CN104106450A

CN104106450A - Method for obtaining pollen by isolated culturing of pear branches

Info

Publication number: CN104106450A
Application number: CN201410252939.7A
Authority: CN
Inventors: 胡红菊; 杨晓平; 陈启亮; 田瑞; 张靖国; 范净; 王春燕
Original assignee: Institute of Fruit and Tea of Hubei Academy of Agricultural Sciences
Current assignee: Institute of Fruit and Tea of Hubei Academy of Agricultural Sciences
Priority date: 2014-06-10
Filing date: 2014-06-10
Publication date: 2014-10-22
Anticipated expiration: 2034-06-10
Also published as: CN104106450B

Abstract

The invention provides a method for obtaining pollen by isolated culturing of pear branches. The method includes the steps that pear flowering branches are collected, after pear trees are dormant in the late December, the pear branches with full blossom buds are collected, sheared openings of bases of the branches can align, and the length of the branches is controlled to be 40 cm; branch cold storage is conducted, the bases of the branches are tightly bound through plastic thin films and then placed into a refrigerator for cold storage at the temperature of 4 DEG C, and the cold storage time is 60 days; branch culture is carried out, the cold storage branches are inserted into a water bucket which is filled with a culture solution 13-15 days before the flowering period of a pear main cultivar, river sand with the thickness being 10 cm is placed into the bottom of the water bucket, the depth of the culture solution is 10 cm higher than the position where the river sand is submerged, and the temperature is controlled to be 26 DEG C; pollen is obtained, after cultivating is performed for 15 days, and at the big bud period of the pears, balloon-shaped pear flower buds are collected for preparing pollen. By the adoption of the method, high quality pear pollen can be obtained, the germination rate of the pear pollen is more than 65%, and thus the pollen viability needs of pears during artificial pollination can be met in scientific research and production.

Description

A kind of cultured in vitro pear tree branch obtains the method for pollen

Technical field

Invention belongs to fruit tree technical field, more specifically relates to a kind of cultured in vitro pear tree branch and obtain the method for pollen, the problem of pollinated variety each other when good pears kind flowering asynchronism in the method processing can effectively solve pears scientific research, breeding and produce.The method is applicable to be engaged in R&D institution, company and the individual of pears scientific research, breeding and production.

Background technology

Pear in the rose family ( rosaceae) Maloideae ( maloideae) pear ( pyrus) plant, for tall this deciduous fruit tree, be that China three one of cultivates apple trees greatly, also occupy very important status in the world.According to FAO (Food and Agriculture Organization of the United Nation) (FAO) statistics, 1,132 thousand hectares of China's pears cultivated areas in 2011,1595.5 ten thousand tons of output, occupy first place in the world.It is the mainstay industry that China's increasing peasant income are got rich that the operatic circle produces always, is also one of China's export main farm produce of earning foreign exchange.

Xenia is the directly-sensing effect of paternal pollen to seed and fruit.The male parent effect difference that different cultivars shows, as the shape of fruit, maturing stage, size, color, local flavor and inherent component content, the shape of seed, size, color etc.Pears researcher finds that in the apolegamy of research pears pollinated variety pears are comparatively significantly fruit trees of Metaxenia both at home and abroad, and pollinated variety not only affects fruit-setting rate, but also affects size, shape, fruit face proterties and the interior quality of pear fruit.Therefore in pears scientific research breeding and production process, select pears kind best in quality directly to have influence on percentage of fertile fruit, the yield and quality of pears production kind as pollinated variety.

The good Sand Pear of promoting is at present as kinds such as kingfisher hat, abundances of water, fine and tender taste and crisp, juice multi-flavor is sweet, fruit quality excellence and pollen amount, Pollen Activity high suitable decide cultivation and pollinated variety, but because its flowering stage is more late, cannot give flowering stage early, colory Sand Pear is as the pollinated variety each other such as No. 2, Hubei Province pears, aqua regin.The main method that solves at present the flowering asynchronisms such as emerald green hat, abundance of water and No. 2, Hubei Province pears, aqua regin is to introduce marginal pollinated variety of florescence to mix and plant as the kind such as Xiang Nan and chrysanthemum.But due to the existence of Metaxenia, the introducing of the kind such as Xiang Nan and Huanghua Pear causes the operatic circle quality of main breed to decline, and the selling price of Xiang Nan and Huanghua Pear is well below the selling price of the improved seeds such as kingfisher hat, abundance of water, No. 2, Hubei Province pears and aqua regin, has affected the economic benefit of pears agricultures.

