CN103548685A - Rooting culture medium for tissue culture seedling of photinia fraseri as well as in-bottle rooting method and outside-bottle rooting method - Google Patents

Rooting culture medium for tissue culture seedling of photinia fraseri as well as in-bottle rooting method and outside-bottle rooting method Download PDF

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CN103548685A
CN103548685A CN201310532712.3A CN201310532712A CN103548685A CN 103548685 A CN103548685 A CN 103548685A CN 201310532712 A CN201310532712 A CN 201310532712A CN 103548685 A CN103548685 A CN 103548685A
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rooting
bottle
seedling
photinia glabra
culture medium
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陈泽雄
刘奕清
黄登艳
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Chongqing University of Arts and Sciences
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Chongqing University of Arts and Sciences
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    • Y02P60/216

Abstract

The invention discloses a rooting culture medium for a tissue culture seedling of photinia fraseri as well as an in-bottle rooting method and an outside-bottle rooting method. The rooting culture medium takes a 1/2MS culture medium as a basic culture medium and is added with 0.2mg/L of naphthylacetic acid, 0.1mg/L of indol-3-butyric acid, 25g/L of sucrose and 4g/L of agar. The outside-bottle rooting method comprises the following steps: cutting a single bud from subculture cluster buds cultured by the tissue culture seedling of the photinia serrulata and immersing the cut single buds into naphthylacetic acid with the concentration being 500mg/L for 5 seconds; and carrying out cuttage on the single bud so as to insert into a turfy soil base material for rooting culture. According to the rooting culture medium, the in-bottle rooting method and the outside-bottle rooting method, researches on a rooting technology of the tissue culture seedling of the photinia serrulata are carried out on the basis of a developed rapid tissue culture propagation technology of the photinia serrulata; the rooting rate of the rooting culture medium is 100% and the average rooting quantity is 5; the rooting rate of the outside-bottle rooting method is 98.3% and the average rooting quantity is 7.6. The rooting culture medium, the in-bottle rooting method and the ex-bottle rooting method have important practical significances on industrial and large-scale rapid seedling growing of the photinia serrulata.

