA+0.1mg/LNAA medium of long 1-1.5cm and breed;

3), when the bud seedling of propagation is while growing to 4-5 sheet leaf, on 1/2 MS+0.2 mg/L NAA medium, carry out culture of rootage;

4), when the group training shoot root of culture of rootage grows to 5-6cm, during average 4-5 bar root, carry out acclimatization and transplants, outdoor cultivation tea tree plant.

Described stem segment with axillary bud is Fuding white tea stem section, and described stem segment with axillary bud is cultivated, and after axillary bud sprouting, upwards comes into leaves, and takes root downwards.

Remarkable advantage of the present invention is:

(1), the construction method of tea tree vitro Regeneration System provided by the invention, the stem section of tea tree breed Fuding white tea that the Comprehensive Traits of usining is good, promote in the whole nation and introduce a fine variety area maximum is as explant, not only be conducive to keep the genetic stability of tea tree breed, also can promote the preservation of Tea Germplasm, and can be the purification and rejuvenation of tea tree in the future and provide fundamental basis.

(2), the construction method of the tea tree vitro Regeneration System set up of the present invention, can be without the direct seedling of evoked callus, cultivation cycle is short, becomes tip rate high, transplant land for growing field crops survival rate high, and method is stable, reliable, through experiment repeatedly, all can obtain good result.

Embodiment

Embodiment 1

A method for building up for tea tree vitro Regeneration System, specifically comprises that step is as follows:

1), by after the Stem of Tea section sterilizing with full axillalry bud, on MS+1.5 mg/L 6-BA+0.3mg/L NAA medium, cultivate 30d;

2), when bud grows to 3 cm, be cut on stem section access MS+1.0mg/L 6-BA+0.1mg/LNAA medium of long 1 cm and breed;

3), when the bud seedling of propagation is while growing to 4 leaves, on 1/2 MS+0.2 mg/L NAA medium, carry out culture of rootage;

4), when the group training shoot root of culture of rootage grows to 5cm, during average 5 roots, carry out acclimatization and transplants, will organize and train transplantation of seedlings extremely after outdoor cultivation 30d, survival rate of plant is 67 %, and plant growth condition is better, blade is emerald green.

Embodiment 2

1), by after the Stem of Tea section sterilizing with full axillalry bud, on MS+1.5 mg/L 6-BA+0.3mg/L NAA medium, cultivate 32 d;

2), when bud grows to 4 cm, be cut on stem section access MS+1.0mg/L 6-BA+0.1mg/LNAA medium of long 1 cm and breed;

3), when the bud seedling of propagation is while growing to 5 leaves, on 1/2 MS+0.2 mg/L NAA medium, carry out culture of rootage;

4), when the group training shoot root of culture of rootage grows to 5.4 cm, during average 5 roots, carry out acclimatization and transplants.To organize training transplantation of seedlings to outdoor cultivation 30d, survival rate of plant is 65 %, and plant growth condition is better, and blade is emerald green.

Embodiment 3

1), by after the Stem of Tea section sterilizing with full axillalry bud, on MS+1.5 mg/L 6-BA+0.3mg/L NAA medium, cultivate 35d;

2), when bud grows to 5cm, be cut on stem section access MS+1.0mg/L 6-BA+0.1mg/LNAA medium of long 1.5cm and breed;

4), when the group training shoot root of culture of rootage grows to 6 cm, during average 4 roots, carry out acclimatization and transplants.To organize training transplantation of seedlings to outdoor cultivation 30d, survival rate of plant is 68 %, and plant growth condition is better, and blade is emerald green.

Utilize method of the present invention to build tea tree vitro Regeneration System, the outdoor transplanting survival rate of group training seedling is higher, average out to 66.7%.

The foregoing is only preferred embodiment of the present invention, all equalizations of doing according to the present patent application the scope of the claims change and modify, and all should belong to covering scope of the present invention.

Claims

1. a construction method for tea tree vitro Regeneration System, is characterized in that: the structure of described tea tree vitro Regeneration System is to adopt stem segment with axillary bud to cultivate as explant, and without the direct seedling of evoked callus; Its method for building up concrete steps are as follows:

1), by after the Stem of Tea section sterilizing with full axillalry bud, on MS medium, cultivate 30-35d;

2), when bud grows to 3-5cm, be cut on the stem section access MS proliferated culture medium of long 1-1.5cm and breed;

3), when the bud seedling of propagation is while growing to 4-5 sheet leaf, on 1/2 MS root media, carry out culture of rootage;

4), when the group training shoot root of culture of rootage grows to 5-6cm, during average 4-5 bar root, carry out acclimatization and transplants.

2. the construction method of tea tree vitro Regeneration System according to claim 1, is characterized in that: described stem segment with axillary bud is Fuding white tea stem section, cultivates, and after axillary bud sprouting, upwards comes into leaves, and takes root downwards.