For this problem, both at home and abroad pear tree expert has carried out the correlative study of pollen storage test, Wang Ling in 2006, and Zhang Shaoling has studied reserve temperature and humidity to the blodynamic impact of abundance of water pear pollen, abundance of water pear pollen is at-73 DEG C, can the long term preserve vitality at-196 DEG C.General pear pollen germination rate more than 55% belongs to high-quality pollen, and the pollen germination rate of Cryopreservation after 540 days all, more than 60%, can meet the requirement of producing upper pollination.But preserve pollen by the method, cost is high and keeper's professional technique is had relatively high expectations, this technology is in the scientific research stage at present, and general peasant household and company are difficult to grasp and use.For many years, we study around this problem always, and the problem that how obtains ahead of time pear pollen has been carried out to a large amount of scientific researches and test, sum up the method that obtains ahead of time pear pollen that invents.

Summary of the invention

The object of the invention is for the problems referred to above, a kind of pollen method that obtains good pears kind indoor cultured in vitro pear tree spray is provided, mutually carries out the requirement of artificial pollination can meet flowering asynchronism between improved seeds time, the method is easily gone, easy and simple to handle, use the method to obtain pear pollen.

Technical scheme of the present invention is:

Cultured in vitro pears branch obtains a method for pollen, comprises step:

A, collection pear tree spray:

After late December pear tree dormancy, gather the pear tree branch with full bud, and make the alignment of branch base portion clip, control branch length is 40cm;

B, branch refrigeration

Branch base portion thickness is to put into 4 DEG C of refrigerations of refrigerator after plastic film wrapping closely, and cold preservation time is 60 days;

C, branch are cultivated

Before pears main breed flowering stage 13～15 days, the branch of refrigeration to be inserted and is equipped with in the bucket of culture fluid, the river sand that 10cm is thick is put in bucket bottom, and the degree of depth of culture fluid is for flooding river sand 10cm; Then bucket is put into air conditioning chamber, temperature control is 26 DEG C;

D, pollen obtain:

Wherein branch is cultivating after 15 days, and in the large flower bud of the pear tree phase, the pear flower bud that gathers balloon-like is prepared pollen.

Wherein change once when culture fluid 5 days, 10 days, while changing culture fluid, branch base portion is cut to 1cm with scissors simultaneously.

Wherein the concentration formula of culture fluid is: KNO ₃2.5g/L, NH ₄nO ₃2g/L, MgSO ₄.7H ₂o 0.5 g/L, CaCl ₂0.8 g/L, H ₃bO ₃0.02 g/L, MnSO ₄. 4H ₂o 0.05 g/L.

For a culture fluid for above-mentioned cultured in vitro pears branch, the concentration formula of culture fluid is: KNO ₃2.5g/L, NH ₄nO ₃2g/L, MgSO ₄.7H ₂o 0.5 g/L, CaCl ₂0.8 g/L, H ₃bO ₃0.02 g/L, MnSO ₄. 4H ₂o 0.05 g/L.

Use said method can obtain the pear pollen of high-quality, and the germination rate of pear pollen is more than 65%, can meet pears scientific research, produce in the needs of Pollen Activity when artificial pollination.In production, general pear pollen germination rate more than 55% belongs to high-quality pollen, and the germination rate of pollen just can meet the needs of high yield more than 20%.

2011, within 2012 and 2013 continuous 3 years, adopt this method and spontaneous pollination method emerald green hat pears and the pollinated variety fruit-setting rate comparison each other of No. 2 pears of Hubei Province pears.

Table 1: adopt this method and spontaneous pollination method emerald green hat pears and the pollinated variety fruit-setting rate comparison each other of No. 2 pears of Hubei Province pears

As can be seen from Table 1, adopt this method can effectively promote bearing fruit of emerald green hat pears and No. 2 pears kinds of Hubei Province pears, within continuous 3 years, keep fruit-setting rate more than 99%; Be significantly higher than the fruit-setting rate of spontaneous pollination situation, can meet flowering asynchronism between improved seeds time, mutually carry out the requirement of artificial pollination, reduce pears agriculture loss, increase the economic benefit of pears agriculture.