Description

Rooting method and outside sprout-cultivating-bottle method in photinia glabra group training seedling rooting medium and bottle
Technical field
The invention belongs to field of plant tissue culture technique, relate to a kind of rooting method and outside sprout-cultivating-bottle method in photinia glabra group training seedling rooting medium and bottle.
Background technology
Photinia glabra (Photinia fraseri) is the crossbreed of rose family Photinia (Photinia), is evergreen dungarunga, because it has cherry young sprout and tender leaf is gained the name.Photinia glabra is as the evergreen color leaf seeds of preciousness, have advantages of that growth rapid, resistance to pruning, strain shape are compact, rudiment power is strong, suitable raw scope is wide, like sun but very resistance to the moon, the acidproof while is Salt And Alkali Tolerance again, drought-enduring, cold-resistant and barren-resistant, on the south the Yellow River, most areas all can be planted, and the antitoxin gas ability that this plant tool is stronger can be in the more serious area plantation of atmospheric pollution.Of many uses on gardens, make hedgerow, Lv Qiang, moulding tree, isolated planting effect all good.Photinia glabra, as a kind of exploitation seeds of new introduction, at present at home still in the seedling breeding stage, only has a small amount of seedling supply market, can not meet the demand of current nursery and landscape engineering application far away.
The modes of reproduction of photinia glabra comprises cottage propagation, tissue culture propagation, propagation by grafiting, seed propagation, and group training or cuttage are its main modess of reproduction.Conventional cottage propagation, due to factors such as reproduction speed are slow, be subject to seasonal restrictions, floor space is large, has limited its promotion rate widely.With respect to cuttage seeding, photinia glabra group training seedling is not subject to seasonal restrictions, and is more suitable for factorial seedling growth, but photinia glabra group training seedling rooting number is few, and rooting rate is low, and improving activity of root system is low, less in the research of the aspects such as batch production production application.
For this reason, on the basis of photinia glabra group culturation rapid propagating technology maturation, carry out the technical research of photinia glabra group training seedling rooting, improve its rooting rate, improve root activity, reduce seedling cost, further improve photinia glabra tissue-culturing rapid propagation and industrial breeding technique system, to realizing the extensive fast seedling growing of photinia glabra batch production, meet the landscape engineering market demand growing to it, the important realistic meaning of tool.
Summary of the invention
In view of this, the object of the present invention is to provide a kind of rooting method and outside sprout-cultivating-bottle method in photinia glabra group training seedling rooting medium and bottle, explore suitable photinia glabra group training seedling rooting method and condition, improve its rooting rate, improve root activity, reduce seedling cost.
For achieving the above object, the invention provides following technical scheme:
The invention discloses a kind of photinia glabra group training seedling rooting medium, described root media is to take 1/2MS medium as minimal medium, and is added with methyl α-naphthyl acetate 0.2mg/L, indole-3-butyric acid 0.1mg/L, sucrose 25g/L and agar 4g/L.
The invention also discloses and use above-mentioned photinia glabra group training seedling rooting medium to carry out the method for taking root in bottle, photinia glabra tissue culture plant inoculation is carried out to culture of rootage to described root media, condition of culture is: cultivation temperature is 25 ± 2 ℃, first astigmatism was cultivated after 5 days, under the condition that to proceed to intensity of illumination and be 1500~2000lx, light application time be 12h/d, cultivated.
The invention also discloses the method for a kind of photinia glabra group training outside sprout-cultivating-bottle radication, the subculture Multiple Buds of photinia glabra group training seedling is cut to simple bud, in the methyl α-naphthyl acetate that is 500mg/L, soak after 5s the simple bud cutting in concentration, cuttage is to culture of rootage in turfy soil matrix.
1/2MS medium described in the present invention is minimal medium conventional in Plant Tissue Breeding, and it specifically fills a prescription as shown in the table:
Beneficial effect of the present invention is: the present invention is on the basis of photinia glabra group culturation rapid propagating technology maturation, carried out the technical research of photinia glabra group training seedling rooting, study by experiment the impact of the factors such as hormon and concentration, kinds of culture medium on photinia glabra group training seedling rooting, found suitable root media to be: 1/2MS+NAA0.2mg/L+IBA0.1mg/L+ sucrose 25g/L+ agar 4g/L, rooting rate reaches 100%, and the number of on average taking root reaches 5; By the research of photinia glabra group training outside sprout-cultivating-bottle radication technology, obtained suitable photinia glabra group training outside sprout-cultivating-bottle radication condition, the growth hormone methyl α-naphthyl acetate that is 500mg/L by concentration (NAA) soaks after 5s, and cuttage is in turfy soil matrix, its rooting rate reaches 98.3%, and the number of on average taking root reaches 7.6.The present invention is further perfect photinia glabra tissue-culturing rapid propagation and industrial breeding technique system, to realizing the extensive fast seedling growing of photinia glabra batch production, meet the landscape engineering market demand growing to it, has important practical significance.