Embodiment

A kind of cultured in vitro in 2013 obtain the method for pear pollen, the steps include:

(1) gather pear tree branch:

After pear tree dormancy on December 25, gather kingfisher hat pears and No. 2 pear tree branches of Hubei Province pears with full bud, and make the alignment of branch base portion clip, control branch length is 40cm;

(2) branch refrigeration

Branch base portion thickness is to put into 4 DEG C of refrigerations of refrigerator after the wrapping closely of 0.027mm plastic film, and cold preservation time is 60 days, to break pear tree dormancy;

(3) branch is cultivated

Be February 26 in No. 2 first 15 days of flowering stage of Hubei Province pears, the kingfisher hat branch of refrigeration is inserted and is equipped with in the bucket of culture fluid, the river sand that 10cm is thick is put in bucket bottom, and the degree of depth of culture fluid is for flooding river sand 10cm, and the formula of culture fluid is KNO ₃2.5g/L, NH ₄nO ₃2g/L, MgSO ₄.7H ₂o 0.5 g/L, CaCl ₂0.8 g/L, H ₃bO ₃0.02 g/L, MnSO ₄. 4H ₂o 0.05 g/L.Then bucket is put into air conditioning chamber, temperature control is 26 DEG C.In incubation, in the time cultivating 5 days, 10 days, change culture fluid once respectively, while changing culture fluid, branch base portion is cut to 1cm with scissors;

On 13～15 days (greatly about March 8) before kingfisher hat pears flowering stage, No. 2 pears branches of Hubei Province pears of refrigeration are cultivated according to the method described above;

(4) pollen obtains

Cultivate after 15 days, in the large flower bud of the pear tree phase, gather the pear flower bud of balloon-like and prepare pollen, just can utilize " a kind of method of quick Fabrication pear pollen " patent No. is that the method that ZL 201310180729.7 makes pollen is made pollen;

(5) pollination

Utilization obtains emerald green hat pear pollen pears No. 2 pears in Hubei Province is carried out to artificial pollination, utilizes the pollen of No. 2 pears of acquisition Hubei Province pears to carry out artificial pollination to kingfisher hat pears, and after pollination, percentage of fertile fruit investigation result is in table 1.

Percentage of fertile fruit investigation result of the present invention is the fruit bearing rate of pear tree.

This method is the pollen that obtains good pears kind indoor cultured in vitro pear tree spray.Adopt this method can obtain ahead of time the pollen of the good pears kind in evening in flowering stage, or postpone the pollen of the improved seeds that obtain evening in flowering stage, can meet flowering asynchronism between improved seeds time, mutually carry out the requirement of artificial pollination, solve scientific research, produce in the problem of pollinated variety each other when good pears kind flowering asynchronism, effect is remarkable.

Claims

1. cultured in vitro pears branch obtains a method for pollen, comprises step:

A, collection pear tree spray

B, branch refrigeration

C, branch are cultivated

D, pollen obtain

Cultivate after 15 days, in the large flower bud of the pear tree phase, the pear flower bud that gathers balloon-like is prepared pollen.

2. the method for claim 1, is characterized in that, when described culture fluid 5 days, 10 days, changes once, while changing culture fluid, branch base portion is cut to 1cm with scissors simultaneously.

3. method as claimed in claim 1 or 2, is characterized in that, the concentration formula of described culture fluid is: KNO ₃2.5g/L, NH ₄nO ₃2g/L, MgSO ₄.7H ₂o 0.5 g/L, CaCl ₂0.8 g/L, H ₃bO ₃0.02 g/L, MnSO ₄. 4H ₂o 0.05 g/L.

4. for a culture fluid for cultured in vitro pears branch as claimed in claim 1, it is characterized in that, the concentration formula of described culture fluid is: KNO ₃2.5g/L, NH ₄nO ₃2g/L, MgSO ₄.7H ₂o 0.5 g/L, CaCl ₂0.8 g/L, H ₃bO ₃0.02 g/L, MnSO ₄. 4H ₂o 0.05 g/L.