Accompanying drawing explanation
In order to make object of the present invention, technical scheme and beneficial effect clearer, the invention provides following accompanying drawing and describe:
Fig. 1 is the seedling of taking root of the photinia glabra after NAA processes;
Fig. 2 is the seedling of taking root of the photinia glabra after IBA processes;
Fig. 3 is the seedling of taking root of the photinia glabra after NAA and IBA combined treatment;
Fig. 4 is that 1/2MS is the photinia glabra of the medium seedling of taking root;
Fig. 5 is that 1/2MS (lacks NH 4nO 3) be the photinia glabra of the medium seedling of taking root;
Fig. 6 is that perlite is the photinia glabra of the medium supporting dielectric seedling of taking root;
Fig. 7 is that turfy soil is the photinia glabra of the medium supporting dielectric seedling of taking root;
Fig. 8 is that perlite is the photinia glabra of cutting medium;
Fig. 9 is that turfy soil is the photinia glabra of cutting medium;
Figure 10 is that cuttage enters photinia glabra after the perlite seedling of taking root;
Figure 11 is that cuttage enters photinia glabra after the turfy soil seedling of taking root.
Embodiment
Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described in detail.
It is research object that the experiment material of the embodiment of the present invention is selected Chongqing higher learning institutions landscape flower ERC of Institute Of Unity and Coherence In Writing Of Chongqing healthy aseptic photinia glabra group training seedling, gets its stem section with 1~2 axillalry bud as supplying examination material.
One. in photinia glabra group training seedling bottle, take root
1. the screening of hormon kind and concentration
The stem section that cuts the photinia glabra group training seedling of 2-3cm, the stem section upgrowth situation cutting is consistent, is seeded to and in root media, carries out culture of rootage.Root media be take 1/2MS as minimal medium, adds sucrose 25g/L, agar 4g/L, and adds different growth hormone (IBA, NAA) and variable concentrations.Every bottle of 5 strains, each processes 10 bottles.Cultivation temperature is 25 ± 2 ℃, and first astigmatism is cultivated 5 days, then take root number and rooting rate of statistics after intensity of illumination is to cultivate 20d under 1500~2000lx, the light application time condition that is 12h/d.
Result of the test is in Table 1, and as can be seen from Table 1, the medium photinia glabra rooting rate that adds NAA is lower, and along with the increase of concentration, the callus of formation is also increasing, not only affects the differentiation of adventive root, also affects transplant survival power, as shown in Figure 1; Add IBA medium bud seedling rooting rate all more than 80% and callus substantially do not have, transplanting survival rate is high, as shown in Figure 2; NAA and IBA combination, rooting rate is the highest, average out to 90%, the combination of low concentration, takes root very fast, and average root reaches 2cm, and callus is also less, as shown in Figure 3.So NAA0.2mg/L+IBA0.1mg/L is suitable hormone combinations.
Table 1 hormon kind and concentration are on the take root impact of seedling of photinia glabra
2. the screening of minimal medium
The stem section that cuts the photinia glabra group training seedling of 2-3cm, the stem section upgrowth situation cutting is consistent, is seeded to and in root media, carries out culture of rootage.Root media lacks NH with MS, 1/2MS, 1/4MS, 1/2MS( 4nO 3) be minimal medium, add sucrose 25g/L, agar 4g/L, and add NAA0.2mg/L, IBA0.1mg/L.Every bottle of 5 strains, each processes 10 bottles.Cultivation temperature is 25 ± 2 ℃, and first astigmatism is cultivated 5 days, then take root number and rooting rate of statistics after intensity of illumination is to cultivate 20d under 1500~2000lx, the light application time condition that is 12h/d.
Result of the test is in Table 2, and as can be seen from Table 2, the MS medium of different proportion has obvious impact to the root induction of photinia glabra, and wherein best with the induction effect of 1/2MS medium, rooting rate reaches 98%, and the number of on average taking root reaches 3.4, as shown in Figure 4; Although 1/2MS (lacks NH 4nO 3) rooting rate, take root number and average root long all higher, but the hardness of root is large, improving activity of root system is low, be difficult for surviving after transplanting, as shown in Figure 5.Experiment shows that MS medium has obviously suppressed the formation of photinia glabra group training seedling adventive root, the rooting rate of group training seedling is very low, and rooting rate and on average the take root number of group training seedling in 1/2MS and 1/4MS medium all increases substantially, on average take root number more than 2, illustrate that in medium, the mineral salt of low concentration are conducive to take root.But the rooting rate of 1/4MS medium and the number of on average taking root be not as good as the height of 1/2MS medium, and root more carefully, a little less than, show that the too low photinia glabra that is not necessarily conducive to of inorganic salt concentration takes root.So be to be applicable to inducing adventitious root for photinia glabra 1/2MS medium.
The comparison of table 2 medium inorganic salt concentration to photinia glabra culture of rootage
3. the screening of medium supporting dielectric
The stem section that cuts the photinia glabra group training seedling of 2-3cm, the stem section upgrowth situation cutting is consistent, is seeded to and in root media, carries out culture of rootage.Root media be take 1/2MS as minimal medium, adds sucrose 25g/L, NAA0.2mg/L, IBA0.1mg/L, and take respectively agar, turfy soil and perlite as medium supporting dielectric.Every bottle of 5 strains, each processes 10 bottles.Cultivation temperature is 25 ± 2 ℃, and first astigmatism is cultivated 5 days, then take root number and rooting rate of statistics after intensity of illumination is to cultivate 20d under 1500~2000lx, the light application time condition that is 12h/d.
Result of the test, in Table 3, as can be seen from Table 3, be take agar as culture medium supports root induction, and effect is better, and root is red, and lateral root is few, and rooting rate is high, can reach 100%.Take perlite and turfy soil as culture medium supports root induction, root tubbiness, base portion is without callus, but rooting rate is all lower, as shown in Figure 6 and Figure 7, its reason may be: turfy soil and perlite are all slant acidity medium, PH adjusting is inaccurate easily departs from 5.8, is unfavorable for the formation of adventive root; The medium that turfy soil and perlite join as matrix of take is unfavorable for high-temperature sterilization, often because sterilizing does not thoroughly cause follow-up pollution, and then greatly affects the induction of adventive root; The medium that the turfy soil of take is matrix, because water content is high, easily makes the group of inoculation train seedling bottom rot and be unfavorable for taking root; Perlite light weight, ventilation are good comparatively favourable to taking root, but in perlite, nutrient is less, and because its proportion is lighter than water, can float over the surface of matrix when trickle is more, cause perlite insecure with contacting of root system and be unfavorable for taking root.So for photinia glabra, agar is suitable medium supporting dielectric.
The impact that the different supporting dielectrics of table 3 are taken root on photinia glabra
4. transition cultivating
The test-tube plantlet of having taken root is moved in seeding room, close uncork hardening 1~2d after bottle hardening 3~4d, carry out adaptive training.During bottle outlet seedling, with clear water, clean the medium that test-tube plantlet root adheres to, with 800 times of tpn immersion bubble test tube base portion 2~3min, aseptic seedling is transplanted in nutrition cup, righting seedling, transplanting medium is turfy soil: peat soil: perlite=3:3:1.In front 7~10d, take to increase relative humidity of atomsphere, sunshade and appropriateness cooling wait measure, the humidity that keeps afterwards environment all the time in 90% left and right, 28 ℃~32 ℃ of temperature, more than illumination 1600lx, foliage spray 0.1% urea or potassium dihydrogen phosphate after 15d, 5~7d/ time, to supplement the nutrients, after 40d, add up test-tube plantlet survival rate.Through 20d, blade increases and has young leaves to grow, and root hair increases, and it is very healthy and strong that nursery stock seems, transplanting survival rate can reach more than 90%.
In sum, the suitable photinia glabra group training seedling rooting medium that the present invention obtains is: 1/2MS+NAA0.2mg/L+IBA0.1mg/L+ sucrose 25g/L+ agar 4g/L, take 1/2MS medium as minimal medium, and be added with methyl α-naphthyl acetate 0.2mg/L, indole-3-butyric acid 0.1mg/L, sucrose 25g/L and agar 4g/L, rooting rate reaches 100%, and the number of on average taking root reaches 5.
Two. photinia glabra group training outside sprout-cultivating-bottle radication
Select the photinia glabra subculture crowd shoots of cultivating more than 40d, put into booth hardening 5d, from blake bottle, take out crowd shoots, with the single plant sprout more than high 3cm of operating scissors clip, in the NAA of 500mg/L, soak after 5s, 30s, 1min respectively, in the flowerpot that cuttage is matrix to turfy soil or perlite, as shown in Figure 8 and Figure 9.Shorten as far as possible and get cuttings to the time of cuttage; The cuttage degree of depth generally will be controlled between l~1.5cm, if cuttage is too shallow, moisture content is easily lost, and is also unfavorable for taking root.After inserting, water immediately permeablely, during cuttage, substantially do not apply fertilizer, after root of hair foliation, spray water-soluble chemical fertilizer (0.2% urea), to promote the healthy and strong growth of cuttage seeding.After 20d, observe the situation of taking root.
Result of the test, in Table 4, as can be seen from Table 4, be take perlite and turfy soil as cutting medium root induction, and lateral root is many, and the elongated and base portion of root is without callus, and its number of on average taking root is all higher, and transplanting survival rate is also high, as shown in Figure 10 and Figure 11.Wherein best to the effect of turfy soil with cuttage soak 5s in the NAA of 500mg/L after, the number of on average taking root is 7.6, and rooting rate is 98.3%.
The impact of table 4 different disposal condition on photinia glabra outside sprout-cultivating-bottle
Therefore, the suitable photinia glabra group training outside sprout-cultivating-bottle radication condition that the present invention obtains is: the subculture Multiple Buds of photinia glabra group training seedling is cut to simple bud, in the methyl α-naphthyl acetate that is 500mg/L in concentration by the simple bud cutting, soak after 5s, cuttage is in turfy soil matrix, its rooting rate reaches 98.3%, and the number of on average taking root reaches 7.6.
Finally explanation is, above preferred embodiment is only unrestricted in order to technical scheme of the present invention to be described, although the present invention is described in detail by above preferred embodiment, but those skilled in the art are to be understood that, can to it, make various changes in the form and details, and not depart from the claims in the present invention book limited range.

Claims (3)

1. photinia glabra group training seedling rooting medium, is characterized in that: described root media is to take 1/2MS medium as minimal medium, and is added with methyl α-naphthyl acetate 0.2mg/L, indole-3-butyric acid 0.1mg/L, sucrose 25g/L and agar 4g/L.
2. right to use requires the photinia glabra group training seedling rooting medium described in 1 to carry out the method for taking root in bottle, it is characterized in that: photinia glabra tissue culture plant inoculation is carried out to culture of rootage to described root media, condition of culture is: cultivation temperature is 25 ± 2 ℃, first astigmatism was cultivated after 5 days, under the condition that to proceed to intensity of illumination and be 1500~2000lx, light application time be 12h/d, cultivated.
3. the method for photinia glabra group training outside sprout-cultivating-bottle radication, is characterized in that: the subculture Multiple Buds of photinia glabra group training seedling is cut to simple bud, in the methyl α-naphthyl acetate that is 500mg/L, soak after 5s the simple bud cutting in concentration, cuttage is to culture of rootage in turfy soil matrix.
CN201310532712.3A 2013-11-01 2013-11-01 Rooting culture medium for tissue culture seedling of photinia fraseri as well as in-bottle rooting method and outside-bottle rooting method Pending CN103548685A (en)

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CN105104144A (en) * 2015-09-01 2015-12-02 张莘蔓 Syzygium hancei water culture cutting propagation method
CN106258958A (en) * 2016-08-08 2017-01-04 天水市果树研究所 A kind of outside sprout-cultivating-bottle method of Fructus Pruni pseudocerasi tissue cultured seedling
CN107197759A (en) * 2017-05-13 2017-09-26 梁钟 A kind of salt tolerant Chinese photinia breeding method
CN108605831A (en) * 2016-12-16 2018-10-02 江苏省中国科学院植物研究所 A kind of method of Wikstroemia indica tissue-cultured seedling Rapid Rooting
CN109952955A (en) * 2019-04-04 2019-07-02 安阳工学院 A method of suitable for color leafed plants tissue culture seedling rooting

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104871945A (en) * 2015-06-02 2015-09-02 北京市花木有限公司 Heuchera micrantha tissue culture propagation ex-vitro rooting seedling hardening method
CN104871945B (en) * 2015-06-02 2017-09-12 北京天卉苑花卉研究所 A kind of method of alum root tissue culture propagation outside sprout-cultivating-bottle hardening
CN105104144A (en) * 2015-09-01 2015-12-02 张莘蔓 Syzygium hancei water culture cutting propagation method
CN106258958A (en) * 2016-08-08 2017-01-04 天水市果树研究所 A kind of outside sprout-cultivating-bottle method of Fructus Pruni pseudocerasi tissue cultured seedling
CN108605831A (en) * 2016-12-16 2018-10-02 江苏省中国科学院植物研究所 A kind of method of Wikstroemia indica tissue-cultured seedling Rapid Rooting
CN107197759A (en) * 2017-05-13 2017-09-26 梁钟 A kind of salt tolerant Chinese photinia breeding method
CN109952955A (en) * 2019-04-04 2019-07-02 安阳工学院 A method of suitable for color leafed plants tissue culture seedling rooting
CN109952955B (en) * 2019-04-04 2022-05-03 安阳工学院 Rooting method suitable for tissue culture seedlings of color-leaf plants

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Application publication date: 20140205